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V Frost

Publications and source records attributed to V Frost.

12 recordsLinked to original sources
PubMed2001

Exploitation of a non-apoptotic caspase to regulate the abundance of the cdkI p27(KIP1) in transformed lymphoid cells.

Expression of the cyclin dependent kinase inhibitor p27(KIP1) is intimately linked to the control of proliferation, and is itself regulated by transcription, translation, phosphorylation, protein stability or sequestration. p27(KIP1) is also regulated during apoptosis; cleavage occurs at DPSD(139)S and ESQD(108)V, by a sub-set of Z-VAD-fmk-sensitive caspases. We have identified a related but distinct mechanism that regulates p27(KIP1) in proliferating lymphoid cell lines. In a B-lymphoid cell line (BJAB), the abundance of p27(KIP1) oscillates inversely to proliferation; loss of full-length p27(KIP1) correlates with the appearance of a truncated version corresponding to cleavage at DPSD(139)S. A direct correlation exists between the appearance of truncated p27(KIP1) and the presence of an activity able to cleave peptides representing DPSD(139)S and a caspase-8 substrate (Ac-IETD-AMC) in vitro. This activity is inhibited by Ac-IETD-CHO but not Z-VAD-fmk in vitro. Furthermore a requirement for caspase-8 has been excluded. The activity differs from the apoptosis related p27(KIP1)-cleaving activity; indeed few cells undergoing apoptosis are present in the population of proliferating cells. The activity is further distinguished by its inability to cleave a peptide based on ESQD(108)V in vitro, together with the lack of a corresponding cleavage product in vivo. Inhibition of the caspase activity in vivo promotes an accumulation of full length p27(KIP1), as well as a decrease in cell proliferation. Together these studies highlight the importance of non-apoptotic caspases in regulating p27(KIP1) in transformed lymphoid cells.

Amino Acid Chloromethyl Ketones↗
PubMed2000

p27KIP1 is down-regulated by two different mechanisms in human lymphoid cells undergoing apoptosis.

The cyclin-dependent kinase inhibitor p27KIP1 is a crucial component of the mammalian restriction point, and as such is subject to multiple regulatory mechanisms. It has recently been shown that the abundance of p27KIP1 is also regulated during apoptosis; p27KIP1 is cleaved by a Z-VAD-fmk-sensitive caspase during apoptosis induced by growth factor deprivation in endothelial cells, and also following exposure of myeloid leukaemia cells to etoposide. Here, we investigate p27KIP1 regulation in B- and T-lymphoid cells undergoing apoptosis. We observe that p27KIP1 is down-regulated following exposure to a variety of apoptotic stimuli including an agonistic anti-Fas antibody, cycloheximide and etoposide. Further investigation revealed the existence of two different routes of p27KIP1 regulation in lymphoid cells undergoing apoptosis. The first pathway is utilized by lymphoid cells stimulated through Fas, is abrogated in a caspase-8-deficient T-cell line, and is blocked by the caspase inhibitors Z-VAD-fmk and Boc-D-fmk. In contrast, the loss of p27KIP1 in cells exposed to cycloheximide and etoposide occurs in the absence of caspase-8 or any Z-VAD-fmk- or Boc-D-fmk-sensitive caspase activities. Thus the down-regulation of p27KIP1 is a common occurrence in lymphoid cells undergoing apoptosis but, depending on the apoptotic trigger, this can be affected by two different mechanisms.

Amino Acid Chloromethyl Ketones↗
PubMed1999

Loss of functional pRB is not a ubiquitous feature of B-cell malignancies.

Human cancers frequently sustain genetic mutations that alter the function of their G1 cell cycle control check point. These include changes to the retinoblastoma gene and to the genes that regulate its phosphorylation, such as the cyclin-dependent kinase inhibitor p16INK4a. Altered expression of retinoblastoma protein (pRb) is associated with non-Hodgkin's lymphoma, particularly centroblastic and Burkitt's lymphomas. pRb is expressed in normal B-cells and its regulatory phosphorylation pathway is activated in response to a variety of stimuli. Since human B-lymphoma-derived cell lines are often used as in vitro model systems to analyse the downstream effects of signal transduction, we examined the functional status of pRb in a panel of human B-cell lines. We identified eleven cell lines which express the hyperphosphorylated forms of pRb. Furthermore, we suggest that the pRb protein appears to be functional in these cell lines.

Burkitt Lymphoma↗
PubMed1996

Hypersensitivity to rapamycin of BJAB B lymphoblastoid cells.

