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Radial artery access is the primary approach for coronary interventions due to higher safety profile in comparison to femoral access. Radial artery occlusion (RAO) is the main complication of transradial catheterization that can lead to severe symptoms and a permanent artery occlusion. The incidence of RAO after transradial access ranges from 5 to 38% and data regarding treatment is scarce. Whether anticoagulation and vasoactive medication provides an additional benefit in recovery of radial artery patency (RAP) after catheterization has not been investigated in detail. The objective was to investigate the impact of anticoagulation and vasoactive medication on regained patency after documented RAO following transradial catheterization. Overall 2635 patients were screened. 2215 (84%) catheterizations were performed by femoral and 420 (16%) by radial access. In 30 patients RAO was observed. In case of RAO patients were classified in three groups: Anticoagulation, anticoagulation added with alprostadil and controls. Follow-up was conducted after 3 months with ultrasound and clinical examination. Eight patients received anticoagulation and 11 patients anticoagulation together with alprostadil. Eleven patients served as controls. Recovery of RAP after catheterization was higher following either treatment (79.5%) compared to controls (0%, p= 0.006). Subgroup analysis yielded a higher RAP recovery in patients treated with anticoagulation (62.5%) as compared to controls (0%, p= 0.002). No effect on regained RAP was found with additional alprostadil therapy (33.3%) compared to anticoagulation therapy (62.5%, p= 0.229). RAO should be treated with anticoagulation to regain patency. Addition of vasoactive medication does not lead to further beneficial effects. Further research is needed regarding preventive and therapeutic strategies following RAO.

Galaxies are surrounded by large reservoirs of gas, mostly hydrogen, that are fed by inflows from the intergalactic medium and by outflows from galactic winds. Absorption-line measurements along the lines of sight to bright and rare background quasars indicate that this circumgalactic medium extends far beyond the starlight seen in galaxies, but very little is known about its spatial distribution. The Lyman-a transition of atomic hydrogen at a wavelength of 121.6 nanometres is an important tracer of warm (about 104kelvin) gas in and around galaxies, especially at cosmological redshifts greater than about 1.6 at which the spectral line becomes observable from the ground. Tracing cosmic hydrogen through its Lyman-a emission has been a long-standing goal of observational astrophysics1–3, but the extremely low surface brightness of the spatially extended emission is a formidable obstacle. A new window into circumgalactic environments was recently opened by the discovery of ubiquitous extended Lyman-a emission from hydrogen around high-redshift galaxies4,5. Such measurements were previously limited to especially favourable systems6–8or to the use of massive statistical averaging9,10because of the faintness of this emission. Here we report observations of low-surface-brightness Lyman-a emission surrounding faint galaxies at redshifts between 3 and 6. We find that the projected sky coverage approaches 100 per cent. The corresponding rate of incidence (the mean number of Lyman-a emitters penetrated by any arbitrary line of sight) is well above unity and similar to the incidence rate of high-column-density absorbers frequently detected in the spectra of distant quasars11–14. This similarity suggests that most circumgalactic atomic hydrogen at these redshifts has now been detected in emission.

Context. The RAVE spectroscopic survey for galactic structure and evolution obtains 8400–8800 Å spectra at 7500 resolving power at the UK Schmidt Telescope using the 6dF multi-fiber positioner. More than 300 000 9 ≤IC≤12 and |b| ≥25° southern stars have been observed to date.

We report optical B-band observations with the Large Binocular Telescope (LBT) of the isolated neutron star RBS1774. The stacked image with a total exposure of $2\fh5$reveals a candidate optical counterpart at mB= 26.96 ±0.20 at position $\alpha {\rm (2000)} = {\rm 21^h43^m03\fs40}$, $\delta{\rm (2000)} = +06\degr54^\prime17\farcs5$, within the joint Chandra and XMM-Newton error circles. We analyse archival XMM-Newton observations and derive revised spectral and positional parameters. The predicted optical flux from the extrapolated X-ray spectrum is likely twice as high as reported before. The measured optical flux exceeds the extrapolated X-ray spectral flux by a factor ${\sim}40$(15-60 at 1σconfidence). We interpret our detection and the spectral energy distribution as further evidence of a temperature structure over the neutron star's surface and present a pure thermal model reflecting both the SED and the pulsed fraction of the light curve.

