Biomedical subjects

T J Woodlock

Publications and source records attributed to T J Woodlock.

16 recordsLinked to original sources

PubMed1999

Active specific immunotherapy for metastatic colorectal carcinoma: phase I study of an allogeneic cell vaccine plus low-dose interleukin-1 alpha.

A vaccine consisting of four allogeneic colon carcinoma cell lines (DLD-1, HCT116, WiDr, and T84) mixed with the adjuvant DETOX (Mycobacterium phlei cell wall and Salmonella minnesota lipid A) was administered to 25 patients with low-volume metastatic colorectal carcinoma. The first eight patients received vaccine only, given intradermally on three occasions at 3-week intervals. Subsequent patients also received subcutaneous interleukin-1 alpha (IL-1 alpha), 0.3-0.5 microgram/m2 per day for 8 days after each vaccination in an outpatient setting. Vaccine alone caused local erythema, induration, and pruritus. IL-1 caused fevers, chills, and rigors that started in 4 h and lasted 1-2 h. One patient developed a brief loss of consciousness with a rigor that resolved without sequelae. One episode of mild hypotension occurred. Fatigue occurred by day 8 of IL-1. A substantial increase in the number of patients with positive skin tests to DLD-1 and HCT116 occurred after vaccine treatment both without and with IL-1 alpha. An allogeneic cell vaccine plus subcutaneous IL-1 was administered safely to outpatients with some evidence of in vivo effect observed.

Adjuvants, Immunologic↗

PubMed1998

Coincident acute myelogenous leukemia and ischemic heart disease: use of the cardioprotectant dexrazoxane during induction chemotherapy.

Treatment of acute myelogenous leukemia is challenging in the setting of ischemic heart disease because anthracycline and anthracenedione drugs used in induction chemotherapy may potentiate myocardial dysfunction. We have managed two patients with coincident acute myelogenous leukemia and ischemic heart disease with the cardioprotectant drug dexrazoxane (ICRF-187), administered before each dose of mitoxantrone or idarubicin. Both patients tolerated their induction chemotherapy, developed marrow hypoplasia from chemotherapy, and achieved clinical remission. Dexrazoxane may have a role as a cardioprotectant in the treatment of select patients with acute myelogenous leukemia.

Adult↗

PubMed1997

Association of HSP60-like proteins with the L-system amino acid transporter.

The carrier protein(s) responsible for mammalian L-system amino acid transport has not been identified. Chronic lymphocytic leukemia (CLL) B-lymphocytes have markedly diminished L-system transport which is restored after treatment with 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Six candidate L-system related plasma membrane proteins were identified in TPA-treated CLL-cells using an L-system photoprobe and ultra-high-resolution two-dimensional gel electrophoresis. In the current studies, two candidate L-system related proteins were obtained from preparative giant two dimensional gels, and the amino acid sequences of peptide fragments from these proteins were determined by Edman degradation. These proteins, forming a doublet on the gels at an apparent size of 40 kDa and pI 5.85, had sequence homology to the mitochondrial heat shock protein 60 (hsp60). The presence of these proteins in CLL lymphocyte plasma membranes was confirmed by immunoblotting with antibodies to hsp60. These observations indicate that an hsp60 homologue is associated with the L-system amino acid transporter.

Amino Acid Sequence↗

PubMed1996

Extrapulmonary sarcoidosis: rapid spontaneous remission of marked splenomegaly.

Splenomegaly, when present as a manifestation of sarcoidosis, may cause pressure symptoms and hypersplenism. Literature reports of giant splenomegaly are few in number and describe the use of aggressive intervention with systemic steroids and splenectomy. However, the natural history of splenic sarcoidosis is not well-defined, particularly the potential for spontaneous remission as seen in pulmonary sarcoidosis. In the case described here, a spontaneous remission of giant splenomegaly from sarcoidosis was observed. Careful follow-up appears to be appropriate management for at least some of these patients.

Adult↗

PubMed1994

Feasibility of recanalization of human coronary arteries using high-intensity ultrasound.

