Multi-component self-assembled anti-tumor nano-vaccines based on MUC1 glycopeptides.

Novel multi-component self-assembled nano-vaccines containing both Pam3CSK4 and CpG were developed for the first time. These multi-component vaccines could effectively activate the macrophages in vitro and elicit strong antibody immune responses and anti-tumor immune responses in vivo.

International medical graduates in the USA: a qualitative study on perceptions of physician migration.

Objectives Physician migration from low-income to high-income nations is a global concern. Despite the centrality of understanding the perspectives of international medical graduates (IMGs) who have experienced migration to understanding the causes and consequences of this phenomenon, empirical literature is limited. The authors sought to characterise the experiences of IMGs from limited resource nations currently practicing primary care in the USA, with a focus on their perspectives on physician migration. Design The authors conducted a qualitative study utilising in-depth, in-person interviews and a standardised interview guide. The sample comprised a diverse, purposeful sample of IMGs (n=25) from limited resource nations (defined as having ≤2 physicians per 1000 population). Results Analyses revealed four recurrent and unifying themes reflecting the perspectives of IMGs in the USA on physician migration: (1) decisions to migrate were pragmatic decisions made in the context of individual circumstance; (2) the act of migration ultimately affected participants' ability to return home in multiple, unpredictable ways; (3) the ongoing process of acclimation was coupled with inherent conflicts surrounding the decision to remain in the USA; and (4) the effects of policies in both the home country and in the USA occurred at multiple levels. Conclusion The perspectives of IMGs who have migrated to the USA are an important addition to the ongoing discussion surrounding the global health workforce. Our findings highlight the effects of workforce policies which are often developed and discussed in abstraction, but have real, measurable impacts on the lives of individuals. Future efforts to address physician migration will need to acknowledge the immediate needs of the health workforce as well as the long-term needs of individuals within health systems.

Anti-leukemic immunity against U937 cells in uremic patients.

To examine anti-tumor immunity in uremic patients undergoing regular hemodialysis, we designed this study using in vitro mononuclear cell (MNC) cultures, with human leukemic U937 cells as the target. MNC were collected and cultured from uremic subjects and age- and gender-matched healthy controls. Conditioned media from the cultures (MNC-CM) were collected after stimulation with various concentrations of phytohemagglutinin (PHA). The proliferation-inhibiting and differentiation-inducing activities of the PHA-MNC-CM on U937 cells were evaluated. The growth inhibition activity of uremic patients' PHA-MNC-CM was lower than that of controls. The differentiation-inducing effects were evaluated by morphological scoring, superoxide production, and monocyte-associated antigen expression (CD14 and CD68). All three parameters demonstrated that the differentiation-inducing effect of MNC-CM increased with increasing doses of PHA. These effects, however, were significantly less in uremic patients compared to controls at higher doses of PHA. The levels of TNF-alpha and IFN-gamma in PHA-MNC-CM increased in a PHA dose-dependent manner and were much higher in the controls. We conclude that the capacity of MNC from uremic hemodialysis patients to produce anti-leukemic immunity is significantly lower than that of healthy controls.

Curcumin, a medicinal herbal compound capable of inducing the heat shock response.

OBJECTIVE: There is interest in developing pharmacologic inducers of the heat shock response as a means to confer cytoprotection in the clinical setting. We propose that a potential strategy for screening novel pharmacologic inducers of the heat shock response is to examine known inhibitors of the transcription factor nuclear factor-kappaB. Curcumin, derived from the tropical herb Curcuma longa, is a recently described inhibitor of nuclear factor-kappaB and is widely used in Eastern medicinal practices. We tested the hypothesis that curcumin can induce expression of heat shock protein 70. DESIGN: Experimental. SETTING: University laboratory. SUBJECTS: HeLa cells. INTERVENTIONS: HeLa cells were exposed to varying concentrations of curcumin and analyzed for expression of heat shock protein 70 by Western blot. MEASUREMENTS AND MAIN RESULTS: Activation of the transcription factor, heat shock factor-1, was analyzed by electromobility shift assays. Curcumin-mediated inhibition of nuclear factor-kappaB activation was measured by transiently transfecting cells with a nuclear factor-kappaB luciferase reporter plasmid. The role of heat shock factor-1 in curcumin-mediated expression of heat shock protein 70 was tested in embryonic fibroblasts derived from heat shock factor-1 knockout mice. Induction of the heat shock response was quantified by transiently transfecting cells with a heat shock protein 70 promoter-luciferase reporter plasmid. Cell viability was measured by using the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Curcumin induced expression of heat shock protein 70, the major inducible heat shock protein in cells undergoing the heat shock response, in a dose-dependent and time-dependent manner. Curcumin induced specific nuclear translocation and activation of heat shock factor-1. Curcumin-mediated expression of heat shock protein 70 was reduced substantially in fibroblasts having genetic ablation of heat shock factor-1. The extent of induction of the heat shock response correlated, in part, with cellular toxicity. CONCLUSIONS: Curcumin, a widely used medicinal compound, induces the heat shock response in vitro as measured by expression of heat shock protein 70. The mechanism of heat shock protein 70 induction depends on activation of heat shock factor-1. Examining known inhibitors of nuclear factor-kappaB for their ability to induce heat shock protein 70 may be a valid screening method to discover novel pharmacologic inducers of the heat shock response.

Basic fibroblast growth factor as a growth factor for SRV-2-infected simian retroperitoneal fibromatosis cells, an animal model for AIDS related Kaposi's sarcoma.