We have used spectra of hot stars from the RAVE Survey in order to investigate the visibility and properties of five diffuse interstellar bands previously reported in the literature. The RAVE spectroscopic survey for Galactic structure and kinematics records CCD spectra covering the 8400–8800 Å wavelength region at 7500 resolving power. The spectra are obtained with the UK Schmidt at the AAO, equipped with the 6dF multi-fiber positioner. The DIB at 8620.4 Å is by far the strongest and cleanest of all DIBs occurring within the RAVE wavelength range, with no interference by underlying absorption stellar lines in hot stars. It correlates so tightly with reddening that it turns out to be a reliable tool to measure it, following the relation $E_{B-V} = 2.72 (\pm 0.03)\times EW~$(Å), valid throughout the general interstellar medium of our Galaxy. The presence of a DIB at 8648 Å is confirmed. Its intensity appears unrelated to reddening, in agreement with scanty and preliminary reports available in the literature, and its measurability is strongly compromised by severe blending with underlying stellar He I doublet at 8649 Å. The two weak DIBs at 8531 and 8572 Å do not appear real and should actually be blends of underlying stellar lines. The very weak DIB at 8439 Å cannot be resolved within the profile of the much stronger underlying hydrogen Paschen 18 stellar line.

Context. The measurement of the scale of the baryon acoustic oscillations (BAO) in the galaxy power spectrum as a function of redshift is a promising method to constrain the equation-of-state parameter of the dark energy w.

We analyze the distribution of G and K type stars towards the Galactic poles using RAVE and ELODIE radial velocities, 2MASS photometric star counts, and UCAC2 proper motions. The combination of photometric and 3D kinematic data allows us to disentangle and describe the vertical distribution of dwarfs, sub-giants and giants and their kinematics.
We identify discontinuities within the kinematics and magnitude counts that separate the thin disk, thick disk and a hotter component. The respective scale heights of the thin disk and thick disk are 225 ±10 pc and 1048 ±36 pc. We also constrain the luminosity function and the kinematic distribution function. The existence of a kinematic gap between the thin and thick disks is incompatible with the thick disk having formed from the thin disk by a continuous process, such as scattering of stars by spiral arms or molecular clouds. Other mechanisms of formation of the thick disk such as “created on the spot” or smoothly “accreted” remain compatible with our findings.

We present a study of the lens properties of quadruply imaged systems, lensed by numerically simulated galaxies. We investigate a simulated elliptical and disc galaxy drawn from high resolution simulations of galaxy formation in a concordance ΛCDM universe. The simulations include the effects of gas dynamics, star formation and feedback processes. Flux-ratio anomalies observed in strong gravitational lensing potentially provide an indicator for the presence of mass substructure in lens galaxies as predicted from CDM simulations. We particularly concentrate on the prediction that, for an ideal cusp caustic, the sum of the signed magnifications of the three highly magnified images should vanish when the source approaches the cusp. Strong violation of this cusp relation indicates the presence of substructure, regardless of the global, smooth mass model of the lens galaxy. We draw the following conclusions: (1) the level of substructure present in simulations produces violations of the cusp relation comparable to those observed; (2) higher-order catastrophes (e.g. swallowtails) can also cause changes of the order of 0.6 in the cusp relation as predicted by a smooth model; (3) the flux anomaly distribution depends on the image parity and flux and both the brightest minimum and saddle-point images are more affected by substructure than the fainter images. In addition, the brightest saddle point is demagnified w.r.t. the brightest minimum. Our results are fully numerical and properly include all mass scales, without making semi-analytic assumptions. They are ultimately limited by the mass resolution of single particles in the simulation determined by current computational limits, however show that our results are not affected by shot-noise due to the finite number of particles.

High-resolution radio measurements of air showers—cascades of secondary particles in the atmosphere initiated by cosmic rays—reveal that cosmic rays with energies of 1017–1017.5electronvolts have a mixed composition, with light elements (protons and helium nuclei) making up 80 per cent of their mass.

LOFAR is a new and sensitive radio interferometer that can be used for dynamic high-resolution imaging spectroscopy at low radio frequencies from 10 to 90 and 110 to 250 MHz. Here we describe its usage for observations of the Sun and in particular of solar radio bursts. We also describe the processing, archiving and accessing of solar LOFAR data, which is accomplished via the LOFAR Solar Imaging Pipeline and the LOFAR Solar Data Center.

In this work we investigate the gravitationally lensed system B1422+231. High-quality VLBI image positions, fluxes and shapes as well as an optical HST lens galaxy position are used. First, two simple and smooth models for the lens galaxy are applied to fit observed image positions and fluxes; no even remotely acceptable model was found. Such models also do not accurately reproduce the image shapes. In order to fit the data successfully, mass substructure has to be added to the lens, and its level is estimated. To explore expectations about the level of substructure in galaxies and its influence on strong lensing, N-body simulation results of a model galaxy are employed. By using the mass distribution of this model galaxy as a lens, synthetic data sets of different four image system configurations are generated and simple lens models are again applied to fit them. The difficulties in fitting these lens systems turn out to be similar to the case of some real gravitationally lensed systems, thus possibly providing evidence for the presence and strong influence of substructure in the primary lens galaxy.

