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Biomedical subjects

K McCall

Publications and source records attributed to K McCall.

At least 19 recordsLinked to original sources

Programmed cell death in the germline.

In many organisms, programmed cell death of germ cells is required for normal development. This often occurs through highly conserved events including the transfer of vital cellular material to the growing gametes following death of neighboring cells. Germline cell death also plays a role in such diverse processes as removal of abnormal or superfluous cells at certain checkpoints, establishment of caste differentiation, and individualization of gametes. This review focuses on the cell death events that occur during gametogenesis in both vertebrates and invertebrates. It also examines the signals and machinery that initiate and carry out these germ cell deaths.

Animals↗

Stage-specific regulation of caspase activity in drosophila oogenesis.

In Drosophila oogenesis, the programmed cell death of germline cells occurs predominantly at three distinct stages. These cell deaths are subject to distinct regulatory controls, as cell death during early and midoogenesis is stress-induced, whereas the cell death of nurse cells in late oogenesis is developmentally regulated. In this report, we show that the effector caspase Drice is activated during cell death in both mid- and late oogenesis, but that the level and localization of activity differ depending on the stage. Active Drice formed localized aggregates during nurse cell death in late oogenesis; however, active Drice was found more ubiquitously and at a higher level during germline cell death in midoogenesis. Because Drice activity was limited in late oogenesis, we examined whether another effector caspase, Dcp-1, could drive the unique morphological events that occur normally in late oogenesis. We found that premature activation of the effector caspase, Dcp-1, resulted in a disappearance of filamentous actin, rather than the formation of actin bundles, suggesting that Dcp-1 activity must also be restrained in late oogenesis. Overexpression of the caspase inhibitor DIAP1 suppressed cell death induced by Dcp-1 but had no effect on cell death during late oogenesis. This limited caspase activation in dying nurse cells may prevent destruction of the nurse cell cytoskeleton and the connected oocyte.

Actins↗

Induction of apoptosis by Drosophila reaper, hid and grim through inhibition of IAP function.

Induction of apoptosis in Drosophila requires the activity of three closely linked genes, reaper, hid and grim. Here we show that the proteins encoded by reaper, hid and grim activate cell death by inhibiting the anti-apoptotic activity of the Drosophila IAP1 (diap1) protein. In a genetic modifier screen, both loss-of-function and gain-of-function alleles in the endogenous diap1 gene were obtained, and the mutant proteins were functionally and biochemically characterized. Gain-of-function mutations in diap1 strongly suppressed reaper-, hid- and grim-induced apoptosis. Sequence analysis of these alleles revealed that they were caused by single amino acid changes in the baculovirus IAP repeat domains of diap1, a domain implicated in binding REAPER, HID and GRIM. Significantly, the corresponding mutant DIAP1 proteins displayed greatly reduced binding of REAPER, HID and GRIM, indicating that REAPER, HID and GRIM kill by forming a complex with DIAP1. These data provide strong in vivo evidence for a previously published model of cell death regulation in Drosophila.

Alleles↗

Strategies for an effective youth counter-marketing program: recommendations from commercial marketing experts.

Intensive and sustained efforts to "counter-market" tobacco among teenagers are necessary to negate the "friendly familiarity" created by tobacco advertising and to communicate the true health and social costs of tobacco use. Counter-marketing campaigns should: highlight a tobacco-free lifestyle as the majority lifestyle of diverse and interesting individuals; explain the dangers of tobacco in a personal, emotional way; offer youth empowerment and control; use multiple voices, strategies, and executions; offer constructive alternatives to tobacco use; and portray smoking as unacceptable and undesirable for everyone. Counter-marketing activities should work in concert with other interventions to alter social norms regarding tobacco.

Adolescent↗

Determination of picomolar concentrations of metal ions using fluorescence anisotropy: biosensing with a "reagentless" enzyme transducer.

Because of their high affinity and selectivity, metalloproteins can be used as transducers in novel sensors, i.e., biosensors, for the determination of trace levels of metal ions in solution. Here, we exploit carbonic anhydrase to determine picomolar to nanomolar concentrations of free transition metal ions by fluorescence anisotropy (polarization) in a reagentless format. Carbonic anhydrase variants engineered with a cysteine replacing a residue chosen near the active site (F131C and H64C) were covalently labeled with derivatives of benzoxadiazole sulfonamide. These labeled variants exhibited changes in anisotropy up to 0.07 upon binding free Cu(II), Co(II), and Zn(II) with apparent Kd's close to the values observed with wild-type apocarbonic anhydrase. The covalent attachment of the label has significant advantages over noncovalent labels we have described previously. Furthermore, the metal ion-dependent anisotropy changes were predictable using simple theory. The results demonstrate that free transition metal ions can be determined at trace levels in aqueous solution using inexpensive instruments.

