Biomedical subjects

J A Williams

Publications and source records attributed to J A Williams.

At least 37 records · Page 2Linked to original sources

PubMed2001

Single nucleotide polymorphisms, metabolic activation and environmental carcinogenesis: why molecular epidemiologists should think about enzyme expression.

This commentary was written to stimulate thoughts on, and consideration of, enzyme expression data in target organs when investigating possible associations between polymorphisms in carcinogen activation enzymes, lifestyle/dietary factors and cancer risk. The lung and breast are taken as examples. There is overwhelming evidence for a genotoxic mechanism in lung cancer development, and compelling evidence for the contribution of genotoxins to breast cancer aetiology. A consistent association has been shown where lung cancer risk is decreased by a G-->A polymorphism in the myeloperoxidase (MPO) gene, which is expressed in neutrophils recruited to the lung after chemical or immunological insults. In the breast, a consistent lack of association has been observed for women who are fast N:-acetyltransferase type 2 (NAT2) acetylators consuming cooked meat. This could be explained by the lack of detectable NAT2-associated sulfamethazine acetylation activity in cytosols prepared from mammary tissue, suggesting a minor contribution to carcinogen activation. The recent identification in mammary cytosols of detectable sulfotransferase isoforms (SULT1A1 and SULT1A3), which have high catalytic efficiency for activating N:-hydroxylated heterocyclic amines (HCAs, mutagens in cooked meat), offers a more important role for these enzymes in the metabolic activation of genotoxins in the breast. The possible contribution of MPO and lactoperoxidase enzymes to carcinogen activation in mammary tissue is also considered. Sulfotransferases and peroxidases have wide substrate specificity in terms of carcinogen activation (HCAs, aromatic amines and polycyclic aromatic hydrocarbons-all present in cooked meat and tobacco smoke) compared with NATs (HCAs and aromatic amines only). For gene-environment interactions, investigations into functional polymorphisms in SULT and peroxidase genes may, therefore, offer new evidence for the involvement of genotoxins in the initiation of carcinogenesis. Identification of the isoforms (if any) of carcinogen activation enzymes that are expressed in the organs of interest will help to determine which genes to investigate in these studies.

Arylamine N-Acetyltransferase↗

PubMed2001

N-Acetyltransferases, sulfotransferases and heterocyclic amine activation in the breast.

Heterocyclic amines are mammary carcinogens in rats and their N-hydroxy metabolites are substrates for subsequent metabolic activation by N-acetyltransferases (NAT) and sulfotransferases (SULT) in man. We investigated the expression of these enzymes in human breast tissue and the relationship between NAT genotype and NAT mRNA expression or enzyme activity. Immunohistochemical staining of sections of breast tissue identified expression of NAT1 and NAT2 protein in human mammary epithelial cells, but not in the stroma. We also measured the formation of DNA adducts of the heterocyclic amines 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine in calf thymus DNA after incubation of their promutagenic N-hydroxy metabolites with mammary cytosols prepared from reduction mammoplasty tissue. Experimental observations gained from use of enzyme cofactors and NAT and/or SULT inhibitors on cytosolic enzyme activity, recombinant NAT1 activity and heterocyclic amine-DNA adduct formation suggest that both NAT1 and SULT1A enzymes contribute significantly to the activation of N-hydroxylated heterocyclic amines in mammary tissue. NAT1 mRNA transcript levels were found to be two- to three-fold higher than mRNA transcripts of the NAT2 gene in reduction mammoplasty tissue and mammary epithelial cells. NAT1-specific p-aminobenzoic acid acetylation activity, but not NAT2-specific sulfamethazine acetylation activity, was detectable in mammary cytosols. There was no association apparent between NAT genotype and the levels of NAT mRNA or NAT enzyme activity, or between NAT1 genotype and IQ-DNA adduct formation mediated by mammary cytosols. Western blot analysis of mammary cytosolic protein showed detectable levels of SULT1A1 and SULT1A3.