We present an inventory ofthe Carte du Ciel CdC plates stored in the Astrophysical Institute Potsdam. The Potsdam CdC zone 32° to 39° was divided into 1232 areas and about 2200 plates from the first and second epochs were obtained within the framework of the CdC project. At present, only 977 plates 45 of all are stored in AIP, the others got lost during the Second World War. The plates for the first epoch measurements had been obtained during the period 1893 May– 1900 February. The plates for the second epoch 1913 August–1924 February can be separated into two time intervals according to the observer and the observing method used: from 1913 August till 1914 July, and from 1916 February to 1924 February. The present work aims to provide online access to the plate information, given in the plate catalogue and is the first step to online access to the plate images digitized with flatbed scanners.

The Hartree−Fock method, two common density functionals (PBE and B3LYP), and two new functionals (B97-D and B2PLYP) together with very large AO basis sets are used to compute the isomerization energies for substituted (RH, F, Cl) branched to linear alkanes and silanes. The results of accurate SCS-MP2 computations are taken as reference. These reactions are an important test of how nonlocal electron correlation effects on medium-range lengths scales in saturated molecules are treated by approximate quantum chemical methods. It is found that the unacceptably large errors observed previously for hydrocarbons persist also for the here considered more polar systems. Although the B97-D and B2PLYP functionals provide improved energetics, the problem is not fully solved, and thus these systems are suggested as mandatory benchmarks for future density functionals.

Visible light irradiation of 5-acyloxymethyl- and 5-phenoxymethyl-2-pyrrolidino-1,4-benzoquinones effects photoisomerization to labile oxazolidines, which undergo elimination of carboxylate or phenolate leaving groups in high yields to generate trappable o-quinone methide intermediates.

Photolysis of α-keto amides bearing 4-YC6H4O leaving groups at the position α to the keto group efficiently produces high yields of phenols when Y is an electron-withdrawing group or H. The photoelimination likely involves cleavage of zwitterionic intermediates produced via excited-state hydrogen transfer. When Y is an electron-donating group, competing excited-state ArO−Cα bond scission to radicals occurs, followed by recombination to give 1,3-photorearrangment products.

Carboxylate groups incorporated at the position α to the keto carbonyl of α-keto amides 1 were photochemically cleaved in aqueous media to give carboxylic acids in 70−90% yields with quantum yields of 0.3. The cleavage coproducts were diastereomeric hemiacetals 2. Prompt release of acetate and γ-aminobutyrate (GABA) in buffer was observed by difference FT-IR spectroscopy upon 355 nm laser flash photolysis. The time-constant for release of GABA was <30 ms.

We previously reported that a helical trigger segment within the GCN4 leucine zipper monomer is indispensable for the formation of its parallel two-stranded coiled coil. Here, we demonstrate that the intrinsic secondary structure of the trigger site is largely stabilized by an intrahelical salt bridge. Removal of this surface salt bridge by a single amino acid mutation induced only minor changes in the backbone structure of the GCN4 leucine zipper dimer as verified by nuclear magnetic resonance. The mutation, however, substantially destabilized the dimeric structure. These findings support the proposed hierarchic folding mechanism of the GCN4 coiled coil in which local helix formation within the trigger segment precedes dimerization.

Um die natürliche ultraviolette (UV) Strahlung der Sonne kontinuierlich erfassen und strahlenhygienisch bewerten zu können, wurde vom Bundesamt für Strahlenschutz gemeinsam mit dem Bundesumweltamt ein UV-Messnetz aufgebaut. Seit 1993 wird an vier Standorten in Deutschland die bodennahe solare UV-Strahlung alle 6 Minuten mit Spektralradiometern im Wellenlängenbereich von 290 bis 400 nm gemessen. Da dem Erythem (Sonnenbrand) unter strahlenhygienischen Gesichtspunkten eine besondere Bedeutung zukommt, wird die erythemale Bestrahlungsstärke als Maß für die biologische Wirkung einer UV-Exposition verwendet. Zu diesem Zweck wird die gemessene spektrale Bestrahlungsstärke mit der relativen erythemalen Wichtungsfunktion multipliziert und die resultierende Funktion Uber den genannten Wellenlängenbereich integriert. In den Sommermonaten Juni und Juli des Jahres 1999 überstieg die erythemale Bestrahlungsstärke im Süden Deutschlands regelmäßig einen Wert von 175 mW/m2. Im Norden wurden in diesem Zeitraum immerhin noch Werte von etwa 160 mW/m2gemessen. Dies entsprach einem UV-Index von mehr als 7 bzw. 6 - einer nach der Klassifikation der Strahlenschutzkommission hohen UV-Belastung, für die Schutzmaßnahmen (unbedingt) empfohlen werden. Zwischen 1995 und 1999 wurde an den vier Standorten ein leichter Anstieg der bodennahen UV-Strahlung von etwa 10 % beobachtet. Allerdings reicht die Messperiode von 5 Jahren aufgrund der vielfältigen Einflussgrößen noch nicht aus, um einen ozonbedingten Effekt sicher nachzuweisen.

