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J L Hudson

Publications and source records attributed to J L Hudson.

At least 55 records · Page 3Linked to original sources
PubMed1986

Lymphocyte subpopulations infiltrating squamous carcinomas of the head and neck: correlations with extent of tumor and prognosis.

Because little is known about the mechanisms involved in local tumor-host immune reactions in squamous carcinoma of the head and neck, a study was undertaken to better characterize the types of immune cells present at the local tumor site and determine their relationship to tumor extent, systemic cellular immune parameters, and clinical outcome. In 40 untreated patients, lymphocyte subsets (LS) at the tumor-host interface were quantitated immunohistologically from serial sections of frozen tumor specimens and correlated with concurrently measured peripheral LS levels and in vitro lymphocyte reactivity to phytohemagglutinin (PHA). The majority of infiltrating lymphocytes were T cells with rare B or Leu 7 cells. Proportions of T4 and T8 were similar in peritumor stroma; however, T8 cells predominated tumor parenchyma. Stromal and parenchymal infiltration by LS were not related to peripheral blood LS levels, lymphocyte reactivity, or tumor site. However, parenchymal T11 and T4 cell infiltration was less in advanced primary tumors (T3, T4) than in early tumors (T1, T2) (P = 0.01, P = 0.067, respectively), as was peripheral lymphocyte reactivity to PHA (P = 0.013). Short-term disease-free interval and actuarial survival differed significantly--according to parenchymal T11 and T4 cell infiltration--and were not related to T8, Leu 7, and B-cell infiltration. The findings extend prior studies of lymphocytic infiltration in head and neck cancer and demonstrate the potential importance of differences in tumor stromal and parenchymal infiltration. Together with recent evidence that T4 cells are critical for lymphokine production and for the proliferation of cytotoxic effector cells, the current results suggest that T4 cells play a critical role in the local immune response in squamous carcinoma of the head and neck.

B-Lymphocytes↗
PubMed1986

Coordination of segments in the vertical jump.

Three general patterns of segmental coordination (i.e., sequential [SEQ], simultaneous [SIM], and modified simultaneous [MSIM]) have been hypothesized for jumping. The purposes of this study were to describe the pattern of segmental coordination used in vertical jumping and to determine if skilled jumpers displayed distinguishing patterns of coordination. Maximum vertical jumps were performed in the counter movement (CMJ) and static jump (SJ) conditions by a heterogeneous group of 20 lean, adult subjects (AS). Smoothed, digitized film records provided the data for four segments: head-arms-trunk, trunk, thighs, and shanks. For each segment the phase of positive contribution was considered to begin with initiation of extension and end with maximum angular velocity. Bisegmental and multisegmental variables were defined to assess the extent of simultaneity. Skill was determined by the effective integration of the legs (ratio of peak upward velocity of CMJ and SJ) and by the use of stored elastic energy. Although 13 AS had MSIM patterns, the amount of flexion was small (less than or equal to 1 degree) so these AS were reclassified. With multisegmental analyses the number of AS with SIM patterns ranged from 13 to 17; about half the time was SIM. Using bisegmental analyses all 20 AS had SIM patterns; about three-fourths of the time was SIM. Skilled AS initiated extension and reached maximum velocity of the segments in proximal to distal order and with very small delays between adjacent segments.

Adult↗
PubMed1985

DNA content of human squamous cell carcinoma cell lines. Analysis by flow cytometry and chromosome enumeration.

Fifteen squamous cell carcinoma cell lines derived from nine patients were examined for DNA content by flow cytometry and chromosome counts. Using human peripheral blood leukocytes and nucleated trout and chicken red blood cells as standards, the DNA indexes of the squamous cell carcinoma cell lines were found to range from 1.1 to 3.3. The DNA content was a stable characteristic of individual cell lines in multiple passages over a seven-month period. Although flow cytometry could detect abnormal DNA content even in diploid tumor lines, the chromosome number correlated well with the DNA content by flow cytometry. In cases in which more than one cell line was established from the same patient, the individual cell lines were found to differ in their DNA content. The cell lines established from metastatic or recurrent tumors usually had a lower DNA content and chromosome number and exhibited a more aggressive in vitro growth pattern than the primary tumor or earlier recurrence. We hypothesize that "streamlined" and aggressive cell populations may evolve in vivo from more slowly growing hyperploid precursor tumor cell populations when in the course of random loss of DNA or chromosomes those that confer no growth advantage are lost, while those that do confer growth advantage are retained.

