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T C Lo

Publications and source records attributed to T C Lo.

At least 37 records · Page 2Linked to original sources
PubMed1995

Use of glucose transport mutants to examine the intrinsic properties of glucose transport processes in rat myoblasts.

Glucose transport mutants were used to examine the intrinsic properties of glucose transport processes in rat myoblasts. Studies with mutants devoid of any functional glucose transporter revealed that substantial amount of sugar analogues was internalized via simple diffusion; however, equilibration of these analogues across the plasma membrane was not achieved after 1 min of incubation at 23 degrees C. The rates of internalization were substantially higher with sugar analogues that were phosphorylated by intracellular kinases. Mutants harbouring only one functional GLUT transporter were also used to examine the intrinsic properties of specific GLUT transporters. The preferred substrate for the GLUT 1 transporter was 2-deoxy-D-glucose (dGlc); the transport affinity for this substrate was reduced by energy uncouplers. Studies with mutants possessing only the GLUT 4 transporter revealed that this transporter existed in a high and a low affinity form. The former was responsible for dGlc uptake; whereas the latter was for the uptake of both 3-O-methyl-D-glucose (MeGlc) and dGlc; only the former was affected by energy uncouplers. These studies illustrated the usefulness of mutants in characterizing glucose transport processes.

Animals↗
PubMed1995

Endobronchial brachytherapy.

Intraluminal endobronchial brachytherapy is a technique in which an encapsulated radioactive source is placed near a tumor for localized irradiation. It is effective, with or without other treatment modalities, in palliating problems caused by endobronchial malignancies, such as dyspnea, hemoptysis, cough, atelectasis, and postobstructive pneumonia. This article describes the different techniques and dosage schemes for brachytherapy, indications and contraindications, reported rates of efficacy and complications, and limitations of the technique.

Brachytherapy↗
PubMed1994

Intestinal spirochaetosis.

Two cases of intestinal spirochaetosis are described. The first case improved with treatment while the second case improved spontaneously without any intervention. Controversy over treatment and pathogenicity of intestinal spirochaetosis is discussed with review of previous publications.

Adolescent↗
PubMed1993

Transcripts for the high and low affinity hexose transporters in rat myoblasts.

The present investigation examined the glucose transporter (GLUT) transcript levels in glucose-grown rat L6 myoblasts, which possessed a high (HAHT) and a low (LAHT) affinity hexose transport system. The most prominent GLUT transcript detected in these cells was a GLUT 3-recognizable 4.1-kilobase transcript. Mutants defective in HAHT contained only residual levels of the GLUT 3 transcript and transporter. Corresponding changes in GLUT 3 transcript levels and HAHT activity were observed in myotubes, confluent cultures, and myogenesis-impaired cells. The other prominent GLUT transcripts detected were the GLUT 4-recognizable 2.8- and 1.4-kilobase transcripts. The close correlation between the GLUT 4 isoform and LAHT activity was indicated by the absence of this isoform in LAHT- mutants, and their corresponding changes in myoblasts grown under different conditions. These findings suggested the HAHT and LAHT transporters might be coded or regulated by the genes responsible for the GLUT 3 and 4 transcripts, respectively. Although relatively high levels of the GLUT 1 transcript and transporter were detected in glucose-grown HAHT-LAHT- mutants, these cells did not exhibit any GLUT 1 transport activity. Since the GLUT 1 transporter was not functional, it was not likely responsible for the HAHT and LAHT activities observed in glucose-grown L6 myoblasts.

Animals↗
PubMed1993

Use of p112-deficient myoblasts to determine the temporal order of the in vitro expression of myogenic components.