To investigate the feasibility of ultrasonic recanalization of obstructed human coronary arteries in vitro, high-intensity ultrasound was applied to 16 coronary arteries obtained at autopsy, using a prototype instrument enabling insonification through a catheter tip. It was a 119 cm long, 0.95 mm thick wire in an 8Fr catheter connected to an external ultrasonic transformer and power generator. A 5 MHz phased-array 2-dimensional echocardiography instrument was used to determine minimal luminal diameter and percent diameter narrowing before and after ultrasound application. The ultrasonic energy was delivered at 21.5 kHz and with a 52 +/- 19 micrometer average amplitude of tip displacement. The mean percent luminal diameter narrowing, flow rate and mean pressure gradient before ultrasound exposure were 74 +/- 11%, 97 +/- 61 ml/min, and 92 +/- 18 mm Hg, respectively. After recanalization, the mean percent luminal diameter narrowing decreased to 45 +/- 17% (p < 0.001), the mean flow rate increased to 84 +/- 92 ml/min (p < 0.001), and the mean pressure gradient was reduced to 45 +/- 24 mm Hg (p < 0.001). Of the debris particles, 95% had a diameter < 9 microns (range 5 to 12). Arterial perforation occurred in 5 of 16 arteries (31%) and all 5 occurred due to stiff wire manipulation and without ultrasound application. Mechanical fracture of the wire occurred in 8 cases (50%). No signs of thermal injury were found on histology. Thus, ultrasonic recanalization of human coronary arteries in vitro is feasible. It may reduce obstruction and improve blood flow. Debris sizes are sufficiently small to minimize the hazard of peripheral embolization.

Adult↗

PubMed1994

Use of cell surface antigen phenotype in guiding therapeutic decisions in chronic myelomonocytic leukemia.

The myelodysplastic syndromes are a heterogeneous group of hematopoietic stem cell diseases in which both diagnosis and prognosis are determined by cellular morphologic criteria. In some patients, prognosis is poor due to proliferation of immature cells, i.e. development of acute leukemia. An important clinical decision for patients with myelodysplastic syndromes is whether to treat with supportive care or to use cytoreductive drugs to control the proliferative component of these illnesses. Two cases of chronic myelomonocytic leukemia are presented where cell surface antigen phenotype analysis showed characteristics suggestive of proliferative disease and the patients were treated and obtained remission with cytoablative therapy. Cell surface marker analysis may be useful in guiding therapeutic decisions in myelodysplasia.

Aged↗

PubMed1993

Phorbol ester-induced membrane proteins in chronic leukemic B-lymphocytes. Candidate proteins for the L-system amino acid transporter.

Chronic lymphocytic leukemia (CLL) B-lymphocytes have markedly diminished membrane L-system amino acid transport as compared with normal mature B- and T-lymphocytes. L-system functional recovery is induced in CLL B-cells by the maturational agent, 12-O-tetradecanoylphorbol-13-acetate (TPA). The studies reported here extend the analysis of CLL B-cell maturation by comparing membrane protein expression in untreated and TPA-treated CLL B-cells, with the identification of candidate proteins for the L-system transporter. Cell membrane proteins of resting and TPA-treated CLL B-lymphocytes were studied using ultrahigh resolution giant two-dimensional gel electrophoresis. Cellular proteins were metabolically labeled with [35S]methionine, and, in separate experiments, membrane proteins were photoaffinity labeled with [125I] iodoazidophenylalanine, an amino acid transporter by the L-system and which binds at or near the L-system transport carrier. In a partially purified membrane preparation, approximately 1400 proteins were identified by metabolic labeling. Following TPA treatment for 17 h, 14 new metabolically labeled membrane proteins were identified, and five of these also were labeled by the L-system photoprobe. Photolabeling of four of these proteins was inhibited by an excess of the L-system prototype amino acid, 2-aminobicyclo(2.2.1)heptane-2-carboxylic acid. Given these labeling characteristics, one or more of these four proteins may be related to the L-system amino acid transport carrier.

Affinity Labels↗

PubMed1991

Ability of high-intensity ultrasound to ablate human atherosclerotic plaques and minimize debris size.

