Syllabic organization and deafness: orthographic structure or letter frequency in reading?

For hearing people, structure given to orthographic information may be influenced by phonological structures that develop with experience of spoken language. In this study we examine whether profoundly deaf individuals structure orthographic representation differently. We ask "Would deaf students who are advanced readers show effects of syllable structure despite their altered experience of spoken language, or would they, because of reduced influence from speech, organize their orthographic knowledge according to groupings defined by letter frequency?" We used a task introduced by Prinzmetal (Prinzmetal, Treiman, & Rho, 1986) in which participants were asked to judge the colour of letters in briefly presented words. As with hearing participants, the number of errors made by deaf participants was influenced by syllable structure (Prinzmetal et al., 1986; Rapp, 1992). This effect could not be accounted for by letter frequency. Furthermore, there was no correlation between the strength of syllable effects and residual speech or hearing. Our results support the view that the syllable is a unit of linguistic organization that is abstract enough to apply to both spoken and written language.

Coleomegilla maculata (Coleoptera: Coccinellidae) predation on pea aphids promoted by proximity to dandelions.

The impact of a predator on its prey may depend on the presence of other species in the community. In particular, if predators are attracted to areas containing one prey species, another prey species may suffer greater predation if it occurs in the same areas. If the predator is omnivorous, this may occur even if one prey species is an animal and the other is a plant. We investigated the role of local dandelion densities on the impact of the predator Coleomegilla maculata on pea aphids in alfalfa fields. At small spatial scales, increased dandelion densities were associated with high C. maculata densities, presumably because these omnivorous ladybird beetles aggregated to pollen resources. In turn, the high C. maculata densities were associated with low aphid densities, presumably because of increased predation. We used laboratory cages to simulate C. maculata foraging in two adjacent patches of alfalfa, one with dandelions and one without. As in the field, the laboratory experiment showed that C. maculata aggregated to alfalfa interspersed with dandelions, which resulted in increased predation on aphids on alfalfa. This study demonstrates that a pollen-producing plant can indirectly decrease nearby herbivore densities by attracting an omnivorous predator.

Connectionist models of neuropsychological disorders.

Artificial neural networks ('connectionist models') embody aspects of real neuronal systems. But does studying the breakdown of performance in such models help us to understand cognitive impairments in humans following brain damage? Here we review recent attempts to capture different neuropsychological disorders using connectionist models with simulated lesions. We show how such lesion studies can be used to evaluate some of the standard assumptions made in neuropsychological research, concerning both double dissociations and associations between patterns of impairment. We also illustrate how lesioned models, like humans, can sometimes be more impaired on the easier of two tasks and demonstrate that connectionist models can incorporate forms of internal structure. Finally we discuss the utility of the models for understanding and predicting the effectiveness of different rehabilitation strategies. Future questions concern the role and possible development of internal structure within these models, whether the models can be generalized to larger-scale simulations, and whether they can accommodate higher-order linguistic disorders.

Structure-function relationships of alkyl-lysophospholipid analogs in selective antitumor activity.

This investigation was initiated in order to delineate the structure-function relationship of the anticancer alkyl-lysophospholipids and assess their degree of selective cytotoxicity toward neoplastic cells. A series of glycerol phosphocholine analogs with varying substitutions in the sn-1 and sn-2 position were tested for their inhibitory activity as measured by thymidine incorporation, clonogenic assays and effects on protein kinase C activity against a series of human leukemic cell lines and healthy bone marrow progenitor cells. The IC50 was determined for each of the compounds in each cell line and healthy bone marrow cells following a 4-h incubation. The data indicated that a 16-18 carbon chain at the sn-1 coupled with a short substitution at sn-2 had the broadest antitumor activity and was the least toxic to normal bone marrow cells. The results provide a number of useful leads toward the design and development of potentially more active phospholipid compounds.

Use of the polymerase chain reaction to detect hypermethylation in the calcitonin gene. A new, sensitive approach to monitor tumor cells in acute myelogenous leukemia.

Based on the recent observations that, in a majority of patients with acute leukemia, the 5' end of the calcitonin gene was hypermethylated and abnormal DNA fragments were observed following HpaII restriction digestion, we have developed a PCR-based method to sensitively detect this abnormal methylation of the calcitonin gene in AML. Applying the concept of competitive PCR, a semi-quantitative correlation was obtained between the amount of hypermethylation and the amount of leukemic cells present. These results suggest that this method will be useful to monitor the amount of tumor cells in bone marrow from patients with AML.

