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STFC are acknowledged for supporting the “Curation and Preliminary Examination of the Winchcombe Carbonaceous Chondrite Fall” project (ST/V000799/1), and Natural History Museum staff for curatorial support. Oxygen isotope studies at the Open University are funded by a consolidated grant from the Science and Technology Facilities Council (STFC), UK GRANT NUMBER: ST/T000228/1 (IAF, RCG, JM, MA), and STFC studentship NUMBER: ST/S505614/1 (RF). As part of an integrated consortium study, we have undertaken O, Cd, Cr, Si, Te, Ti, and Zn whole rock isotopic measurements of the Winchcombe CM2 meteorite. δ66Zn values determined for two Winchcombe aliquots are +0.29 ± 0.05‰ (2SD) and +0.45 ± 0.05‰ (2SD). The difference between these analyses likely reflects sample heterogeneity. Zn isotope compositions for Winchcombe show excellent agreement with published CM2 data. δ114Cd for a single Winchcombe aliquot is +0.29 ± 0.04‰ (2SD), which is close to a previous result for Murchison. δ130Te values for three aliquots gave indistinguishable results, with a mean value of +0.62 ± 0.01‰ (2SD) and are essentially identical to published values for CM2s. ε53Cr and ε54Cr for Winchcombe are 0.319 ± 0.029 (2SE) and 0.775 ± 0.067 (2SE), respectively. Based on its Cr isotopic composition, Winchcombe plots close to other CM2 chondrites. ε50Ti and ε46Ti values for Winchcombe are 3.21 ± 0.09 (2SE) and 0.46 ± 0.08 (2SE), respectively, and are in line with recently published data for CM2s. The δ30Si composition of Winchcombe is −0.50 ± 0.06‰ (2SD, n = 11) and is essentially indistinguishable from measurements obtained on other CM2 chondrites. In conformity with petrographic observations, oxygen isotope analyses of both bulk and micromilled fractions from Winchcombe clearly demonstrate that its parent body experienced extensive aqueous alteration. The style of alteration exhibited by Winchcombe is consistent with relatively closed system processes. Analysis of different fractions within Winchcombe broadly support the view that, while different lithologies within an individual CM2 meteorite can be highly variable, each meteorite is characterized by a predominant alteration type. Mixing of different lithologies within a regolith environment to form cataclastic matrix is supported by oxygen isotope analysis of micromilled fractions from Winchcombe. Previously unpublished bulk oxygen isotope data for 12 CM2 chondrites, when combined with published data, define a well‐constrained regression line with a slope of 0.77. Winchcombe analyses define a more limited linear trend at the isotopically heavy, more aqueously altered, end of the slope 0.77 CM2 array. The CM2 slope 0.77 array intersects the oxygen isotope field of CO3 falls, indicating that the unaltered precursor material to the CMs was essentially identical in oxygen isotope composition to the CO3 falls. Our data are consistent with earlier suggestions that the main differences between the CO3s and CM2s reflect differing amounts of water ice that co‐accreted into their respective parent bodies, being high in the case of CM2s and low in the case of CO3s. The small difference in Si isotope compositions between the CM and CO meteorites can be explained by different proportions of matrix versus refractory silicates. CMs and COs may also be indistinguishable with respect to Ti and Cr isotopes; however, further analysis is required to test this possibility. The close relationship between CO3 and CM2 chondrites revealed by our data supports the emerging view that the snow line within protoplanetary disks marks an important zone of planetesimal accretion. Publisher PDF

The second-phonon ( n ω = 2 ) wobbling band has been established in the nucleus 135Pr. Conclusive evidence for its wobbling nature comes from the Δ I = 1 , E2 character of the transitions between the new band and the previously identified transverse wobbler band ( n ω = 1 ) in this nucleus. Theoretical calculations in the framework of the quasiparticle triaxial rotor and triaxial projected shell models are found to be in good agreement with the experimental results.

The CNO cycle is the main energy source in stars more massive than our sun, it defines the energy production and the cycle time that lead to the lifetime of massive stars, and it is an important tool for the determination of the age of globular clusters. One of the largest uncertainties in the CNO chain of reactions comes from the uncertainty in the $^{14}$N$(p,\gamma)^{15}$O reaction rate. This uncertainty arises predominantly from the uncertainty in the lifetime of the sub-threshold state in $^{15}$O at $E_{x}$ = 6792 keV. Previous measurements of this state's lifetime are significantly discrepant. Here, we report on a new lifetime measurement of this state, as well as the excited states in $^{15}$O at $E_{x}$ = 5181 keV and $E_{x}$ = 6172 keV, via the $^{14}$N$(p,\gamma)^{15}$O reaction at proton energies of $E_{p} = 1020$ keV and $E_{p} = 1570$ keV. The lifetimes have been determined with the Doppler-Shift Attenuation Method (DSAM) with three separate, nitrogen-implanted targets with Mo, Ta, and W backing. We obtained lifetimes from the weighted average of the three measurements, allowing us to account for systematic differences between the backing materials. For the 6792 keV state, we obtained a $\tau = 0.6 \pm 0.4$ fs. To provide cross-validation of our method, we measured the known lifetimes of the states at 5181 keV and 6172 keV to be $\tau = 7.5 \pm 3.0$ and $\tau = 0.7 \pm 0.5$ fs, respectively, which are in good agreement with previous measurements., Comment: 10 pages, 13 figures