Amines↗

PubMed2001

Arginine induced acute pancreatitis alters the actin cytoskeleton and increases heat shock protein expression in rat pancreatic acinar cells.

Arginine induced acute pancreatitis was evaluated as a novel and distinct form of experimental pancreatitis with particular attention to the actin cytoskeleton and expression of heat shock or stress proteins. Arginine induced a dose related necrotising pancreatitis in rats, as shown by histological evaluation, and an increase in serum amylase. Severe pancreatitis induced by 4.5 g/kg arginine was accompanied by dramatic changes in the actin cytoskeleton, as visualised with rhodamine phallodin. Intermediate filaments were also disrupted, as visualised by cytokeratin 8/18 immunocytochemistry. Arginine pancreatitis was accompanied by a stress response with a large increase in the small heat shock protein HSP27, as well as HSP70, peaking at 24 hours and localised to acinar cells. There was a lower increase in HSP60 and HSP90 and no effect on GRP78. HSP27 was also shifted to phosphorylated forms during pancreatitis. A lower dose of arginine (3.0 g/kg) induced less pancreatitis but a larger increase in HSP70 and HSP27 expression and phosphorylation of HSP27. Thus HSP expression can be overwhelmed by severe damage. The present work in conjunction with earlier work on caerulein induced pancreatitis indicates that changes in the actin cytoskeleton are an early component in experimental pancreatitis.

Actin Cytoskeleton↗

PubMed2001

Molecular components of the circadian system in Drosophila.

Much of our current understanding of how circadian rhythms are generated is based on work done with Drosophila melanogaster. Molecular mechanisms used to assemble an endogenous clock in this organism are now known to underlie circadian rhythms in many other species, including mammals. The genetic amenability of Drosophila has led to the identification of some genes that encode components of the clock (so-called clock genes) and others that either link the clock to the environment or act downstream of it. The clock provides time-of-day cues by regulating levels of specific gene products such that they oscillate with a circadian rhythm. The mechanisms that synchronize these oscillations to light are understood to some extent. However, there are still large gaps in our knowledge, in particular with respect to the mechanisms used by the clock to control overt rhythms. It has, however, become clear that in addition to the brain clock, autonomous or semi-autonomous clocks occur in peripheral tissues where they confer circadian regulation on specific functions.

Animals↗

PubMed2001

Intracellular signaling mechanisms activated by cholecystokinin-regulating synthesis and secretion of digestive enzymes in pancreatic acinar cells.

The intracellular signaling mechanisms by which cholecystokinin (CCK) and other secretagogues regulate pancreatic acinar function are more complex than originally realized. CCK couples through heterotrimeric G proteins of the Gq family to lead to an increase in intracellular free Ca2+, which shows spatial and temporal patterns of signaling. The actions of Ca2+ are mediated in part by activation of a number of Ca2+-activated protein kinases and the protein phosphatase calcineurin. By the process of exocytosis the intracellular messengers Ca2+, diacylglycerol, and cAMP activate the release of the zymogen granule content in a manner that is poorly understood. This fusion event most likely involves SNARE and Rab proteins present on zymogen granules and cellular membrane domains. More likely related to nonsecretory aspects of cell function, CCK also activates three MAPK cascades leading to activation of ERKs, JNKs, and p38 MAPK. Although the function of these pathways is not well understood, ERKs are probably related to cell growth, and through phosphorylation of hsp27, p38 can affect the actin cytoskeleton. The PI3K (phosphatidylinositol 3-kinase)-mTOR (mammalian target of rapamycin) pathway is important for regulation of acinar cell protein synthesis because it leads to both activation of p70S6K and regulation of the availability of eIF4E in response to CCK. CCK also activates a number of tyrosyl phosphorylation events including that of p125FAK and other proteins associated with focal adhesions.