The present investigation examines the function and site(s) of involvement of an ecto-protein kinase and its substrate protein (a cell surface 112 kDa protein) in the in vitro myogenic pathway. The phosphorylated 112 kDa protein (p112) has recently been shown to be involved in myogenesis. Not much information is currently available on the role of the ecto-protein kinase and the 112 kDa protein in modulating the expression of the myogenic factors and various muscle-specific proteins. Five different p112-deficient rat myoblasts were used to examine the temporal order of the in vitro expression of the myogenic components; namely, L6 myoblasts treated with BrdUrd or phloretin, a conditional p112-defective mutant (clone D1), an ecto-protein kinase-deficient mutant (clone F72), and a mutant defective in the 112 kDa protein (clone D1/S4). All these p112-deficient myoblasts were also impaired in myogenesis. The absence of p112, ecto-protein kinase, and/or the 112 kDa protein was found to have no effect on the Myf-5 mRNA level. However, the expected increase in NCAM and Myf-4 mRNAs was not observed in any of the p112-deficient myoblasts examined. This suggests that the p112 site of action is probably located upstream of the Myf-4 and NCAM sites in the myogenic pathway. While 7-28 fold increases in the MLC, MHC, and TnT transcripts were observed during myogenesis, such increases were not observed in the p112-deficient myoblasts. However, when mutant D1/S4 was transfected with the myf-4 cDNA, expression of Myf-4 in the transfectant resulted in increased level of the MLC, MHC, and TnT mRNAs, and in myotube formation, even though the Myf-5 and NCAM mRNA levels and p112 were not altered. This suggests that p112 may function by activating transcription of Myf-4, which will subsequently promote the expression of muscle-specific proteins and myotube formation. In the absence of p112, Myf-5 cannot activate the expression of Myf-4, NCAM, MLC, MHC, TnT, and myotube formation. If all these components are involved in the same myogenic pathway, then p112 may be acting downstream from Myf-5, and upstream from NCAM and Myf-4.

Animals↗
PubMed1992

Intraluminal low-dose rate brachytherapy for malignant endobronchial obstruction.

From October 1985 through October 1989, 87 patients underwent 105 intraluminal brachytherapy treatments for endobronchial or endotracheal malignant tumors. Low-dose rate iridium-192 seeds were used. Of the 60 patients treated for primary lung carcinoma, 52 patients (87%) had previously received full-course external beam radiotherapy to the tumor sites. Ten patients were treated for symptomatic metastatic disease, and one patient had extension of tumor into the trachea from carcinoma of the cervical esophagus. Clinical or bronchoscopic improvement was noted in 42 patients (59%). No significant difference in the response rate was observed between various types of tumor. Patients who were treated with a radiation dose larger than 2500 cGy at a 2 cm radius had a significantly greater response rate (77%) than patients treated with a dose less than 2500 cGy (38%) (p = 0.001). A trend toward better results was apparent in patients who had undergone Nd:YAG laser bronchoscopy in the 2 weeks before brachytherapy. The treatments were well tolerated, and the incidence of serious complications was low and acceptable.

Brachytherapy↗
PubMed1991

Phosphorylation of a cell surface 112 kDa protein by an ecto-protein kinase in rat L6 myoblasts.

Studies with subconfluent day 2 cultures of rat myoblasts revealed that a cell surface 112 kDa protein could be phosphorylated by extracellular ATP. Analysis of the phosphorylated 112 kDa protein suggested the involvement of a serine protein kinase. The following evidence indicated the cell surface location of this protein kinase: (i) extracellular ATP was unable to penetrate the cell membrane under our experimental conditions; (ii) the phosphorylated protein profile of intact cells differed significantly from that of broken cells; (iii) the phosphorylation of the 112 kDa protein could be abolished by pretreatment of cells with very low concentrations of trypsin; (iv) the phosphorylated 112 kDa protein could be dephosphorylated by exogenously added alkaline phosphatase; (v) the phosphorylation of the 112 kDa protein was inhibited by exogenously added proteins; and (vi) exogenously added proteins could be phosphorylated by intact cells under similar experimental conditions. The phosphorylated 112 kDa protein was detected only when the reaction was carried out in the presence of Ca2+, Mg2+, and F- ions. Kinetic analysis that revealed that the Km value of the ecto-protein kinase for ATP was 0.04 microM, and the Vmax. value for phosphorylation of the 112 kDa protein was 1.67 x 10(-4) pmol/min per 10(5) cells. Data presented in the accompanying paper [Chen & Lo (1991) Biochem. J. 279, 475-482] show that there was a constant and adequate supply of ATP on the cell surface of rat myoblasts for efficient functioning of this protein kinase, and that mutants defective in either the ecto-protein kinase or the 112 kDa protein were also impaired in myogenic differentiation. This and other biochemical studies suggest that the ecto-protein kinase and the 112 kDa protein might play important roles in myogenic differentiation.