Background Initiation of @b-blocker therapy is often limited by worsening congestive heart failure, which may manifest as worsening hemodynamics. Deleterious hemodynamic effects might be mitigated with the vasodilation of combined calcium channel/@b-blocker therapy. Methods and Results This prospective, randomized study assessed the safety and efficacy of metoprolol alone or combined with amlodipine on hemodynamic parameters at baseline, 2 hours after the first dose of study medication, and after 12 weeks of therapy in patients receiving background triple therapy for mild to severe heart failure. Functional, exercise, and hormonal status were assessed at baseline and end of study. Twenty-nine patients (mean age 50 +/- 12.1 years) were enrolled; 21 completed 12 weeks of treatment. Mean ejection fraction at baseline was 13.4% +/- 5.7%; 79% of patients had heart failure classified as New York Heart Association class III, and 66% had heart failure of idiopathic origin. Heart rate and blood pressure did not change with short-term therapy in either group. The first dose of both regimens produced significant increases in systemic vascular resistance and significant decreases in cardiac output and index and stroke volume and stroke work indexes; combination therapy acutely yielded small but statistically significant increases in pulmonary artery, pulmonary capillary wedge, and right atrial pressures. Long-term therapy with both regimens produced significant decreases in heart rate, systemic vascular resistance, and pulmonary capillary wedge pressure and significant increases in cardiac output and index and stroke volume and stroke work indexes. Combination therapy produced significant long-term decreases in blood pressure. Conclusions There was no further measurable benefit with the addition of amlodipine to metoprolol compared with the effects of metoprolol alone. Therapy with metoprolol alone and the combination of metoprolol and amlodipine was well tolerated in patients with mild to severe heart failure, as evidenced by a lack of adverse effects on hemodynamic parameters over the short term and clinical and hemodynamic improvement with long-term treatment. (Am Heart J 1999;138:261-8.)

A clone bearing the structural gene sacB, coding for the exoenzyme levansucrase, was isolated from a library of Bacillus subtilis DNA that was cloned in phage lambda charon 4A on the basis of the transforming activity of the chimeric DNA. This lambda clone also was found to contain the sacR and smo loci. Subcloning the sacB-sacR region in plasmid pBR325 resulted in a clone which directed levansucrase synthesis in Escherichia coli. The nucleotide sequence coding for the secreted protein was localized on the physical map of the cloned DNA.

Mutations affecting the genes involved in B. subtilis D-glucitol catabolism were mapped either by PBS1-mediated transduction or DNA-mediated transformation. It was shown that the genes gutA and gutB coding for the D-glucitol permease and the D-glucitol dehydrogenase, respectively, and regulatory locus gutR are clustered in a gut operon localized between purB and dal close to the pha marker. A mutation affecting fructokinase activity (fruC) was mapped near the gut markers. The fruC gene does not belong to the operon. A mutation affecting phosphofructokinase activity (pfk) was mapped between the leuA and aroG markers.

The K, I and S regions of the mouse major histocompatibility complex (MHC) are composed of long tracts of DNA which differ in sequence divergence. A correlation exists between the location of an MHC gene in a variable or conserved chromosomal tract and the degree of polymorphism and diversity of the proteins encoded by its alleles. Variable tracts appear to be the result of mechanisms which mutate certain coding and non‐coding sequences to the same extent and selective pressures operating on the genes.

Several inbred strains and a certain percentage of wild mice bear a deletion in the E alpha gene of the mouse major histocompatibility complex (H‐2). This mutation prevents transcription of the E alpha gene and hence functional expression of the E alpha E beta dimer on the cell surface. Two strains were selected for a more precise localization of this deletion. BALB.B is a congenic line carrying the H‐2b haplotype on the BALB/c background. CRO435 is an outbred stock derived from a wild mouse captured near Cairo, Egypt; it carries the H‐2w37 haplotype including a null Ew28 alpha allele, as well as semi‐lethal mutations in the H‐2 linked t complex (tTuw7). From these two strains, we have isolated genomic clones that contain fragments spanning the E alpha deletion, and have sequenced the breakpoint region. The deletions in the two strains are identical, spanning 627 bp which include the promoter region and the signal peptide exon of the E alpha gene. Limited sequence comparison suggests that the Eb alpha allele of BALB.B is more closely related to the Ew28 alpha allele of CRO435 than both of these are to an E alpha‐expressor allele, Ed alpha. It is therefore likely that the Eo deletions in the various inbred strains and wild mice are of the same origin, and we propose that they have been disseminated throughout the mouse population because of linkage to the t complex.

