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Biomedical subjects

R M Siegel

Publications and source records attributed to R M Siegel.

At least 19 recordsLinked to original sources
PubMed2003

Cell responsiveness in macaque superior temporal polysensory area measured by temporal discriminants.

The firing-rate data from 341 cells from two macaques' superion temporal polysensory area (STPa) were subjected to three different analyses to determine the temporal firing-rate patterns in response to visual optic flow patterns. The data were collected while the monkey viewed four types of optic flow and responded to the change in the display. The mean firing rate (MFR) analysis considered the mean change in firing rate for 500 ms after stimulus onset; the discriminant (DIS) analysis and the principal components (PCA+DIS) analysis considered the change in time-binned firing rate over 1000 ms after stimulus presentation, using bin sizes of 30 to 500 ms. The DIS analysis used a step-down discriminant analysis to find temporal windows in which the cell's firing rate could discriminate among the stimuli; the PCA+DIS analysis extracted the principal components of the cell's firing rates without regard for the stimulus type and then applied a step-down discriminant analysis to the PCA scores to determine whether any of the principal components could discriminate among the stimuli. The two temporal analyses found cells sensitive to the optic flows that the MFR analysis missed. A small proportion of cells showed multiple selectivities under the temporal analyses. Thus, the temporal analyses give a more complete representation of the information encoded by the firing properties of STPa neurons. Finally, this approach incorporates temporal approaches with classical statistical techniques in order to select tuned neurons from a population in an unbiased manner.

Action Potentials↗
PubMed2001

Immunophenotypic profiles in families with autoimmune lymphoproliferative syndrome.

Autoimmune lymphoproliferative syndrome (ALPS) type Ia is caused by inherited defects in apoptosis and is characterized by nonmalignant lymphoaccumulation, autoimmunity, and increased alpha/beta(+) double-negative T cells (alpha/beta(+)-DNT cells). This study reports immunophenotypic findings in 166 members of 31 families with ALPS type Ia, associated with genetic mutations in the TNFRSF6 gene encoding Fas. The ALPS type Ia probands (n = 31) and relatives having both a Fas mutation and clinically proven ALPS (n = 28) showed significant expansion of CD8(+) T cells, alpha/beta(+)-DNT cells, gamma/delta(+)-DNT cells, CD3(+)/ HLA-DR(+) T cells, CD8(+)/CD57(+) T cells, and CD5(+) B cells. Relatives with Fas mutations, but without all the required criteria for ALPS (n = 42), had expansions of CD8(+) T cells, alpha/beta(+)-DNT cells, and gamma/delta(+)-DNT cells. Interestingly, relatives without a Fas mutation and with no features of ALPS (n = 65) demonstrated a small but significant expansion of CD8(+) T cells, both DNT cell subsets, and CD5(+) B cells. As compared to unrelated healthy controls, lymphocyte subset alterations were greatest in the probands, followed by the relatives with mutations and ALPS. Probands and relatives with mutations and ALPS also showed a lower number of CD4(+)/CD25(+) T cells that, in combination with an independent increase in HLA-DR(+) T cells, provided a profile predictive of the presence of clinical ALPS. Because quantitative defects in apoptosis were similar in mutation-positive relatives regardless of the presence of clinical ALPS, factors, other than modifiers of the Fas apoptosis pathway, leading to these distinctive immunophenotypic profiles most likely contribute to disease penetrance in ALPS.

Adult↗
PubMed2001

Fluorescence resonance energy transfer analysis of cell surface receptor interactions and signaling using spectral variants of the green fluorescent protein.

BACKGROUND: Fluorescence resonance energy transfer (FRET) is a powerful technique for measuring molecular interactions at Angstrom distances. We present a new method for FRET that utilizes the unique spectral properties of variants of the green fluorescent protein (GFP) for large-scale analysis by flow cytometry. METHODS: The proteins of interest are fused in frame separately to the cyan fluorescent protein (CFP) or the yellow fluorescent protein (YFP). FRET between these differentially tagged fusion proteins is analyzed using a dual-laser FACSVantage cytometer. RESULTS: We show that homotypic interactions between individual receptor chains of tumor necrosis factor receptor (TNFR) family members can be detected as FRET from CFP-tagged receptor chains to YFP-tagged receptor chains. Noncovalent molecular complexation can be detected as FRET between fusions of CFP and YFP to either the intracellular or extracellular regions of the receptor chains. The specificity of the assay is demonstrated by the absence of FRET between heterologous receptor pairs that do not biochemically associate with each other. Interaction between a TNFR-like receptor (Fas/CD95/Apo-1) and a downstream cytoplasmic signaling component (FADD) can also be demonstrated by flow cytometric FRET analysis. CONCLUSIONS: The utility of spectral variants of GFP in flow cytometric FRET analysis of membrane receptors is demonstrated. This method of analyzing FRET allows probing of noncovalent molecular interactions that involve both the intracellular and extracellular regions of membrane proteins as well as proteins within the cells. Unlike biochemical methods, FRET allows the quantitative determination of noncovalent molecular associations at Angstrom level in living cells. Moreover, flow cytometry allows quantitative analyses to be carried out on a cell-by-cell basis on large number of cells. Published 2001 Wiley-Liss, Inc.

