Biomedical subjects

S A McCarthy

Publications and source records attributed to S A McCarthy.

At least 19 recordsLinked to original sources

PubMed2005

The immediate effects of Hurricane Iniki on intertidal fauna on the south shore of O'ahu.

When Hurricane Iniki struck the Hawaiian Islands in September 1992, it provided a rare opportunity to examine the immediate effects of a hurricane on two intertidal benthic communities off the reefs of O'ahu, Hawai'i. The Niu Beach site contained large, obvious aggregations of the tube building polychaete Diopatra dexiognatha, and the Wailupe Beach site was without obvious tubiculous fauna at the surface. Ten replicate sediment cores were taken before and after the hurricane with a 7.6 cm PVC corer and organisms were identified to family and enumerated. There were no substantial depletions or loss of taxa after the hurricane. Oligochaetes were the most dominant taxa pre-and post-hurricane. The abundance of all dominant polychaete families increased post-hurricane. The three most abundant polychaetes were capitellids and D. dexiognatha (Onuphidae) at Niu Beach and Pygospio muscularis (Spionidae) at Wailupe Beach. We suggest that D. dexiognatha and P. muscularis help stabilize the sediments since they both form dense tube mats while capitellids and oligochaetes are considered highly adaptive surface burrowers that can take advantage of newly disturbed sediments. Overall, there was no substantial effect observed on the intertidal fauna exposed to this severe disturbance. It is suggested here that invertebrate communities in this area are adapted to survive and thrive in high-energy environments and possibly benefit from dense aggregations of tube building polychaetes.

Animals↗

PubMed2002

Teaching medical students how to take a sexual history and discuss sexual health issues.

BACKGROUND: Taking a sexual history and discussing sexual health issues with patients form an important part of a medical consultation. These specific communication skills can be acquired through various teaching methods. OBJECTIVE: This paper describes the communication skill workshops conducted for undergraduate medical students on how to talk to patients about sex. METHODOLOGY: 198 medical students participated in a series of workshops conducted in the University of Malaya in 2001-2002. Pre- and post-workshop evaluations of the programme were carried out to find out the students' difficulties and to assess the usefulness of the workshop. The workshop consisted of a short lecture, role-plays and discussion. RESULTS: Only 34% of the participants had received some informal training during their clinical years. The main barriers encountered were gender and age differences, language and choice of words, patients and doctors feeling shy, and cultural differences. The workshop was felt to be useful (mean score 4.38, maximum 5.0), most students felt comfortable during the workshop (mean score 4.10, maximum 5.0) and there was significant improvement in the "comfort level" when talking to patients about sex after attending the workshop (P < 0.001). CONCLUSION: Gender, language and cultural differences were the main barriers in taking a sexual history and discussing sexual health issues among the medical students. Communication workshop was felt to be a useful and comfortable method of learning these specific.

Communication↗

PubMed2001

Local high-capacity adenovirus-mediated mCTLA4Ig and mCD40Ig expression prolongs recombinant gene expression in skeletal muscle.