We have analyzed the T-cell-receptor repertoire expressed in the synovial fluid of a patient with rheumatoid arthritis by using an inverse polymerase chain reaction. Total RNA was isolated from Ficoll-purified mononuclear cells and converted into circularized double-stranded cDNA. Specific amplification of alpha- and beta-chain variable regions (V alpha and V beta) was achieved with inverted alpha- and beta-chain constant region (C alpha and C beta) primer pairs, and the amplification products were cloned into phage vectors. A total of 78 alpha and 76 beta clones were sequenced, and 67 and 72 productively rearranged alpha and beta genes were identified, respectively. Thirty-one V alpha, 33 alpha-chain joining region (J alpha), 29 V beta, and 12 beta-chain joining region (J beta) gene segments were found in the productively rearranged clones, indicating that the T-cell repertoire expressed in the synovial fluid of this RA patient is highly heterogenous and polyclonal. Comparison of peripheral blood and synovial fluid repertoires showed that the most abundant V beta sequences, V beta 2.1 and V beta 3.1, were enriched in the inflamed joint by a factor of 2 to 3. It is possible that T cells expressing these V beta gene segments, which recognize bacterial superantigens, play a role in the disease.

Human gamma heavy chain disease (HCD) is characterized by the presence in serum of a short monoclonal Ig gamma chain unattached to light chains. Although most HCD proteins have internal deletions, in some the defect is NH2-terminal. The OMM gamma 3 HCD serum protein is of the latter type, having undergone an extensive NH2-terminal deletion with a sequence starting within the hinge. A cell line synthesizing the OMM protein has enabled us to study the biogenesis of the abnormal molecule. In vitro translation of isolated mRNA yields a protein containing a hydrophobic NH2-terminal leader sequence. In the intact cell, the precursor molecule is processed normally to yield a protein with an NH2-terminal sequence homologous to the beginning of the variable (V) region. The nucleotide sequence of cDNA prepared from the OMM mRNA encodes a 19-amino acid leader followed by the first 15 residues of the V region. An extensive internal deletion encompasses the remainder of the V and the entire CH1 domain. Immediately following the short V region, there is information in the cDNA for the entire normal hinge. The primary synthetic product is thus an internally deleted molecule that undergoes postsynthetic degradation to yield the NH2-terminally deleted serum protein. The structure of the OMM mRNA suggests that the protein abnormality results from a partial gene deletion rather than defective splicing.

A TL+ leukemia of a (B6 X A)F1 hybrid mouse (H-2b/H-2a) was previously subjected to immunoselection against H-2a by passage in (B6 X A.SW)F1 mice (H-2b/H-2s). A variant leukemia line was obtained that serologically lacked not only the H-2a phenotype but also the TL phenotype determined by the linked cis Tlaa allele of strain A. The H-2b phenotype and the TL phenotype of the Tlab allele of the B6 strain, which is expressed only by leukemia cells, were retained by the variant. Southern blotting with an H-2 cDNA probe that identifies restriction fragment polymorphisms distinguishing alleles of the H-2 and Tla regions of the B6 and A strains indicates that both the H-2a and Tlaa alleles are missing from the genome of this H-2a:Tlaa negative variant. Since the variant has two apparently unaltered chromosomes 17, where the H-2:Tla complex is situated, and since the intensity of bands in Southern blotting is suggestive of H-2b homozygosity, it is considered that loss of the H-2a:Tlaa haplotype by the variant was accompanied by duplication of the H-2b:Tlab haplotype. The implied change from heterozygosity to homozygosity that the variant has undergone with respect to H-2:Tla was not paralleled by a similar change at the three other loci tested, since the variant retained heterozygosity for Pep-3 (chromosome 1), Gpi-1 (chromosome 7), and Es-1 (chromosome 8).