Quasi-elastic scattering measurements have been performed using 16O and 24Mg projectiles off 90Zr at energies around the Coulomb barrier. Experimental data have been analyzed in the framework of coupled channels (CC) calculations using the code CCFULL. The quasi-elastic scattering excitation function and derived barrier distribution for 16O + 90Zr reaction are well reproduced by the CC calculations using the vibrational coupling strengths for 90Zr reported in the literature. Using these vibrational coupling strengths, a Bayesian analysis is carried out for 24Mg + 90Zr reaction. The β 2 and β 4 values for 24Mg are determined to be + 0.43 ± 0.02 and − 0.11 ± 0.02 , respectively. The β 2 parameter determined in the present work is in good agreement with results obtained using inelastic scattering probes. The hexadecapole deformation of 24Mg has been measured very precisely for the first time. Present results establish that quasi-elastic scattering could provide a useful probe to determine the ground state deformation of atomic nuclei.

Carbon and oxygen burning reactions, in particular, $^{12}\mathrm{C}+^{12}\mathrm{C}$ fusion, are important for the understanding and interpretation of the late phases of stellar evolution as well as the ignition and nucleosynthesis in cataclysmic binary systems such as type Ia supernovae and x-ray superbursts. A new measurement of this reaction has been performed at the University of Notre Dame using particle-$\ensuremath{\gamma}$ coincidence techniques with SAND (a silicon detector array) at the high-intensity 5U Pelletron accelerator. New results for $^{12}\mathrm{C}+^{12}\mathrm{C}$ fusion at low energies relevant to nuclear astrophysics are reported. They show strong disagreement with a recent measurement using the indirect Trojan Horse method. The impact on the carbon burning process under astrophysical scenarios will be discussed.

Background Oral squamous cell carcinomas (OSCC) secrete exosomes into the surrounding extracellular environment to promote the horizontal transfer of bioactive molecules including microRNA (miRNA). The primary objective of this study was to explore potential differences in miRNA content between OSCC and OSCC stem cells. Methods The OSCC cell lines SCC4, SCC15, SCC25 and CAL27 were used in these studies. The corresponding OSCC stem cells that demonstrated phenotypic adhesion independent tumor spheres (AiTS) were manually isolated. All cells were cultured in DMEM containing 10% exosome-free fetal bovine serum. Exosomes were isolated using Total Exosome Isolation reagent (Invitrogen) and RNA was purified using Total Exosome RNA isolation kit (Invitrogen). Exosome miRNA content was evaluated using miRNA Advanced Taqman Assays for miR-21, − 155, − 133, − 34, − 31, − 32, and − 365. The fold change of miRNA content was calculated using the comparative CT (ΔΔCT) method using miR-16 as an endogenous control. Results After successful cell cultures were established, AiTS (cancer stem cells) were manually separated and confirmed using CD133 and Sox-2 biomarkers. Exosomes and extracellular vesicles were successfully isolated from all cell lines and AiTS isolates for miRNA screening. All isolates exhibited miR-16 expression (positive control), but none contained mir-31, − 32, or 133a. Differential expression of miR-21, miR-34 and miR-155 were observed with patterns observed among the cancer cell lines which were distinct from the corresponding AiTS isolates. Conclusions Exosomes isolated from these different OSCC stem cell populations displayed nearly consistent downregulation/loss of miR-21 and miR-34 suggesting the possibility of a unique miRNA profile characteristic of oral cancer stem cells. These findings highlight the need to investigate the comprehensive functions of miR-21 and miR-34 in tumor progression and continued research to refine a miRNA profile that could aid in distinguishing tumors with poor prognosis.

Carbon and oxygen burning reactions, in particular, $^{12}$C+$^{12}$C fusion, are important for the understanding and interpretation of the late phases of stellar evolution as well as the ignition and nucleosynthesis in cataclysmic binary systems such as type Ia supernovae and x-ray superbursts. A new measurement of this reaction has been performed at the University of Notre Dame using particle-$\gamma$ coincidence techniques with SAND (a silicon detector array) at the high-intensity 5U Pelletron accelerator. New results for $^{12}$C+$^{12}$C fusion at low energies relevant to nuclear astrophysics are reported. They show strong disagreement with a recent measurement using the indirect Trojan Horse method. The impact on the carbon burning process under astrophysical scenarios will be discussed., Comment: accepted for publication in Phys. Rev. Lett