Multiple forms of muscular dystrophy are due to the absence of cytoskeletal muscle proteins that normally protect the integrity of muscle cells. The lack of any adequate treatments for these devastating diseases propels research toward the development of strategies for gene delivery to skeletal muscle. High-capacity adenoviral vectors (HC-AdV) devoid of all viral coding sequences have been developed to avoid expression of viral proteins by the gene therapy vector. However, the capsid proteins that are an essential component of the input viral vector and any residual helper virus in the vector preparation could induce an immune response. Furthermore, the therapeutic protein provided by a gene transfer vector presents the potential to induce an immune response in a patient who does not express a normal cellular protein due to genetic mutation. Therefore, we hypothesize that some immune suppression will be required with therapeutic gene delivery designed for the treatment of patients with inherited muscle diseases. In this study, we constructed and rescued three HC-AdVs expressing murine CTLA4Ig, murine CD40Ig, or both. The backbone vector without a gene insert was rescued as a negative control vector. The production of relevant proteins from each vector was determined in vitro. In vivo function of each of the immunosuppressant vectors was assayed by co-injection with an enhanced green fluorescent protein (EGFP)-expressing first-generation adenoviral vector (AdEGFP) into the tibialis anterior muscle of C57BL/10 mice. Higher levels of muscle EGFP expression were observed in animals receiving an immunosuppressant vector. Furthermore, the production of total anti-AdV and anti-EGFP antibodies was reduced in mice treated with each of the three immunosuppressant vectors. A second intramuscular administration of AdEGFP alone 4 weeks after the initial co-injection was successful in all immunosuppressant vector-treated groups, but not in the negative control vector-treated group. All groups had a high antibody response to adenoviral proteins after the second injection of AdEGFP alone, indicating that the initial co-injection did not tolerize against vector capsid antigens.

Abatacept↗

PubMed2001

Nitric oxide induces murine thymocyte apoptosis by oxidative injury and a p53-dependent mechanism.

Previously, we showed that NO induces thymocyte apoptosis via a caspase-1-dependent mechanism [(1) ]. In the present study, we investigated the role of heme oxygenase, catalase, bax, and p53 in this process. The NO donor, S-nitroso-N-acetyl penicillamine (SNAP), induced DNA fragmentation in thymocytes in a time- and concentration-dependent way. SNAP (100 microM) induced 50--60% apoptosis; higher doses did not increase the rate of apoptosis significantly. SNAP decreased catalase and heme iron (Fe) levels without affecting superoxide dismutase, glutathione, or total Fe stores in thymocytes. SNAP significantly increased the expression of heme oxygenase 1 (HSP-32), p53, and bax but not bcl-2. Treatment with the heme oxygenase inhibitor, tin protoporphyrin IX inhibited SNAP-induced thymocyte apoptosis. Furthermore, thymocytes from p53 null mice were resistant to NO-induced apoptosis. Our data suggest that NO may induce its cytotoxic effects on thymocytes by modulating heme oxygenase and catalase activity as well as up-regulating pro-apoptotic proteins p53 and bax.

Animals↗

PubMed2000

Generation and regeneration of a novel anti-CD8-resistant cytolytic T lymphocyte population.

We recently reported that the in vivo development of a novel CD8(+), but anti-CD8 mAb-resistant, CTL population is complex and distinct from that of conventional anti-CD8 mAb-sensitive CD8(+) CTL. In this study, we explored the role of the thymus in the generation of anti-CD8-resistant pCTL and in their maintenance once they are generated. We also investigated the capacities of the adult periphery and thymus to support the regeneration of anti-CD8-resistant pCTL after peripheral lymphocyte and/or thymocyte depletion. These studies indicate that the thymus is necessary for the generation but not the maintenance of peripheral anti-CD8-resistant pCTL. These studies also indicate that the adult thymus can produce these pCTL and the adult periphery can support their regeneration, if a new wave of thymic maturation is experimentally induced. These results may have implications for immune reconstitution after treatment for cancer or HIV infection.

Animals↗

PubMed2000

A conditionally-active form of MEK1 results in autocrine tranformation of human and mouse hematopoietic cells.

