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Biomedical subjects

M A Clark

Publications and source records attributed to M A Clark.

At least 127 records · Page 7Linked to original sources

Spatial planning deficits in limb apraxia.

Geschwind (1975) proposed a disconnection model in which an apraxic subject is unable to carry out movements to command because the left hemisphere that comprehended the verbal command is disconnected from the right premotor and motor areas which controls the left hand. An alternate model, however, proposes that apraxia results from destruction of spatiotemporal representations of learned movement stored in the left hemisphere (Heilman, 1979). The disconnection hypothesis would predict that apraxic subjects should be able to correctly imitate gestures and correctly use actual tools since these tasks do not require language. The movement representation model predicts that imitation and actual tool use would also be impaired. Motion analyses were performed on the trajectories of repetitive 'slicing' gestures made in a series of conditions in which contextual cues were introduced in a graded fashion. Four cue conditions were presented: no cues (verbal command), object present, tool present and both object and tool present. Positions of the hand, wrist, elbow, and shoulder were digitized from neighbouring views, reconstructed in three dimensions and analysed with respect to specific spatiotemporal features of the trajectories. Three subjects with limb apraxia, who had lesions that included left parietal cortex, and four neurologically intact subjects participated. The apraxic subjects showed disturbances in planning the movement of the hand in space across the cue conditions. For example, they showed deficits in the plane of motion, the shape of the trajectory and in the coupling of hand speed and trajectory shape even when given full contextual cues. These data support the hypothesis that apraxia can result from the destruction of spatiotemporal representations of learned movement, rather than from a disconnection between the receptive language areas in the left hemisphere and the contralateral motor cortices.

Aged↗

Differential surface characteristics of M cells from mouse intestinal Peyer's and caecal patches.

The mouse caecal patch is located near the blind end of the caecum, and consists of a group of lymphoid follicles. In common with the Peyer's patches, the follicle-associated epithelium overlying these follicles is largely composed of enterocytes, goblet cells and membranous epithelial (M) cells. Each of these types of cell was readily identified by electron microscopy, although caecal patch enterocytes and M cells were morphologically distinct from those of the Peyer's patches. Staining for alkaline phosphatase activity demonstrated that the majority of caecal follicle-associated epithelial cells were alkaline phosphatase-negative, positive cells consisting of a mixture of enterocytes and M cells. In contrast, it has previously been found that Peyer's patch enterocytes are positive for alkaline phosphatase while the M cells are relatively lacking in alkaline phosphatase activity. Lectin histochemistry revealed that surface glycoconjugate expression differs between the caecal and Peyer's patch follicle-associated epithelial cells; in particular, the characteristic staining of Peyer's patch M cells by Ulex europaeus agglutinin 1 was absent on the caecal patch follicle-associated epithelium. These altered surface characteristics indicate that the development of the caecal patch follicle-associated epithelial cells is influenced by the local environment, and these altered properties may be indicative of modified functional roles for the cells at this site.

Alkaline Phosphatase↗

Stage IB carcinoma of the cervix: are all staging tests and procedures necessary?

The prognostic value and cost effectiveness of generally recommended ancillary tests and staging procedures in 115 patients with clinical Stage IB carcinoma of the cervix were retrospectively reviewed. All 112 intravenous pyelograms, 108 barium enemas, 102 cystoscopies and 98 sigmoidoscopies were normal. No malignant cells were found in pelvic washings. Of 111 patients who had paraaortic lymph node biopsies, only one had a positive node. This node was grossly enlarged and clinically suspicious. Paraaortic lymph node status was not influenced by tumor grade, prior conization, lymphovascular space involvement, depth of stromal invasion, positive pelvic lymph nodes or number of paraaortic nodes biopsied. The tests evaluated in this study are unnecessary and should no longer be performed in patients with clinical Stage IB carcinoma of the cervix. In addition, paraaortic lymph node biopsies in the absence of clinically suspicious nodes are not warranted. Eliminating these tests and procedures would result in substantial savings in health care cost.

Adenocarcinoma↗

Investigation of the 1987 Indianapolis Airport Ramada Inn incident.