Isoscalar giant resonances, being the archetypal forms of collective nuclear behavior, have been studied extensively for decades with the goal of constraining bulk nuclear properties of the equation of state, as well as for modeling dynamical behaviors within stellar environments. An important such mode is the isoscalar electric giant monopole resonance (ISGMR) that can be understood as a radially symmetric density vibration within the saturated nuclear volume. The field has a few key open questions, which have been proposed and remain unresolved. One of the more provocative questions is the extra high-energy strength in the $A\approx 90$ region, which manifested in large percentages of the $E0$ sum rule in $^{92}$Zr and $^{92}$Mo above the main ISGMR peak. The purpose of this article is to introduce these questions within the context of experimental investigations into the phenomena in the zirconium and molybdenum isotopic chains, and to address, via a discussion of previously published and preliminary results, the implications of recent experimental efforts on extraction of the nuclear incompressibility from this data., 9 pages, 7 figures, invited to be submitted to a special issue of EPJA honoring Prof. P. F. Bortignon

"Why are the tin isotopes soft?" has remained, for the past decade, an open problem in nuclear structure physics: models which reproduce the isoscalar giant monopole resonance (ISGMR) in the "doubly-closed shell" nuclei, $^{90}$Zr and $^{208}$Pb, overestimate the ISGMR energies of the open-shell tin and cadmium nuclei, by as much as 1 MeV. In an effort to shed some light onto this problem, we present results of detailed studies of the ISGMR in the molybdenum nuclei, with the goal of elucidating where--and how--the softness manifests itself between $^{90}$Zr and the cadmium and tin isotopes. The experiment was conducted using the $^{94,96,98,100}$Mo($\alpha,\alpha^\prime$) reaction at $E_\alpha = 386$ MeV. A comparison of the results with relativistic, self-consistent Random-Phase Approximation calculations indicates that the ISGMR response begins to show softness in the molybdenum isotopes beginning with $A=92$., Comment: Accepted for publication to Physics Letters B

The isoscalar giant monopole, dipole, and quadrupole strength distributions have been deduced in $^{90,92}\mathrm{Zr}$ and $^{92}\mathrm{Mo}$ from ``background-free'' spectra of inelastic $\ensuremath{\alpha}$-particle scattering at a beam energy of 385 MeV at extremely forward angles, including ${0}^{\ensuremath{\circ}}$. These strength distributions were extracted by a multipole-decomposition analysis based on the expected angular distributions of the respective multipoles. All these strength distributions for the three nuclei practically coincide with each other, affirming that giant resonances, being collective phenomena, are not influenced by nuclear shell structure near $A\ensuremath{\sim}90$, contrary to the claim in a recent measurement.

Past research suggests that senior citizens often face challenges related to deteriorating physical and mental health, and the quality of their lives may suffer as a result. Past research also suggests that volunteering can improve the health and quality of life for seniors. In the present study, 451 volunteers enrolled in the Retired and Senior Volunteer Program (RSVP) completed surveys including questions regarding their volunteer experiences and how these experiences have affected their health and quality of life. The results suggest that volunteering through RSVP is associated with improvements in health and quality of life across a variety of dimensions. Furthermore, these improvements may be particularly greater for women, current volunteers, and older seniors. These findings may help guide interventions designed to enhance the health and well-being of senior citizens in a variety of settings.

"Background-free" spectra of inelastic $\alpha$-particle scattering have been measured at a beam energy of 385 MeV in $^{90, 92}$Zr and $^{92}$Mo at extremely forward angles, including 0$^{\circ}$. The ISGMR strength distributions for the three nuclei coincide with each other, establishing clearly that nuclear incompressibility is not influenced by nuclear shell structure near $A\sim$90 as was claimed in recent measurements., Comment: 5 pages, 4 figures; accepted for publication in Phys. Lett. B

First estimates of sex allocation patterns and body size-at-sexual maturity and at protogynous sex change are presented for the five major (including one endemic) species of parrotfishes of Hawaii. Median body size at initial maturation as a female (LM50) and at protogynous sex change from adult female to adult male (LΔ50) varied greatly among the five species. Estimates of LM50 were about 14, 17, 24, 34 and 35 cm fork length (LF) in palenose Scarus psittacus, Pacific bullethead Chlorurus spilurus, stareye Calotomus carolinus, spectacled Chlorurus perspicillatus and redlip parrotfish Scarus rubroviolaceus. Values of LΔ50 were c. 23, 27, 37, 46 and 47 cm LF in the respective species. Length at female maturation was proportional to maximum body size (Lmax) of the respective species, ranging from 50 to 72% and averaging 62% of Lmax across species. LΔ50 was also proportional to Lmax, ranging from 82 to 97% and averaging 92%. Males of both pairs of Scarus and Chlorurus spp. reported here are diandric. Only one of the five major species (C. carolinus) is functionally monandric, with either all or nearly all males secondarily derived from adult females. The broadly differing absolute body sizes at sexual maturation and at sex change among the five species have important implications for improving regulatory size limits for parrotfishes in the State of Hawaii, where parrotfish species have historically been managed based on a single minimum size limit of 30·5 cm LF. This study provides a model demonstration of why catch data for parrotfishes, and other size-structured reef-fish populations, should be recorded either by species or by functional size-groups of species that allow setting more meaningful minimum size limits.

