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D Carpenter

Publications and source records attributed to D Carpenter.

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Megakaryocytopenia in W/Wv mice is accompanied by an increase in size within ploidy groups and acceleration of maturation.

Megakaryocytopoiesis was evaluated in W/Wv mice and their normal +/+ littermates to analyze the mechanisms by which normal platelet production is maintained in W/Wv mice even though numbers of megakaryocytes are low. Relative sizes of megakaryocytes, and their nuclei and cytoplasm, were measured microscopically in bone marrow smears, and the ploidy of the same cells was measured by two-wavelength microspectrophotometry. Maturation rate of megakaryocytes was estimated after they were labeled with tritiated thymidine. W/Wv megakaryocytes were macrocytic: average cell size was increased in each ploidy group. The increase in cytoplasmic area exceeded that of the nucleus. Further analysis of the predominant 16N ploidy group revealed that the increase in average cell size was due to depletion of cells of small size. Megakaryocytes matured more rapidly than normal in W/Wv mice. These results showed that megakaryocyte size and ploidy can be regulated separately. They suggest that alterations in cell growth and maturation may be mechanisms by which the organism can compensate for a deficiency in numbers of megakaryocytes, but they do not define the mechanism by which the deficiency may be sensed or by which the compensatory changes may be mediated. This is a US government work. There are no restrictions on its use.

Animals

Morphological and kinetic abnormalities of platelets in hypercholesterolemic rabbits.

Hypercholesterolemia (HC = hypercholesterolemia or hypercholesterolemic) was produced in rabbits by feeding them diets supplemented with cholesterol and peanut oil. Platelet counts and volumes, white cell counts, reticulocyte counts, and hematocrits were determined at intervals for 8-12 weeks in blood from HC animals and controls on a normal rabbit diet. Microthrombocytosis was a consistent occurrence in the presence of HC, developing as early as 2 weeks into the diet. Microthrombocytosis was generally associated with normal platelet counts, but mild thrombocytosis occurred late in the diet at the time of the highest levels of serum cholesterol (greater than 1300 mg/dl). Platelets from HC rabbits were morphologically normal by transmission electron microscopy. Survivals of 51Cr-labeled platelets from HC and non-HC rabbits were measured in HC and non-HC recipients. The results identified an intrinsic defect in the ability of HC platelets to survive in the circulation. They also confirmed previous findings of an environmental defect in HC that causes shortened platelet survival.

Animals

Variation in indirect cell-mediated lympholysis.

The ability of an allograft recipient to respond to donor mononuclear cells in an indirect cell-mediated lympholysis (ICML) assay is an in vitro correlate of allograft rejection, but the value of this correlation depends upon the assay's reliability. We had observed inconsistency in the cytotoxic response of normal human mononuclear cells (MNC) to the same allogeneic stimulator MNC when cytotoxicity was measured repeatedly on different occasions by micro-ICML. We, therefore, investigated the extent and reasons for this inconsistency. Method variation, determined by duplicate ICML of 18 stimulator: responder MNC, was not statistically significant. Variation in cytotoxicity over time was greater but still not statistically significant. The contribution to method variation of 51Cr release from 3 different sets of target cells, cultured and labeled in duplicate, was minimal (6.33%). We then asked if in vitro generation of effector MNC under laboratory conditions was a major cause of ICML variation. We tested this using a stable transplant's in vivo sensitized effector cells against donor MNC in a direct CML (DCML) and obtained consistent results. Finally, to gain an understanding of some of the factors which might influence the generation of in vitro cytotoxicity, we measured the frequencies of cell surface antigens (DR, TAC, transferrin, Leu 2 and 3) concomitantly with ICML on day 6 of culture. Statistical analysis of the results led us to conclude that the micro-ICML is reproducible. The magnitude of lysis depends upon activated target cells (TAC- and transferrin-positive) and an increase in the proportion of helper/inducer to cytotoxic/suppressor T-lymphocytes during effector cell generation.

Antigens, Surface

Megakaryocytes increase in size within ploidy groups in response to the stimulus of thrombocytopenia.

Experiments were done to determine if sizes of megakaryocytes within a defined maturation stage were strictly determined by amount of nuclear DNA. Normal mice, mice recovering from an acute episode of thrombocytopenia induced by a single injection of heterologous antiplatelet serum (APS), and mice with sustained thrombocytopenia from daily injections of APS were examined. Areas of mature megakaryocytes were measured in bone marrow smears stained with polychromatic stains. The nuclear DNA content of the same cells was then measured microspectrophotometrically after staining by the Feulgen reaction. Normal megakaryocytes showed a trimodal, lognormal distribution of nuclear chromophore, corresponding to 8n, 16n, and 32n with smaller numbers of 4n and 64n cells; 16n was the predominant ploidy class. In response to thrombocytopenia, ploidy values shifted: the proportions of 8n and 16n cells decreased; 32n and 64n cells increased; 128n megakaryocytes occasionally appeared. These shifts were accompanied by an increase in the average size of all megakaryocytes. In addition to shifts to higher ploidy values, megakaryocytes within ploidy groups became larger than normal megakaryocytes of the same ploidy especially in the mice with sustained thrombocytopenia. These findings show that megakaryocyte size in thrombocytopenic mice is influenced by factors other than the ploidy and maturity of the cell.