On October 20, 1987, a military reserve aircraft lost power during a transcontinental flight and attempted an emergency landing at The Indianapolis International Airport. The pilot ejected and the disabled and pilotless aircraft struck a bank building. It then skidded across the street and entered the lobby of The Airport Ramada Inn where it exploded. This incident was unusual in that the fatal injuries occurred in individuals on the ground and not in the occupant of the aircraft. Seven people were killed in the lobby area and two were trapped in a laundry where they died of smoke inhalation. A tenth person died of burns ten days later. Minor injuries were reported among four hotel guests, two firefighters and the Air Force pilot. A multiagency mass disaster-plan had been formulated and rehearsed in preparation for the Panamerican Games, which had been held in Indianapolis in August 1987. A number of volunteers arrived before a security perimeter was established. They began an undocumented removal of the bodies from the scene and were about to remove valuables for "safekeeping" when stopped by coroners' office personnel. Fatalities resulted from smoke inhalation, burns or a combination. Bodies were identified by a combination of dental records, personal effects and visual means within 24 hours. The problems encountered in managing this disaster scene will also be compared with previously reported incidents.

Accidents, Aviation↗

Disinterment from paving materials: use of heavy equipment for exhumation and examination of bodies.

Common paving materials--concrete and asphalt--pose an unusual and complex barrier to disinterment and examination of human remains. Although not commonly encountered, these materials are seen with sufficient frequency to justify consideration of the procedures and equipment necessary for disinterment. Over a seven year period we have encountered paving materials seven times. In each of these cases, the material was slightly different in quality, and different tools--including heavy construction equipment--were necessary. Sometimes the paving material could be managed without heavy power tools, but occasionally specialized construction tools have proven very useful.

Autopsy↗

Firearm suicides during confrontations with police.

We reviewed the case records of suicides in Marion County, Indiana (Indianapolis) and in surrounding counties from 1984 through 1992. Out of 1203 suicides, there were 14 in which armed, on-duty police officers were confronting, pursuing, or apprehending the subject of the death investigation. All subjects were male, and the peak age range was 30 to 34 years. The head, especially the right temple, was the usual site of the fatal wound. More than half of the incidents started as domestic disputes with a wife or girlfriend. Many of the others occurred when police officers pursued or arrested a suspect wanted for a previous felony. Ethanol was involved in less than half of the cases, and drugs were not a factor. In at least four cases, the presence of police was a factor that precipitated the suicide. The histories of the 14 cases demonstrate the potential for controversy and the challenges for forensic scientists investigating suicides during police confrontations.

Adolescent↗

Elevated levels of glycoprotein gp200 in progeria fibroblasts.

The glycosylation of proteins in fibroblasts from people with the premature ageing disease Hutchinson-Gilford Progeria Syndrome (progeria) was investigated. Protein was prepared from fibroblast cell lines established from skin biopsy taken from progeria patients and control donors. Glycoproteins were labelled by the covalent attachment of the steroid hapten digoxygenin to the sugar group. After separation of total protein by SDS-PAGE and electroblotting onto Immobilon-PTM, glycoproteins were detected by enzyme immunoassay. We have observed a glycoprotein of M(r) 200 kDa which is consistently present in protein preparations from progeria fibroblasts and which is absent, or markedly reduced, in preparations from control fibroblasts. This suggests that it may be useful as a marker for progeria. Similar analysis of progeria lymphoblast and control lymphoblast cultures did not show this altered pattern of glycosylated proteins, indicating that it may be cell-type specific. Glycoproteins were also detected by labelling fibroblasts in vitro with D-[6-3H]glucosamine hydrochloride followed by SDS-PAGE of isolated protein and subsequent fluorography. Profiles of glycoproteins from progeria and control fibroblasts were consistent with those obtained from labelling of carbohydrate groups with digoxygenin. Protease digestion of cell protein verified that the band at M(r) 200 kDa contains a protein core. Characteristic features of progeria primarily involve the connective tissue and include wrinkled and loose skin, loss of soft tissue, thin limbs and stiff joints. Death of progeria patients is usually a result of cardiovascular abnormalities. The most consistent manifestations thus involve the connective tissue. The glycoprotein of M(r) 200 kDa which we have observed in progeria fibroblasts in vitro could reflect a perturbation in glycosylation which may underly the connective tissue defects seen in progeria.

Adult↗

Responses of purified phospholipases A2 to phospholipase A2 activating protein (PLAP) and melittin.