The Raf/MEK/MAP kinase cascade plays a critical role in transducing growth signals from activated cell surface receptors. Using deltaMEK1:ER, a conditionally-active form of MEK1, we demonstrate the ability of this dual specificity protein kinase to abrogate the cytokine-dependency of the human and murine hematopoietic cells lines TF-1, FDC-P1 and FL5.12. Cytokine-independent cells were obtained from TF-1, FDC-P1 and FL5.12 cells at frequencies of 2.5 x 10(-3), 5 x 10(-5) and 10(-7) respectively, indicating that not all cells expressing deltaMEK1:ER were factor-independent. In general, cells that were converted to a cytokine-independent phenotype displayed a higher level of MAP kinase activity in response to deltaMEK1:ER activation than those that remained cytokine-dependent. deltaME-K1:ER-responsive cells could be maintained long-term in the presence of beta-estradiol as well as the estrogen-receptor antagonist 4-Hydroxy-Tamoxifen and the anti-estrogen ICI 164383. Removal of hormone led to the rapid cessation of cell growth in a manner similar to that observed when cytokine is withdrawn from the parental cells. Treatment of deltaMEKI:ER-responsive cells with a specific and selective inhibitor, PD98059, prevented growth in response to beta-estradiol. GM-CSF mRNA transcripts were detected in the MEK1-responsive cells indicating that the activated deltaMEK1:ER may induce a pathway leading to autocrine proliferation. Treatment of MEK1-responsive cells with an anti-GM-CSF antibody, but not a control antibody, suppressed cell growth. The cell lines described here will be useful for elaborating the ability of the MAP kinase pathway to regulate cell proliferation in hematopoietic cells.

Animals↗

PubMed2000

Evaluation of nonisotopic DNA hybridization methods for detection of the tdh gene of vibrio parahaemolyticus.

Production of the thermostable direct hemolysin (TDH) by Vibrio parahaemolyticus is associated with pathogenicity of the organism and is encoded by the tdh gene. The timely resolution of seafood-associated outbreaks requires rapid and accurate detection of pathogenic V. parahaemolyticus. The specificity of alkaline phosphatase- and digoxigenin-labeled tdh gene probes was evaluated against 61 strains of V. parahaemolyticus (including isolates from recent outbreaks involving oysters from the Pacific Northwest, Texas, and New York), 85 strains of other vibrios, and 7 strains of non-vibrio species from clinical and environmental sources. The probes were specific for detection of the V. parahaemolyticus tdh gene.

Alkaline Phosphatase↗

PubMed1999

Cytokine requirements for production of a novel anti-CD8-resistant CTL population.

A population of CD8+ CTL can be generated in vitro in the presence of anti-CD8 mAb. Due to their apparent high avidity characteristic, these anti-CD8-resistant CD8+ CTL may have important functional in vivo roles in graft rejection, and may be important in antiviral and antitumor responses. We have previously reported that this anti-CD8-resistant subset of CD8+ CTL demonstrates functional differences from anti-CD8-sensitive CD8+ CTL. One important difference between the subsets is the markedly greater dependence of anti-CD8-resistant CTL upon exogenous cytokines for their generation in vitro. In this study, we have investigated in detail the cytokine requirements for the generation of allospecific CD8+ CTL in vitro and have found that IL-4 can augment the generation of anti-CD8-sensitive but not anti-CD8-resistant CTL, whereas IL-2 or IL-12 can augment the generation of both anti-CD8-sensitive and anti-CD8-resistant CTL. However, anti-CD8-resistant CTL require at least 10-fold higher concentrations of IL-2 than do anti-CD8-sensitive CTL. This more stringent IL-2 requirement precludes the efficient generation of anti-CD8-resistant CTL in vitro in the absence of exogenous IL-2 because they cannot produce sufficient IL-2 to meet their needs, in contrast to anti-CD8-sensitive CTL. By providing exogenous cytokines to allospecific CTL generation cultures, we further demonstrate that anti-CD8-resistant CTL can be functionally skewed to the Tc1 subset, but differ from anti-CD8-sensitive conventional CTL in that they cannot be skewed to the Tc2 subset.

Animals↗

PubMed1999

Functional and structural diversity of the human Dickkopf gene family.