Adaptor Proteins, Signal Transducing↗
PubMed2001

Barriers to domestic violence screening in the pediatric setting.

OBJECTIVE: By surveying practitioners in our community, we hoped to determine what pediatricians and family physicians (FPs) perceive as barriers to the American Academy of Pediatrics (AAP) Recommendation on Domestic Violence Screening. BACKGROUND: When screened in the pediatric setting, as many as 40% of mothers will disclose domestic violence (DV) by their partner. Recognizing the profound effects of DV on children, the AAP recently recommended that all practitioners incorporate DV screening as a part of routine anticipatory guidance. Yet, there is little information about whether pediatricians have the training, the time to screen, or understand the magnitude of this problem. DESIGN/METHODS: A 22-question survey about attitudes, training, and current DV screening practice was sent to all general pediatricians and FPs with admitting privileges to Children's Hospital Medical Center in Cincinnati, Ohio. A copy of the AAP recommendation on screening was included. The vast majority of practitioners with an appreciable pediatric practice in the surrounding tri-state area of 1.8 million people have privileges at the institution. RESULTS: After 2 mailings, 310 (57%) of 547 of questionnaires were returned. The majority of practitioners (64%) were unaware of the AAP recommendation, but 51% of practitioners screened at least high-risk families for DV and 49% had identified a case of DV in their practice. Still, only 8.5% routinely screened for DV and 74% had received no specific DV training. Fifty-eight percent of practitioners estimated the incidence of DV to be <5% in their practice. The most commonly perceived barriers to screening were lack of education (61%), office protocol (60%), time (59%), and support staff (55%). FPs were significantly more likely to have DV training (64% vs 21%), more likely to screen at least high-risk women (79% vs 56%), and more likely to have identified a case of DV (92% vs 40%) than pediatricians. FPs were less likely to cite lack of education (46% vs 65%) and lack of time (50% vs 61%) than pediatricians. Physicians licensed in Ohio were less likely to have specific domestic violence training (23% vs 60%) as compared with Kentucky physicians, where domestic violence education is required for licensing. Kentucky physicians were less likely to cite lack of education as barrier to DV screening (20% vs 62%). When comparing the characteristics of those who screen to those who do not, those with DV training were 10.9 times (odds adjusted ratio) more likely to screen. CONCLUSIONS: Practitioners grossly underestimate the incidence of DV in their practices. Lack of education including knowledge of screening recommendations is a barrier to DV screening by pediatricians. Greater efforts are needed to educate pediatricians on DV for the AAP recommendations to be accepted.domestic violence, child abuse, screening, physician attitude.

Adult↗
PubMed2000

A domain in TNF receptors that mediates ligand-independent receptor assembly and signaling.

A conserved domain in the extracellular region of the 60- and 80-kilodalton tumor necrosis factor receptors (TNFRs) was identified that mediates specific ligand-independent assembly of receptor trimers. This pre-ligand-binding assembly domain (PLAD) is physically distinct from the domain that forms the major contacts with ligand, but is necessary and sufficient for the assembly of TNFR complexes that bind TNF-alpha and mediate signaling. Other members of the TNFR superfamily, including TRAIL receptor 1 and CD40, show similar homotypic association. Thus, TNFRs and related receptors appear to function as preformed complexes rather than as individual receptor subunits that oligomerize after ligand binding.

Amino Acid Substitution↗
PubMed2000

Fas preassociation required for apoptosis signaling and dominant inhibition by pathogenic mutations.

Heterozygous mutations encoding abnormal forms of the death receptor Fas dominantly interfere with Fas-induced lymphocyte apoptosis in human autoimmune lymphoproliferative syndrome. This effect, rather than depending on ligand-induced receptor oligomerization, was found to stem from ligand- independent interaction of wild-type and mutant Fas receptors through a specific region in the extracellular domain. Preassociated Fas complexes were found in living cells by means of fluorescence resonance energy transfer between variants of green fluorescent protein. These results show that formation of preassociated receptor complexes is necessary for Fas signaling and dominant interference in human disease.

Animals↗
PubMed2000

Measurement of molecular interactions in living cells by fluorescence resonance energy transfer between variants of the green fluorescent protein.