The role of the phospholipase A2 (PLA2) stimulating protein PLAP in the regulation of PLA2 activity was assessed by determination of the effects of PLAP on two purified PLA2s. An approx. 14 kDa enzyme was purified from mouse thymoma cells, EL-4 cells, by cation ion exchange HPLC and immunoaffinity HPLC (with antiserum to the N-terminal sequence of an inflammatory exudate PLA2). An approx. 110 kDa enzyme was purified from mouse mammary carcinoma derived cells by sequential hydrophobic, anion exchange, hydroxyapatite and gel filtration HPLC. Neither PLAP nor melittin, an immunologically related PLA2 stimulating peptide from bee venom, increased the activity of the high molecular weight enzyme. In contrast, there was more than a 20-fold stimulation of the low molecular weight PLA2 by PLAP and an approx. 5-fold stimulation by melittin. The stimulation of enzyme activity by PLAP was observed at a protein to phospholipid ratio of 1:10(6) while the ratio of melittin to phospholipid was 1:3. Thus, PLAP mediated stimulation of PLA2 activity may include an interaction between PLAP and the enzyme, in contrast to melittin stimulation, which involves interactions between melittin and phospholipid.

Animals↗

Phospholipase A2 and arthritis.

Over the last 30 years, interest in PLA2 has grown beyond its enzymatic capacity to cleave phospholipids. It has been recognized as the rate-limiting step in the release of arachidonic acid and subsequent formation of prostaglandins, leukotrienes, and other bioactive lipids. Subsequently, PLA2 has not only been found to be present in high concentrations in inflammatory arthritis, but also to induce inflammation when injected into animals. At the same time, investigators into mechanisms of signal transduction demonstrated that a variety of cytokines including IL-1 and TNF, which are found in high concentrations in synovial fluid from patients with RA, stimulate PLA2 activity. These investigations demonstrated further the central role for PLA2 in inflammatory events, especially inflammatory arthritis. Numerous other PLA2 proteins, in addition to the low molecular weight synovial fluid/platelet enzyme, also have been characterized. Their clinical role in arthritis is yet to be elucidated. Human proteins which either inhibit or stimulate PLA2 have also been identified, characterized, and cloned. More recently, exciting investigations, primarily from biotechnology and pharmaceutical companies, into inhibitors of PLA2 have been reported. New PLA2-regulating compounds, which will hopefully move from the laboratory and through clinical trials and then be used to treat patients with arthritis, are on the horizon.

Arthritis↗

Epithelial M cells in the rabbit caecal lymphoid patch display distinctive surface characteristics.

The follicle-associated epithelium (FAE) in the rabbit caecal lymphoid patch is characterized by the presence of membranous (M) cells, which are believed to be functionally equivalent to those present at other sites of gut-associated lymphoid tissue (GALT). Caecal patch M cells display distinctive features compared with those of other GALT sites, despite similar general morphology and expression of the M cell marker vimentin, suggesting marked heterogeneity in the apical surface of M cells at discrete GALT sites. Electron microscopy reveals that rabbit caecal patch M cells differ from those in the small intestinal Peyer's patch FAE: the former have a prominent aspect within the epithelium and possess microvilli which are longer than those of adjacent enterocytes. Many of the M cells in peripheral regions of the caecal patch FAE are not associated with leucocytes and may thus represent an immature M cell population. The M cells are also histochemically distinct from adjacent enterocytes and from Peyer's patch M cells, showing greater expression of brush-border alkaline phosphatase activity and affinity for certain lectins (peanut and wheat germ agglutinins, Bandeiraea simplicifolia agglutinin II). The differences in the brush-border morphology and glycocalyx structure between M cells at different GALT sites may affect their function at these sites by influencing the interaction of luminal antigens and microorganisms with the M cell surface. The present data also support the hypothesis that M cells arise directly from differentiation of crypt stem cells and not from the transformation of existing fully differentiated enterocytes.

Alkaline Phosphatase↗

Aspects of energy-yielding metabolism in the aphid, Schizaphis graminum, and its endosymbiont: detection of gene fragments potentially coding for the ATP synthase beta-subunit and glyceraldehyde-3-phosphate dehydrogenase.

Specialized cells within the aphid, Schizaphis graminum, contain intracellular, vesicle-enclosed eubacterial endosymbionts (Buchnera aphidicola). Using oligonucleotide probes derived from conserved sequences of the ATP synthase beta-subunit and glyceraldehyde-3-phosphate dehydrogenase, and the polymerase chain reaction (PCR), we have amplified, cloned, and sequenced three DNA fragments. Amino acid sequence similarity indicated that two of these fragments corresponded to endosymbiont and host genes potentially coding for the beta-subunit of ATP synthase. The host gene fragment contained two putative introns. The third DNA fragment corresponded to a portion of a gene coding for a glyceraldehyde-3-phosphate dehydrogenase that was highly related to one of the enzymes from Escherichia coli (GapA). These results indicate that B. aphidicola may have an ATP synthase and consequently could synthesize ATP from a proton motive force generated within the intracellular vesicles of host cells containing the endosymbionts. The detection of a gene fragment coding for a protein similar to glyceraldehyde-3-phosphate dehydrogenase suggests the presence of this glycolytic enzyme in the endosymbiont and its involvement in energy-yielding metabolism.