Autologous bone marrow transplantation in acute leukemia with marrow purged with alkyl-lysophospholipid.

Alkyl-lysophospholipids are anticancer agents that are selectively toxic to leukemic cells and relatively sparing of normal bone marrow cells. Thus, they would be likely candidates for purging remission marrows before autologous bone marrow transplant. One of the more promising agents is edelfosine, which could be safely used for purging without prolonging marrow recovery. Assays for marrow progenitor cells were performed before and after purging and cryopreservation in 64 patients. There was no significant reduction in colony formation after purging when compared with unpurged cryopreserved marrow, but there was a significant reduction after cryopreservation. Twenty-four patients with acute leukemia in second (16 patients) or third remission (3 patients), early relapse (3 patients), or in first remission with successfully treated extramedullary relapse (2 patients) received marrow-ablative chemotherapy and total body irradiation followed by infusion of marrow purged for 4 hours with 50 to 100 micrograms/mL of edelfosine. There were 9 lymphoblastic and 15 myelogenous leukemia patients. The median time to granulocyte recovery to 500/microL was 26 and 33 days for the 50 and 75 microgram/mL doses, respectively. The patient whose marrow was purged at the dose of 100 micrograms/mL failed to engraft. The median time to platelet recovery to 25,000/microL was 45 and 37 days for the 50 and 75 micrograms/mL doses, respectively. Twenty-nine percent of the patients remain disease free from 131 to 1,291 days, with a median of 356 days. These results have established that purging with 75 micrograms/mL of edelfosine is a safe dose and is recommended for a phase II trial.

Comparison of selective cytotoxicity of alkyl lysophospholipids.

Alkyl lysophospholipids have been shown to be cytooxic to a number of neoplastic tissues. One, ET-18-OCH3, has been used to selectively purge leukemic cells from mixtures with normal marrow progenitor cells, in vitro and in vivo. We have measured the 50% inhibitory (IC50) effect of a series of ether lipids (EL) on leukemic cells (HL60, K562, Daudi, KG-1, KG-1a) and normal marrow progenitor cells. Cells were incubated with varying concentrations of EL for 4 hr and assayed for viability, [3H]thymidine incorporation and clonogenicity in semi-solid media. The effect on protein kinase C (PKC) activity was assayed for each compound. Compounds tested included three glycerophosphocholine analogs--ET-18-OCH3, ET-16-NHCOCH3, and BM 41.440. In addition, a lipoidal amine, CP 46665, an ethyleneglycolphospholipid, AEPL, and four single chain alkylphosphocholine analogs, HePC2, HePC3, HePC4 and HePC6 were also tested. During the period of incubation, the cells remained viable (greater than 70%) as judged by trypan blue dye exclusion. The glycerophosphocholines were the most active and showed the highest therapeutic index. The lipoidal amine was active, but toxic to normal marrow progenitor cells. The ethyleneglycolphospholipid was active against HL60, but not against the other cell lines. The single chain alkylphosphocholine analogs were less active. All of the compounds inhibited PKC activity; however, the glycerophosphocholines were the most inhibitory.

Purging leukemia remission marrows with alkyl-lysophospholipids, preclinical and clinical results.

Alkyl-lysophospholipids are unique compounds which have selective anti-cancer activity with relative sparing of normal bone marrow stem cells. Thus, they would appear to be ideal candidates for marrow purging. In contrast to most anti-cancer drugs, these compounds act on the cell membrane resulting in inhibition of phosphatidylcholine biosynthesis. In addition, these compounds inhibit protein kinase C activity and may interfere with signal transmission. In low doses in in vitro systems, they have been shown to induce differentiation in leukemic cell lines. Lethally irradiated Balb/c mice were injected with normal bone marrow cells containing 1-2% leukemic cells (WEHI III-B) to simulate a remission marrow after the cells were incubated in vitro for 24 hours with 0-100 ug/ml of 1-0-octadecyl-2-0-methyl-rac-glycero-3-phosphocholine (ALP). All the mice given cells not treated with ALP succumbed to leukemia, whereas there was a dose-response increase in survival in those given ALP treated cells. Similar results were observed when the cells were cryopreserved and thawed before injection. Very little effect of ALP was noted on stem cells as measured by the spleen colony assay. In vitro studies using human marrow progenitor assays (CFU-GEMM) were undertaken in which marrow was mixed with 0.1% HL60 cells and exposed for 1 or 4 hours to ALP at 0, 25 and 50 ug/ml. The cells were plated and assayed for CFU-GEMM and leukemic colonies. At 50 ug/ml, HL60 colonies were eliminated and there was no significant reduction in normal marrow progenitor cells. Clinical trials were initiated. Eleven patients with acute leukemia in remission who were candidates for autologous bone marrow transplantation had marrow harvested, incubated with 50 ug/ml of ALP and cryopreserved. CFU-GEMM assays before and after purging showed no differences. There was a similar significant reduction after cryopreservation regardless of whether marrows were purged. Two patients were transplanted with purged marrow, one in early relapse and one in a marrow remission, but had evidence of CNS leukemia. In both, ablative therapy consisted of cytosine arabinoside, 3 gm/m2 x 12 doses, followed by fractionated TBI to a total of 12 Gy prior to reinfusion of thawed marrow. The patient in early relapse had progression of leukemia by day 28. The patient in marrow remission is disease-free 9 months after transplantation. These studies indicate that purging marrow with ALP is a promising approach and further clinical studies are indicated.

