Sweepouts of amalgamated 3-manifolds

We show that if two 3-manifolds with toroidal boundary are glued via a `sufficiently complicated' map then every Heegaard splitting of the resulting 3-manifold is weakly reducible. Additionally, if Z is a manifold obtained by gluing X and Y, two connected small manifolds with incompressible boundary, along a closed surface F. Then the genus g(Z) of Z is greater than or equal to 1/2(g(X)+g(Y)-2g(F)). Both results follow from a new technique to simplify the intersection between an incompressible surface and a strongly irreducible Heegaard splitting.Comment: This is the version published by Algebraic & Geometric Topology on 24 February 200

Rethinking synchronization of mammalian cells for cell cycle analysis

An analysis of different classes of forced or batch synchronization methods reveals why these methods, in theory, do not produce synchronized cultures. Cells may be aligned for a particular property after specific treatments, but these aligned cells do not correspond to any particular cell age during the normal cell cycle. The experimental methods analyzed are those that arrest cells with a G1 phase amount of DNA, those that inhibit DNA synthesis, and those that arrest cells at mitosis. Release of arrested cells from inhibition does not produce cells reflecting cells during the normal division cycle. Thus, cells produced by batch or forcing methods are not experimental models for analysis of the normal cell cycle.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41836/1/30601099.pd

Role differentiation as a dimension in community diagnosis

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/44036/1/10464_2004_Article_BF00878038.pd

Revisiting retinoblastoma protein phosphorylation during the mammalian cell cycle

It is widely accepted that phosphorylation of the retinoblastoma (Rb) protein during the G1 phase of the mammalian division cycle is a major control element regulating passage of cells into S phase and through the division cycle. The experiments supporting G1-phase-specific Rb phosphorylation and the historical development of this idea are reviewed. By making a rigorous distinction between 'growth cessation' and the phenomena of 'cell cycle exit' or 'G1-phase arrest', the evidence for the G1-phase-specific phosphorylation of Rb protein is reinterpreted. We show that the evidence for G1-phase phosphorylation of Rb rests on few experiments and a chain of reasoning with some weak links. Evidence is reviewed that growth conditions regulate the phosphorylation of Rb. A growth-regulated control system that is independent of the cell cycle explains much of the evidence adduced to support cycle-specific phosphorylation of Rb. We propose that additional experimental evidence is needed to decide whether there is a G1-phase-specific phosphorylation of Rb protein.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/41833/1/18-58-4-580_10580580.pd

Replication of mini-F plasmids during the bacterial division cycle

The cell-cycle replication patterns of two mini-F plasmids have been examined using the membrane-elution technique (to produce cells labelled at different times during the division cycle) and scintillation counting (for quantitative analysis of radioactivity incorporated into plasmid DNA). The mini-F plasmid pML31, which contains the oriV and oriS origins of replication, replicates in a cell-cycle-specific manner with a pattern and cell-cycle timing similar to the parental F plasmid. The mini-F plasmid pMF21, deleted for the region containing the oriV origin of replication, replicates more randomly throughout the division cycle. These results suggest that the oriV origin of replication may be related to cell-cycle-specific replication of the F plasmid.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30355/1/0000757.pd

Effect of mecillinam on peptidoglycan synthesis during the division cycle of Salmonella typhimurium 2616

The effects of mecillinam, ampicillin and cephalexin on peptidoglycan synthesis in Salmonella typhimurium 2616 have been studied at equivalent concentrations or "isoactivities". Using antibiotics at isoactivities allows a direct comparison of the biochemical effects of different antibiotics. When mecillinam was added at different times during the division cycle at a concentration that produced 50 % inhibition of peptidoglycan synthesis in an exponential culture over a short period of time, the inhibition of synthesis was greatest in the newborn cells and least in the dividing cells. Antibiotic competition experiments showed that mecillinam preferentially bound to penicillin-binding protein 2 in S. typhimurium 2616. High performance liquid chromatography analysis of the residual peptidoglycan synthesized in the presence of mecillinam showed an unexpected increase in pentapeptides and a significant increase in cross-linking. Other antibiotics added at equivalent activities did not show an increase in cross-linking.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/30717/1/0000363.pd