Adenosine Triphosphate↗
PubMed1991

Involvement of a cell surface protein and an ecto-protein kinase in myogenesis.

Myogenic differentiation is composed of a sequential cascade of multiple steps leading to the formation of multinucleated myotubes. The interference with any one step would abolish myogenesis. The present investigation examined the cell surface components which might be involved in myogenesis. Studies with subconfluent day 2 cultures of rat L6 myoblasts revealed that a cell surface 112 kDa protein was phosphorylated by a Ca(2+)-, F(-)- and Mg(2+)-dependent ecto-protein kinase [Chen & Lo (1991) Biochem. J. 279, 467-474]. We have shown in the present investigation that adequate ATP was present on the cell surface for efficient functioning of this ecto-protein kinase. The phosphorylation of the 112 kDa protein by this ecto-protein kinase was decrease dramatically in confluent cells and in multinucleated myotubes. The following evidence suggests that both the 112 kDa protein and the ecto-protein kinase may play important roles in myogenesis. (i) The highest phosphorylation activity was observed in subconfluent cultures, i.e. before the onset of morphological differentiation. (ii) Treatment of cells with chemical reagents resulted in a corresponding decrease in the ecto-protein kinase, the 112 kDa protein, the phosphorylated 112 kDa protein (p112) and the ability to form myotubes. (iii) The level of p112 in a conditional myogenesis-defective mutant corresponded with the cells' eventual ability to differentiate. (iv) A mutant defective in the ecto-protein kinase was impaired in the phosphorylation of the 112 kDa protein and in myogenesis. (v) A mutant containing only residual levels of the 112 kDa protein was deficient in both p112 and myogenesis. (vi) Since the level of p112 was normal in another myogenesis-defective mutant, the phosphorylation of this protein was not likely to be a consequence of myogenic differentiation. The above findings suggest that the ecto-protein kinase and the 112 kDa protein may directly or indirectly be associated with the myogenic pathway. Since the levels of the ecto-protein kinase, the 112 kDa protein and p112 decreased dramatically upon the formation of myotubes, these proteins were probably not required once morphological differentiation had been initiated.

Adenosine Triphosphate↗
PubMed1990

Use of a genetic variant to study the hexose transport properties of human skin fibroblasts.

Human skin fibroblasts from 'normal' subjects were found to possess at least two hexose transport systems. One system was responsible for the uptake of 2-deoxy-D-glucose (dGlc), D-glucose and D-galactose, whereas the other was responsible primarily for the uptake of 3-O-methyl-D-glucose (MeGlc). The transport of dGlc was the rate-limiting step in the uptake process; over 97% of the internalized dGlc was phosphorylated and the specific activity of hexokinase was several times higher than that for dGlc transport. The dGlc transport system was activated by glucose starvation, and was very sensitive to inhibition by cytochalasin B and energy uncouplers. Fibroblasts isolated from a patient with symptoms of hypoglycaemia were found to differ from their normal counterparts in the dGlc transport system. They exhibited a much higher transport affinity for dGlc, D-glucose and D-galactose, with no change in the respective transport capacity. Transport was not the rate-limiting step in dGlc uptake by these cells. Moreover, the patient's dGlc transport system was no longer sensitive to inhibition by cytochalasin B and energy uncouplers. This suggested that the intrinsic properties of the patient's dGlc transport system were altered. It should be noted that the patient's dGlc transport system could still be activated by glucose starvation. Despite the changes in the dGlc transport system, the MeGlc transport system in the patient's fibroblasts remained unaltered. The observed difference in the properties of the two hexose transport systems in the 'normal' and the patient's fibroblasts strongly suggests that the two transport systems may be coded or regulated by different genes. The present finding provides the first genetic evidence from naturally occurring fibroblasts indicating the presence of two different hexose transport systems.