Wnt proteins influence many aspects of embryonic development, and their activity is regulated by several secreted antagonists, including the Xenopus Dickkopf-1 (xDkk-1) protein. xDkk-1 inhibits Wnt activities in Xenopus embryos and may play a role in induction of head structures. Here, we characterize a family of human Dkk-related genes composed of Dkk-1, Dkk-2, Dkk-3, and Dkk-4, together with a unique Dkk-3 related protein termed Soggy (Sgy). hDkks 1-4 contain two distinct cysteine-rich domains in which the positions of 10 cysteine residues are highly conserved between family members. Sgy is a novel secreted protein related to Dkk-3 but which lacks the cysteine-rich domains. Members of the Dkk-related family display unique patterns of mRNA expression in human and mouse tissues, and are secreted when expressed in 293T cells. Furthermore, secreted hDkk-2 and hDkk-4 undergo proteolytic processing which results in cleavage of the second cysteine-rich domain from the full-length protein. Members of the human Dkk-related family differ not only in their structures and expression patterns, but also in their abilities to inhibit Wnt signaling. hDkk-1 and hDkk-4, but not hDkk-2, hDkk-3 or Sgy, suppress Wnt-induced secondary axis induction in Xenopus embryos. hDkk-1 and hDkk-4 do not block axis induction triggered either by Xenopus Dishevelled (Xdsh) or Xenopus Frizzled-8 (Xfz8), both of which function to transduce signals from Wnt ligands. Thus, hDkks 1 and 4 may inhibit Wnt activity by a mechanism upstream of Frizzled. Our findings highlight the structural and functional heterogeneity of human Dkk-related proteins.

Amino Acid Sequence↗

PubMed1999

Evidence of positive cross-regulation on Th1 by Th2 and antigen-presenting cells: effects on Th1 induced by IL-4 and IL-12.

The response of Th cells to cytokines is normally strictly regulated, such that following antigenic stimulation, Th cells respond for only a short period of time, after which they become refractory to cytokine-mediated effects. IL-12, a costimulator of Th1 having no proliferation-inducing capacity of its own, allows Th1 clones and lines to respond to IL-4 when they would otherwise be unable to respond to this cytokine. Cells that have proliferated in response to IL-4 plus IL-12 are fully able to be subsequently activated by specific Ag and APC. Additionally, the response to IL-4 of Th1 effector cells derived from normal murine spleen is enhanced significantly by IL-12. Furthermore, in the presence of IL-12, stimulated Th2 can induce proliferation of Th1 via IL-4 production, in a dual chamber culture system. We hypothesize that the effects of IL-4 and IL-12 represent a novel, positive cross-regulatory pathway that acts on Th1, and is mediated by Th2 (the IL-4 source) and APC (the IL-12 source). We propose this as a way for a Th2 immune response to positively influence an ongoing or waning Th1 response.

Animals↗

PubMed1999

Evaluation of alkaline phosphatase- and digoxigenin-labelled probes for detection of the thermolabile hemolysin (tlh) gene of Vibrio parahaemolyticus.

The biochemical identification and enumeration of Vibrio parahaemolyticus as described in the FDA Bacteriological Analytical Manual is expensive and labour-intensive. To reduce the time and effort necessary to verify the identity of V. parahaemolyticus, the use of a thermolabile haemolysin (tlh) gene probe is proposed. An alkaline phosphatase (AP)-labelled probe was evaluated for specificity against 26 strains of V. parahaemolyticus, 88 strains of other Vibrio species and 10 strains of non-vibrio species. Of the 124 isolates tested, the probe hybridized only with the 26 strains of V. parahaemolyticus, indicating species specificity. Two hundred and six suspect V. parahaemolyticus isolates from oysters were tested by this probe and API-20E diagnostic strips; there was 97% agreement between results. A digoxigenin (DIG)-labelled probe for detection of the tlh gene fragment was prepared by PCR and compared with the AP-labelled probe. When tested on 584 suspect V. parahaemolyticus isolates, results obtained with the AP- and DIG-labelled probes were in 98% agreement. These results suggest that the probes are equivalent for detection of the V. parahaemolyticus tlh gene.