Animals↗

PubMed2001

We make foot exams a priority.

A simple foot exam takes less than five minutes, but it can help save a limb--or a life. So why doesn't it happen more often?

Diabetic Foot↗

PubMed2000

Activation of genotoxins to DNA-damaging species in exfoliated breast milk cells.

Exfoliated cells, isolated from breast milk samples donated by UK-resident women (n=15), were incubated, either immediately or after culture for 7 days, with one of a series of genotoxins, either in the presence or absence of the DNA-repair inhibitors, hydroxyurea (HU), and cytosine arabinoside (ara-C). The numbers of DNA single-strand breaks induced were then assessed as comet tail length (CTL) (microm) using the alkaline single cell-gel electrophoresis ('Comet') assay; cell viability was measured by trypan blue exclusion. The heterocyclic aromatic amines (HAAs) (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) (0.4 mM), 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) (1.67 mM), 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) (1.77 mM)), a polycyclic aromatic hydrocarbon (benzo[a]pyrene (B[a]P) (0.36 mM)), a nitro-polycyclic aromatic hydrocarbon (1-nitropyrene (1-NP) (1.84 mM)) and aromatic amines (o-toluidine (0.85 mM), p-chloroaniline (0. 71 mM)) each induced statistically significant (P<0.0001, Mann-Whitney test) increases in median CTLs in breast milk cells from all the donors examined when incubated (30 min, 37 degrees C) in the presence of HU/ara-C. In some cases, these compounds were also active in the absence of the repair inhibitors. There were marked variations in comet formation between donors and between genotoxins. Cell culture appeared to increase the epithelial cell proportion and cultured cells retained their ability to activate genotoxins. The results suggest that breast milk is a valuable source of human mammary cells for the study of the metabolic activation of possible carcinogens.

Biotransformation↗

PubMed2000

Programmed cell death in zebrafish rohon beard neurons is influenced by TrkC1/NT-3 signaling.

Rohon Beard (RB) cells are embryonic primary sensory neurons that are removed by programmed cell death during larval development in zebrafish. RB somatosensory functions are taken over by neurons of the dorsal root ganglia (DRG), suggesting that RB cell death may be triggered by the differentiation of these ganglia, as has been proposed to be the case in Xenopus. However, here we show that the timing of RB cell death correlates with reduced expression of trkC1, the receptor for neurotrophin NT-3, but not with the appearance of DRG, which differentiate only after most RB cells die. trkC1 is expressed in subpopulations of RB neurons during development, and cell death is initiated only in trkC1-negative neurons, suggesting a role for TrkC1 and its ligand, NT-3, in RB cell survival. In support of this, antibodies that deplete NT-3 induce RB cell death while exogenous application of NT-3 reduces death. In addition, we show that RB cell death can be prevented using a caspase inhibitor, zVADfmk, showing that during normal development, RB cells die by a caspase-dependent programmed cell death pathway possibly triggered by reduced signaling via TrkC1.

Animals↗

PubMed2000

A clinicopathologic reappraisal of brain stem tumor classification. Identification of pilocystic astrocytoma and fibrillary astrocytoma as distinct entities.