Anemia

Survival of rabbit platelets labeled with gallium 67.

The viability of rabbit platelets labeled with radioactive gallium was determined to analyze the feasibility of using platelets labeled with gallium 68 as an imaging reagent for positron emission tomography. Platelets were labeled with a complex of the longer lived gallium 67 and mercaptopyridine-N-oxide (MPO) or with sodium chromate Cr 51. Their survival after transfusion was measured. Labelling efficiency of 67Ga-MPO was 6.5% to 45.8% (26.8% +/- 2.8%) when platelets were suspended in saline solution, but was much lower (1.6% +/- 0.8%) in plasma. Platelets labeled with either radioisotope in a saline medium survived as well as platelets labeled with 51Cr in plasma. Recovery values 1 hour after transfusion and mean platelet survivals were 68.6% +/- 4.9% and 3.4 +/- 0.2 days for 67Ga in saline solution, 76.5% +/- 6.8% and 3.8 +/- 0.5 days for 51Cr in saline solution, and 73.7% +/- 7.4% and 3.6 +/- 0.5 days for 51Cr in plasma. Labeled platelet concentrates always contained extra radioactivity not firmly bound to viable platelets. A postlabeling wash in saline solution did not reduce this contamination and resulted in reduction of the number of viable platelets. The results showed that rabbit platelets labeled with 67Ga-MPO survived in the circulation as well as those labeled by a standard protocol with sodium chromate Cr 51.

Animals

A note on the effect of gamma-rays on cefamandole and oxacillin.

The feasibility of the radiation sterilization of two beta-lactam antibiotic powders, cefamandole nafate and oxacillin sodium, has been examined by subjecting them to a range gamma-radiation doses, followed by chemical and microbiological analyses. It would appear feasible to radiation sterilize oxacillin sodium. The radiation sterilization of cefamandole nafate may be realistic at low doses or under conditions that minimize radiolysis.

Cefamandole

Effects of intermittent pneumatic calf compression on postoperative thrombin and plasmin activity.

A previous study of neurosurgical patients demonstrated an imbalance between thrombin and plasmin action following surgery. The present study was designed to determine the effect of intermittent pneumatic calf compression on postoperative enzyme activity. Fibrinopeptide A (FPA) and B beta 1-42 levels, reflecting thrombin and plasmin action respectively, were measured daily in patients undergoing elective craniotomy. Two of 9 patients not receiving calf compression developed positive fibrinogen leg scans, while none of 5 patients receiving prophylaxis had positive scans. Calf compression was associated with a markedly altered pattern of changes in the fibrinopeptide values following surgery. Without compression, there was perturbation of the balance between thrombin and plasmin action on the day after surgery as reflected by an increase in the FPA/B beta 1-42 ratio. In contrast, in those receiving prophylaxis there was no change in this ratio on the first postoperative day. Calf compression both blunted the mean postoperative increase in the FPA level (1.8 nM vs 4.7 nM; p less than .05) and augmented the mean B beta 1-42 value (3.0 nM vs 0.2 nM; p less than .05) so that the mean increase in the FPA/B beta 1-42 ratio was only 0.1 with calf compression as compared to 2.2 without it (p less than .05). Systemic modulation of both the coagulation and fibrinolytic pathways thus occurred in association with calf compression.

Adult

Functionally abnormal stromal cells and megakaryocyte size, ploidy, and ultrastructure in Sl/Sld mice.

The first goal of the present studies was to determine if Sl/Sld megakaryocytes have features in common with the macrocytic megakaryocytes that genetically normal mice produce in response to acute platelet depletion. The second was to test the hypothesis that megakaryocyte abnormalities in Sl/Sld mice are due to genetically determined hemopoietic stromal cell abnormalities. Sizes and ploidies of mature Sl/Sld megakaryocytes were measured. Macrocytosis and a shift to higher ploidy values were found compared with normal. Within ploidy groups 16N-64N, Sl/Sld megakaryocytes were larger than normal megakaryocytes of the same ploidy. Transmission electron microscopy revealed that Sl/Sld megakaryocyte nuclei contain more and larger nucleoli, and the chromatin was more dispersed than in normal megakaryocyte nuclei of comparable maturity. Asynchronous megakaryocyte cytoplasmic maturation was found. Sl/Sld macrophages were also ultrastructurally abnormal. Megakaryocytic macrocytosis was reproduced in long-term bone marrow cultures in which the adherent layer was formed by Sl/Sld cells. It was the same if cultures were recharged with Sl/Sld or +/+ hemopoietic cells. Previously reported ambiguities in mixed cell cultures were avoided by recharging the adherent layers with only a million cells. These results were correlated with previously published observations. Sl/Sld megakaryocytes have features in common with megakaryocytes from acutely thrombocytopenic animals. One feature, macrocytosis, appears to be due to abnormal Sl/Sld stromal cells that are reproduced as adherent layer cells in long-term cultures. The responsible stromal cells in Sl/Sld mice may be counterparts of megakaryocytopoietic regulatory cells in the marrow stroma of normal animals.