Many signal transduction pathways operate through oligomerization of proteins into multi-subunit complexes. Although biochemical assays can identify potential protein-protein interactions, studying these interactions in living cells is more challenging. Fluorescence resonance energy transfer (FRET) has been used as a "spectroscopic ruler" to measure molecular proximity, but these methods have been limited by the need for chemical labeling of target proteins or labeled antibodies. We present methods for examining interactions between target proteins molecularly fused to cyan and yellow variants of the green fluorescent protein (GFP) by FRET in living cells. Flow cytometric and microscope-based methods are described that have been applied to a variety of interacting proteins.

Cell Line↗
PubMed2000

The multifaceted role of Fas signaling in immune cell homeostasis and autoimmunity.

Originally identified as a cell surface receptor that triggered the death of lymphocytes and tumor cells, it is now recognized that Fas (also known as CD95 or Apo-I) has distinct functions in the life and death of different cell types in the immune system. Fas signaling may also be involved in T cell costimulation and proliferation. Although Fas deficiency in humans and mice predisposes them towards systemic autoimmunity, Fas-FasL interactions can also facilitate organ-specific immunopathology. Proximal signaling by Fas and related receptors depends on subunit preassembly, which accounts for the dominant-negative effect of pathogenic receptor mutants and natural splice variants.

Animals↗
PubMed2000

Speed selectivity for optic flow in area 7a of the behaving macaque.

Area 7a, in the inferior parietal lobe, has been implicated in optic flow processing to obtain spatial information about the environment. Optic flow, angle-of-gaze and center-of-motion dependencies are already documented, but the selectivity of area 7a to speed is unknown. Such information is crucial as area 7a provides the final step in visual motion analysis that begins at the lateral geniculate nucleus and passes through MT, MST and LIP/VIP. Macaque area 7a neurons were tested with optic flows with speeds of 0.5-128 degrees /s. Of 161 neurons tested in four hemispheres of two adult male macaques, 53% (86/161) were speed selective at either the time of stimulus onset, at the end of the trial, or at both times. Speed selec- tivities resembling the basic filter types (band-pass, band-reject, high-pass, low-pass, broadband) were found. Area 7a neurons exhibited two novel properties not previously reported elsewhere. Speed selectivity was found to be dynamic in that many cells gained, lost or changed speed tuning over the course of a trial. In addition, speed dependence and optic flow selectivity interacted. For example, a cell could preferentially respond to one type of naviga- tional optic flow at a slow speed and a different navigational optic flow at a fast speed. The presence of speed selectivity combined with other properties of area 7a neurons indicates that these neurons may have a role in the concurrent representation of heading as well as multiple object speeds and directions.

Animals↗
PubMed2000

Generation of an apoptotic intracellular peptide by gamma-secretase cleavage of Alzheimer's amyloid beta protein precursor.

The amyloid beta protein precursor (AbetaPP) is sequentially processed by beta- and gamma-secretases to generate the Abeta peptide. The biochemical path leading to Abeta formation has been extensively studied since extracellular aggregates of amyloidogenic forms of Abeta peptide (Abeta42) are considered the culprit of Alzheimer's disease. Aside from its pathological relevance, the biological role of AbetaPP proteolysis is unknown. Although never previously described, cleavage of AbetaPP by gamma-secretase should release, together with Abeta, a COOH-terminal AbetaPP Intracellular Domain, herein termed AID. We have now identified AID-like peptides in brain tissue of normal control and patients with sporadic Alzheimer's disease and demonstrate that AID acts as a positive regulator of apoptosis. Thus, overproduction of AID may add to the toxic effect of Abeta42 aggregates and further accelerate neurodegeneration.

Journal Article↗
PubMed1999

Defective CD95/APO-1/Fas signal complex formation in the human autoimmune lymphoproliferative syndrome, type Ia.

Heterozygous mutations in the CD95 (APO-1/Fas) receptor occur in most individuals with autoimmune lymphoproliferative syndrome (ALPS) and dominantly interfere with apoptosis by an unknown mechanism. We show that local or global alterations in the structure of the cytoplasmic death domain from nine independent ALPS CD95 death-domain mutations result in a failure to bind the FADD/MORT1 signaling protein. Despite heterozygosity for the abnormal allele, lymphocytes from ALPS patients showed markedly decreased FADD association and a loss of caspase recruitment and activation after CD95 crosslinking. These data suggest that intracytoplasmic CD95 mutations in ALPS impair apoptosis chiefly by disrupting death-domain interactions with the signaling protein FADD/MORT1.

Adaptor Proteins, Signal Transducing↗
PubMed1999

Optic flow selectivity in the anterior superior temporal polysensory area, STPa, of the behaving monkey.