BACKGROUND: Brain stem tumors in children have been classified pathologically as low grade or high grade gliomas and descriptively as diffuse gliomas, intrinsic gliomas, midbrain tumors, tectal gliomas, pencil gliomas, dorsal exophytic brain stem tumors, pontine gliomas, focal medullary tumors, cervicomedullary tumors, focal gliomas, or cystic gliomas. METHODS: To search for a simplified and prognostic clinicopathologic scheme for brain stem tumors, the authors reviewed a consecutive cohort of patients younger than age 21 years with tumors diagnosed from 1980 through 1997. Pathology specimens and neuroimaging were classified by masked review. Statistical and survival analysis along with Cox proportional hazards regression was performed. RESULTS: Seventy-six patients were identified, with initial diagnostic magnetic resonance imaging available for 51 and pathology specimens for 48 patients. Twenty cases were classified histologically as pilocytic astrocytoma (PA), 14 as fibrillary astrocytoma (FA), and 14 as other tumors or indeterminate pathology. For all tumors, characteristics significantly associated with a worse survival rate were: symptom duration less than 6 months before diagnosis (P = 0.004); abducens palsy at presentation (P < 0.0001); pontine location (P = 0.0002); and engulfment of the basilar artery (P = 0.006). Pilocytic astrocytoma was associated with location outside the ventral pons (P = 0.001) and dorsal exophytic growth (P = 0.013); Fibrillary astrocytoma was associated with symptoms less than 6 months (P = 0. 006), abducens palsy (P < 0.001), and engulfment of the basilar artery (P = 0.002). Pilocytic astrocytoma showed 5-year overall survival (OS) of 95% (standard error [SE], 5%) compared with FA 1-year OS of 23% (SE, 11%;P < 0.0001). CONCLUSIONS: Brain stem tumors can be succinctly and better biologically classified as diffusely infiltrative brain stem gliomas-generally FA located in the ventral pons that present with abducens palsy, often engulf the basilar artery, and carry a grim prognosis-and focal brain stem gliomas-frequently PA arising outside the ventral pons, often with dorsal exophytic growth, a long clinical prodrome, and outstanding prognosis for survival. Our findings emphasize the individuality of PA as a distinct clinicopathologic entity with an exceptional prognosis.

Adolescent↗

PubMed2000

Mammary expression of xenobiotic metabolizing enzymes and their potential role in breast cancer.

Breast cancer is the major cause of cancer death in women worldwide. High penetrance genes account for only 5% of cases, whereas polymorphic low penetrance genes acting in concert with lifestyle/environmental risk factors are likely to account for a much higher proportion. Genotoxic compounds implicated in human breast carcinogenesis include endogenous compounds, estrogens, and dietary or environmental xenobiotics-heterocyclic amides, aromatic amines, polycyclic aromatic hydrocarbons, and nitropolycyclic aromatic hydrocarbons. Here we review evidence for a role of mammary-expressed enzymes that metabolically activate and/or detoxify potential genotoxic breast carcinogens: cytochrome P-450s, catechol-O-methyltransferase, epoxide hydrolase, peroxidases, glutathione S-transferases, N-acetyltransferases, sulfotransferases, and other enzymes catalyzing conjugation reactions. This information is particularly relevant in the light of evidence for the presence of genotoxic agents that require metabolic activation in mammary lipid, in nipple aspirates and in breast milk, and for the presence of DNA adducts in human mammary epithelial cells (from which most breast carcinomas originate). The effect of polymorphisms in the genes encoding these enzymes on breast cancer risk are also considered. The evidence for the role of genotoxic carcinogens in the etiology of breast cancer is compelling, but mammary-specific enzyme expression should be taken into account when considering the contribution of polymorphisms to risk.

Animals↗

PubMed2000

Scatter factor/hepatocyte growth factor protects against cytotoxic death in human glioblastoma via phosphatidylinositol 3-kinase- and AKT-dependent pathways.