The mechanism of heat extraction from the lower oceanic crust near the ridge axis is poorly constrained despite its importance for understanding both the process of accretion of the plutonic complex and the mass fluxes associated with ridge hydrothermal systems. We have investigated the role of zones of focussed fluid flow in the plutonic complex of the Oman ophiolite in the near-axis cooling of the oceanic crust. Lineaments identified on aerial photographs, that occur at 1 km spacing, show evidence for extensive hydrothermal fluid flow through regions 10 to 50 m wide. Fluid flow is initiated in these regions at 800°C and continues at least into the lower greenschist facies. Strontium-isotope analyses indicate that the fluid flux through these zones is sufficient to transport a metasomatic front from the base of the sheeted dike complex to close to the Moho. Computed minimum fluid fluxes to transport a metasomatic front through the focussed fluid flow zones are 1x108 kgm2. Modeling of diffusive exchange of calcium from olivine to clinopyroxene indicates enhanced cooling rates adjacent to the focussed fluid flow zones. Heat fluxes estimated from the enhanced cooling rates are broadly consistent with the fluid fluxes determined from modeling the Sr-isotopic composition of samples from the focussed fluid flow zones. The combination of independent estimates of the fluid and heat fluxes, such as these, can provide more rigorous constraints on the thermal history than either approach used in isolation. Our results show that focussed fluid flow could play a major role in the cooling in the lower oceanic crust. Significant focussed fluid flow in the lower oceanic crust has important implications for predicting the total mass flux associated with hydrothermal circulation at mid-ocean ridges. This is because fluids flowing through channels become chemically rock-buffered at smaller fluid fluxes than those flowing pervasively through a rock mass. Thus, if focussed fluid flow is an important mechanism of heat loss from the lower oceanic crust the chemical fluxes from ridge hydrothermal systems into the oceans may be smaller than currently thought.

The Workers' Compensation Board of British Columbia requested a retrospective analysis of all fishermen's deaths from immersion in water in British Columbia.To identify the underlying cause of drowning and make recommendations to improve safety in the fishing industry.Eighty-nine inshore and offshore fishing accidents were analysed. Where possible, deaths were classified into the four stages of cold-water immersion: cold shock, swimming failure, hypothermia and post-rescue collapse. Other factors that led up to the drowning were also identified.One hundred and thirty fishermen died from immersion between 1976 and 2002. One hundred and twenty-eight drownings were certified by the coroner as drowning or drowning/hypothermia and two were certified as cardiac event after immersion. The underlying causes of drownings were reclassified as: cold shock (5.4%), swimming failure (5.4%), hypothermia (5.4%), post-rescue collapse (0.8%), cardiac event (0.8%) and drowning/other (10%). In the remaining 72.2% of deaths, there was insufficient information to determine an underlying cause. All deaths occurred in water below 17.5 degrees C but 95% were in water less than 15 degrees C.Immersion in water below 15 degrees C is dangerous and this should be emphasized on marine survival courses. Accident investigators, coroners and pathologists need a common checklist to record vital data. A recommended format is included as Supplementary data available at Occupational Medicine Online. Fishermen should be educated about the dangers of sudden, unexpected immersion in cold water. Consideration should be given to making marine survival courses mandatory for fishermen.

San Diego is facing significant challenges to its future development. How do the some of the region's leading lights think it will cope? Ken Howard finds out.

Patient injuries are thought to have a substantial financial impact on the health care system, but recent studies have been limited to estimating the costs of adverse drug events in teaching hospitals. This analysis estimated the costs of all types of patient injuries from a representative sample of hospitals in Utah and Colorado. We detected 459 adverse events (of which 265 were preventable) by reviewing the medical records of 14,732 randomly selected 1992 discharges from 28 hospitals. The total costs (all results are discounted 1996 dollars) were $661,889,000 for adverse events, and $308,382,000 for preventable adverse events. Health care costs totaled $348,081,000 for all adverse events and $159,245,000 for the preventable adverse events. Fifty-seven percent of the adverse event health care costs, and 46% of the preventable adverse event costs were attributed to outpatient medical care. Surgical complications, adverse drug events, and delayed or incorrect diagnoses and therapies were the most expensive types of adverse events. The costs of adverse events were similar to the national costs of caring for people with HIV/AIDS, and totaled 4.8% of per capita health care expenditures in these states.