Animals↗
PubMed1985

Flow cytometric analysis of DNA in diagnostic cytology.

Flow cytometric analysis of nuclear DNA content and cell surface immunologic phenotyping were used in the evaluation of cytologic samples obtained from four patients. In each sample, a lymphoid cell population was present, which was difficult to evaluate by traditional cytopathologic methods. In two of the cases, the flow cytometric demonstration of monoclonal populations of lymphoid cells characterized by abnormal amounts of nuclear DNA gave support to the cytologic interpretation of malignancy. In a third sample, a lymphoid cell population that could not be cytologically distinguished from malignant lymphoma cells of small lymphoid cell type was shown to be composed of euploid, polyclonal cells. In the fourth case, the demonstration of euploidy in morphologically distinctive cell populations was helpful in interpreting the fine-needle aspirate of the thyroid gland. The authors conclude that flow cytometry can be used to great advantage in the evaluation of cytologic samples as well as in predicting biologic behavior.

Adult↗
PubMed1985

Plant growth-promoting hormones activate mammalian guanylate cyclase activity.

In vivo injections of plant growth-promoting hormones increase the growth of animals as well as plants. Plant growth-promoting hormones and positive plant growth regulators are known to increase RNA and protein synthesis. Since cyclic GMP also increases RNA and protein synthesis, the object of the present investigation was to determine whether physiological levels of plant growth-promoting hormones and positive plant growth regulators have part of their mechanism(s) of action through stimulation of the guanylate cyclase (EC 4.6.1.2)-cyclic GMP system. Representatives of the three classes of growth-promoting hormones were investigated. Thus, auxins (indole-3-acetic acid, indole-3-butyric acid, beta-naphthoxyacetic acid, and 2,4,5-trichlorophenoxy acetic acid), gibberellins (gibberellic acid), and cytokinins [N6-benzyl adenine, kinetin (6-furfuryl aminopurine), and beta-(2-furyl) acrylic acid] all increased rat lung, small intestine, liver, and renal cortex guanylate cyclase activity 2- to 4-fold at the 1 microM concentration. Dose response curves revealed that maximal stimulation of guanylate cyclase by these plant growth regulators was at 1 microM; there was no augmented cyclase activity at 1 nM. The guanylate cyclase cationic cofactor manganese was not essential for augmentation of guanylate cyclase by these plant growth-promoting regulators. The antioxidant butylated hydroxytoluene did not block the enhancement of guanylate cyclase by these plant growth-promoting factors. These data suggest that guanylate cyclase may play a role in the mechanism of action of plant growth-promoting hormones and even of positive plant regulators at the cellular level.

Animals↗
PubMed1985

Effects of guanine ribonucleotide accumulation on the metabolism and cell cycle of human lymphoid cells.