Basic fibroblast growth factor (bFGF) and vascular endothelial cell growth factor (VEGF) were demonstrated to be important factors sustaining the growth of Kaposi's sarcoma. RF cells were used to provide a model to study the pathogenesis of Kaposi's sarcoma. In this paper, we demonstrated that bFGF is present in the RF cells, cultured media, and tissues from monkey. The biological activities of bFGF on RF cells were also studied in vitro with serum-free media. The bFGF from serum-free-conditioned media is biologically active to stimulate RF cells in certain media condition. The mitogenic effect was abrogated by sheep neutralizing anti-bFGF antibody. Furthermore, the effect of antibody was reversed by the addition of exogenous bFGF. ELISA measurements indicating the growth potency of conditioned media correlated with the amount of bFGF in the conditioned media. The data from flow cytometry demonstrated the co-existence of SRV-2 and bFGF among RF cells and RF tissues. Immunohistochemical staining of RF tissue blocks for bFGF revealed that bFGF was present in the tumor and the presence of bFGF was not caused by the artifact of tissue culture. These results indicate that bFGF is an important growth factor to promote RF cell growth in vitro and RF tumor in vivo. Further studies are required to determine the relationship between the interaction of bFGF, SRV-2, and VEGF. This model also provides an adequate alternative to the model induced by simian immunodeficiency virus (SIV) to study the Kaposi's sarcoma.

Replication of hepatitis C virus in ascitic mononuclear cells with development of distinct viral quasispecies.

To investigate whether distinct compositions of viral quasispecies developed in the ascitic fluid of patients with late-stage chronic hepatitis C virus (HCV) infection, samples of various origins, including ascitic fluid and ascitic mononuclear cells (AMCs), were analyzed by the method of single-strand conformation polymorphism (SSCP). Subsequently, the major species were isolated, sequenced, and subjected to phylogenetic analysis. Of 42 patients analyzed, HCV-RNA was detectable in the AMCs of 25 patients. SSCP analysis indicated that the compositions of viral quasispecies among samples of different origins were markedly different in this group of patients. Phylogenetic analysis revealed that the ascitic-fluid-derived clones were most closely related to the AMC-derived clones. Minus-strand HCV-RNA was detectable in 5 of them. Our data suggest that HCV can replicate in the AMCs of patients with late-stage chronic HCV, which results in the development of distinct viral quasispecies.

In vitro effect of granulocyte-colony stimulating factor and all-trans retinoic acid on the expression of inflammatory cytokines and adhesion molecules in acute promyelocytic leukemic cells.

Differentiation therapy with all-trans retinoic acid (ATRA) represents a landmark approach in the treatment of acute promyelocytic leukemia (APL). However, a potentially fatal complication of retinoic acid (RA) syndrome occurs in about a quarter of patients and its pathophysiology is still unclear. In order to investigate whether or not the treatment with ATRA leads to increased elaboration of inflammatory cytokines and adhesion molecules by the APL cells, the expression of interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, IL-8, L-selectin and intercellular adhesion molecule-1 (ICAM-1) was examined in the APL cells after induction of differentiation with ATRA in the presence or absence of granulocyte-colony stimulating factor (G-CSF) or IL-3 in the present study. Cytokine elaboration by the treated cells was detected using both Northern blotting and enzyme-linked immunosorbent assay. Our results have shown that ATRA induces an increased expression of IL-8, IL-1beta, TNF-alpha and ICAM-1 in APL cells, which can be amplified by the addition of G-CSF. These data imply that the induction of inflammatory cytokines in APL cells may play an important role in the pathogenesis of RA syndrome. Furthermore, G-CSF, through its potent differentiating activity, may increase the risk of such complications during ATRA treatment.

Comparison of antileukemic immunity against U937 cells in atopic asthmatics versus healthy controls.

To assess the antitumor effects in atopic asthmatics versus healthy adults, we designed this study using in vitro mononuclear cells (MNC) culture as an immunity model with human leukemic U937 cells as the target. MNCs were collected from asthmatic subjects and healthy controls. Conditioned media from the MNC cultures (MNC-CM) were collected after stimulation with various concentrations of phytohemagglutinin (PHA). We treated U937 cells with these MNC-CMs, then assayed their proliferation and differentiation after 5 days of culture. At lower PHA doses (1.25 microg/ml), as well as in absence of PHA, the asthmatic MNC-CMs inhibited U937 cells growth to a slightly greater extent than did the MNC-CMs from controls. In contrast, when higher doses of PHA were used (5, 10 microg/ml), this growth-inhibiting effect was dramatically reversed. The dual effect of MNC-CM in these two groups was also shown in U937 cell differentiation assay, assessed as follows: morphological change by Liu's staining, functional change by NBT reduction test and CD 14 expression by flow cytometric detection. We suggest that the antileukemic effects of MNCs from asthmatic patients result from a slightly immunopotentiated status. This immunity may be dramatically reversed, however, after marked activation of MNCs.

Molecular physiology of carbamoylation under extreme conditions: what can we learn from extreme thermophilic microorganisms?

The importance of protein-protein interactions in the physiology of extreme thermophiles was investigated by analyzing the enzymes involved in biosynthetic carbamoylation in Thermus ZO5 and by comparing the results obtained with already available or as yet unpublished information concerning other thermophilic eu- and archaebacteria such as Thermotoga, Sulfolobus, and Pyrococcus. Salient observations were that (i) the highly thermolabile and reactive carbamoylphosphate molecule appears to be protected from thermodegradation by channelling towards the synthesis of citrulline and carbamoylaspartate, respectively precursors of arginine and the pyrimidines; (ii) Thermus ornithine carbamoyltransferase is clearly a thermophilic enzyme, intrinsically thermostable and showing a biphasic Arrhenius plot, whereas aspartate carbamoyltransferase is inherently unstable and is stabilized by its association with dihydroorotase, another enzyme encoded by the Thermus pyrimidine operon. Possible implications of these results are discussed.