The acridonecarboxamide GF120918 potently reverses P-glycoprotein-mediated resistance in human sarcoma MES-Dx5 cells

The doxorubicin-selected, P-glycoprotein (P-gp)-expressing human sarcoma cell line MES-Dx5 showed the following levels of resistance relative to the non-P-gp-expressing parental MES-SA cells in a 72 h exposure to cytotoxic drugs: etoposide twofold, doxorubicin ninefold, vinblastine tenfold, taxotere 19-fold and taxol 94-fold. GF120918 potently reversed resistance completely for all drugs. The EC50s of GF120918 to reverse resistance of MES-Dx5 cells were: etoposide 7 ± 2 nM, vinblastine 19 ± 3 nM, doxorubicin 21 ± 6 nM, taxotere 57 ± 14 nM and taxol 91 ± 23 nM. MES-Dx5 cells exhibited an accumulation deficit relative to the parental MES-SA cells of 35% for [3H]-vinblastine, 20% for [3H]-taxol and [14C]-doxorubicin. The EC50 of GF120918, to reverse the accumulation deficit in MES-Dx5 cells, ranged from 37 to 64 nM for all three radiolabelled cytotoxics. [3H]-vinblastine bound saturably to membranes from MES-Dx5 cells with a KD of 7.8 ± 1.4 nM and a Bmax of 5.2 ± 1.6 pmol mg–1 protein. Binding of [3H]-vinblastine to P-gp in MES-Dx5 membranes was inhibited by GF120918 (Ki = 5 ± 1 nM), verapamil (Ki = 660 ± 350 nM) and doxorubicin (Ki = 6940 ± 2100 nM). Taxol, an allosteric inhibitor of [3H]-vinblastine binding to P-gp, could only displace 40% of [3H]-vinblastine (Ki = 400 ± 140 nM). The novel acridonecarboxamide derivative GF120918 potently overcomes P-gp-mediated multidrug resistance in the human sarcoma cell line MES-Dx5. Detailed analysis revealed that five times higher GF120918 concentrations were needed to reverse drug resistance to taxol in the cytotoxicity assay compared to doxorubicin, vinblastine and etoposide. An explanation for this phenomenon had not been found. © 1999 Cancer Research Campaig

Fatal hepatitis B reactivation following discontinuation of nucleoside analogues for chronic hepatitis B

Background: Nucleoside analogues such as lamivudine for chronic hepatitis B have an excellent safety profile while patients are on therapy but reactivation flares occur in 19–50% of patients after stopping therapy, some of whom develop liver decompensation. Aims: To describe and report three cases who developed fatal hepatitis B reactivation after stopping nucleoside analogue therapy. Subjects and results: Three patients are described who developed hepatitis B reactivation and liver decompensation after stopping therapy. One of the three patients was participating in a famciclovir trial and the other two were receiving lamivudine therapy for active hepatitis B infection. All three patients had documented hepatitis B flares, and all had hepatitis B virus DNA detected at that time. All patients developed decompensated liver disease despite one patient having had a prior liver biopsy showing absence of cirrhosis. Reintroduction of lamivudine therapy failed to halt progression of liver decompensation even after hepatitis B virus DNA had been demonstrated to be absent. Sequencing for lamivudine resistant mutants in two cases where serum was available failed to show evidence of mutations associated with lamivudine resistance. Conclusion: Hepatitis B virus reactivation, leading to decompensation and death, are possible complications of treatment withdrawal and patients should be monitored closely if therapy is ceased

Sex and Trait Anxiety Differences in Psychological Stress are Modified by Environment