3-O-Methylglucose↗
PubMed1990

Involvement of hexose transport in myogenic differentiation.

A high (HAHT) and a low (LAHT) affinity hexose transport system are present in undifferentiated rat L6 myoblasts; however, only the latter can be detected in multinucleated myotubes. This suggests that HAHT is either down-regulated or modified as a result of myogenesis. The present investigation examined the relationship between HAHT and myogenic differentiation. While myogenesis could be inhibited by the potent hexose transport inhibitor phloretin, it was not affected by phlorizin which had no effect on hexose transport. This relationship was further explored using six different HAHT-defective mutants. All six mutants, altered in either the HAHT transport affinity (Type I mutants) or capacity (Type II mutants), were impaired in myogenesis. Since these mutants were selected from both mutagenized and non-mutagenized cells with different reagents, or with different concentrations of the same reagent, the deficiency in myogenesis was likely due to changes in HAHT properties. This notion was confirmed by the observation that growth of Type I mutants in high D-glucose concentrations could rectify the defect in myogenesis. D-glucose was unlikely to rectify the defect in myogenesis, if this defect was due to a second unrelated mutation that may have arisen during isolation of the mutants. Since both types of mutants were not altered in LAHT, D-glucose should still be taken up into the cells. The fact that the glucose-mediated increase in fusion could not be observed in Type II mutants (deficient in the HAHT transporter) suggested that myogenesis was dependent on the presence of D-glucose or its metabolites in specific HAHT-accessible compartments. It is tempting to speculate that trans-acting regulators involved in myogenesis may be synthesized from the glucose metabolites in these specialized HAHT-accessible compartments.

Animals↗
PubMed1990

Single-dose electron beam irradiation in treatment and prevention of keloids and hypertrophic scars.

Low megavolt electron beam irradiation was used on 354 sites in 199 patients at the Lahey Clinic either for palliation of symptomatic hypertrophic scars or as post-operative irradiation in an attempt to prevent formation or recurrence of hypertrophic scars. Electron energies used ranged from 1.5 to 3.5 MeV. The median age of the 59 male patients was 22 years and of the 140 female patients, 35 years. All patients had at least one follow-up visit, and the median follow-up was 35 months. Of the 294 sites treated for the first time, 272 (93%) were irradiated with a single fraction with a skin dose ranging from 2 to 20 Gy. Of the 85 sites in 63 patients without excision of symptomatic hypertrophic scars, single-dose electron beam irradiation was of clinically significant value in only 41 sites (48%). No patients have been treated without surgical excision since 1973. Because of a history of formation of hypertrophic scars elsewhere in the body, 13 patients with 19 incisions were treated prophylactically after operation for other diseases. All sites were irradiated with single doses ranging from 8 to 20 Gy, and hypertrophic scars did not subsequently develop in any patient. Altogether, 119 patients with 174 sites were irradiated after surgical excision of hypertrophic scars to prevent recurrence; 168 sites (97%) received single-fraction irradiation, and 161 received a dose of 8 Gy or greater, up to 15 Gy. No statistically significant differences were observed in complete success rates, ranging from 82 to 90% with doses of 9 Gy or greater.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1990

Single-dose radiation therapy for prevention of heterotopic ossification after total hip arthroplasty.