A deficiency of purine nucleoside phosphorylase activity is associated with marked depletion of T-lymphocytes which is felt to be mediated by accumulation and further metabolism of the purine nucleoside phosphorylase substrate, 2'-deoxyguanosine. Human T-lymphoblasts incubated in the presence of 2'-deoxyguanosine and the purine nucleoside phosphorylase inhibitor 8-aminoguanosine accumulate deoxyguanosine 5'-triphosphate whereas B-lymphoblasts and mature T4+-cell lines accumulate GTP under identical conditions. We have compared the effects of guanine ribo- and deoxyribonucleotide accumulation on the metabolism and cell cycle of the respective cell lines. Deoxyguanosine 5'-triphosphate elevations in T-lymphoblasts are associated with inhibition of [3H]uridine incorporation into DNA and a complete block at the G1-S interface of the cell cycle. In contrast 3- to 5-fold increases in guanosine 5'-triphosphate pools in B-lymphoblasts and mature T-cell lines do not inhibit [3H]uridine incorporation into DNA or RNA but do cause a pronounced slowing in the progression of cells through S phase. B-lymphoblasts deficient in the salvage enzyme hypoxanthine guanine phosphoribosyltransferase do not accumulate guanosine 5'-triphosphate from 2'-deoxyguanosine and progress normally through the cell cycle, demonstrating a requirement for guanine salvage to inhibit cell growth. Guanine ribonucleotide accumulation was also associated with inhibition of de novo purine biosynthesis and a moderate decline in adenine nucleotide pools but not with inhibition of protein synthesis or alterations in basal levels of 3':5'-cyclic adenosine monophosphate or 3':5'-cyclic guanosine monophosphate. We conclude that the accumulation of guanine ribonucleotides by actively cycling human lymphoid cells is associated with an increase in S-phase cells and inhibition of growth. This effect is distinctly different from that produced by 2'-deoxyguanosine 5'-triphosphate and should be taken into account in pharmacological studies with 2'-deoxyguanosine and its analogues.

Cell Cycle↗
PubMed1985

Functional defects in phagocytic cells following thermal injury. Application of flow cytometric analysis.

Defective phagocytic cell function may partially account for the morbidity and mortality associated with thermal injury. In experimental thermal injury in the rat, small circulating blood volumes increase the difficulty in obtaining significant data. Furthermore, purification and or elicitation procedures have the potential for altering the cell surface characteristics and/or the functional response of the cell in question. We have examined the circulating neutrophils and pulmonary alveolar macrophages of anesthetized rats following a 16-20% body surface area scald injury to the shaved back. The circulating neutrophils of thermally injured rats were examined by flow cytometry following stimulation with phorbol myristate acetate (PMA) (100 ng/ml) in terms of the change in fluorescence intensity of the potentiometric cyanine dye, dipentyloxocarboxyanine and the formation of the oxidized product of 2',7'-dichlorofluorescin diacetate-loaded cells. The alveolar macrophages were examined after stimulation with PMA (100 ng/ml) in terms of the change in fluorescence intensity of the potentiometric dye, dipropylthiodicarbocyanine and the generation of superoxide production, as assessed by the superoxide dismutase inhibitable reduction of cytochrome c. Both cells exhibited a profound inhibition of cell function 4 hours after the insult, with partial return toward control values at later time points. Furthermore, the plasma of thermally injured rats, 4 hours after the burn was inhibitory to normal rat neutrophils. Fluorescent compounds suggestive of in vivo lipid peroxidation were maximally detectable at this time point. Further research is needed to establish the role of these products in the induction of phagocytic cell dysfunction.

Animals↗
PubMed1985

DNA content of murine fibrosarcoma cell lines with varying metastatic potential.

The DNA content of murine fibrosarcoma cell lines of various metastatic potential was the subject of the current investigation. The cell lines were derived from methylcholanthrene-induced tumors as described previously (J. Varani et al., J. Natl. Cancer Inst., 71: 1281-1287, 1983). Cells were maintained in vitro and used for DNA studies no more than 48 h after passage. DNA staining was accomplished using propidium iodide and flow cytometry was used to quantitate relative amounts of DNA. Trout and chicken erythrocytes and mouse thymocytes were used as internal DNA standards for each cell line. DNA indices were calculated as the ratio of the G0-G1 peak channel number of the tumor cells to the G0-G1 peak channel number of the thymocytes. Manual chromosome counts were also obtained from each cell line using Giemsa-stained preparations. All cell lines demonstrated a single aneuploid population. The two tumor lines with the highest metastatic potential were slightly hyperdiploid whereas three low metastatic lines were near tetraploid. A sixth line of moderate metastatic potential was also found to be near tetraploid. Chromosome counts and flow cytometric analyses were in close agreement indicating that DNA content was largely due to chromosome replication. These data suggest that, in this model, metastatic potential and DNA content are inversely related once diploidy is exceeded.