HeLa X human skin fibroblast hybrid cells have been developed into a model for radiation-induced neoplastic transformation of human cells. Previous studies indicate that the appearance of neoplastically transformed foci in this system is delayed for several population doublings after irradiation and appears to involve the loss of putative tumor suppressor loci on fibroblast chromosomes 11 and 14. We now show that after treatment with 7 Gy of X-rays, transformed foci initiation correlates with delayed apoptosis initiated in the progeny of the irradiated cells after 10-12 cell divisions and with reduced plating efficiency (delayed death). The cells develop classic apoptotic morphology, positive terminal deoxynucleotidyl transferase-mediated nick end labeling and phosphatidylserine (annexin V) staining, and cleavage of poly(ADP-ribose) polymerase. In addition, a delayed induction of the p53 protein and the proapoptotic Bax protein is evident over a week after radiation exposure. We propose that a delayed build-up of mitosis-dependent genomic DNA damage or a loss of genetic material over time (10-12 cell divisions postirradiation) has two relevant outcomes: (a) cell death due to the delayed induction of a p53-dependent apoptosis; and (b) neoplastic transformation of a minor subset of survivors that has lost fibroblast chromosomes 11 and 14 (tumor suppressor loci for this system) and has either evaded apoptosis or not acquired enough genetic damage to induce apoptosis. It is postulated that both phenomena result from X-ray-induced, translesion-mediated genomic instability.

Platinum-DNA adducts can be measured in peripheral blood leukocytes during platinum-based chemotherapy, and high adduct levels have been correlated with favorable clinical response in patients with germ cell cancer. Twenty-five patients with advanced germ cell cancer were treated with platinum-based chemotherapy regimens using the same dose and schedule of cisplatin. Platinum-DNA adducts were measured by atomic absorption spectrometry on the first and fifth days of the first cycle of cisplatin-based therapy. The patients were followed prospectively for 6-35 months (median, 26 months). Twenty-two patients had adduct levels measured 24 h after the first dose of cisplatin. There was no difference in the mean adduct levels of those who were alive and without progression of disease compared to those who were dead or progressing (P = 0.65). Twenty-three patients had day 5 adduct levels measured. The mean day 5 adduct level in the 15 patients who were alive and without progression was 62.133 fmol/microgram compared to 153.50 fmol/microgram in the patients who were dead or progressing (two-sided P = 0.02). Contrary to previous reports, these data indicate that high platinum-DNA adduct levels do not correlate with favorable outcome in patients with advanced germ cell cancer.

Research suggests that underage drinking is a serious problem in the United States, and that many of the factors that influence minors to consume alcohol are environmental in their nature. One such factor is youth-oriented alcohol advertisements. Although many adults support restrictions on such advertisements, others do not or do so to a lesser degree. In this study, we explored one factor that may influence how strongly adults support restrictions on youth-oriented alcohol advertisements: The frequency with which adults themselves report consuming alcoholic beverages. A total of 767 adult Idahoans completed a survey asking about a variety of perceptions related to underage drinking, including whether they supported five types of restrictions on youth-oriented alcohol advertisement. They also answered a question about their own drinking behavior. The results revealed a significant effect of adults’ self-reported alcohol consumption behavior on their support for all five types of advertising restrictions. Adults who reported not consuming alcohol were significantly more supportive of all five types of advertising restrictions than those who reported consuming at least one alcoholic beverage in an average week. Significant differences in adults’ levels of support for all five types of advertising restriction were also revealed as a function of frequency with which adults consumed alcoholic beverages. Adults who reported consuming alcohol on one day in an average week reported significantly more support for all five types of advertising restrictions than those who reported consuming alcohol four or more days in an average week. These findings identify a population—heavier drinking adults—for education about problems associated with underage drinking and advertising alcohol to minors.

The rate of administration of an intravenous anesthetic induction agent is an important variable determining the total dose required to reach a given endpoint, such as loss of consciousness (LOC). The influence of infusion rate on the dose-response relationship has not been described rigorously. In this study we characterized the effect of different thiopental infusion rates on the times and doses required to reach a clinical (induction) endpoint.Fifty-six healthy, non-premedicated men, aged 19-59 yr, were randomly assigned to receive one of seven different thiopental infusion rates (40, 60, 75, 150, 300, 600, and 1,200 mg/min). The infusion was continued until the patient dropped a held object, indicating LOC. The infusion rates were selected using a simulation which predicted the relationship between the rate of administration and cumulative dose administered at the time of LOC. Average population pharmacokinetic parameters from a three-compartment thiopental model were combined with an effect-site rate constant for thiopental equilibration of 0.58 min-1 and a median effect-site concentration of 13.8 mg/l from previously published pharmacokinetic and pharmacodynamic models for thiopental. This derived model was used to predict the total amount of thiopental required, at each infusion rate, to produce LOC.The observed median effective doses for infusion rates of 40-150 mg/min were similar and ranged from 296 to 318 mg. Dose requirements increased significantly with increasing infusion rates greater than 150 mg/min; median effective doses for infusion rates of 300, 600, and 1,200 mg/min were significantly different from each other (436, 555, and 711 mg, respectively). The original simulation underestimated the observed thiopental doses at all but the lowest infusion rate. A new simulation was performed using a recently developed combined pharmacokinetic-pharmacodynamic model. This model incorporated a four-compartment thiopental pharmacokinetic model with quantal dose-response data to derive an effect-site rate constant for thiopental equilibration of 0.29 min-1 and a median effect-site concentration for LOC of 11.3 mg/l. The median thiopental doses predicted by this new simulation under the extreme conditions of a 30-fold range of infusion rates were within 13% of the observed doses.In this study we quantified the relationship between the rate of thiopental administration and the resultant cumulative thiopental dose necessary to produce LOC. This study validated a novel pharmacokinetic-pharmacodynamic model based on a four-compartment pharmacokinetic model and infusion quantal dose-response data. Finally, we demonstrated that thiopental dose-response relationships are dependent on drug administration rate, and found that the ability to predict this dependence accurately is influenced by the pharmacokinetics, pharmacodynamics, and median effect-site concentration used to simulate the dose-response relationships.