Purging of acute myeloid leukemic cells by ether lipids and hyperthermia.

Ether lipids (EL) and hyperthermia have been shown to possess a relatively selective cytotoxicity to leukemic cells. In this study, the combined effects of EL (ET-18-OCH3, ET-16-NHCOCH3, or BM 41.440) and hyperthermia on the growth of hematopoietic progenitors, myeloid leukemic cell lines, and leukemic cells obtained from patients with acute myeloid leukemia (AML) were examined to determine if this combination resulted in a greater selective killing of leukemic cells than that achieved by either EL or heat alone. When the cells were treated simultaneously with EL (50 micrograms/mL) and hyperthermia (42 degrees C) for one hour, the killing of leukemic cell line cells was enhanced considerably. Among the three EL, however, the combination of ET-18-OCH3 and heat seemed to be the most cytotoxic to leukemic cell line cells with no effect on the growth of hematopoietic progenitors. An increase in the duration of treatment with ET-18-OCH3 to four hours with heat added during the last hour resulted in a further reduction of leukemic cell line cells while sparing 50% of hematopoietic progenitors after cryopreservation. The combined treatment with ET-18-OCH3 and heat also inhibited the growth of leukemic progenitors obtained from AML patients by 97% to 100%. These data indicate that the combined treatment with EL and hyperthermia might offer an efficient means to eliminate myeloid leukemic cells in vitro.

Experimental studies on the role of alkyl lysophospholipids in autologous bone marrow transplantation.

The selective cytocidal effect of alkyl lysophospholipids against neoplastic cells while sparing normal cells make these ideal candidates for purging leukemic cells from bone marrows obtained during remission. To test the feasibility of such an approach, a murine model and an in vitro human cell model were developed. In the murine system a mixture of normal bone marrow cells and WEHI IIIB myelomonocytic leukemic cells was incubated with varying doses of 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-Me) for 24 hr before being injected into tail veins of lethally irradiated Balb/c mice. At doses of 20 and 100 micrograms/ml, long-term survivors were noted. The additional steps of freezing and thawing following incubation resulted in significantly longer survival with doses of 10 to 50 micrograms/ml, but were toxic to marrow stem cells at 100 micrograms/ml. In the in vitro model, normal marrow progenitor cells and leukemic cells (the promyelocytic cell line HL60) were exposed to varying concentrations of ET-Me for 1 and 4 hr alone or mixed, and clonogenicity was assayed by colony formation in semisolid medium during 7-14 days' incubation. At doses up to 100 micrograms/ml exposed for 4 hr normal progenitor cells were spared and HL60 colonies eliminated. Other phospholipids analogues were less effective in eliminating leukemic cells, but spared normal progenitor cells. A survey of fresh leukemic cells found varying degrees of sensitivity to ET-Me, indicating the need for testing a variety of compounds. These studies clearly indicated the potential usefulness of alkyl lysophospholipid compounds in selectively purging leukemic cells from remission marrows for autologous bone marrow transplantation.

Purging leukemic cells from simulated human remission marrow with alkyl-lysophospholipid.