Earlier studies of neurons in the anterior region of the superior temporal polysensory area (STPa) have demonstrated selectivity for visual motion using stimuli contaminated by nonmotion cues, including texture, luminance, and form. The present experiments investigated the motion selectivity of neurons in STPa in the absence of form cues using random dot optic flow displays. The responses of neurons were tested with translation, rotation, radial, and spiral optic flow displays designed to mimic the types of motion that occur during locomotion. Over half of the neurons tested responded significantly to at least one of these displays. On a cell by cell basis, 60% of the neurons tested responded selectively to rotation, radial, and spiral motion, whereas 20% responded selectively to translation motion. The majority of neurons responded maximally to single-component optic flow displays but was also significantly activated by the spiral displays that contained their preferred component. Moreover, there was a bias in the selectivity of the neurons for radial expansion motion. These results suggest that neurons within STPa are contributing to the analysis of optic flow. Furthermore, the preponderance of cells selective for radial expansion provides evidence that this area may be specifically involved in the processing of forward locomotion and/or looming stimuli. Finally, these results provide carefully controlled physiological evidence for an extension and specialization of the motion-processing pathway into the anterior temporal lobe.

Analysis of Variance↗
PubMed1999

The role of Fas and related death receptors in autoimmune and other disease states.

The Fas receptor, also known as APO-1 or CD95, has emerged as a key initiator of apoptotic programmed cell death in a variety of cell types. CD4(+) T cells are unique in their ability to commit "suicide" by stimulating their own Fas receptors with secreted or membrane-bound Fas ligand. This takes place in the setting of repeated stimulation with T-cell antigens and is thought to be a mechanism for controlling the expansion of T cells during viral infections and autoimmune disease states. T cells can also trigger apoptosis in B cells, macrophages, and other cell types through Fas ligand. These interactions negatively regulate the immune system but can also contribute to immunopathology, as occurs in Fas-mediated damage of target tissues in hepatitis and other organ-specific autoimmune diseases. The dual role of Fas in the immune response complicates the understanding of its role in disease states and may limit its potential as a therapeutic target. Despite the many roles of Fas in immunoregulation, findings in experimental mouse strains and human patients with genetic deficiencies in the Fas pathway have shown that the main result of disrupting this pathway in vivo is systemic autoimmunity and a predisposition toward lymphoid malignancies. The role of Fas in various cell types and the lessons we have learned from Fas-deficient patients with the autoimmune lymphoproliferative syndrome will be discussed.

Animals↗
PubMed1999

Stored representations of three-dimensional objects in the absence of two-dimensional cues.

Object recognition was studied in human subjects to determine whether the storage of the visual objects was in a two-dimensional or a three-dimensional representation. Novel motion-based and disparity-based stimuli were generated in which three-dimensional and two-dimensional form cues could be manipulated independently. Subjects were required to generate internal representations from motion stimuli that lacked explicit two-dimensional cues. These stored internal representations were then matched against internal three-dimensional representations constructed from disparity stimuli. These new stimuli were used to confirm prior studies that indicated the primacy of two-dimensional cues for view-based object storage. However, under tightly controlled conditions for which only three-dimensional cues were available, human subjects were also able to match an internal representation derived from motion of that of disparity. This last finding suggests that there is an internal storage of an object's representations in three dimensions, a tenet that has been rejected by view-based theories. Thus, any complete theory of object recognition that is based on primate vision must incorporate three-dimensional stored representations.

Analysis of Variance↗
PubMed1999

Screening for domestic violence in the community pediatric setting.

OBJECTIVE: Children exposed to domestic violence (DV) can experience a variety of adverse effects such as behavior disorders, developmental delay, and child abuse. Recently, the American Academy of Pediatrics recommended that all pediatricians incorporate screening for DV as a part of anticipatory guidance. To date, however, there is little information on how likely women are to disclose DV or whether there are any benefits to screening in the pediatric office setting. The purpose of our pilot study was to gain an understanding of whether screening for DV in the pediatric office setting could be helpful to abused women and their children. METHODS: During a 3-month period, 92% of the women who accompanied their children for a well-child visit to a hospital-based suburban pediatrician were asked about violence in the home with a six-question screening tool. RESULTS: Of the 154 women screened, 47 (31%) revealed DV at some time in their lives. Twenty-five women (17%) reported DV within the past 2 years and were reported to the mandated state agency. There were 5 episodes of child abuse reported of which two had not been previously reported. Interestingly, there were 5 women injured during their most recent pregnancy and who had separated from their abusive partner, but no legal action had been taken to protect them from their partner's return. There was no significant difference in the incidence of DV reported in families with Medicaid (37%) versus private insurance (20%). Before routine DV screening in our office, only one previous DV report had been made in 4 years. CONCLUSIONS: Our preliminary results suggest that many women will reveal DV when screened in the pediatric office setting. Also, there is a subgroup of women, those with young children who have recently separated from their partners, who may particularly benefit from DV screening.

Adolescent↗