To ascertain the likelihood that perivascular leukocyte infiltrates are sites for replication and dissemination of HIV-1.
We measured the ability of HIV patients' peripheral blood mononuclear ...leukocytes to migrate through confluent endothelial monolayers in vitro and infect phytohemagglutinin-stimulated allogeneic lymphoblasts. We also measured the ability of migratory leukocytes to transmit virus to uninfected leukocytes that have localized outside an endothelial barrier, and the subsequent ability of these newly infected cells to reverse-migrate back across the endothelial barrier - a process that might facilitate reentry of infected cells into the circulation and dissemination of the virus to distant sites.
Leukocytes from 27 out of 63 unselected patients spontaneously carried infectious virus across endothelial barriers. On follow-up, these 27 patients were frequently observed to develop uncontrolled viremia, despite treatment, and be hospitalized for secondary infections. Migratory leukocytes transmitted HIV to both T lymphocytes and non-T cells that had previously crossed the endothelial barrier. Either cell type could subsequently reverse-migrate carrying virus back across this barrier.
Reverse-migration of HIV-1 infected leukocytes out of perivascular reservoirs may provide a way to disseminate HIV-1 and expand the body burden of virus in some patients receiving highly active antiretroviral therapy.
Radial velocity (RV) measurements and sine curve fits to the orbital RV variations are presented for the last eight close binary systems analyzed in the same way as in the previous papers of this ...series: QX And, DY Cet, MR Del, HI Dra, DD Mon, V868 Mon, ER Ori, and Y Sex. For another seven systems (TT Cet, AA Cet, CW Lyn, V563 Lyr, CW Sge, LV Vir, and MW Vir), phase coverage is insufficient to provide reliable orbits but RVs of individual components were measured. Observations of a few complicated systems observed throughout the David Dunlap Observatory (DDO) close binary program are also presented; among them is an especially interesting multiple system V857 Her which-in addition to the contact binary-very probably contains one or more subdwarf components of much earlier spectral type. All suspected binaries which were found to be most probably pulsating stars are briefly discussed in terms of mean RVs and projected rotation velocities (vsin i) as well as spectral-type estimates. In two of them, CU CVn and V752 Mon, the broadening functions show a clear presence of nonradial pulsations. The previously missing spectral types for Paper I are given here in addition to such estimates for most of the program stars of this paper.
Radial velocity (RV) measurements and sine curve fits to the orbital RV variations are presented for 10 close binary systems: TZ Boo, VW Boo, EL Boo, VZ CVn, GK Cep, RW Com, V2610 Oph, V1387 Ori, AU ...Ser, and FT UMa. Our spectroscopy revealed two quadruple systems, TZ Boo and V2610 Oph, while three stars showing small photometric amplitudes, EL Boo, V1387 Ori, and FT UMa, were found to be triple systems. GK Cep is a close binary with a faint third component. While most of the studied eclipsing systems are contact binaries, VZ CVn and GK Cep are detached or semidetached double-lined binaries, and EL Boo, V1387 Ori, and FT UMa are close binaries of uncertain binary type. The large fraction of triple and quadruple systems found in this sample supports the hypothesis of formation of close binaries in multiple stellar systems; it also demonstrates that low photometric amplitude binaries are a fertile ground for further discoveries of multiple systems.
Telomere 3' overhang-specific DNA oligonucleotides (T-oligos) induce cell death in cancer cells, presumably by mimicking telomere loop disruption. Therefore, T-oligos are considered an exciting new ...therapeutic strategy. The purpose of this study was to elucidate how T-oligos exert antitumor effects on human malignant glioma cells @@iin vitro@ and @@iin vivo.@ We demonstrated that T-oligos inhibited the proliferation of malignant glioma cells through induction of nonapoptotic cell death and mitochondria hyperpolarization, whereas normal astrocytes were resistant to T-oligos. Tumor cells treated with T-oligos developed features compatible with autophagy, with development of autophagic vacuoles and conversion of an autophagy-related protein, microtubule-associated protein 1 light chain 3 from type I (cytoplasmic form) to type II (membrane form of autophagic vacuoles). A reverse-phase protein microarray analysis and Western blotting revealed that treatment with T-oligos inhibited the mammalian target of the rapamycin (mTOR) and the signal transducer and activator of transcription 3 (STAT3). Moreover, pretreatment with T-oligos significantly prolonged the survival time of mice inoculated intracranially with malignant glioma cells compared with that of untreated mice and those treated with control oligonucleotides (@iP=0.0065 and @iP=0.043, respectively). These results indicate that T-oligos stimulate the induction of nonapoptotic autophagic also known as type II programmed cell death and are thus promising in the treatment of malignant glioma.--Aoki, H., Iwado, E., Eller, M. S., Kondo, Y., Fujiwara, K., Li, G.-Z., Hess, K. R., Siwak, D. R., Sawaya, R., Mills, G. B., Gilchrest, B. A., Kondo, S. Telomere 3' overhang-specific DNA oligonucleotides induce autophagy in malignant glioma cells.