Versatile Diphosphine Chelators for Radiolabeling Peptides with ^99mTc and ⁶⁴Cu

We have developed a diphosphine (DP) platform for radiolabeling peptides with 99mTc and 64Cu for molecular SPECT and PET imaging, respectively. Two diphosphines, 2,3-bis(diphenylphosphino)maleic anhydride (DP Ph) and 2,3-bis(di- p-tolylphosphino)maleic anhydride (DP Tol), were each reacted with a Prostate Specific Membrane Antigen-targeted dipeptide (PSMAt) to yield the bioconjugates DP Ph-PSMAt and DP Tol-PSMAt, as well as an integrin-targeted cyclic peptide, RGD, to yield the bioconjugates DP Ph-RGD and DP Tol-RGD. Each of these DP-PSMAt conjugates formed geometric cis/ trans-[MO 2(DP X-PSMAt) 2] + (M = 99mTc, 99gTc, natRe; X = Ph, Tol) complexes when reacted with [MO 2] + motifs. Furthermore, both DP Ph-PSMAt and DP Tol-PSMAt could be formulated into kits containing reducing agent and buffer components, enabling preparation of the new radiotracers cis/ trans-[ 99mTcO 2(DP Ph-PSMAt) 2] + and cis/ trans-[ 99mTcO 2(DP Tol-PSMAt) 2] + from aqueous 99mTcO 4 - in 81% and 88% radiochemical yield (RCY), respectively, in 5 min at 100 °C. The consistently higher RCYs observed for cis/ trans-[ 99mTcO 2(DP Tol-PSMAt) 2] + are attributed to the increased reactivity of DP Tol-PSMAt over DP Ph-PSMAt. Both cis/ trans-[ 99mTcO 2(DP Ph-PSMAt) 2] + and cis/ trans-[ 99mTcO 2(DP Tol-PSMAt) 2] + exhibited high metabolic stability, and in vivo SPECT imaging in healthy mice revealed that both new radiotracers cleared rapidly from circulation, via a renal pathway. These new diphosphine bioconjugates also furnished [ 64Cu(DP X-PSMAt) 2] + (X = Ph, Tol) complexes rapidly, in a high RCY (&gt;95%), under mild conditions. In summary, the new DP platform is versatile: it enables straightforward functionalization of targeting peptides with a diphosphine chelator, and the resulting bioconjugates can be simply radiolabeled with both the SPECT and PET radionuclides, 99mTc and 64Cu, in high RCYs. Furthermore, the DP platform is amenable to derivatization to either increase the chelator reactivity with metallic radioisotopes or, alternatively, modify the radiotracer hydrophilicity. Functionalized diphosphine chelators thus have the potential to provide access to new molecular radiotracers for receptor-targeted imaging. </p

Hα star formation main sequence in cluster and field galaxies at z ∼ 1.6

We calculate Hα-based star formation rates and determine the star formation rate–stellar mass relation for members of three Spitzer Adaptation of the Red-Sequence Cluster Survey (SpARCS) clusters at z ∼ 1.6 and serendipitously identified field galaxies at similar redshifts to the clusters. We find similar star formation rates in cluster and field galaxies throughout our range of stellar masses. The results are comparable to those seen in other clusters at similar redshifts, and consistent with our previous photometric evidence for little quenching activity in clusters. One possible explanation for our results is that galaxies in our z ∼ 1.6 clusters have been accreted too recently to show signs of environmental quenching. It is also possible that the clusters are not yet dynamically mature enough to produce important environmental quenching effects shown to be important at low redshift, such as ram-pressure stripping or harassment

Phase 1 Trial of AMA1-C1/Alhydrogel plus CPG 7909: An Asexual Blood-Stage Vaccine for Plasmodium falciparum Malaria

Apical Membrane Antigen 1 (AMA1), a polymorphic merozoite surface protein, is a leading blood-stage malaria vaccine candidate. This is the first reported use in humans of an investigational vaccine, AMA1-C1/Alhydrogel, with the novel adjuvant CPG 7909.A phase 1 trial was conducted at the University of Rochester with 75 malaria-naive volunteers to assess the safety and immunogenicity of the AMA1-C1/Alhydrogel+CPG 7909 malaria vaccine. Participants were sequentially enrolled and randomized within dose escalating cohorts to receive three vaccinations on days 0, 28 and 56 of either 20 microg of AMA1-C1/Alhydrogel+564 microg CPG 7909 (n = 15), 80 microg of AMA1-C1/Alhydrogel (n = 30), or 80 microg of AMA1-C1/Alhydrogel+564 microg CPG 7909 (n = 30).Local and systemic adverse events were significantly more likely to be of higher severity with the addition of CPG 7909. Anti-AMA1 immunoglobulin G (IgG) were detected by enzyme-linked immunosorbent assay (ELISA), and the immune sera of volunteers that received 20 microg or 80 microg of AMA1-C1/Alhydrogel+CPG 7909 had up to 14 fold significant increases in anti-AMA1 antibody concentration compared to 80 microg of AMA1-C1/Alhydrogel alone. The addition of CPG 7909 to the AMA1-C1/Alhydrogel vaccine in humans also elicited AMA1 specific immune IgG that significantly and dramatically increased the in vitro growth inhibition of homologous parasites to levels as high as 96% inhibition.The safety profile of the AMA1-C1/Alhydrogel+CPG 7909 malaria vaccine is acceptable, given the significant increase in immunogenicity observed. Further clinical development is ongoing.ClinicalTrials.gov NCT00344539

Human Immunity and the Design of Multi-Component, Single Target Vaccines

BACKGROUND: Inclusion of multiple immunogens to target a single organism is a strategy being pursued for many experimental vaccines, especially where it is difficult to generate a strongly protective response from a single immunogen. Although there are many human vaccines that contain multiple defined immunogens, in almost every case each component targets a different pathogen. As a consequence, there is little practical experience for deciding where the increased complexity of vaccines with multiple defined immunogens vaccines targeting single pathogens will be justifiable. METHODOLOGY/PRINCIPAL FINDINGS: A mathematical model, with immunogenicity parameters derived from a database of human responses to established vaccines, was used to predict the increase in the efficacy and the proportion of the population protected resulting from addition of further immunogens. The gains depended on the relative protection and the range of responses in the population to each immunogen and also to the correlation of the responses between immunogens. In most scenarios modeled, the gain in overall efficacy obtained by adding more immunogens was comparable to gains obtained from a single immunogen through the use of better formulations or adjuvants. Multi-component single target vaccines were more effective at decreasing the proportion of poor responders than increasing the overall efficacy of the vaccine in a population. CONCLUSIONS/SIGNIFICANCE: Inclusion of limited number of antigens in a vaccine aimed at targeting a single organism will increase efficacy, but the gains are relatively modest and for a practical vaccine there are constraints that are likely to limit multi-component single target vaccines to a small number of key antigens. The model predicts that this type of vaccine will be most useful where the critical issue is the reduction in proportion of poor responders