We have shown recently that the multifunctional growth factor, scatter factor/hepatocyte growth factor (SF/HGF), and its receptor c-met enhance the malignancy of human glioblastoma through an autocrine stimulatory loop (R. Abounader et al., J. Natl. Cancer Inst., 91: 1548-1556, 1999). This report examines the effects of SF/HGF:c-met signaling on human glioma cell responses to DNA-damaging agents. Pretreating U373 human glioblastoma cells with recombinant SF/HGF partially abrogated their cytotoxic responses to gamma irradiation, cisplatin, camptothecin, Adriamycin, and Taxol in vitro. This cytoprotective effect of SF/HGF occurred at least in part through an inhibition of apoptosis, as evidenced by diminished terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling index and reduced DNA laddering. Anti-c-met U1/ribozyme gene transfer inhibited the ability of SF/HGF to protect against single-strand DNA breakage, DNA fragmentation, and glioblastoma cell death caused by DNA-damaging agents, demonstrating a requirement for c-met receptor function. Phosphorylation of the cell survival-promoting kinase Akt (protein kinase B) resulted from SF/HGF treatment of U373 cells, and both Akt phosphorylation and cell survival induced by SF/HGF were inhibited by phosphatidylinositol 3-kinase inhibitors but not by inhibitors of mitogen-activated protein kinase kinase or protein kinase C. Cytoprotection by SF/HGF in vitro was also inhibited by transient expression of dominant-negative Akt. Transgenic SF/HGF expression by intracranial 9L gliosarcomas reduced tumor cell sensitivity to gamma irradiation, confirming the cytoprotective effect of SF/HGF in vivo. These findings demonstrate that c-met receptor activation by SF/HGF protects certain glioblastoma cells from DNA-damaging agents by activating phosphoinositol 3-kinase-dependent and Akt-dependent antiapoptotic pathways.

Animals↗

PubMed2000

Oxidative stress interferes with cancer chemotherapy: inhibition of lymphoma cell apoptosis and phagocytosis.

Many antineoplastic drugs kill tumor cells by inducing apoptosis. This highly controlled mechanism of cell death is thought to be physiologically advantageous because apoptotic cells are removed by phagocytosis before they lose their permeability barrier, thus preventing induction of an inflammatory response to the dying cells. In contrast, necrotic cells lyse and release their contents into the extracellular space, thus inducing inflammation. In this report, we examine the effects of oxidative stress on chemotherapy-induced cell killing. We find that H(2)O(2) inhibits the ability of 4 different chemotherapy drugs (VP-16, doxorubicin, cisplatin, and AraC) to induce apoptosis in human Burkitt lymphoma cells. H(2)O(2) shifts the form of cell death from apoptosis to pyknosis/necrosis, which occurs after a significant delay compared with chemotherapy-induced apoptosis. It can also lower the degree of cell killing by these drugs. These effects of H(2)O(2) can be prevented by the antioxidant agents Desferal, Tempol, and dimethylsulfoxide. Phagocytosis by monocyte-derived macrophages of VP-16-treated lymphoma cells is also inhibited by H(2)O(2). Cells killed with H(2)O(2) (with or without VP-16) do ultimately undergo phagocytosis, but this occurs only after they have lost their permeability barrier. Thus, membrane-intact apoptotic cells are recognized and phagocytosed by monocyte-derived macrophages, but membrane-intact pyknotic/necrotic cells are not. The results suggest that chemotherapy-induced apoptosis and phagocytosis of cancer cells may be enhanced by including certain antioxidant agents in the treatment protocol.

Antineoplastic Agents↗

PubMed2000

Optimization of multiple-isocenter treatment planning for linac-based stereotactic radiosurgery.

OBJECTIVE: Computer-assisted treatment planning for linac-based radiosurgery is still an open research problem, especially for multiple-isocenter procedures, primarily due to its high complexity and computational requirements. This paper focuses on the optimization of multiple-isocenter treatment planning for linac systems, and addresses several important issues associated with multiple isocenters, such as dose conformality, homogeneity, and optimization of isocenter position and dose. METHODS: The key idea behind our approach is that the desired dose distribution can be decomposed into a number of fundamental components. In the current paper, an analytical form, the so-called Ellipsoidal Dose Distribution Estimation (EDDE) model, represents each component. We establish ways (arc configurations) to achieve such ellipsoidal doses of arbitrary position, orientation, and size. Since the EDDE model is described by relatively few parameters, it allows very quick estimation of the dose distribution corresponding to a particular isocenter and thus makes the optimization of isocenter position very efficient. It is further used in a framework for optimal treatment planning, in which a number of ellipsoidal dose distributions, each corresponding to a different isocenter, are optimally placed to cover the target while sparing healthy tissue. RESULTS: The general ellipsoidal dose distribution of linac-based radiosurgery is summarized as a mathematical model with the aid of supporting experiments. Comparisons between the EDDE-optimized and clinically implemented plans are made, revealing the superior performance of the former. In addition, a dramatic reduction in planning time is achieved using the EDDE model. CONCLUSION: The proposed EDDE model is a useful and effective dose model in multiple-isocenter treatment planning for linac-based radiosurgery.