Evidence-based research has revealed how physiological and emotional responses to acute stress are adaptive. However, under conditions of unpredictable or protracted stress, health and drug vulnerability can be compromised. In this study, we examined anxiety-like behavioral responses of 4th generation adolescent male and female Long Evans rats selectively bred for high (HAn) and low (LAn) anxiety-like behavior when housed in an isolated environment (IE) versus a social environment (SE). After 35 days in IE or SE, animals were tested in the elevated plus maze (EPM), injected with amphetamine (AMPH: 0.5 mg/kg, IP) in the locomotor activity (LMA) chamber, measured for basal and post air puff-stressor core body temperature and blood pressure. Following select rearing, SE reduced the anxiogenic response in HAn rats with females displaying the lowest anxiety-like behavior in the EPM. During habituation in the LMA, IE rats remained active, while post-AMPH injection HAn females were hyperactive, followed closely by LAn females. Our findings from the post-stressor physiological measurements indicate that temperature differences due to environment are observed only in the SE females. We also observed group differences for diastolic (DBP) and systolic (SBP) blood pressure. HAn IE males experienced higher DBP and SBP but LAn IE females only experienced higher SBP. Not only do our findings corroborate earlier work on HAn/LAn lines but the findings obtained from this research offer new insights about the role of environment and the role of sex in (1) modulation of anxiety-like behavior, (2) AMPH sensitivity, and (3) basal and stress-induced physiological changes

Polarized integrin mediates human keratinocyte adhesion to basal lamina.

Epithelial cell interactions with matrices are critical to tissue organization. Indirect immunofluorescence and immunoprecipitations of cell lysates prepared from stratified cultures of human epidermal cells showed that the major integrins expressed by keratinocytes are alpha E beta 4 (also called alpha 6 beta 4) and alpha 2 beta 1. The alpha E beta 4 integrin is localized at the surface of basal cells in contact with the basement membrane, whereas alpha 2 beta 1/alpha 3 beta 1 integrins are absent from the basal surface and are localized only on the lateral surface of basal and spinous keratinocytes. Anti-beta 4 antibodies potently inhibited keratinocyte adhesion to matrigel or purified laminin, whereas anti-beta 1 antibodies were ineffective. Only anti-beta 4 antibodies were able to detach established keratinocyte colonies. These data suggest that alpha E beta 4 mediates keratinocyte adhesion to basal lamina, whereas the beta 1 subfamily is involved in cell-cell adhesion of keratinocytes

Annexin-1-deficient mice exhibit spontaneous airway hyperresponsiveness and exacerbated allergen-specific antibody responses in a mouse model of asthma

Background Glucocorticoids are the mainstream drugs used in the treatment and control of inflammatory diseases such as asthma. Annexin-1 (ANXA1) is an anti-inflammatory protein which has been described as an endogenous protein responsible for some anti-inflammatory glucocorticoid effects. Previous studies have identified its importance in other immune diseases such as rheumatoid arthritis and cystic fibrosis. ANXA1-deficient((-/-)) mice are Th2 biased, and ANXA1 N-terminus peptide exhibits anti-inflammatory activity in a rat model of pulmonary inflammation. Objective ANXA1 protein is found in bronchoalveolar lavage fluid from asthmatics. However, the function of ANXA1 in the pathological development of allergy or asthma is unclear. Thus, in this study we intended to examine the effect of ANXA1 deficiency on allergen‐specific antibody responses and airway responses to methacholine (Mch). Methods ANXA1−/− mice were sensitized with ovalbumin (OVA) and challenged with aerosolized OVA. Airway resistance, lung compliance and enhanced pause (PenH) were measured in naïve, sensitized and saline or allergen‐challenged wild‐type (WT) and ANXA1−/− mice. Total and allergen‐specific antibodies were measured in the serum. Results We show that allergen‐specific and total IgE, IgG2a and IgG2b levels were significantly higher in ANXA1−/− mice. Furthermore, naïve ANXA1−/− mice displayed higher airway hypersensitivity to inhaled Mch, and significant differences were also observed in allergen‐sensitized and allergen‐challenged ANXA1−/− mice compared with WT mice. Conclusions In conclusion, ANXA1−/− mice possess multiple features characteristic to allergic asthma, such as airway hyperresponsiveness and enhanced antibody responses, suggesting that ANXA1 plays a critical regulatory role in the development of asthma. Clinical Relevance We postulate that ANXA1 is an important regulatory factor in the development of allergic disease and dysregulation of its expression can lead to pathological changes which may affect disease progression