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Biomedical subjects

C C Wang

Publications and source records attributed to C C Wang.

At least 289 records · Page 16Linked to original sources

Collagen synthesis in idiopathic and dilantin-induced gingival fibromatosis.

Gingival fibromatosis is an uncommon but benign oral disease which causes progressive and non-hemorrhagic enlargement of the gingiva. There are two types of gingival fibromatosis: idiopathic gingival fibromatosis (GF), which is of unknown cause and may be associated with hereditary factors, and drug-induced gingival fibromatosis which is caused primarily by phenytoin intake. In cases of gingival fibromatosis, either the teeth are delayed in emergence or most of the crowns are embedded in the soft tissue even after full eruption. The objective of this study was to examine the basis of excess collagen formation in the two types of gingival fibromatosis in four patients admitted to the dental clinic at Kaohsiung Medical College Hospital, Kaohsiung, Taiwan. There were two male patients, aged ten (Case 1) and sixteen (Case 2), with idiopathic gingival fibromatosis and hypertrichosis, and two female patients, aged sixteen (Case 3) and eleven (Case 4), with Dilantin-induced gingival hyperplasia (DGH). Cultures of gingival fibroblast were established either from clinically excised hyperplastic tissues or from pre-orthodontic surgical normal gingiva. The synthesis of collagen and levels of prolyl hydroxylase, a key enzyme in collagen synthesis, were examined in the healthy and affected fibroblasts. Our studies showed that after two days in culture, fibroblasts from all four patients multiplied faster than healthy gingival fibroblasts, though the amount of DNA and protein per cell remained unchanged. In addition, all cultures (except Case 1) had a 2- to 3- fold increase of prolyl hydroxylase activity over that of the controls. As in the cases of prolyl hydroxylase activity, Case 1 did not show any change in collagen synthesis when compared to the control. However, Cases 2, 3, and 4 showed appreciable collagen increases in the cell and medium: 61% and 60% for Case 2; 16% and 36% for Case 3; and 21% and 80.7% for Case 4 respectively.

Adolescent↗

Liposomes-coated hydroxyapatite and tricalcium phosphate implanted in the mandibular bony defect of miniature swine.

Hydroxyapatite and tricalcium phosphate have been used as bone implants for some period of time. Now unfortunately, these materials have failed to become the nucleation sites for bone regeneration. We hypothesized that coating hydroxyapatite and tricalcium phosphate with negatively charged liposomes may improve the nucleation process for new bone formation. The present study was designed to test this hypothesis. Experiments were carried out in 15 miniature swines' mandibular angle with artificial bony defects. In each of the swine, the bony defects on one side were implanted with either liposomes coated with hydroxyapatite or liposomes coated with tricalcium phosphate, while the other side served as control. At the end of the third and sixth weeks following the operation, we observed result, took histology and radiographs of the operated area. The results showed that liposomes-coated materials were biocompatible and their clinical endpoint was enhanced. At the end of the third week, the implant material was surrounded by dense connective tissues. At the end of the sixth week, there were new bone formations near the implanted material. In addition, liposomes which were immobilized in agarose gel and implanted in the defects showed new bony bridge formation. These observations suggest that liposomes have the ability in promoting repair of osseous deficiencies.

Alveolar Process↗

[Multiple myeloma with oral manifestations--report of two cases].

Multiple myeloma is a neoplastic disorder of bone that originates from cells of bone marrow. It is most commonly seen within the age range of 50-80 years, cases under the age of 40 being rare. Men are affected more frequently than are women. Bone pain is the cardinal clinical symptom in multiple myeloma. Because of the destruction of bone, pathologic fracture is fairly common. In the oral manifestations, the mandible is involved far more frequently than the maxilla, especially the most active hematopoietic areas-the remus, angle and molar region of the mandible. Other signs and symptoms of jaw involvement include swelling, pain, and increased tooth mobility. Extraosseous lesions may result in paresthesia of soft tissue and gingival enlargement with bleeding tendency. Roentgenographic examination will usually reveal numerous punched-out lesions in a variety of bones. In addition, blood examination will reveal hyperglobulinemia and Bence-Jones protein may be present in the urine of myeloma patients. The histological features of myeloma are closely packed cells resembling plasma cells. Case 1 in this report is a 64-year-old female, who has been diagnosed as having multiple myeloma (IgG, lambda). She was referred to our hospital because of gingival swelling, bleeding and pain. Case 2 is a 60-year-old female suffering from spontaneous gingival bleeding. After blood, urine examination and bone marrow biopsy, multiple myeloma was diagnosed (IgG, lambda). This paper reports the clinical manifestations and treatment courses of these two cases, and the concerns of treatment of multiple myeloma are also discussed.

Female↗

Validating targets for antiparasite chemotherapy.