We have cloned and sequenced a meiotic recombinational hotspot between the A beta 3 and A beta 2 genes in the major histocompatibility complex (MHC) of the mouse. This recombinational hotspot in the Mus musculus castaneus cas3 haplotype was previously localized to a region of 9.5 kb of DNA in which five independent crossing‐over events occurred at the unusually high frequency of 0.6%. Aside from cas3, the hotspot appears to be absent in many other MHC haplotypes. We have now confined the five recombinational breakpoints to a stretch of 3.5 kb of DNA. From the nucleotide sequence around the recombinational breakpoints, determined in the parental cas3 and b haplotypes as well as for two recombinant haplotypes, we show that the two recombinant haplotypes were generated by homologous equal crossing‐over and place the breakpoints within two non‐overlapping stretches of 10 and 36 bp, respectively. Comparison of the DNA sequences of the hotspot‐positive cas3 and the hotspot‐negative b haplotypes reveals a number of differences, in particular, a CAGA‐repeat sequence which is present in CAS3 in six, but only four copies in C57BL/6 DNA. This repeat sequence is reminiscent of one in a previously characterized hotspot in the E beta gene.

Clones coding for the mouse neural cell adhesion molecule (N‐CAM) were isolated from a cDNA library prepared in the expression vector lambda gt 11 from mRNA extracted from a mouse neuroblastoma cell line. This library was screened with two anti‐N‐CAM monoclonal antibodies directed against different sites on the molecule and with rabbit anti‐N‐CAM serum. Two clones were identified with the first monoclonal antibody, three with the second one, none reacted with both. The relevance of these cDNA clones to N‐CAM was confirmed by several observations. First, cDNA sequences detected with one monoclonal antibody cross‐hybridized with those identified by the other antibody. Second, the different fusion proteins all bound the rabbit serum in addition to one monoclonal antibody. Finally, the probes hybridized to discrete mRNA species of sufficient lengths to code for the very large N‐CAM polypeptides in RNA preparations from N‐CAM‐expressing, but not from N‐CAM‐negative cells. An additional mRNA species not seen in embryonic brain was expressed in adult mouse brain. Genomic blot experiments indicated that sequences corresponding to one of our probes are present only a few times in the mouse genome.

Over the past few years six gene clusters have been isolated from the major histocompatibility complex (MHC) of the BALB/c mouse encompassing a total of 1600 kb of DNA and 48 genes. The molecular distances between these gene clusters and the orientation of four of the six clusters on chromosome 17 is not known. Here we use pulse‐field gradient gels and Southern blot hybridization to establish large‐scale genomic restriction maps covering several hundreds of kb surrounding the three gene clusters located in the K, I, S, and D regions of the MHC. Comparison of the maps orients the complement gene clusters in the S region with the 21‐OHB gene pointing towards the K end and the C2 gene pointing towards the D end of the MHC. The distances between the E alpha and 21‐OHB genes is 430 kb and between the C2 and TNF‐alpha genes at least 420 kb.

The murine major histocompatibility antigens are cell surface glycoproteins which play an important role in the recognition of foreign antigens by cytotoxic T lymphocytes. Modulation of the level of expression of histocompatibility antigens could therefore be useful for the study of the interaction between the antigen presenting cells and T lymphocytes. The glucocorticoid hormone‐inducible promoter, located in the long terminal repeat of mouse mammary tumor virus, was used to replace the promoter region of a cloned H‐2Ld class I gene. The chimeric gene was introduced into cultured cells. Glucocorticoid induction of MMTV LTR H‐2Ld mRNA could be shown by blot analysis. An S1 nuclease protection assay indicated that the transfected cells accurately initiate the chimeric mRNA. Immunoprecipitation of H‐2Ld protein with a specific monoclonal antibody showed inducibility also at the cellular protein level. Fluorescence‐activated cell sorter analysis monitored a 3‐fold increase of H‐2Ld on the cell surface when the transfected cells were grown in the presence of dexamethasone. This increase of H‐2Ld expression was accompanied by a corresponding decrease on the cell surface of the endogenous H‐2Kk.

To study rearrangement of T cell receptor (TCR) genes, transgenic mice were generated with a TCR beta minilocus in germline configuration, containing three V beta, two D beta, fourteen J beta and two C beta gene segments and the TCR beta enhancer. Using the polymerase chain reaction as an analytical tool both partial DJ as well as complete VDJ rearrangements were seen, indicating that the minilocus contained all sequence elements required for rearrangment. Rearrangements of minilocus gene segments were restricted to T cells in the thymus and the periphery and did not occur in B cells. V beta 8.3 and V beta 5 sequences encoded by the minilocus were expressed on the surface of peripheral T cells at high frequencies. Transgenic mice with TCR minilocus genes will be a useful system to identify DNA sequence elements required for regulation of rearrangement in vivo.