The presence of the Philadelphia chromosome in patients with acute lymphoblastic leukemia (Ph+ALL) is a negative prognostic indicator. Tyrosine kinase inhibitors (TKI) that target BCR/ABL, such as ...imatinib, have improved treatment of Ph+ALL and are generally incorporated into induction regimens. This approach has improved clinical responses, but molecular remissions are seen in less than 50% of patients leaving few treatment options in the event of relapse. Thus, identification of additional targets for therapeutic intervention has potential to improve outcomes for Ph+ALL. The human epidermal growth factor receptor 2 (ErbB2) is expressed in ~30% of B-ALLs, and numerous small molecule inhibitors are available to prevent its activation. We analyzed a cohort of 129 ALL patient samples using reverse phase protein array (RPPA) with ErbB2 and phospho-ErbB2 antibodies and found that activity of ErbB2 was elevated in 56% of Ph+ALL as compared to just 4.8% of Ph-ALL. In two human Ph+ALL cell lines, inhibition of ErbB kinase activity with canertinib resulted in a dose-dependent decrease in the phosphorylation of an ErbB kinase signaling target p70S6-kinase T389 (by 60% in Z119 and 39% in Z181 cells at 3 mu M). Downstream, phosphorylation of S6-kinase was also diminished in both cell lines in a dose-dependent manner (by 91% in both cell lines at 3 mu M). Canertinib treatment increased expression of the pro-apoptotic protein Bim by as much as 144% in Z119 cells and 49% in Z181 cells, and further produced caspase-3 activation and consequent apoptotic cell death. Both canertinib and the FDA-approved ErbB1/2-directed TKI lapatinib abrogated proliferation and increased sensitivity to BCR/ABL-directed TKIs at clinically relevant doses. Our results suggest that ErbB signaling is an additional molecular target in Ph+ALL and encourage the development of clinical strategies combining ErbB and BCR/ABL kinase inhibitors for this subset of ALL patients.
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Background: Hormone receptor-positive breast cancer (HR+BC) is heterogeneous. Attempts to use transcriptional analysis such as Oncotype Dx facilitate therapy planning but fail to ...identify patients who will relapse or potential therapy targets. Methods: We used reverse phase protein lysate arrays and Microvigene software to quantify expression of estrogen receptor (ERα) and 36 total/activated components of the HER2, phosphatidylinositol-3-kinase (PI3K), mitogen-activated protein kinase (MAPK), and STAT pathways in 64 HR+BCs and 40 breast cancer cell lines. Clustering was performed with Xcluster and Treeview. Results: 47/64 HR+BC patients were treated with adjuvant hormone therapy and 43/64 with chemotherapy. There were 12 recurrences including 5 patients diagnosed with metastases within 0–3 months of diagnosis. Unsupervised analysis using the expression of all 37 proteins revealed two large subclusters of HR+BCs. One large cluster was composed of tumors with lower ERα expression levels and was driven by an antibody group composed mostly of phosphoproteins indicative of activated growth factor signaling pathways. Thus, there were significant inverse correlations between ERα expression and the expression and activation of components of the PI3K/MAPK pathways including EGFR, src, AKT, 4EBP1, and PKCα (p≤0.05 for each). Similar inverse correlations were seen in 40 assayed breast cancer cell lines. The clinicoproteomic predictors of relapse among HR+BCs were nuclear grade (p=0.001), low expression of ERα (p=0.04), low p38 phosphorylation (p=0.02), and high p53 (p=0.02). There also was a trend (p≤0.1) to the association of low MAPK and S6 phosphorylation, low p27, and high cyclin B1 with relapse. Using these antibodies to perform a supervised analysis, we found a small group of p53-high, cyclin B1-high, ERα-low HR+BCs with a 75% likelihood of relapse, significantly greater than in other tumors (p<0.003). Since 10/12 relapses occurred in 26 grade 3 tumors, we searched for and found a ‘grade 3’ HR+BC 6 antibody signature associated with a recurrence-free survival at 20 months of 17% compared to 100% in other patients (p=0.002). Conclusion: Although patient numbers are small and the findings require validation, kinase signaling profiles have potential in breast cancer prediction.