The enzymes and receptors in parasites that can be qualified as targets for antiparasite chemotherapy should perform essential functions in the parasites and demonstrate some feasibility for selective inhibition. They can be tentatively identified through detailed analysis of various aspects of metabolisms in the parasites or elucidation of the mechanisms of action among proven antiparasitic agents. Preliminary verifications of these putative targets can be indicated by in vitro antiparasite activity of an inhibitor of the target. However, before a major long-term effort to pursue in-depth structure-activity analysis of the target is to be committed for specific inhibitor design, further validations of the target are essential to insure that future studies are not misguided. One old-fashioned approach to validate a target in the pharmaceutical industry is by correlating target inhibitions with antiparasitic activities among large numbers of drug derivatives. The results are often indicative but hardly ever conclusive. Another method is by comparing the putative drug targets between the drug-sensitive and the drug-resistant parasites for potential discrepancies. Unfortunately, the latter often result from indirect causes, such as reduced drug transport, instead of an alteration of the drug target itself. The third experimental approach is by disrupting the gene encoding the putative target in parasite, which can provide the most conclusive evidence on whether the target plays an indispensible role in the parasite. But special conditions are needed for the gene knockout mutants to survive to exhibit their phenotypes and to allow genetic complementation studies for further verifications. Furthermore, gene knockout experiments are often difficult to perform on cells of multiple ploidy or genes of multiple copies, and are currently applicable only to a limited number of protozoan parasites. In the current article I have tried to take a cursory look at some eleven putative drug targets among various parasites, each supported by well-established antiparasitic agents identified as its inhibitors. I have also considered the evidence for validity of each of them and the potential means of further verifying their validity.

Animals↗

[The shear bond strength of porcelain and base metal alloys for metal-ceramic crown (VI)].

The popularity of base metal alloy for porcelain fused to a metal crown and bridges has increased recently because of lower price, superior yield strength and modulus of elasticity (rigidity). The use of these alloys give them the potential advantage of thinner coping with less material and the required rigidity for long-span fixed partial dentures. The addition of beryllium to base metal alloys increases fluidity and improves casting fit. Beryllium also controls surface oxidation and bonding strength. Oxidation heat treatment of the metal is used to remove entrapped air, eliminate organic material and form the metal oxidized layer. The bonding agent and gold bonding agent also affect the bonding strength between porcelain and metal alloys. Four commercially available ceramic base alloys (two alloys contain beryllium element, another two do not) are studied. The purposes of this investigation are to test the shear bond strength between porcelain and metal alloy under different conditions (oxidized layer removed or not, gold bonding agent and uniseal bonding agent used in combination, separately or not at all). The following results were obtained: 1. Whether the oxidized layer was removed or not did not have a significant effect on the bond strength of porcelain (p > 0.05). 2. When different metal alloys was used, Wiron 88 alloy showed significantly the best bond strength of all alloys tested (p < 0.05). 3. When different bonding agent were used, the combination of bonding agent and gold bonding agent showed the best bond strength of all bonding agents tested. It showed a significantly higher bonding strength than opaque layer and gold bonding agent. 4. Vita porcelain powder showed a significantly higher bond strength than did Unibond porcelain (p < 0.05). 5. When opaque layer was used and when no bonding agent was used, the porcelain powder significantly affected the bond strength (p < 0.0001). The Vita porcelain showed a significantly higher bond strength than did Unibond porcelain. Wiron 88 alloy showed significantly the high bond strength than Rexillium III alloy (p < 0.05). 6. When bonding agent was used, the porcelain powder and alloy significantly affected bond strength. The Vita porcelian showed a significantly higher bond strength than did Unibond porcelain. Wiron 88 alloy showed significantly the best bond strength than other alloys Wiron (p < 0.05). 7. When bonding agent and gold bonding agent were used, the porcelain powder significantly affected the bond strength (p < 0.05). The Vita porcelain showed a significantly higher bond strength than did Unibond porcelain (p < 0.05). 8. When gold bonding agent was used, there was not any bond strength between metal and powder.

Crowns↗

Stable coexpression of a drug-resistance gene and a heterologous gene in an ancient parasitic protozoan Giardia lamblia.

Manipulation of gene expression in Giardia lamblia, one of the most ancient eukaryotes, may provide insights into the evolutionary transition from prokaryotes to eukaryotes. Two recent successes in transient expression of the firefly luciferase (luc) gene in G. lamblia were mediated by a 5'-untranslated region (UTR) of the Giardia glutamate dehydrogenase (gdh) gene and a giardiavirus (GLV) genomic transcript, respectively. We now report a stable coexpression of luc gene with a neomycin phosphotransferase (neo(r)) gene in G. lamblia. An in vitro transcript of the construct pC670-Neo; containing the neo(r) encoding region flanked with the 5'670 nucleotides (nt) and the 3'2022 nt portion of GLV positive strand RNA, was electroporated into G. lamblia trophozoites that were infected with GLV. G418-resistant Giardia trophozoites were cloned, and the neo(r) mRNA in these clones was found to increase with increasing G418 pressure. This drug resistance remained stable upon continuous in vitro cultivation in the absence of G418 for over 15 days. Another plasmid pNeo/GDH/Luc, was constructed by inserting luc gene downstream from the neo(r) gene and the 193 nt 5' portion of gdh gene in pC670-Neo, and its bicistronic in vitro transcript was introduced into GLV-infected G. lamblia by electroporation. The transfectants demonstrated G418-resistance and persistent luciferase activity at levels parallel to the amount of G418 used for selection, peaking at a level of several thousand-fold above the background. Taken together, these data indicate that the neo(r) gene provides an effective selection marker for transformation of Giardia trophozoites, and the bicistronic RNA transfection vector may open the way for functional analysis of other genes in Giardia.