Proliferation of the BJAB B-lymphoblastoid cell line was rapidly and almost completely suppressed by picomolar concentrations of the immunosuppressive macrolide rapamycin (50% inhibitory concentration 10-20 pM for cells reactivated from stationary phase). This cell line was considerably more sensitive to rapamycin than any other B-lymphoblastoid cell line tested, the Jurkat T-cell line or the HL60 promyelocytic cell line. BJAB cell proliferation was not affected by the related immunosuppressive macrolides FK506 or L-685,818, which bind to the immunophilin FKBP12 competitively with rapamycin and also inhibit its peptidylprolyl cis-trans isomerase activity. Excess FK506 or L-685,818 added simultaneously competitively antagonized rapamycin's anti-proliferative action. Levels of FKBP12 and uptake of rapamycin from the culture medium were also normal in BJAB cells. The hypersensitivity to rapamycin of BJAB cells thus reflects an unusual dependence on the intracellular signalling system targeted by the rapamycin-FKBP12 complex, and may provide a model system for elucidating the role played by this pathway in lymphocyte activation. The proliferation of BJAB cells reactivated from stationary phase can also be used as the basis for a highly sensitive bioassay for the presence of rapamycin in culture media or other biological fluids.

B-Lymphocytes↗
PubMed1995

The phosphodiesterase inhibitor SQ 20006 selectively blocks mitogen activation of p70S6k and transition to S phase of the cell division cycle without affecting the steady state phosphorylation of eIF-4E.

In quiescent cells high levels of protein synthesis are required in order to re-enter the cell cycle upon stimulation. Initiation of polypeptide synthesis is the step most often subject to regulation, controlled in part by phosphorylation of 40 S ribosomal protein S6 and a number of initiation factors. The kinase responsible for S6 phosphorylation is p70S6k. We now show that the p70S6k pathway can be selectively blocked by the aminopurine analogue, SQ 20006. This agent is known to raise cAMP levels, resulting in activation of protein kinase A. We present evidence that the increase in cAMP is not responsible for the inhibitory effect observed. We also show that SQ 20006 can prevent the activation of p70S6k in a rapid and reversible manner. The compound does not exert its inhibitory activity on p70S6k but can inhibit in vitro two protein kinase C isozymes (alpha and gamma). In a B lymphoblastoid cell line, treatment with SQ 20006 results in inhibition of protein synthesis at the initiation stage. In contrast, when tested directly upon the translational machinery in the reticulocyte lysate, inhibition is manifest at both the level of initiation and elongation. The role of protein kinase A in the modulation of p70S6k and the rate of translation is discussed.

1-Methyl-3-isobutylxanthine↗
PubMed1994

Effects of extracellular divalent cations on responses of human blood platelets to adenosine 5'-diphosphate.

The effects of extracellular divalent cations on the responses of human platelets to adenosine 5'-diphosphate (ADP) and on its inhibition by the competitive antagonist adenosine 5'-triphosphate (ATP) were investigated. Two responses were studied, shape change and the inhibition of prostaglandin E1 (PGE1)-stimulated adenylate cyclase, and experiments were carried out in the presence of divalent cations (Ca2+ and Mg2+, 1 mM) or in their absence. For each response there was a small leftward shift of the concentration-response curve to ADP in the absence of divalent cations compared to that in their presence, and this leftward shift disappeared when the results were plotted in terms of ADP3- rather than total ADP concentration. The shape change results were, however, complicated by a reduction in the maximal response to ADP in the absence of divalent cations. For each response there was also a marked increase in the pA2 value of ATP in the absence of divalent cations compared to that in their presence, and this difference disappeared if the results were calculated in terms of ATP4- instead of total ATP. These results suggest that the human platelet ADP receptor, in common with other receptors for adenine nucleotides, recognises predominantly the uncomplexed forms of ADP and ATP as ligands.

Adenosine Diphosphate↗
PubMed1991

Psychological aspects of orthognathic surgery: how people respond to facial change.

This study was undertaken to explore the motivations and problems shared by adult orthognathic surgical patients over the age of 25. Objective findings included demographic information and reasons for seeking surgery. The majority of 65 respondents cited functional problems as their primary reason for seeking treatment. Eighty-nine percent were pleased with esthetic changes, and 83% responded that the functional problem had been corrected. For most, the greatest discomfort related to the surgery was the postoperative intensive care unit. Two of the most common side effects of the surgery were the loss of sensation in the lips and chin area and a short period of depression. Women had depression more often than men but were more enthusiastic about the final results of the procedure. Two major areas of interest to surgeons emerged from the research. First, although women have functional problems, the majority seem to have a desire for cosmetic improvement. Having a functional problem seemed to provide the psychological permission necessary to spend the time and money for a cosmetic change. The second point focused on the need for good communication between surgeon and patient. The patients who were more positive toward the procedure and more satisfied with the results were those who were better informed and who thought they had a good system of communication with the orthodontist, surgeon, and their respective staffs.

Adult↗