To investigate whether high-intensity ultrasound can destroy atherosclerotic plaques while sparing the normal arterial wall, 279 normal human aortic sites and 119 fibrous and 193 calcified plaques, obtained from 24 necropsies, were insonified in a water tank, at 20 kHz and at 5 different power intensities, ranging from 68 W/cm2 (P1) to 150 W/cm2 (P5). These intensities were associated with a total excursion of the ultrasound irradiation apparatus tip from 90 to 268 microns, respectively. Time to perforate normal aortic sites and fibrous and calcified plaques was recorded at each intensity. There was no difference in perforation time between normal aortic sites and fibrous and calcified plaques when high-power levels (P2 to P5) were used. However, at the lowest power (P1), perforation time for the normal aortic wall was significantly longer than for fibrous and calcified plaques: 30 +/- 18 seconds (166 observations), 14 +/- 7 seconds (p less than 0.001) (78 observations) and 12 +/- 8 seconds (p less than 0.001) (115 observations), respectively. When perforation times for normal vessel wall versus fibrous plaque and normal vessel wall versus calcified plaque from the same necropsy specimen were compared in a pairwise manner, the results were: 29 +/- 13 vs 16 +/- 7 (p less than 0.001) (48 paired observations) and 26 +/- 9 vs 10 +/- 5 seconds (p less than 0.001) (55 paired observations), respectively. Regardless of whether paired or unpaired comparison was applied, no significant difference was found in perforation time between fibrous and calcified plaques. The debris did not differ in size as measured separately for normal sites and fibrous and calcified plaques by a computer-interfaced Channelizer and Coulter Counter system.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1990

Decreased L system amino acid transport and decreased gamma-glutamyl transpeptidase are independent processes in human chronic lymphocytic leukemia B-lymphocytes.

The L system of amino acid transport is markedly diminished in chronic lymphocytic leukemia (CLL) B-lymphocytes, with a maximal velocity less than 15% that of normal B-lymphocytes. Another membrane-associated function, the activity of the ectoenzyme, gamma-glutamyl transpeptidase (GGT), is diminished in CLL B-cells to 30% that of normal B-cells. In addition to its transpeptidase activity, a role for GGT has been postulated in the transport of amino acids. In the present report, the possible relationship of these two physiologic functions CLL B-cells was studied. The L system transport defect in CLL is restored by phorbol ester-induced cell maturation; following incubation with 0.15 microM tetradecanoyl phorbol acetate (TPA) for 17 hours, the L system initial velocity showed a 20-fold increase. In contrast, there was no significant effect on GGT activity with cell maturation. Furthermore, an antibody which diminished GGT activity by 50% in lymphoid cells did not inhibit L system transport. Thus, the impaired L system amino acid transport and GGT activity appear to be independent processes in CLL B-cells.

Amino Acids↗

PubMed1990

Signal transduction in N-formyl-methionyl-leucyl-phenylalanine and concanavalin A stimulated human neutrophils: superoxide production without a rise in intracellular free calcium.

Changes in intracellular ionized free calcium ([Ca]i), inositol triphosphate (IP3), and sn-1,2-diacylglycerol (DAG) were determined in relation to agonist-induced human neutrophil superoxide (O2-) production. With 0.1 microM N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation, generation of IP3 and a peak rise in [Cai] occurred at 30 sec, preceding maximal O2- production (1.5 min) and the maximal rise in DAG mass (4 min). FMLP-induced O2- production was inhibited by pertussis toxin. In cytochalasin B-primed, concanavalin A (Con A) stimulated neutrophils, a peak rise in [Ca]i but not IP3 proceeded O2- production, and pertussis toxin did not inhibit O2- production. EGTA inhibited the cytochalasin B/fMLP-induced increment in [Ca]i and O2- production by 75% and 50%, respectively, and completely ablated the response to cytochalasin B/Con A, suggesting a role for extracellular as well as intracellular calcium in the respiratory burst. However, three types of experiments indicate that an increase in [Ca]i is neither sufficient nor always required for O2- production. First, treatment with ionomycin resulted in a marked increase in [Ca]i but did not cause O2- production. Second, pertussis toxin inhibited both fMLP-induced IP3 generation and O2- production but did not inhibit the rise in [Ca]i. Third, following neutrophil priming with dioctanoylglycerol (diC8), maximal O2- production occurred in response to 0.015 microM fMLP or Con A without a rise in [Ca]i, and diC8/fMLP-induced O2- production was not inhibited by EGTA. Taken together, these data suggest that 1) an increment in [Ca]i is not strictly essential for neutrophil O2- production, 2) unlike fMLP, Con A-induced O2- production does not proceed through a pathway involving the pertussis toxin-sensitive G protein, and 3) regulation of neutrophil [Ca]i involves mechanisms independent of IP3 concentration.

Alkaloids↗

PubMed1989

Photoinhibition of 2-amino-2-carboxybicyclo[2,2,1]heptane transport by O-diazoacetyl-L-serine. An initial step in identifying the L-system amino acid transporter.