Animals↗
PubMed1985

Effects of the tricyclic nucleoside 6-amino-4-methyl-8-(beta-D-ribofuranosyl)- pyrrolo[4,3,2-de]pyrimido[4,5-c]pyridazine on the viability and cell cycle distribution of L1210 cells in vitro.

TCN (1 microM) totally inhibited the growth of L1210 cells in culture and caused progressive loss of cellular viability, as indicated by a decreased clonogenicity and nigrosin dye exclusion. After 24 h or more of TCN treatment, a significant fraction of the cells had shrunk in size but did not fragment into dye-impermeable vesicles as reported previously for N1S1-67 hepatoma cells (P. G. W. Plagemann, J. Natl. Cancer Inst., 57: 1283-1295, 1976). TCN-induced growth inhibition was accompanied by a block of cell cycle progression in G1 or at the G1-S boundary. At all TCN concentrations studied, progression of cells out from behind this block was evident as a depletion of the early S-phase population in comparison to controls, while increasing the concentration of TCN (0.1 to 1 microM) led to a progressive retention of cells in S phase, suggesting a slowing of progression through S phase. The fraction of S-phase cells incorporating [methyl-3H]thymidine and the amount of [methyl-3H]thymidine incorporated per labeled cell were both decreased by TCN treatment. Increasing the concentration of TCN (0.1 to 1 microM) progressively decreased DNA synthesis and increased cell lethality. Thus it appeared that inhibition of DNA synthesis might cause the retention of cells in S phase which is associated with TCN lethality.

Animals↗
PubMed1984

A rapid method for preparation of urinary bladder epithelium for flow cytometric analysis.

A rapid and reliable method is described for preparation of urinary bladder epithelium for flow cytometric analysis. This method has proven useful for measurement of cell cycle kinetics in bladder epithelium from normal fetal, neonatal and adult mice as well as animals that have been exposed to urinary bladder toxicants. A modified Vindelov 's propidium iodide stain was instilled directly into the urinary bladder through the urethra, allowing the propidium iodide to lyse the epithelial cells and stain the DNA in the liberated nuclei. The stained epithelial cell nuclei were then removed from the lumen and analyzed by flow cytometry without centrifugation or filtration techniques. This method allows examination of epithelial cell nuclei without contamination from other tissue elements due to the fact that the propidium iodide did not act beyond the epithelial basement membrane under our experimental conditions.

Animals↗
PubMed1984

The preparation of electronically sorted hematopoietic cells for visual analysis.

The authors describe a simple method of collecting and preparing electronically sorted hemic cells for visual analysis. Wright-stained preparations were obtained easily by this method and offered excellent preservation of morphologic features. Flow cytometer-derived data thus can be correlated with the morphologic characteristics of immunologically distinctive subsets of hemic cells.

Cell Separation↗
PubMed1983

Preparation and stability of sixteen murine tissues and organs for flow cytometric cell cycle analysis.

Three different technical protocols were used to prepare samples for flow cytometric (FCM) analysis. Each protocol developed worked best for only certain organs. Protocol I involved mincing small pieces of fresh tissue in the propidium iodide (PI) staining solution and filtering through packed glass wool. The organs that were prepared by protocol I were: submandibular gland, urinary bladder, liver, thymus, bone marrow, spleen, lung, kidney and testis. Protocol II involved exposure of the organ to 0.5% acetic acid for 48 h prior to mincing in the PI. The organs that were prepared by protocol II were: uterus, rectum, colon, ileum, and heart. Protocol III utilized an exposure to 0.5% acetic acid, pepsinization, and then staining with PI. The tissues that were prepared by protocol III were the epithelium of the anterior surface of the cornea and the epithelium of the surface of the tongue. A total of 16 different organs and tissues were successfully prepared. For each organ, averaged DNA histograms were analyzed by nonparametric and parametric programs and the results (phase fractions) are presented in tabular form. Several of the organs used came from animals exposed to 1.0 mg/kg vincristine (VC) for 5-6 h to test the capability of the different protocols to detect the enlargement of the G2 + M compartment by the accumulation of VC-arrested mitotic figures. The stability of the many different sample preparations was tested by comparing averaged DNA histograms obtained on the day of sample preparation to averaged DNA histograms of the same set of samples after storage at 4 degrees C, with or without fixation in 10% phosphate-buffered formalin, for days to weeks. After staining with propidium iodide, fixation of the sample with a final concentration of 2-3% phosphate-buffered formalin, was the procedure adopted to assure sample stability. The demonstration of sample stability permits sample preparation to occur at one site followed by transport of the samples to the FCM laboratory at another geographical location. The major findings of this work were a) technical protocols were developed which resulted in acceptable nuclear suspensions for FCM from 16 different murine organs or tissues, b) the stability of these samples can be assured by fixing the PI stained nuclear suspension with formalin, and c) each different protocol was capable of detecting and preserving at least some of the mitotic figures arrested and collected by vincristine.