Brain Neoplasms↗

PubMed2000

Effect of monovalent ions in glass ionomer cements on their interaction with sodium fluoride solution.

The effects on surface morphology of glass ionomer cements following exposure to 0.2% NaF solution were studied. The effect of cement on the solution was also evaluated. The four cements were chosen to contain Na and F, Na alone, F alone and neither Na nor F to show any interactions produced by having the same ion in both the cement and solution. Four glass ionomer cements were formulated so that they differed only in respect of the glass component. AH2 (a glass used in dental restorative cement) contained both Na and F, MP4 (a glass used in orthopaedic cement) contained Na only, LG26 (a glass used in surgical cement) contained F only and LG30 (an experimental control glass) contained neither F nor Na. Discs of cement were set in moulds at 37 degrees C for 1 h, then matured in water for 3 d. Each test disc was then immersed in 10 ml 0.2% NaF for 24 h at 37 degrees C whereas control discs remained in water. The test and control disc surfaces were assessed qualitatively using electron microscopy and quantitatively by linear profilometry generating roughness values (Ra). Test solution pH was measured before and after cement immersion. Inspection of the electron micrographs showed considerable disruption of AH2 and LG26 test surfaces compared to their controls whereas MP4 and LG30 showed similar surfaces for test and control. Statistical analysis of the Ra values showed that AH2 and LG26 test surfaces were significantly rougher than their controls as well as LG30 and MP4 test surfaces, which were not significantly different from their controls. All NaF solutions show pH increases; those for AH2 and MP4 were significantly higher than those for LG26 and LG30. The F-containing cements were subject to surface disruption whereas F-free cements were not. The Ra values of test surfaces correlated strongly (r = 0.998) with the F uptake of the cements (data from a previous study) but it was not possible to ascribe the causality to this association. The pH changes appear to be influenced by whether or not Na is present in the cement. The resultant pH values are too near to neutral for pH alone to explain the surface disruption observed. In addition, it is concluded that the changes in OH ion concentration are too low to permit F-/OH- interchange as a possible explanation for F uptake by these cements.

Anions↗

PubMed2000

Effects of adding sodium and fluoride ions to glass ionomer on its interactions with sodium fluoride solution.