A comparative study of recombinant 51- and 66-kDa subunits comprising equine infectious anemia virus reverse transcriptase (EIAV RT) is reported. Both polypeptides sedimented as stable homodimers (molecular mass, 102 and 132 kDa, respectively) when analyzed by rate sedimentation through glycerol gradients. Consistent with their dimer composition, each preparation displayed considerable levels of both RNA- and DNA-dependent DNA polymerase activity on different homopolymeric template/primer combinations. However, a detailed analysis of the polymerization products indicated qualitative differences. Whereas p66 EIAV RT proceeded essentially unimpaired along both RNA and DNA templates, p51-catalyzed DNA synthesis was interrupted close to or in the immediate vicinity of the primer. A series of "programmed" 2-step polymerization reactions suggests that p51 EIAV RT enters an abortive mode of polymerization. Duplication of this observation with p51 human immunodeficiency virus-1 RT, together with recent observations from murine RT, suggests that lack of a ribonuclease H domain and loss of contact with the nascent product from the polymerase active center have profound consequences on the mode of polymerization.

A contribution of the 51-kDa subunit of human immunodeficiency virus type-1 reverse transcriptase to activities of the parental heterodimer (p66/p51) was assessed in "selectively deleted" heterodimers whose p51 component contained C-terminal truncations of 13, 19, or 25 residues. Analyses included (i) efficiency of reconstitution into heterodimer, (ii) retention of polymerase and ribonuclease H (RNase H) function, and (iii) interaction with the HIV replication primer, tRNA(Lys,3). Our data suggest that these features of heterodimer reverse transcriptase can be modulated by the extent of the C-terminal p51 deletion. Severely impaired tRNA binding in a selectively deleted heterodimer whose 51-kDa subunit lacks 13 residues, despite retention of enzymatic functions, strengthens arguments for p51 involvement in tRNA binding.

The interaction of several forms (p51, p66, and p66/p51) of recombinant human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) with a synthetic derivative of its cognate replication primer, tRNA(Lys-3), has been determined by gel-mobility shift analysis. While p66/p51 RT is proficient in tRNA binding, preparations of p66 and p51 display only weak binding at elevated protein:tRNA ratios, despite the former containing both RNA-dependent DNA polymerase and ribonuclease H (RNase H) activity. Gel permeation analysis of purified p66 RT indicate this to be predominantly monomeric, suggesting that dimerization may be a prerequisite for efficient tRNA binding. Prolonged incubation of a mixture of the 66- and 51-kDa polypeptides results in heterodimer reconstitution, restoration of tRNA binding, and recovery of appreciable levels of RNA-dependent DNA polymerase activity. Under the same conditions, both the tRNA binding and RNA-dependent DNA polymerase activities of the 66- and 51-kDa polypeptides are unaffected, suggesting that they remain in the monomeric conformation.

Metal chelate affinity chromatography has been used to follow reconstitution of the 66- and 51-kDa human immunodeficiency (HIV)-1 and HIV-2 reverse transcriptase (RT) subunits into heterodimer, as well as chimeric enzymes comprised of heterologous subunits. By adding a small N-terminal polyhistidine extension to the 51-kDa subunit of either enzyme, reconstituted RT could be recovered from a cell lysate by chromatography on Ni(2+)-nitrilotriacetic acid-Sepharose. Homologous RT subunits rapidly associated to form the respective heterodimers (1-p66.1-p51 and 2-p66.2-p51) when bacterial lysates containing the individual components were mixed. Under the same conditions, association of p66 HIV-2 and p51 HIV-1 RT was inefficient and could be improved slightly by prolonged incubation of the respective p66 and p51 subunits. In contrast, HIV-1 p66 RT rapidly associated with the 51-kDa subunit of the HIV-2 enzyme. RNA-dependent DNA polymerase activity was associated with all reconstituted enzymes, and the response of each chimeric RT to an inhibitor selective for the HIV-1 enzyme indicated that sensitivity to inhibition was determined by the source of its 66-kDa subunit.