Single-dose radiation therapy was prospectively evaluated for its efficacy in prevention of heterotopic ossification in patients at high risk after total hip arthroplasty. Thirty-one patients (34 hips) were treated between 1981 and 1988. Risk factors for inclusion in the protocol included prior evidence of heterotopic ossification, ankylosing spondylitis, and diffuse idiopathic skeletal hyperostosis. Patients with hypertrophic osteoarthritis or traumatic arthritis with osteophytes were not included. Operations on 34 hips included 19 primary total and 11 revision total hip arthroplasties and 4 excisions of heterotopic ossification. All patients received radiotherapy to the hip after operation with a single dose of 700 centigray. Radiotherapy is recommended on the first postoperative day. After this single-dose radiation treatment, no patient had clinically significant heterotopic ossification. Recurrent disease developed in two hips (6%), as seen on radiography (grades 2 and 3). This series documents a 100% clinical success rate and a 94% radiographic success rate in preventing heterotopic ossification in patients at high risk after total hip arthroplasty. Single-dose radiotherapy is as effective as other radiation protocols in preventing heterotopic ossification after total hip arthroplasty. It is less expensive and easier to administer than multidose radiotherapy.

Adult↗
PubMed1990

Hexose transport properties of myoblasts isolated from a patient with suspected muscle carnitine deficiency.

The human primary carnitine deficiency syndromes are potentially fatal disorders affecting children and adults. The molecular etiologies of these syndromes have not been fully determined. Muscle carnitine deficiency syndrome is characterized by mild to severe muscle weakness, lipid accumulation in muscle, and reduced muscle carnitine concentration. In the present investigation, the hexose transport properties of muscle cells isolated from a patient with suspected muscle carnitine deficiency (MCD) were examined. We have previously shown that myoblasts from normal human subjects possessed at least two hexose transport systems, the low (LAHT) and the high (HAHT) affinity hexose transport systems. Their preferred substrates were 3-O-methyl-D-glucose and 2-deoxyglucose (dGlc), respectively; HAHT, but not LAHT, was sensitive to inhibition by carbonyl cyanide m-chlorophenylhydrazone (CCCP). Here we show that the kinetic properties of HAHT in the MCD myoblasts differ significantly from those of normal myoblasts and that the rates of dGlc transport by MCD myoblasts are restored to normal by growth in 40 microM L-carnitine. We also demonstrate that the kinetic properties of LAHT are quite similar in both normal and MCD myoblasts. It can be inferred from these findings that HAHT and LAHT may be coded or regulated by different genes. Based on the finding that the dGlc transport system in L-carnitine grown cells is no longer sensitive to inhibition by CCCP, it is thought that L-carnitine may play a regulatory role in HAHT, viz., by maintaining the HAHT transporter in a functional state, even in energy-uncoupled cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1990

Genetic evidence indicating the identity of the cytochalasin B photolabelled components in rat myoblasts.

While photolabelling with cytochalasin B (CB) has been widely used in the identification of eukaryotic glucose transporters, there is presently no unequivocal evidence indicating that the CB-labelled components are indeed the glucose transporters. A combination of biochemical, physiological and genetic manipulations was used in the present investigation to demonstrate that the plasma membrane hexose transporters can indeed by photolabelled by CB. In this study, plasma membranes from glucose-grown and glucose-starved hexose transport mutant D23 and its parental L6 cells were photolyzed in the presence of 3H-CB. The amount of CB bound to the 40-60 kDa region (CB50) was found to be differentially inhibited by D-glucose, 2-deoxy-D-glucose (dGlc) and 3-O-methyl-glucose (MeGlc). Mutant D23 exhibited not only reduced hexose transport activity but also significantly lower level of CB50. Glucose-starvation resulted not only in elevated hexose transport activity but also increased level of CB50. It should be noted glucose-starvation did not have much effect on the hexose transport activity and on the level of CB50 in mutant D23. The present study provides the first genetic evidence indicating that the CB-labelled component(s) are indeed associated with the hexose transport systems.

Animals↗
PubMed1989

Hexose transport in human myoblasts.