This investigates the effects of the addition of Na and F ions to a glass ionomer cement in which those ions are not inherently present on its interactions with dilute (0.2%) NaF solution. Both the effect of the solution on the cement's surface morphology and the effect of the cement on the solution in terms of take up of Na+ and F- and of change in pH are to be investigated. These results are to be compared to previous results obtained with glasses which contained both, one, or neither of the ions as components of their glasses. NaF (1.3% by weight in the mixed cement) was added to the powder components of a glass ionomer based on LG30 glass (which contains Al, Si, Ca, P, and O only). Discs of cement were set in moulds at 37 degrees C for 1 h then stored in water at 37 degrees C for 3 days. Each test disc was then immersed in 10 ml 0.2% NaF solution whereas controls remained immersed in water (N = 3 for test and control). Test and control disc surfaces were assessed both qualitatively by electron microscopy and quantitatively by linear profilometry (Ra values). Potentiometry was used to measure solution pH and Na and F concentrations using a pH electrode and suitable ion selective electrodes both before and after cement immersion. The surface of test specimens was subject considerable disruption with the polysalt cement matrix being removed and residual glass particles being disclosed. The controls showed no such disruption. This effect was reflected in a significant difference of Ra. Such an effect was not shown by test and control surfaces of LG30 but a similar effect was to that shown by LG26 (which contains F as a glass component). Solution pH changed by 1 unit which was much more than the change shown by LG30 or LG26 but is similar to that of AH2 and MP4 cements which both contain Na. The Na and F uptake was much lower than for LG30 whereas that of LG26 was higher than LG30. The Na:F ratio was 0.29:1 compared to 1.26:1 for LG30 (LG26 = 1.01:1, AH2 = 1.02:1, MP4 = 1.04:1). Fluoride addition to a F-free glass ionomer renders it vulnerable to surface disruption by NaF solution showing that fluoride complexes produced in glass dissolution are not necessarily involved in this process. Sodium addition to a Na-free glass ionomer confirms the role of this cement in enhancing pH change in NaF solution. The level of uptake of F- from a NaF solution in much lower than that for the F-free glass ionomer which shows there is no direct relationship between F- uptake and surface disruption. The ratio of Na:F uptake is below 0.3:1, but the pH change is similar to cements where the ratio is close to unity which indicates that F-/OH- interchange is not a significant mechanism even when anion/cation uptake is not balanced.

Anions↗

PubMed2000

Molecular modelling and drug design.

Drug design is a creative act of the same magnitude as composing, sculpting, or writing. The results can touch the lives of millions, but the creator is rarely one scientist and the rewards are distributed differently in the arts than in the sciences. The mechanisms of creativity are the same, i.e., incremental (plodding from darkness to dawn) or sudden (the "Eureka" effect) realization, but both are poorly understood. Creativity remains a human characteristic, but it is directly related to the tools available, especially computer software and hardware. While modelling software continues to mature, very little new has evolved in terms of hardware. Here, we discuss the history of molecular modelling and describe two novel modelling tools, a haptic device and a program, SCULPT, to generate solid molecular models at atomic resolution.

Databases, Factual↗

PubMed2000

Cell turnover in neuromasts of zebrafish larvae.

The numbers and positions of cells undergoing cell death and proliferation in the neuromasts of 10 day old zebrafish larvae were assessed to investigate the ability of supporting cells to differentiate into hair cells. Evaluations of cell death and proliferation showed that a subpopulation of cells located in the centre of the neuromast undergo cell death, and a different subpopulation located at the periphery proliferate. This suggests that cell death of hair cells and proliferation of mantle supporting cells occurs as part of normal development, creating constant turnover of hair cells. We show that the caspase inhibitor zVADfmk reduces cell death while the aminoglycoside neomycin specifically induces an increased amount of cell death in the central population of cells. Both of these treatments affect the rate of proliferation of the peripheral subpopulation of cells in the neuromast suggesting that a feedback mechanism occurs regulating cell death and proliferation. We propose that the dying population of cells are hair cells and the proliferating cells are 'mantle' supporting cells, which is in agreement with previous observations suggesting that supporting cells can give rise to hair cells following hair cell death.

Amino Acid Chloromethyl Ketones↗

PubMed2000

Rest in Drosophila is a sleep-like state.

To facilitate the genetic study of sleep, we documented that rest behavior in Drosophila melanogaster is a sleep-like state. The animals choose a preferred location, become immobile for periods of up to 157 min at a particular time in the circadian day, and are relatively unresponsive to sensory stimuli. Rest is affected by both homeostatic and circadian influences: when rest is prevented, the flies increasingly tend to rest despite stimulation and then exhibit a rest rebound. Drugs acting on a mammalian adenosine receptor alter rest as they do sleep, suggesting conserved neural mechanisms. Finally, normal homeostatic regulation depends on the timeless but not the period central clock gene. Understanding the molecular features of Drosophila rest should shed new light on the mechanisms and function of sleep.

Animals↗