Amino Acid Sequence↗

Guanine nucleotides and pertussis toxin reduce the affinity of histamine H3 receptors on AtT-20 cells.

Agonist occupancy of high affinity histamine H3 receptors on AtT-20 cells induces increased ACTH release. However, the signal transduction process by which this occurs is presently unknown. As a first step in characterizing this pathway, we have examined the effects of a variety of nucleotides and nucleotide analogs on Na-methylhistamine binding to these receptors. Nonhydrolyzable guanine nucleotide analogs inhibit up to 40% of the [3H]Na-methylhistamine binding by increasing the dissociation rate of the ligand from the receptor and, thereby, reducing receptor affinity. Pertussis toxin also decreases the affinity of the H3 receptors and ADP ribosylates a 41 kDa protein. Neither GTP gamma S nor pertussis toxin change Bmax. These data indicate that the H3 receptors on these cells are coupled to a G protein of the Gi subclass.

Adenosine Diphosphate Ribose↗

Digitalis-like factor and ouabain-like compound in plasma of volume-expanded dogs.

Previous studies demonstrated that ouabain-like compound (OLC) is widely distributed in mammalian species and is found in a variety of different tissues. Although much evidence suggests that OLC is endogenous to mammals, little information is available concerning physiological and/or pathophysiological roles for OLC. In this study, generic endogenous digitalis-like factor (E-DLF) was measured using an enzyme bioassay and the more specific OLC was determined using ouabain antisera in plasma drawn from dogs before and 30 and 120 min after massive volume expansion with isotonic saline. Plasma OLC was not changed by the saline load, whereas Na excretion was significantly elevated at the 30-min blood draw and remained elevated at the 120-min blood draw. Because renal exposure to OLC did not change with saline loading, it is unlikely that any portion of the profuse natriuresis in these animals could be attributed to OLC. In contrast, plasma E-DLF was higher after the saline load than before in each of four dogs at 30 and 120 min after the infusion. What portion of the profuse natriuresis can be attributed to E-DLF is unknown, although it is reasonable to assume that nanomolar levels of pump inhibitor contributed to the natriuresis to some degree.

Animals↗

Actin accumulation at sites of attachment of indigenous apathogenic segmented filamentous bacteria to mouse ileal epithelial cells.

Selective attachment of indigenous, apathogenic, segmented filamentous bacteria to follicle-associated epithelia of terminal ileal Peyer's patches involves both the specialized M cells and normal enterocytes. A striking, highly localized cytoplasmic accumulation of polymerized actin, which resembles that associated with adherence of enteropathogenic Escherichia coli to cultured epithelial cells, occurs at attachment sites of segmented filamentous bacteria. The epithelial response to the attachment of microorganisms may thus involve mechanisms which are the same for diverse microorganisms.

Actins↗

Differential expression of lectin-binding sites defines mouse intestinal M-cells.

We investigated the binding of four lectins to the follicle-associated epithelium (FAE) overlying fixed mouse small intestinal Peyer's patches to identify M-cell-specific surface markers. Wheat germ agglutinin and peanut agglutinin displayed heterogeneous staining patterns, binding most avidly to the intestine goblet cells. In contrast, the lectins Ulex europaeus 1 (UEA 1) and Psophocarpus tetragonolobus (winged bean; WBA) were almost exclusively M-cell specific. When confocal laser scanning images of tissues stained with fluorescein isothiocyanate (FITC)-conjugated UEA1 or WBA were compared with the appearance of the same tissues under the scanning electron microscope (SEM), UEA1 strongly stained 97.2% (106/109) of M-cells, 0.6% (3/516) enterocytes, and 0% (0/28) goblet cells, whereas WBA stained 100% (83/83) M-cells, 1.7% (6/361) enterocytes, and 5.3% (1/19) goblet cells. The M-cell specificity of the lectin binding was further demonstrated by localization of horseradish peroxidase (HRP)-conjugated lectins under the transmission electron microscope (TEM). This is the first demonstration of carbohydrates in the glycocalyx of M-cells that are not expressed elsewhere on the FAE surface. These carbohydrates not only provide a means to identify mouse M-cells by LM but may also contribute to the occurrence of specific interactions between microorganisms and the M-cell apical membrane.