Neutral amino acid uptake into mammalian cells occurs predominantly through the L, A, and ASC carrier-mediated transport systems. The proteins responsible for transport by these systems have not been isolated, and the three pathways presently are defined by their amino acid specificity and physiologic parameters. We have found that the amino acid derivative, O-diazoacetyl-L-serine (azaserine), is a potentially useful probe for identification of the L-(leucine-favoring) system transporter in human T-lymphocytes. Uptake of azaserine competitively inhibits the uptake of the prototype L-system amino acid, 2-amino-2-carboxybicycloheptane (BCH). Azaserine undergoes photolytic cleavage with 365 nm incident light to yield a highly reactive carbene intermediate and free N2. Following photolysis of [14C]azaserine in a suspension of lymphocytes, the 14C label is detectable within a crude cytoplasmic membrane preparation, and this process is inhibited by a 50-fold excess of unlabeled azaserine or 2-amino-2-carboxybicycloheptane, suggesting that the 14C-product is associated with the membranes at or near the L-system transport site. Furthermore, photolysis of azaserine in the presence of lymphocytes results in specific irreversible inhibition of L-system transport. Thus, photolysis of azaserine provides an initial step toward the identification of the L-system transporter.

Amino Acid Transport Systems↗

PubMed1989

Diacylglycerol and calcium induce rapid enhancement of A-system amino acid transport by independent mechanisms in human T-lymphocytes.

Sn-1,2-diacylglycerols (DAG) and ionized-free calcium can act as intracellular second messengers for cell activation. Traditionally, T-lymphocyte activation is assessed by measurements of DNA synthesis or lymphokine production, but these responses require several days to occur and involve multiple intermediary regulatory steps. In contrast, we have found that T-lymphocytes demonstrate rapid enhancement of A-(alanine-favoring) system amino acid uptake when treated with DAG or ionomycin. A 30-40% increase in the initial velocity of uptake (vi) of the synthetic A-system specific amino acid, methylamino-isobutyric acid (MeAIB), was measured following 5 min of exposure to DAG or ionomycin. The vi was enhanced 60% from 12 to 19 mumol/liter cell water per min after 30 min exposure of T-cells to optimal concentrations of dioctanoylglycerol (30 microM), oleoylacetylglycerol (30 microM), or ionomycin (5 microM) (P less than .01 for each agent). A 50-fold excess of non-radioactive MeAIB inhibited 80% of [14C]MeAIB uptake in both unstimulated and stimulated cells, indicating that uptake remained largely carrier-mediated on treatment with these agents. Cycloheximide, 100 micrograms/ml, inhibited protein synthesis but did not block the A-system amino acid transport enhancement induced by DAG or ionomycin. The DAG-induced increase in the vi was blocked 40% with 100 microM H-7, an inhibitor of protein kinase C. H-7 treatment did not inhibit the ionomycin-induced A-system enhancement. A marked increase in cytoplasmic free calcium was measured when T-lymphocytes were exposed to ionomycin but not on DAG exposure, and the A-system effect of ionomycin but not DAG was blocked by extracellular EGTA. These data are compatible with two pathways for rapid enhancement of A-system amino acid uptake in T-lymphocytes. DAG stimulation is mediated via protein kinase C whereas ionomycin produces an A-system effect of similar magnitude independent of protein kinase C by an increase in cytoplasmic calcium.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1988

Prolonged remission after life-threatening gastrointestinal hemorrhage from coexistent angiodysplasia and acquired bleeding diathesis.

A patient with angiodysplasia of the stomach and duodenum developed exceptionally severe and protracted gastrointestinal hemorrhage. Bleeding was intensified by a coexistent acquired hemostatic disorder characterized by decreased platelet aggregation with adenosine diphosphate, collagen, epinephrine and ristocetin, and a decrease in both plasma ristocetin cofactor activity and high-molecular-weight von Willebrand factor multimers. Cryoprecipitate infusion corrected the von Willebrand factor defect but did not improve platelet aggregation. Bleeding stopped after prolonged aggressive combined medical and surgical therapy, and the patient had no recurrence of bleeding while followed for 27 months. Hemorrhage from the vascular lesions themselves dominated the clinical picture of this patient and other reported patients with coexistent angiodysplasia and congenital or acquired hemostatic abnormalities.