Alkyl-lysophospholipids (ALP) are analogues of 2-lysophosphatidylcholine that have been reported to have selective antitumor activity. These compounds could potentially be useful in purging bone marrow of leukemic cells in autologous marrow transplantation in acute leukemia. To determine the efficacy of pharmacological purging by ALP, we have designed a human assay system to mimic the conditions expected in the clinical setting of autotransplantation using remission marrow. A simulated remission marrow (SRM) was prepared by mixing normal marrow cells and HL60 cells in a ratio of 1,000:1. The effect of cryopreservation on ALP-treated normal, HL60, and SRM cells was examined. In separate experiments, ALP significantly reduced the number of clonogenic HL60 cells with no effect on normal marrow progenitors. The effect of ALP was more apparent after cryopreservation. Incubation of HL60 cells with 50 micrograms/mL ALP for four hours followed by cryopreservation resulted approximately in a 3 log reduction of clonogenic HL60 cells. ALP also selectively purged the small number of leukemic cells from SRM. In SRM, the data suggested that ALP had indirect cytotoxic activity on leukemic cells by enhancing the cytotoxic activity of monocytes in addition to its direct effect. We found no evidence that clonogenic HL60 cells decreased because of induction of differentiation by ALP. These data indicated that treatment of marrow cells with ALP offers an efficient means to eliminate leukemic cells from the graft.

Elimination of leukemic cells by the combined use of ether lipids in vitro.

Two ether lipids, CP-46,665-1 (4-aminomethyl-1-[2,3-(di-n-decyloxy)-n- propyl]-4-phenylpiperidine) and ET-18-OCH3 (racemic 1-O-octadecyl-2-O-methylglycero-3-phosphocholine) have been shown to possess antileukemic activity in vitro. To explore the possible use of these compounds for purging remission bone marrow cells of leukemic cells, we examined the cytotoxic effect of these compounds on normal hematopoietic progenitor cells and leukemic cell line cells (HL-60, K-562, KG-1a, KG-1, and Daudi) by using the clonogenic assay. When cells were treated with CP-46,665-1 or ET-18-OCH3 (50 micrograms/ml for 1 h), these compounds did not inhibit the growth of normal progenitors, whereas the growth of the clonogenic leukemic cells was inhibited with differences in their sensitivities to the cytotoxic effect of CP-46,665-1 and ET-18-OCH3. Incubation of leukemic cells (HL-60 and Daudi cells) with both CP-46,665-1 (50 micrograms/ml) and ET-18-OCH3 (50 micrograms/ml) for 1 h resulted in a greater reduction of clonogenic leukemic cells than treated with each compound alone. Approximately a 3 log killing of clonogenic HL-60 cells and a 5 log killing of Daudi cells was achieved; however, the combined treatment of normal bone marrow cells with CP-46,665-1 and ET-18-OCH3 did not alter the growth of normal progenitors. This combined treatment also selectively eliminated the leukemic cells (HL-60 and Daudi cells) from a mixture (1000:1) of normal bone marrow cells and leukemic cells. It is conceivable that the pronounced difference in sensitivity to this combined treatment can be exploited for the elimination of residual leukemic cells in autologous remission marrow grafts.

Effect of alkyl-lysophospholipids on phosphatidylcholine biosynthesis in leukemic cell lines.

Alkyl-lysophospholipids are ether analogues of lysophospholipids that have tumoricidal activity mediated through activation of macrophages or by direct effect on tumor cells by disturbance of phospholipid metabolism. The effect of racemic 1-octadecyl-2-methyl-sn-glycero-3 phosphocholine on phosphatidylcholine synthesis was investigated in sensitive (HL-60) and resistant (K-562) human leukemic cell lines. Radiolabeled lysophosphatidyl-choline, choline, and methionine incorporation into phosphatidylcholine was measured in intact cells exposed for 24 h to varying concentrations of the compound. In HL-60 cells, marked inhibition of phosphatidylcholine synthesis was demonstrated using lysophosphatidylcholine or choline as precursors, but no effect was observed on methionine incorporation. No effects were observed in K-562 cells. These investigations suggest that alkyl-lysophospholipids inhibit phosphatidylcholine synthesis via the acyltransferase reaction and from choline, but not from methionine.

Proper identification of anisakine worms.

Luminal, gastric, intestinal and mesenteric forms of anisakiasis are known and can be encountered where raw or undercooked marine fish or squid are eaten. Although the anisakine nematodes which cause infection in humans are usually identified after surgical removal, laboratory personnel should be aware of their similarities to other nematodes. Cases have been reported of detection of larval nematodes in the throats or mouths of patients who have vomited or coughed. When such specimens are submitted to the clinical laboratory, problems in identification can be minimized by proper fixation and clearing. Systems for study involving clearing in phenol-ethanol and dissection to observe presence or absence of a ventricular appendix or intestinal cecum to distinguish Anisakis-, Phocanema- and Contracaecum-type larvae are described. Distinguishing characteristics are illustrated. The recovery of a Phocanema-type larva from a California woman is reported; the presence of the larvae in fish sold for human consumption in San Diego is exemplified.