Anisotropy↗

The Drosophila gene hid is a direct molecular target of Ras-dependent survival signaling.

Extracellular growth factors are required for the survival of most animal cells. They often signal through the activation of the Ras pathway. However, the molecular mechanisms by which Ras signaling inhibits the intrinsic cell death machinery are not well understood. Here, we present evidence that in Drosophila, activation of the Ras pathway specifically inhibits the proapoptotic activity of the gene head involution defective (hid). By using transgenic animals and cultured cells, we show that MAPK phosphorylation sites in Hid are critical for this response. These findings define a novel mechanism by which growth factor signaling directly inactivates a critical component of the intrinsic cell death machinery. These studies provide further insights into the function of ras as an oncogene.

Animals↗

Mahogany (mg) stimulates feeding and increases basal metabolic rate independent of its suppression of agouti.

The mahogany (mg) locus originally was identified as a recessive suppressor of agouti, a locus encoding a skin peptide that modifies coat color by antagonizing the melanocyte-stimulating hormone receptor or MC1-R. Certain dominant alleles of agouti cause an obesity syndrome when ectopic expression of the peptide aberrantly antagonizes the MC4-R, a related melanocyte-stimulating hormone receptor expressed in hypothalamic circuitry and involved in the regulation of feeding behavior and metabolism. Recent work has demonstrated that mg, when homozygous, blocks not only the ability of agouti to induce a yellow coat color when expressed in the skin of the lethal yellow mouse (AY), but also the obesity resulting from ectopic expression of agouti in the brain. Detailed analysis of mg/mg AY/a animals, presented here, demonstrates that mg/mg blocks the obesity, hyperinsulinemia, and increased linear growth induced by ectopic expression of the agouti peptide. Remarkably, however, mg/mg did not reduce hyperphagia in the AY/a mouse. Furthermore, mg/mg induced hyperphagia and an increase in basal metabolic rate in the C57BL/6J mouse in the absence of AY. Consequently, although mahogany is broadly required for agouti peptide action, it also appears to be involved in the control of metabolic rate and feeding behavior independent of its suppression of agouti.

Agouti Signaling Protein↗

Requirement for DCP-1 caspase during Drosophila oogenesis.

Caspases, a class of cysteine proteases, are an essential component of the apoptotic cell death program. During Drosophila oogenesis, nurse cells transfer their cytoplasmic contents to developing oocytes and then die. Loss of function for the dcp-1 gene, which encodes a caspase, caused female sterility by inhibiting this transfer. dcp-1- nurse cells were defective in the cytoskeletal reorganization and nuclear breakdown that normally accompany this process. The dcp-1- phenotype suggests that the cytoskeletal and nuclear events in the nurse cells make use of the machinery normally associated with apoptosis and that apoptosis of the nurse cells is a necessary event for oocyte development.

Actins↗

DCP-1, a Drosophila cell death protease essential for development.

Apoptosis, a form of cellular suicide, involves the activation of CED-3-related cysteine proteases (caspases). The regulation of caspases by apoptotic signals and the precise mechanism by which they kill the cell remain unknown. In Drosophila, different death-inducing stimuli induce the expression of the apoptotic activator reaper. Cell killing by reaper and two genetically linked apoptotic activators, hid and grim, requires caspase activity. A Drosophila caspase, named Drosophila caspase-1 (DCP-1), was identified and found to be structurally and biochemically similar to Caenorhabditis elegans CED-3. Loss of zygotic DCP-1 function in Drosophila caused larval lethality and melanotic tumors, showing that this gene is essential for normal development.

Amino Acid Sequence↗

Facing death in the fly: genetic analysis of apoptosis in Drosophila.