SWR/J transgenic (tg) mice were generated expressing the TCR beta chain derived from an anticollagen type II (CII) arthritogenic T cell clone. The SWR/J strain was selected because it is resistant to collagen-induced arthritis (CIA) and lacks the V beta gene segment used by the T cell clone. Expression of the tg beta chain on all thymocytes and peripheral lymph node T cells led to a more efficient anti-CII immune response, but did not confer CIA susceptibility to SWR/J mice. Nevertheless, this tg beta chain enhanced predisposition to CIA as (DBA/1 x SWR) F1 beta tg mice were more susceptible than normal F1 littermates. Our results demonstrate that the expression of the tg beta chain contributes to CIA susceptibility, but by itself it is not sufficient to overcome CIA resistance in the SWR/J strain.

Labeled cDNA probes highly enriched for B-cell-specific (B-T) and T-suppressor-cell-specific (Ts-B) sequences were used to screen a set of genomic cosmid clones spanning 230 kilobases of the murine immune response (I) region. With the B-cell derived probe, four I-region genes were detectable (A beta, E beta, E beta 2, and E alpha), as well as an additional (fifth) region of hybridization. The T-cell probe, prepared from a putative I-J positive suppressor cell hybridoma, was negative in a parallel experiment. A genomic fragment corresponding to the new region of hybridization seen with the B-cell cDNA probe identified a discrete mRNA species in RNA blotting analysis that had a pattern of expression strikingly similar to E alpha mRNA in a variety of lymphoid tumor lines. This fragment was also used to isolate cDNA clones from a library highly enriched (20-40 times) for B-cell-specific sequences (selected cDNA cloning). DNA sequence analysis of one of these cDNA clones indicates that this gene encodes the A alpha molecule of the d haplotype (Ad alpha). These findings establish that the order of the class II genes in the Ia complex is as follows: centromere-A beta-A alpha-E beta-E beta 2-E alpha.

Induction of the Bacillus subtilis sacB gene and sacPA operon and Escherichia coli bgl operon is mediated by structurally homologous antiterminators encoded by the sacY, sacT, and bglG genes, respectively. When activated, these proteins prevent early transcription termination at terminators located in the leader regions of the three operons. BglG was previously shown to bind in vitro to an imperfectly palindromic 29-nucleotide RNA sequence located upstream of the terminator and partially overlapping with it [Houman, F., Diaz-Torres, M.R. & Wright, A. (1990) Cell 62, 1153-1163]. Similar motifs, here termed ribonucleic antiterminators (RATs), strongly conserved in sequence and in position, are found in the leader of both sacB and sacPA. Mutations were created in sacB RAT and tested in B. subtilis; this showed that sacB RAT is the target for SacY-mediated induction of sacB and that a stem-loop structure in the mRNA is required for regulatory function. Mutations increasing the similarity of the sacB RAT with those of sacPA or bgl rendered sacB inducible by SacT or BglG, respectively; most of these changes did not strongly affect induction by SacY, suggesting that the nucleotides at these variable positions act as negative specificity determinants.

From a genomic library constructed from sperm DNA of the inbred BALB/c mouse, we previously isolated 54 cosmid clones that contain 36 class I genes and can be divided by restriction map analyses into 13 gene clusters. We have isolated single- and low-copy DNA probes from each of these clusters to visualize restriction enzyme site polymorphisms in the DNAs from various congeneic and recombinant congeneic mice. These polymorphisms permit us to map each of the 13 cosmid clusters to a precise location in the major histocompatibility complex of the mouse. Thirty-one of 36 class I genes map into the Tla complex of the major histocompatibility complex whereas the remaining 5 genes map to the H-2 complex. Thus, all 36 class I genes are located in the major histocompatibility complex. Analysis of the number of restriction enzyme fragments visualized by the single- and low-copy DNA probes suggests that the class I genes in different inbred strains of mice probably undergo gene duplications and deletions, presumably by homologous but unequal crossing-over.

The I-J subregion of the mouse major histocompatibility complex has been reported to encode antigenic determinants expressed by suppressor T cells. Previously, cosmid clones were obtained from mouse sperm DNA that contain all of the sequences between the I-A and I-E subregions, where I-J has been mapped genetically. However, hybridization of these sequences to RNA prepared from several I-J-positive suppressor T-cell hybridomas did not reveal the presence of a transcript. In addition, no rearrangements in this DNA were detected in the suppressor T cells that we have analyzed. Our results indicate that the I-J polypeptides are not encoded between the I-A and I-E subregions of the major histocompatibility complex. We discuss several hypotheses concerning the possible location and expression of I-J genes.