In animal cells, unliganded steroid receptors are complexed with a 90-kDa heat shock protein, HSP90; hormone binding by the receptor leads to the release of HSP90. We found that the 795-amino acid rat glucocorticoid receptor protein formed oligomeric complexes in vitro upon synthesis in rabbit reticulocyte lysates; these oligomers also dissociated in the presence of hormone. Similar complexes formed when X795, a receptor derivative containing only the C-terminal half (amino acids 407-795) of the protein, was translated in vitro. Moreover, X795 was co-immunoadsorbed from the reticulocyte lysates together with HSP90 by three different anti-HSP90 monoclonal antibodies, indicating that the in vitro translated receptor binds HSP90 and that the interaction occurs within the C-terminal half of the receptor. To localize the HSP90 binding region in greater detail, various deletion mutants of X795 were translated in vitro and assayed for oligomer formation and for co-immunoadsorption with HSP90. The results indicated that HSP90 interacted with the receptor within a subregion of the hormone binding domain, between amino acids 568 and 616. These findings are consistent with the proposal that HSP90 may participate in the mechanism of signal transduction by steroid receptors.

During development, the precursors for eggs and sperm — the germ cells — migrate to the gonads to mature. But what attracts them there? Results from a fruitfly screen indicate that one component in this pathway is a gene called columbus, which encodes HMG-CoA reductase. In humans, this enzyme is involved in cholesterol biosynthesis, raising the intriguing possibility that the attractant may be a lipid.

Alternaria crassa (Sacc.) Rands. and Alternaria cassiae Jurair & Khan are important fungal pathogens of jimsonweed (Datura stramonium) and sicklepod (Cassia obtusifolia), respectively (Walker, 1982; Boyette, 1986). Commercial production of these Alternaria spp. depends on numerous factors including cost-effective sporulation techniques. Efforts to elicit sporulation of Alternaria spp. in submerged culture have largely been unsuccessful. Other fungi produce far more new conidia than were initially present in the inoculum after growth in a bioreactor. However, in Alternaria spp. there is no morphological difference between newly formed conidia and old conidia and the inability to distinguish the two has hampered efforts to increase or improve conditions for conidia production. Consequently, a staining procedure was sought to identify newly formed spores in fungi, such as A. crassa and A. cassiae, where spore production may be limited by numerous factors. Studies were conducted in column bioreactors containing a minimal medium. Conidial inoculum was stained with Nile red fluorescent dye and added to the column bioreactors yielding final concentrations of 1 x 103 to 1 x 104 conidia/ml. Between 430 and 3.5 x 103 conidia/ ml were recovered from each fungus after 96 h in minimal media and approximately 20% of the total spores were identified as new conidia. Nile red, 9-diethylamino-5H-benzo(a)phenoxazine-5one (Sigma Co.), is a hydrophobic fluorescent dye which is highly selective for intracellular lipid droplets (Greenspan et al., 1985). It is nontoxic to fungi at concentrations up to 100 Ag/ml, stable, intensely fluorescent only in a hydrophobic environment and very soluble in lipids (Butt et al.

The first hydrothermal field on the northern Gorda Ridge, the Sea Cliff hydrothermal field, was discovered and geologic controls of hydrothermal activity in the rift valley were investigated on a dive series using the DSV Sea Cliff. The Sea Cliff hydrothermal field was discovered where predicted at the intersection of axis-oblique and axis-parallel faults at the south end of a linear ridge at mid-depth (2700 m) on on the east wall. Preliminary mapping and sampling of the field reveal: a setting nested on nearly sediment-free fault blocks 300 m above the rift valley floor 2.6 km from the axis; a spectrum of venting types from seeps to black smokers; high conductive heat flow estimated to be equivalent to the convective flux of multiple black smokers through areas of the sea floor sealed by a caprock of clastic breccia primarily derived from basalt with siliceous cement and barite pore fillings; and a vent biota with Juan de Fuca Ridge affinities. These findings demonstrate the importance of off-axis hydrothermal activity and the role of the intersection of tectonic lineations in controlling hydrothermal sites at sea-floor spreading centers.

Mutations of the hairy locus in Drosophila may affect both adult chaeta differentiation and embryonic segmentation. In an effort to understand this phenotypic complexity, we have analyzed 30 mutant alleles of the locus. We find that the alleles fall into four groups according to their complementation properties, suggesting a structurally complex locus in which two distinct functions share a common coding region.

The vacuum expectation value of the stress-energy tensor for the Hartle-Hawking state in Schwarzschild space-time has been calculated for the conformal scalar field. $〈{T}_{\ensuremath{\mu}}^{\ensuremath{\nu}}〉$ separates naturally into the sum of two terms. The first coincides with an approximate expression suggested by Page. The second term is a "remainder" that may be evaluated numerically. The total expression is in good qualitative agreement with Page's approximation. These results are at variance with earlier results by Fawcett whose error is explained.