Aged↗

PubMed1988

Trans-stimulation of L-system amino acid transport in normal and chronic leukemic human lymphocytes: phorbol ester restores function in CLL.

Chronic lymphocytic leukemia (CLL) B-lymphocytes have a unique and specific diminution of L-system (leucine favoring) amino acid uptake; the maximal velocity is approximately 10% of normal B-lymphocytes. Treatment of CLL B-cells with the maturational agent, tetradecanoyl phorbol acetate, results in restoration of L-system amino acid uptake to normal velocity. To further characterize the effect of phorbol ester on the L-system of CLL B-cells, we have examined the ability of normal and CLL lymphocytes to exchange intracellular for extracellular amino acids by the L-system (trans-stimulation). A 60% increase in L-system uptake was noted in normal B- and T-lymphocytes in the presence of a high intracellular concentration of 2-amino-2-carboxy-bicycloheptane (BCH), a largely L-system-specific substrate. L-system transport was not trans-stimulated in CLL B-lymphocytes. Phorbol ester treatment restored L-system uptake in CLL to a normal Vmax of 900 mumol/liter cell water per minute in the absence of BCH loading. The Vmax could be increased further to 2,400 if phorbol ester-treated CLL cells were loaded with BCH. Hence, phorbol esters result not only in a normalization of L-system uptake in CLL B-cells but the transport system demonstrates exchange rates comparable to normal lymphocytes.

Amino Acids↗

PubMed1988

Phorbol ester restores L-system amino acid transport of B lymphocytes in chronic lymphocytic leukemia.

L (leucine-favoring)-system amino acid transport is uniquely and selectively diminished in chronic lymphocytic leukemia B lymphocytes: the maximal velocity of transport is 10% of normal B lymphocytes. We examined L-system transport in chronic leukemic B lymphocytes after incubation with tetradecanoyl phorbol acetate to determine if the transport abnormality can be corrected by the apparent cell maturation induced by this agent. Amino acid uptake was measured using 2-amino-2-carboxy-bicycloheptane, an L-system specific synthetic amino acid. Marked enhancement of L-system transport occurred in each of 12 leukemic cell populations; the initial velocity of transport in phorbol ester-treated cells increased 8-fold and 14-fold at 16 and 40 h, respectively, compared with untreated cells. The Vmax of the L-system in phorbol ester-treated leukemic cells was similar to that of phorbol ester-treated normal B lymphocytes. The L-system enhancement of the leukemic cells paralleled the development of plasmacytoid features at 40 h. Uptake of leucine, a naturally occurring L-system amino acid, was also increased by tetradecanoyl phorbol acetate. Cycloheximide, 100 micrograms/ml, which inhibited over 90% of protein synthesis in phorbol ester-treated chronic leukemic cells, blocked completely the phorbol ester-induced L-system enhancement. Phorbol ester treatment restores the selective L-system transport defect in chronic lymphocytic leukemia B lymphocytes, and this process coincides with in vitro maturation of the leukemic cells.

Aminoisobutyric Acids↗

PubMedDate not supplied

Prohibitin expression is increased in phorbol ester-treated chronic leukemic B-lymphocytes.

Chronic lymphocytic leukemia (CLL) is characterized by the gradual accumulation of immature B-lymphocytes. CLL B-lymphocytes mature to a plasmacytoid phenotype when treated in vitro with phorbol esters. CLL B-cell apparent maturation is associated with altered expression of specific plasma membrane and mitochondrial proteins including heightened expression of a 30-kDa heat shock protein 60 (hsp60) analog. During our efforts to further characterize this hsp60 analog by mass spectrometry, we detected the mitochondrial protein prohibitin in phorbol-ester-matured CLL B-lymphocytes. Prohibitin modulates cell proliferation and inhibits cell cycle traverse in several systems, although few data are available for lymphocytes. A twofold increase in prohibitin concentration was observed in phorbol-ester-matured compared to resting CLL B-cells as determined by quantitative Western immunoblot analysis. A similar increase in prohibitin was observed in phorbol-ester-treated normal human B-lymphocyte populations. An antisense oligonucleotide complementary to the 5' coding region of the prohibitin gene blunted the increase in prohibitin protein in phorbol-ester-treated CLL B-cells by 42%. These data suggest that increased prohibitin expression is associated with and may facilitate B-cell maturation.

Antineoplastic Agents↗