Alkaline Phosphatase↗

PubMed1998

Characterization of a novel Bax-associated protein expressed in hemopoietic tissues and regulated during thymocyte apoptosis.

Members of the Bcl-2 protein family have been implicated as critical intracellular regulators of apoptosis. Most studies of this protein family have utilized transformed and/or transfected cell lines expressing high levels of these proteins. In the current study, we have analyzed normal murine lymphoid cells and tissues and have detected a previously unreported protein of approximately 16 kDa recognized by an anti-Bax Ab. This 16-kDa protein is abundant in hemopoietic tissues of both wild-type and Bax knock-out mice, it can heterodimerize with Bax in normal lymphocytes, and it is dramatically down-modulated in thymocytes in response to apoptotic stimuli. These results suggest that this protein may have antiapoptotic activity and may participate in the regulation of apoptosis in normal lymphocytes.

Animals↗

PubMed1998

Down-modulation of a novel Bax-associated protein during apoptosis in normal mature B lymphocytes.

We have recently characterized a novel 16-kDa Bax-associated protein. In this study, we investigate the regulation of this protein's expression during in vitro induction of apoptosis in mature splenic B cells. A panel of biochemically distinct apoptotic stimuli induced the dramatic down-modulation of the 16-kDa protein in B cells; this down-modulation was rapid, and did not require DNA fragmentation. Reciprocally, stimuli that induced protection from apoptosis prevented down-modulation of the 16-kDa protein. These regulatory effects were specific, since Bcl-2 and Bax protein levels were not similarly modulated. Stimuli that reduce expression of the 16-kDa protein may therefore act indirectly to increase the proapoptotic activity of Bax, perhaps by altering Bax binding to other cellular proteins.

Animals↗

PubMed1998

In vitro thymocyte maturation is associated with reduced cellular susceptibility to Fas-mediated apoptosis.

We have developed a novel system in which the susceptibility of murine thymocytes to Fas-mediated apoptosis can be modulated. Thymocyte susceptibility to Fas decreases under in vitro culture conditions that promote aspects of thymocyte maturation. The hyporesponsive state is specific for the Fas pathway, since cellular susceptibility to other apoptotic stimuli is not reduced. Hyporesponsiveness is not associated with alterations in the thymocyte subset distribution, decreased expression of full-length Fas protein, or alterations in FADD, Bcl-2, or Bcl-XL expression. Hyporesponsiveness is overcome by increasing the strength of the Fas cross-linking stimulus, leading us to propose that reduced thymocyte susceptibility to apoptosis results from altered Fas signaling. The block in Fas signaling resides proximal to ceramide generation, since Fas-hyporesponsive thymocytes are susceptible to ceramide-induced apoptosis. Further characterization of Fas signaling in these in vitro cultured thymocytes may facilitate the identification of factors regulating the susceptibility of wild-type lymphocytes to Fas.

Adaptor Proteins, Signal Transducing↗

PubMed1998

Activation requirements, lytic mechanism, and development of a novel anti-CD8-resistant CTL population.

Almost all conventional CD8+ CTL and their CD8+ precursors are inhibited by anti-CD8 mAb. This requirement for CD8 function reflects both an avidity-augmentation role and a signal-transduction role for CD8 on T cells. We have, however, previously identified and partially characterized a novel functional population of CD8+, but anti-CD8-resistant, MHC class I-allospecific CTL. These CTL have unusual activation requirements in that their efficient generation in vitro requires inhibition of the CD8 avidity contribution (but not the CD8 signaling contribution), by anti-CD8 mAb. In this study, we have investigated the relationship of anti-CD8-sensitive and anti-CD8-resistant CTL by several criteria. These CTL populations share the phenotypic markers we have tested to date, they have similar but not identical Ag-specific repertoires, and they both appear to be generated from naive unprimed T cells. However, anti-CD8-sensitive and anti-CD8-resistant CTL populations exhibit important functional differences. They differ in their kinetics of activation in vitro, their dependence on exogenous cytokines, their use of lytic effector mechanisms, and their tissue distribution during ontogeny. Based on these results, we favor the hypothesis that these CTL populations represent distinct T cell lineages or subsets, and not merely different TCR avidity ranges within a single T cell lineage or subset.