Acetates↗
PubMed1983

Flow cytometric analysis of the effect of phenytoin and its major metabolite on mitogen stimulated mouse spleen cells.

The in vitro immunopharmacological effects of phenytoin (PHT) and its major metabolite 5-(4-hydroxyphenyl)-5-phenylhydantoin (HPPH) were investigated by flow cytometric analysis of the DNA content of mitogen stimulated mouse spleen cells. The qualitative effects of PHT and HPPH were similar in concanavalin A stimulated mouse spleen cells with both compounds causing an increase in the percentage of S phase cells. The data suggests that this effect is due to an augmentation of cell cycling as demonstrated by the significant increase in 4N cells in PHT treated cultures relative to control cultures following colcemid treatment. A PHT time course study revealed an increase in S phase cells and a subsequent increase in 4N cells. PHT had no significant effect on congenitally athymic nude mouse spleen cells stimulated with lipopolysaccharide (LPS) except at the highest concentration tested (80 micrograms/ml) where a depression of cell cycling was observed. HPPH caused a colcemid-like accumulation of 4N cells in the LPS stimulated nude mouse spleen cell cultures. PHT and HPPH were found to be effective in enhancing cell cycling in cultures containing a significant population of T-cells stimulated with a T-cell mitogen whereas an inhibitory effect was observed in cultures without T-cells stimulated with a B-cell mitogen. The capacity of PHT to enhance the mitogenic action of concanavalin A may relate to its capacity to induce immunologic abnormalities and lymphadenopathy in humans.

Animals↗
PubMed1983

Fatty acids and the inhibition of mitogen-induced lymphocyte transformation by leukemic serum.

The effects of sera from 23 patients with acute lymphocytic leukemia on mitogen-induced transformation of normal human lymphocytes were examined. All sera (100%) at diagnosis and 70% of those obtained during the induction of remission demonstrated inhibition of mitogen-induced lymphocyte transformation as evidenced by decreased uptake of [3H]thymidine. The inhibition could not be overcome by an increase in the mitogen concentration. Eighteen sera demonstrating a mean inhibition of 44.2% had elevated levels of free fatty acids (FFA) compared with five sera showing no inhibition (p value = 0.001). Inhibition, within the range produced by leukemic sera, could be achieved by the purified methyl esters of linoleic and linolenic acid in this system. Flow cytometric analysis of mitogen-induced cycling of normal human lymphocytes suggested that deoxyribonucleic acid (DNA) replication was inhibited by leukemic sera containing elevated levels of FFA after the G1 phase of cell cycling. Similar inhibition could be achieved by the methyl ester of linolenic acid in this system. Gas chromatographic analysis revealed that sera demonstrating inhibition contained linolenic (C18:3) acid (p value = 0.015), and the majority showed one or more of the following: 1) a concentration of oleic (C18:1) acid greater than 2 standard deviations (SD) above the mean of the control leukemic sera (i.e., those not demonstrating inhibition); 2) an arachidonic (C20:4) to C18:1 ratio that is reduced to greater than 2 SD below the mean of the control group; or 3) the presence of an unexpected fatty acid fraction. The data raise the question of an influence of FFA and, specifically, an interplay of unsaturated fatty acids on immune function during the natural history of acute lymphocytic leukemia.

Fatty Acids↗