Mouse liver cDNA clones related to the C4 and C4-Slp isoforms of the fourth component of complement differ by few nucleotide changes within a region of substantial divergence from human C4. It is suggested that the mouse C4 gene duplication is an evolutionarily recent event with respect to the time of mammalian radiation. This conclusion is reinforced by the presence of a single C4 gene in the Syrian hamster. Most H-2 haplotypes, including those characterized by an undetectable C4-Slp protein, possess two C4 gene copies which, in contrast to the neighboring factor B, show a marked restriction site polymorphism. The genetic variation of this region is emphasized by the presence in the mouse of a rare "polymorphism" for C4 gene number. Multiple C4-related gene copies characterize those exceptional wild-derived H-2 haplotypes, H-2w7, H-2w16, and H-2w19, that determine the expression of the C4-Slp protein in female animals.

The adjacent sacX and sacY genes are involved in sucrose induction of the Bacillus subtilis sacB gene by an antitermination mechanism. sacB, encoding the exoenzyme levansucrase, is also subject to regulation by the DegS-DegU signalling system. Using sacXY'-lacZ and sacX'-lacZ fusions, we show that the transcription of the sacX and sacY genes is both inducible by sucrose and regulated by DegU. sacX and sacY appear to constitute an operon, since the deletion of the sacX leader region abolished the expression of a sacXY'-lacZ fusion. The degU-dependent promoter was located by deletion analysis and reverse transcriptase mapping 300 nucleotides upstream from the sacX initiator codon. Sucrose induction of the sacX'-lacZ fusion requires either SacY or the homologous SacT antiterminator, which is involved in sucrose induction of the intracellular sucrase gene (sacPA operon). Sequence analysis of the sacX leader region revealed (20 nucleotides downstream from the transcription start site) a putative binding site for these regulators; however, no structure resembling a rho-independent terminator could be found overlapping this site, unlike the situation for sacPA and sacB. Deletion of a segment of the leader region located 100 nucleotides downstream from this site led to constitutive expression of the sacXY'-lacZ and sacX'-lacZ fusions. These results suggest that the mechanism of sucrose induction of sacXY is different from that of sacPA and sacB.

The genes ptsI and ptsH, which encode, respectively, enzyme I and Hpr, cytoplasmic proteins involved in the phosphoenolpyruvate:sugar phosphotransferase system, were cloned from Bacillus subtilis. A plasmid containing a 4.1-kilobase DNA fragment was shown to complement Escherichia coli mutations affecting the ptsH and ptsI genes. In minicells this plasmid expressed two proteins with the molecular weights expected for Hpr and enzyme I. Therefore, ptsH and ptsI are adjacent in B. subtilis, as in E. coli. In E. coli a third gene (crr), involved in glucose translocation and also in catabolite repression, is located downstream from the ptsHI operon. The 4.1-kilobase fragment from B. subtilis was shown to contain a gene that enables an E. coli crr mutant to use glucose. This gene, unlike the E. coli crr gene, was located to the left of ptsH.

The target of the induction by sucrose of the levansucrase gene is a transcription terminator (sacRt) located upstream from the coding sequence, sacB. The two-gene locus sacX-sacY (formerly sacS) and the ptsI gene were previously shown to be involved in this induction. ptsI encodes enzyme I of the phosphoenolpyruvate-dependent phosphotransferase system. SacX is strongly homologous to sucrose-specific phosphotransferase system-dependent permeases. SacY is a positive regulator of sacB. Here we show that SacY is probably an antiterminator interacting directly with sacRt, since in Escherichia coli the presence of the sacY gene stimulates the expression of a reporter gene fused downstream from sacRt. Missense mutations affecting sacY were sequenced, and the sacB regulation was studied in isogenic strains carrying these mutations or in vitro-generated mutations affecting sacX, sacY, or ptsI. The phenotype of double mutants suggests a model in which SacX might be a sucrose sensor that would be phosphorylated by the phosphotransferase system and, in this state, could inhibit the SacY antiterminator. Exogenous sucrose, or a mutation inactivating the phosphotransferase system, would dephosphorylate SacX and allow antitermination at sacRt.

Sucrose induces two saccharolytic enzymes in Bacillus subtilis, an intracellular sucrase and an extracellular levansucrase, encoded by sacA and sacB, respectively. It was previously shown that the sacY gene encodes a positive regulator involved in a sucrose-dependent antitermination upstream from the sacB coding sequence. We show here that the sacY product is not absolutely required for sacB induction: a weak but significant induction can be observed in strains harboring a sacY deletion. The sacY-independent induction was altered by mutations located in the sacP and sacT loci but was observed in both sacU+ and sacU32 genetic backgrounds. These results suggest that B. subtilis has two alternative systems allowing sacB induction by sucrose. Both systems also seem to be involved in sacA induction.