Animals↗

PubMed1997

Allospecific cytotoxic T cells generated from beta 2m-/- mice in primary MLC: analysis of activation requirements, specificity, and phenotype.

It has been demonstrated by several investigators that beta 2m-/- knockout mice are deficient in the expression of MHC Class I molecules but can nevertheless generate CD8(+) allospecific cytotoxic T cells following vigorous in vivo priming. We demonstrate here that in vivo priming is not necessary to generate MHC Class I allospecific CTL from beta 2m-/- mice. When splenocytes from naive unprimed beta 2m-/- mice were provided exogenous cytokines in MHC Class I disparate primary MLC, allospecific cytolytic effectors were generated. beta 2m-/- MHC Class I allospecific CTL that were CD3+ and Thy1.2+ were otherwise heterogeneous in phenotype, including CD8+, CD4+, CD8-CD4-, TCR alpha beta+, and TCR gamma delta+ T cells. This phenotypic variability of beta 2m-/- CTL generated in primary MLC reveals the diversity of CTL precursors that develop in vivo in the absence of MHC Class I.

Animals↗

PubMed1997

Modeling the proliferative response of T cells to IL-2 and IL-4.

Interleukin (IL) -2 and IL-4 are growth factors for both T and B cells. When both cytokines are present, synergy is observed in some cases and antagonism in others. The studies presented here describe the use of a detailed mathematical model for the proliferative response of the T cell line, HT-2. This cell line responds to IL-2 and to IL-4 and shows a synergistic response when both cytokines are present. This model incorporates the observed synergy between these two cytokines while at the same time incorporating the known down-regulatory effect of IL-4 on the number of IL-2 receptors (IL-2R) at the cell surface, and it is able to reproduce a variety of experimental data. The major results from these studies include the following: the observation that the binding of IL-4 to its receptor is 1/10 as effective in delivering a proliferative signal as IL-2 binding to its receptor, the determination of the threshold number of bindings required to signal proliferation stimulated by IL-2 and IL-4, the demonstration that many different sets of experimental data can be accurately modeled, and the use of simple parameter terms to model the synergy between IL-2 and IL-4.

Cell Division↗

PubMed1997

Crossregulation between T helper cell (Th)1 and Th2: inhibition of Th2 proliferation by IFN-gamma involves interference with IL-1.

The Th1-derived cytokine IFN-gamma inhibits the proliferation of Th2 lymphocytes, but the mechanism of inhibition is not known. Under certain disease conditions, an established Th2-mediated immune response is undesirable and a Th1-mediated response is beneficial. However, established Th2 cells appear to be phenotypically stable. Thus, learning more about cytokine-mediated regulation of established Th2 cells is important if deleterious immune responses are to be altered. We studied the effects of IFN-gamma on a panel of recently derived Th2 lines and clones, as well as a previously established Th2 clone, 13.26. Inhibition by IFN-gamma was observed only when there was a concomitant response to IL-1, a known costimulator of Th2. Clone 13.26 was particularly sensitive to both IL-1 and IFN-gamma, so it was studied in greater detail. We examined cytokine responses using stimulation by anti-TCR mAb-coated plates, or Ag presented by APC populations that do or do not produce IL-1. All IL-1-mediated proliferative responses of 13.26 were inhibited by IFN-gamma, whereas IL-1-independent (IL-4-associated) responses were unaffected. Our data suggest that IFN-gamma inhibits Th2 proliferation through an IL-1-dependent mechanism, and furthermore, that the costimulatory pathways used by APCs may be critical for subsequent Th cell responses to cytokines.

Animals↗