A series of plasmids encoding various Klebsiella pneumoniae nif (nitrogen fixation) genes were constructed to determine which were required to produce active iron (Fe) protein in Escherichia coli, a species which does not normally fix nitrogen. The greatest success was achieved with binary plasmid systems that produced nifA regulatory protein under the control of a tac promoter on one plasmid, which then induced synthesis of nifH and nifM proteins from their native promoter sites on a second plasmid. nifH protein, the monomeric subunit of Fe protein, produced in the presence of nifM constituted nearly 10% of the whole cell protein and exhibited the corresponding amount of C2H2-reducing activity in nitrogenase assays conducted in vitro. nifH protein formed in the absence of nifM constituted 4.7% of the whole cell protein and exhibited no detectable activity in assays of whole cell extracts. The plasmid-encoded Fe protein was purified to homogeneity and was found to be indistinguishable from that isolated from derepressed wild type K. pneumoniae, having a similar specific activity, approximately 4 Fe/dimer of 68 kDa, and similar epr features. Although these experiments do not exclude the participation of other E. coli gene products in the maturation of nifH protein, they limit the nif-specific genes required for active Fe protein production to nifA, nifH, and nifM. Since nifA is thought to be the required activator protein involved in nif operon transcription, the simplest explanation for these observations is that nifH codes for the peptide of the Fe protein, while nifM acts to convert this nifH peptide to the functioning Fe protein of nitrogenase. In the absence of nifM, only an inactive nifH polypeptide is produced.

Segmentation of the Drosophila embryo requires expression of the pair-rule genes, mutations of which cause reiterated deletions in alternate segments along the antero-posterior body axis. We find that transcripts of one such gene, hairy, accumulate in eight distinct regions of the early embryo. This pattern of expression is compared with that of another pair-rule gene, fushi tarazu, and its dependence on maternally expressed genes is described.

Rhodopseudomonas viridis ATCC 19567 grows by means of nitrogen fixation in yeast extract-N2 or nitrogen-free medium when sparged with 5% CO2 and 95% N2 in the light at 30 degrees C. Acetylene reduction assays for nitrogenase activity revealed an initially high level of activity during early-logarithmic growth phase, a lower plateau during mid- to late-logarithmic phase, and a dramatic reduction of activity at the beginning of the stationary phase. When viewed by electron microscopy, nitrogen-fixing R. viridis cells appeared to be morphologically and ultrastructurally similar to cells grown on nitrogen-rich media. Whole cells prepared under reducing conditions in the dark for electron spin resonance spectroscopy yielded g4.26 and g3.66 signals characteristic of the molybdenum-iron protein of nitrogenase. During growth on N2 in the absence of fixed-nitrogen sources, the nitrogenase activity of R. viridis measured by acetylene reduction stopped rapidly in response to the addition of NH4Cl as has been observed in other Rhodospirillaceae. However, unlike the nitrogenase of Rhodopseudomonas palustris or Rhodospirillum rubrum, which recover from this treatment within 40 min, the nitrogenase activity of R. viridis was not detectable for nearly 4 h.

The first hint of segmentation in the Drosophila embryo is the appearance of shallow transverse grooves that divide the main part of the germ band into 14 uniform domains1. These grooves are conspicuous in the 5–7-h-old embryo when viewed in the scanning electron microscope2 and can also be seen in longitudinal sections. Since their discovery in 1950, the domains delineated by the grooves haye been thought to be segments1,2, but now several lines of evidence suggest that they are ‘parasegments’—units consisting of a posterior compartment3 of one segment and an anterior compartment of the next4,5. If this is the case, the posterior compartments would lie in the anterior parts, of the domains delineated by the grooves. By using the engrailed gene product as a marker, we have now mapped the posterior compartments and report here that they are indeed located in the anterior quarter of the domains and that their anterior borders coincide with the grooves, establishing that the domains are parasegments.

A kinetic pulse-chase labeling technique was used to measure the intracellular half-life of the glucocorticoid receptor in S49 mouse lymphoma cells. Cells were pulse-labeled with [35S]methionine for 30 min and then cultured in the presence of unlabeled methionine (chase). Labeled receptors were quantitated at periodic time points during the chase by immunoadsorption to protein A-Sepharose using the BuGR2 monoclonal antireceptor antibody. The decay of labeled receptors during the chase was linear on a semilog plot, consistent with first order kinetics. Receptor half-life was 9 h when cells were cultured in either phenol red-containing medium supplemented with fetal calf serum or in phenol red free-medium supplemented with charcoal extracted serum, indicating that endogenous steroids do not affect receptor half-life. Receptor half-life was also unchanged when cells were cultured in the presence of 0.1 microM dexamethasone, a glucocorticoid hormone, or 0.1 microM RU486 (11 beta-(4-dimethylamino-phenyl)-17 beta-hydroxy-17 alpha-(propynylestra-4,9- diene-3-one), an antiglucocorticoid hormone. We conclude that the intracellular half-life of the glucocorticoid receptor in S49 mouse lymphoma cells is not regulated by either glucocorticoid or antiglucocorticoid hormones.