Animals

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Administrative Personnel

Tension pneumoperitoneum. A cause of acute aortic occlusion.

Massive accumulation of intraperitoneal air may result in an entity known as tension pneumoperitoneum. The patient usually complains only of abdominal fullness, and the abdomen becomes ballooned, barrel-shaped, and tympanic in all quadrants. Upward displacement of the diaphragm may cause respiratory embarrassment. There may also be diminished venous return due to compression of the inferior vena cava. Abdominal paracentesis may be necessary to improve respiratory exchange before laparotomy. In one patient with a perforated peptic ulcer and tension pneumoperitoneum, an enormous increase in intra-abdominal pressure apparently led to acute aortic occlusion. This particular complication of tension pneumoperitoneum has not previously been reported, to our knowledge. The clinical findings of aortic occlusion were reproduced in a canine model by insufflation of air into the abdomen. An intra-abdominal pressure of 100 mm Hg resulted in loss of the femoral pulse wave measured by an indwelling arterial catheter.

Acute Disease

Smoking, ignorance, and peripheral vascular disease.

The effect of the surgeon's advice to give up smoking was studied in a group of 43 patients with peripheral vascular disease (PVD), using blood carboxyhemoglobin levels to detect deception. Only seven of 40 patients (17.5%) actually stopped smoking, but the remainder appeared fairly honest in reporting their continued habits. Few patients were aware of the possible harmful effect of smoking on the peripheral arteries. The 43 patients with PVD who smoked had significantly higher carboxyhemoglobin levels than a group of 25 smokers without PVD although their cigarette consumption was the same. This suggests that carboxyhemoglobin levels may provide a better indication of the risk of smoking than overall cigarette consumption alone in the development of PVD.

Adult

Application of recategorized WISC-R scores for adjudicated adolescents.

Bannatyne's recategorization of WISC-R scores was applied to 97 juveniles who had been adjudicated by the juvenile court system to a diagnostic and evaluation unit in Eastern Alabama. Mean ranks werre analyzed using the Friedman test for repeated-measures analysis of variance. The total sample exhibited a significant mean rank pattern of scores in the following descending order: Spatial, Conceptual, Sequential, and Acquired Knowledge. Subgroup patterns varied for those youths with IQs 90 and above (Conceptual, Spatial, Sequential, and Acquired Knowledge) and IQs below 90 (Spatial, Sequential, Conceptual, and Acquired Knowledge). Pairwise comparisons using the Scheffé procedure indicated significant differences between the Spatial score and Acquired Knowledge score for the total group and those youths with IQs below 90. The study extended the application of recategorized scores for adjudicated youth suggested by Bannatyne (1968, 1971, 1974). This investigation affirms the need for further studies of the differential effect of IQ on the patterns of WISC-R scaled scores.

Adolescent

Heterogeneity of Staphylococcus aureus enterotoxin B as a function of growth stage: implications for surveillance of foods.

Staphylococcus aureus was grown in a fermentor under controlled conditions of pH, oxygenation, and temperature, while the higher-molecular-weight products of its growth were continuously removed across ultrafiltration membranes. These products were examined by single and double gel diffusion and immunoelectrophoresis against a variety of available anti-enterotoxin B antisera. All antisera examined were polyvalent for S. aureus antigens. However, two electrophoretically distinct proteins were the major reactants with the antisera. One of these was present in early- to mid-log-phase cultures. After mid-log growth was achieved, both were present but in continuously changing proportions. This observation was repeated with a variety of growth conditions and media. A significant part of the physicochemical heterogeneity of enterotoxin B observed over the past 20 years is thus correlated with the growth phase of the organism. Taken together, these facts are used to argue for a two-step rationale for the detection of food-borne staphylococcal disease: (i) screening for a presumptive hazard by analysis for any antigen, toxin, or enzyme of S. aureus in a foodstuff and (ii) confirmation of the hazard by identifying the presence of an enterotoxin using a combination of physicochemical and serological techniques.

Antigens, Bacterial