Animals↗

Asphyxial deaths due to hanging in children.

A review of the records for five years from the Coroner's Offices of Marion County, Indiana and Franklin County, Ohio, which have a combined population of 2.4 million, yielded 12 cases of hanging deaths in children 13 years of age or younger. The age range was 2 1/2 to 13 years and only two of the 12 victims were female. There were three unequivocal suicides, five accidental deaths and four cases where, despite extensive investigation, the manner of death remained undetermined. The scene and autopsy findings are presented and the cases discussed with regard to determination of manner of death. Additionally, the significance and pathophysiology of petechiae are discussed.

Accidents↗

Unique natriuretic properties of the ATP-sensitive K(+)-channel blocker glyburide in conscious rats.

Small-conductance, ATP-sensitive K(+)-channels (KATP) localized in apical membranes of both thick ascending limb of the loop of Henle and cortical collecting duct cells may be involved in Na+ reabsorption and K+ secretion in the mammalian kidney. Possible pharmacologic tools to evaluate such an hypothesis may be the antidiabetic sulfonylureas which block K(+)-channels in pancreatic beta-cells. In saline-loaded conscious rats, glyburide (GLY) dose-dependently increased urinary Na+ excretion with little change in urinary K+ excretion after i.p. administration (10-100 mg/kg). In renal clearance studies, GLY at 25 mg/kg i.v. increased Na+ excretion 350% during the first hour post-treatment without affecting K+ excretion, glomerular filtration rate, mean arterial pressure or heart rate. GLY at 50 mg/kg was no more natriuretic than the 25 mg/kg dose, whereas 12.5 mg/kg of GLY increased Na+ excretion 200%. The change in Na+ excretion produced by 25 mg/kg of GLY in streptozotocin-induced diabetic rats was significantly greater than the change after drug vehicle in these animals. It is unlikely that the natriuresis produced by GLY is secondary to changes in plasma insulin and/or glucose because the doses used were far above GLY's insulin-releasing action (i.e., all natriuretic doses would have produced maximal insulin release) and GLY was natriuretic in streptozotocin-induced diabetic rats. It is possible that GLY interferes with reabsorption of Na+ by blocking KATP and thereby interrupting K+ recycling and Na(+)-2Cl(-)-K+ cotransport in the loop of Henle.

Adenosine Triphosphate↗

Butyric acid-induced differentiation of HL-60 cells increases the expression of a single lysophospholipase.

Treatment of HL-60 cells with 0.5 mM-butyric acid resulted in morphological changes, including the formation of cytoplasmic granules, nuclear condensation and segmentation. These differentiated cells had an elevated phospholipase A2 activity and an increased capacity to synthesize a variety of eicosanoids, including both lipoxygenase and cyclooxygenase products. Phospholipase A2-mediated release of arachidonic acid is accompanied by an equimolar production of potentially cytotoxic lysophospholipid. In association with the differentiation process, there was a 2-3-fold increase in lysophospholipase activity. Subsequent studies were undertaken to identify and characterize the lysophospholipases in this cell system, with 1-[1-14C]palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine as substrate. Hydrophobic chromatography of both undifferentiated and differentiated cell extracts revealed three peaks of enzyme activity. Extracts of differentiated cells contained a dramatic increase in activity contained in peak 2. The increase in enzymic activity of peak 2 appeared to account for the increase in total lysophospholipase activity found in the differentiated cell homogenates. The lysophospholipases contained in peaks 2 and 3 were purified to homogeneity and were 20 and 22 kDa respectively, as determined by denaturing polyacrylamide-gel electrophoresis. Peaks 2 and 3 were similar on the basis of amino acid composition, but had distinctive C-terminal peptide amino acid sequences. Enzymic characterization of these proteins demonstrated that there was no detectable level of non-specific esterase, acyltransferase or transacylase activity associated with these proteins. We concluded that peak 2 lysophospholipase is regulated by differentiation in HL-60 cells and may play an important role in protecting these cells from the cytolytic effects of the lysophospholipids produced by the activation of phospholipase A2.

Amino Acid Sequence↗