No significant financial relationships to disclose.
Environmental survival of human immunodeficiency virus type 1 (HIV-1) is an important public health concern. Survival of HIV in waste water is of particular interest to those who work at treatment ...facilities and to the general public who have contact with rivers or ocean water receiving treated sewage effluent. Other researchers have reported that HIV can be detected in waste water. Their studies, however, detected homologous nucleic acid sequences but did not attempt to determine infectivity. The current study tested primary and secondary effluent from a major metropolitan sewage agency for the presence of HIV-1 using reverse transcriptase polymerase chain reaction (RT-PCR), HIV-1 p24 antigen enzyme-linked immunosorbent assay, and infectivity testing. For RT-PCR, primers SK38/SK39 and M667/AA55 were used to identify HIV-1 RNA sequences from concentrated and extracted sewage samples. Infectivity assays employed donor peripheral blood mononuclear cells (PBMCs) stimulated with phytohemagglutinin. Coxsackievirus B4, echovirus 7, and poliovirus 1, enteroviruses normally present in sewage, were tested for replication in PBMCs. Poliovirus 1 was found to infect the PBMCs. To eliminate other enteroviruses that may also infect the PBMCs and interfere with HIV-1 testing, concentrated sewage was treated with human immunoglobulin (free of HIV antibodies) and poliovirus antisera before infectivity assays were performed. All treated sewage samples tested negative for HIV-1 by all methods used. HIV-1 seeded into sewage, however, remained infectious in the assay, indicating that the sewage water sample did not interfere with HIV infectivity nor was it toxic to the PBMCs.
To identify factors that cause HIV-1 to establish perivascular foci of infected cells, we studied the transendothelial migration of blood mononuclear leukocytes (MNL) from 76 HIV+ patients and 41 ...controls. The fraction of patients' lymphocytes that migrated across endothelial cell monolayers in vitro was significantly increased (p < or = 0.03) compared with that of control donors. Migration of patients' CD4+ T cells was particularly enhanced, whereas the migration of monocytes did not differ between patients and controls. Lymphocyte migration correlated with expression of CD11a/CD18 and CD49d/CD29 and with the quantity of TNF-alpha produced as MNLs migrated through the endothelium. Measurement of HIV-1 proviral DNA copies in the patients' MNLs (n = 26) suggested that in half the cases virus-infected cells accumulated preferentially amidst the migratory leukocytes. We observed the same behavior with normal donor MNLs infected, in vitro, with each of 4 strains of HIV-1. The number of HIV-1 proviral DNA copies per million MNLs was 40 to 178 times higher in the migratory population than in the original population added to the endothelium. To test whether only certain strains of HIV-1 stimulate transendothelial migration of infected cells, we used single strand conformation polymorphism analysis to identify quasispecies of HIV-1 in the MNLs. If all strains of HIV-1 were equal in their ability to stimulate transendothelial migration, we expected to find no differences in the quasispecies present in the original and migratory cell populations. In fact the quasispecies differed in 14 of 19 paired samples, suggesting that only certain HIV-1 quasispecies promote transendothelial migration of infected cells.
Context. Gaia18cjb is one of the Gaia-alerted eruptive young star candidates which has been experiencing a slow and strong brightening during the last 13 years, similar to some FU Orionis-type ...objects. Aims. The aim of this work is to derive the young stellar nature of Gaia18cjb, determine its physical and accretion properties to classify its variability. Methods. We conducted monitoring observations using multi-filter optical and near-infrared photometry, as well as near-infrared spectroscopy. We present the analysis of pre-outburst and outburst optical and infrared light curves, color-magnitude diagrams in different bands, the detection of near-IR spectral lines, and estimates of both stellar and accretion parameters during the burst. Results. The optical light curve shows an unusually long (8 years) brightening event of 5 mag in the last 13 years, before reaching a plateau indicating that the burst is still on-going, suggesting a FUor-like nature. The same outburst is less strong in the infrared light curves. The near-infrared spectra, obtained during the outburst, exhibit emission lines typical of highly accreting low-intermediate mass young stars with typical EXor features. The spectral index of Gaia18cjb SED classifies it as a Class I in the pre-burst stage and a Flat Spectrum young stellar object (YSO) during the burst. Conclusions. Gaia18cjb is an eruptive YSO which shows FUor-like photometric features (in terms of brightening amplitude and length of the burst) and EXor-like spectroscopic features and accretion rate, as V350 Cep and V1647 Ori, classified as objects in between FUors and EXors
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