Apoptosis, a gene-directed form of cell death, occurs normally during development and plays a major role in many diseases, including cancer and neurodegenerative disorders. Molecular genetic studies in Drosophila have revealed the existence of three novel apoptotic activators, reaper, head involution defective and grim. Additionally, Drosophila homologs of evolutionarily conserved IAPs (inhibitor of apoptosis proteins) and CED-3/ICE-like proteases have been identified and characterized. Through the combined use of genetic, molecular, biochemical and cell biological techniques in Drosophila it should now be possible to elucidate the precise mechanism by which apoptosis occurs, and how the death program is activated in response to many distinct death-inducing signals.

Animals↗

Probes of chromatin accessibility in the Drosophila bithorax complex respond differently to Polycomb-mediated repression.

The Polycomb group (PcG) of genes are required for maintenance of the repressed state of the homeotic genes in Drosophila. There are similarities between the PcG repression and mating-type silencing in yeast or heterochromatic position effect in Drosophila, which suggest that PcG repression may involve a highly compacted chromatin structure. To test for such a structure, heterologous DNA- binding proteins were used as probes for DNA accessibility in Drosophila embryos. Binding sites for the yeast transcriptional activator GAL4 and for bacteriophage T7 RNA polymerase were inserted into the bithorax (bx) regulatory region of the endogenous Ultrabithorax (Ubx) gene, which is regulated by the PcG. Ubiquitously expressed GAL4 protein directs transcription through its binding sites only in the posterior segments where the bx region is active. The block to GAL4 activation in the more anterior segments is dependent on Polycomb (Pc) function. In contrast, T7 RNA polymerase can transcribe from its target promoter in all segments of the embryo. Thus, Pc-mediated repression blocks activated polymerase II transcription, but does not simply exclude all proteins.

Animals↗

Importance of emergency physicians as referral sources for academic medical centers. The Gallup Organization Healthcare Group.

STUDY OBJECTIVE: To identify physician referral patterns and factors involved in the patient referral process. DESIGN: A telephone survey conducted on a systematic sample of physicians who referred at least one patient a month to a tertiary care center. SETTING: A major southeastern tertiary care academic medical center. TYPE OF PARTICIPANTS: Self-identified emergency physicians, family physicians, general surgeons, internists, obstetrician-gynecologists, and pediatricians. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Emergency physicians comprised 6% of the surveyed sample. Emergency physicians referred the largest number of patients each month to a tertiary care center (40 patients per month) compared with the other target specialists, who referred an average of 16 patients per month (P < .05). Specialty services most often requested by emergency physicians were internal medicine, trauma, and cardiology. CONCLUSION: Emergency physicians refer more patients to academic medical centers than any other specialty. Academic medical centers should target emergency physicians if they wish to increase their patient referral base.

Academic Medical Centers↗

Enhancer traps in the Drosophila bithorax complex mark parasegmental domains.

Eight P elements carrying a beta-galactosidase (lacZ) reporter have been mapped to sites within the Drosophila bithorax complex. The bithorax complex contains three homeotic genes, and at least nine regulatory regions which control their expression in successive parasegments of the fly. The enhancer traps inserted at the promoter of one of the genes, Ultrabithorax, express lacZ in patterns which mimic the Ultrabithorax protein pattern. Enhancer traps in the regulatory regions do not mimic the endogenous genes, but express lacZ globally in the relevant parasegments. Some P elements carry large DNA fragments upstream of the lacZ promoter but internal to the P element. In cases where these internal sequences specify a lacZ pattern, that pattern is generally suppressed when the element is inserted in the bithorax complex. In embryos mutant for genes of the Polycomb group, the lacZ expression from the enhancer traps spreads to all segments. Thus, the enhancer traps reveal parasegmental domains that are maintained by Polycomb-mediated repression. Such domains may be realized by parasegmental differences in chromatin structure.

Animals↗

Generating lineage-specific markers to study Drosophila development.

To generate cell- and tissue-specific expression patterns of the reporter gene lacZ in Drosophila, we have generated and characterized 1,426 independent insertion strains using four different P-element constructs. These four transposons carry a lacZ gene driven either by the weak promoter of the P-element transposase gene or by partial promoters from the even-skipped, fushi-tarazu, or engrailed genes. The tissue-specific patterns of beta-galactosidase expression that we are able to generate depend on the promoter utilized. We describe in detail 13 strains that can be used to follow specific cell lineages and demonstrate their utility in analyzing the phenotypes of developmental mutants. Insertion strains generated with P-elements that carry various sequences upstream of the lacZ gene exhibit an increased variety of expression patterns that can be used to study Drosophila development.

Animals↗