The present investigation reports on the hexose transport properties of human myoblasts isolated from normal subjects and from patients with Duchenne muscular dystrophy (DMD). Similar to rat myoblast L6, normal human myoblasts possess a high- (HAHT) and a low- (LAHT) affinity hexose transport system. The non-metabolizable hexose analogue, 2-deoxyglucose, is preferentially taken up by HAHT. The transport of this analogue is the rate-limiting step in the uptake process. This human myoblast HAHT is also similar to that of the rat myoblast in its substrate specificity and in response to the energy uncouplers, cytochalasin B and phloretin. The human myoblast LAHT resembles that of rat myoblast in its insensitivity to energy uncouplers, and in its transport affinity and capacity for 3-O-methyl-D-glucose. Although DMD myoblasts resemble their normal counterpart in their ability to differentiate, they differ significantly in their hexose transport properties. In addition to HAHT and LAHT present in normal human myoblast, DMD myoblasts contain a super-high-affinity hexose transport system (SHAHT). SHAHT can be detected only at very low substrate concentrations. It differs from HAHT not only in its much higher transport affinity, but also in its response to the traditional hexose transport inhibitors. For example, SHAHT can be activated by cytochalasin B and phlorizin, whereas it is more sensitive to inhibition by phloretin. Unlike HAHT, energy uncouplers are found to be ineffective in inhibiting SHAHT. It should be mentioned that SHAHT cannot be detected in myoblasts isolated from patients with other types of myopathy. The present study serves to demonstrate that more than one hexose transport system is operating in human skeletal muscle cells, as found in other cell types.

2,4-Dinitrophenol↗
PubMed1989

Regulation of hexose transport in rat myoblasts during growth and differentiation.

We report here the effects of growth conditions and myogenic differentiation on rat myoblast hexose transport activities. We have previously shown that in undifferentiated myoblasts the preferred substrates for the high (HAHT)- and low (LAHT)-affinity hexose transport systems are 2-deoxyglucose (2-DG) and 3-O-methyl-D-glucose (3-OMG), respectively. The present study shows that at cell density higher than 4.4 x 10(4) cells/cm2, the activities of both transport processes decrease with increasing cell densities of the undifferentiated myoblasts. Since the transport affinities are not altered, the observed decrease is compatible with the notion that the number of functional hexose transporters may be decreased in the plasma membrane. Myogenic differentiation is found to alter the 2-DG, but not the 3-OMG, transport affinity. The Km values of 2-DG uptake are elevated upon the onset of fusion and are directly proportional to the extent of fusion. This relationship between myogenesis and hexose transport is further explored by using cultures impaired in myogenesis. Treatment of cells with 5-bromo-2'-deoxyuridine abolishes not only myogenesis but also the myogenesis-induced change in 2-DG transport affinity. Similarly, alteration in 2-DG transport affinity cannot be observed in a myogenesis-defective mutant, D1. However, under myogenesis-permissive condition, the myogenesis of this mutant is also accompanied by changes in its 2-DG transport affinity. The myotube 2-DG transport system also differs from its myoblast counterpart in its response to sulfhydryl reagents and in its turnover rate. It may be surmised from the above observations that myogenesis results in the alteration of the turnover rate or in the modification of the 2-DG transport system. Although glucose starvation has no effect on myogenesis, it is found to alter the substrate specificity and transport capacity of HAHT. In conclusion, the present study shows that hexose transport in rat myoblasts is very sensitive to the growth conditions and the stages of differentiation of the cultures. This may explain why different hexose transport properties have been observed with myoblasts grown under different conditions.

3-O-Methylglucose↗
PubMed1989

Multiple distinct intracranial tumors: association of pinealoma and craniopharyngioma. Case report.

We report the case of a patient with multiple primary brain neoplasms. A pinealoma was treated with radiotherapy and surgical resection in 1983. In 1987, a craniopharyngioma was resected surgically. Various possible determinants of multiple brain tumors have been described in the literature and are discussed. The possibility of an unknown underlying pathologic mechanism predisposing to multiple malignancies must be considered, but we are unable to conclude that this particular occurrence is more than a random chance finding.

Adult↗