Role of pH in production of hydrogen from carbohydrates by colonic bacterial flora. Studies in vivo and in vitro.

Hydrogen produced by colonic bacteria and excreted in breath is a useful index of carbohydrate malabsorption. Since colonic contents are often acidic in individuals with carbohydrate malabsorption and in normal newborns, we determined the effect of colonic acidification on H2 production. Acidification of colonic contents by dietary means significantly reduced excess breath H2 excretion from 55.4 +/- 11.1 (SEM) to 12.2 +/- 3.1 ml/4 h (P less than 0.05) after administration of 0.3 g/kg of the nonabsorbable sugar lactulose to five normal adult subjects. Similarly, the breath H2 response to lactose was reduced or eliminated in two proven lactose malabsorbers after acidification. The correlation between pH and H2 production from carbohydrate was further investigated in adults and neonates, using an in vitro fecal incubation system. Glucose disappearance and H2 production were pH dependent and highly correlated (r = 0.94) in the pH range 5.5-7.6. Maximal production of H2 from glucose by fecal incubates occurred at pH 7.0-7.45. Inhibition of H2 production from carbohydrate occurred at acid pH. H2 per hour from glucose at pH 6.2 and 5.5 averaged 60.2% and 24.2%, respectively, of that produced at neutral pH. Rapid reversal of pH-induced inhibition by neutralization indicated a metabolic, rather than a bactericidal process. The observations indicate that the breath H2 response to malabsorbed carbohydrate is affected by colonic pH. It appears that the efficiency of bacterial carbohydrate metabolism in the colon is pH dependent.

Classification of cultured mammalian cells by shape analysis and pattern recognition.

We have developed a method for classifying cultured cells on the basis of shape characteristics. High-resolution optical information on three-dimensional shape was obtained by anodic oxide interferometry. Each interference order formed in a cell was considered as a closed figure; measurement of 37 mathematical descriptors was carried out for each figure. The individual cells were classified according to the values of their descriptors. We used standard principles of pattern recognition, such as hierarchical cluster analysis and nearest neighbor analysis, as a basis for ordering the cells into groups. Alternatively, linear discriminant functions could be used, but they provided only a slight improvement in correct classification of the cells. We anticipate that the method will be appropriate for classification of cultured cell lines and for determination of the magnitude and direction of cell shape changes implicated in various biological processes.

Quantitative analysis of disturbed cell maturation in dysplastic lesions of the respiratory tract epithelium.

Autoradiographic patterns of [3H]thymidine incorporation, nuclear/cytoplasmic ratios (N/C), and the percentage of dark epithelial cells were analyzed in a group of epithelial lesions induced by 7,12-dimethylbenz[a]anthracene (DMBA) in rat tracheal transplants. It was found that similar lesions of different age exhibit the same labeling indices (LIs), therefore the lesions of different age were subsequently pooled in the following groups and studied by high resolution light microscopic autoradiography: squamous metaplasia without or with only mild atypia, squamous metaplasia with moderate atypia, squamous metaplasia with severe atypia, carcinoma in situ, and microinvasive carcinoma. Normal tracheal and esophageal epithelia were also analyzed. Whereas the normal tracheal basal layer exhibited an LI smaller than 1%, a clear difference between the carcinomas (in situ and invasive) on one hand (LI approximately 32%) and all the remaining epithelia on the other hand (LI approximately 18%) was detected. The LIs of the suprabasal cells exhibited a statistically significant difference between the squamous epithelia without atypia (LI approximately 2%) and the group comprising all the atypical lesions (LI approximately 9%). Gradients of increasing N/C (nucleus-cytoplasm ratios) values could be observed as the lesions increased in severity, especially in the middle and surface layers (e.g., in the surface layer regular metaplasia N/C = 0.08, squamous metaplasia with moderate atypia N/C = 0.26, and carcinoma in situ N/C = 0.50). Dark cells were absent in the normal esophageal epithelium, were present in moderate numbers in the basal layer of regular squamous metaplasia (18%), and increased markedly in the atypical epithelial lesions (approximately 50% in the atypical squamous metaplasias and 70% in carcinoma in situ). In the suprabasal layer dark cells increased from 3% in squamous metaplasia with moderate atypia to 28% in metaplasia with severe atypia and 56% in carcinoma in situ. The results confirm in a quantitative fashion that disturbances of cell maturation and cell proliferation are key features of dysplastic lesions induced by chemical carcinogens, and suggest the use of objective parameters for evaluation and classification of preneoplastic alterations.