Amebicides↗

The role of brachytherapy in early-stage nasopharyngeal carcinoma.

PURPOSE: To present the treatment results and assess the optimal radiation dose and the role of brachytherapy in early stage nasopharyngeal cancer (NPC). METHODS AND MATERIALS: One hundred eighty-three patients with Stage I and II (American Joint Committee on Cancer Staging System, 1987) NPC completed the planned radiotherapy in our institution from 1979 to 1991. In 133 patients, radiotherapy was given to the nasopharynx by external beam to 64.8-68.4 Gy. Further boost was done by high dose rate (HDR) brachytherapy for 5-16.5 Gy in one to three fractions. For the remaining 50 patients, a course of external radiotherapy to the nasopharynx for 68.4-72 Gy was given to nasopharynx. Age (>40 or not), sex, neck boost or not, brachytherapy, and irradiation dose were analyzed to determine significant factors that influence the probabilities of local control and actuarial survival. RESULTS: The 5-year disease-specific survival was 85.8% and local control was 83%. Only the brachytherapy and irradiation dose significantly affected the results. The use of the brachytherapy had significant impact on overall survival and local control. Furthermore, we compared the prognostic effect of various radiation dosage among Group I of 50 patients (<72.5 Gy, no brachytherapy, excluding four patients who received brachytherapy), Group II of 71 patients (72.5-75 Gy; one to two fractions of brachytherapy), and Group III of 58 patients (>75 Gy; three fractions of brachytherapy). Five-year disease-specific survival rates of Group I, Group II, and Group III were 77, 95.5, and 82.4%, respectively. Five-year local control rates were: 73.7, 93.9, and 79.5%. We found that the Group II had the best actuarial survival and local control rate (log-rank test,p < 0.05). Most patients receiving brachytherapy encountered foul odor because of nasopharynx crust; 12 of them had palate or sphenoid sinus floor perforation or nasopharynx necrosis. None of the patients without brachytherapy experienced the same complications. CONCLUSIONS: The optimal radiotherapy dose to the nasopharynx area in early stage NPC may be within 72.5 to 75 Gy by our treatment protocol. A dose of more than 75 Gy did not have significant local control or survival advantage. The use of brachytherapy to elevate radiation dose had significant local control and survival benefit for early stage NPC patients, but the fractionation size should be decreased to reduce the complications.

Adult↗

Prevention of graft-versus-host disease by intrathymic injection of recipient-type splenocytes into donor.

We have prevented graft-versus-host disease (GVHD) by tolerizing graft donors to host antigens by intrathymic injection of recipient-type splenocytes into donors. A unidirectional GVHD model was used in which intravenous injection of 3-4 x 10(8) Lewis rat (donor) lymphocytes into (Lewis x Brown Norway)F1 rats (recipients) causes lethal GVHD. The donor animals were divided into five treatment groups. The group 1 donor animals received no treatment. The group 2 donors received a single intraperitoneal injection of 1 ml of antilymphocyte antiserum (ALS). The group 3 donors received an intrathymic injection of 50x10(6) host splenocytes. The group 4 donors received both ALS (intraperitoneally) and intrathymic allograft. The group 5 donors received both ALS (intraperitoneally) and intravenous allograft. Two weeks after these treatments, 3-4x10(8) lymphocytes from each of these donors were injected (intravenously) into the recipients. The clinical signs of GVHD, as measured by profound weight loss, hair loss, inflammation of foot pads and ears, and profound splenomegaly, were evident in recipients of groups 1, 2, and 3 between days 9 and 10 and in the recipients (two of four) of group 5 on day 17. No GVHD was observed by histopathology in all 14 hosts that received lymphocyte injection from the group 4 donor animals (up to 300 days). These results demonstrate that GVHD can be eliminated by tolerizing donors toward host by intrathymic injection of the recipient-type lymphocytes into the donor. A single injection of ALS is necessary to possibly eliminate antihost response from the donor for the tolerance induction. The thymic route appears to be superior to the intravenous route for tolerance induction.

Adoptive Transfer↗

Prolonged circulatory arrest in moderate hypothermia with retrograde cerebral perfusion. Is brain ischemic?

BACKGROUND: Circulatory arrest (CA), which provides a bloodless field and good visualization without the need of aortic cross-clamp, is commonly used to facilitate repair of aortic lesions. However, extended periods of CA may impair cerebral metabolism and cause ischemic injury. Studies were performed to evaluate the efficacy of retrograde cerebral perfusion (RCP) in protecting the brain from ischemic injury during a prolonged period of CA in moderate hypothermia. METHODS AND RESULTS: Twenty-three patients (18 men and 5 women) were operated on for aortic lesions (17 acute type A aortic dissection and 6 chronic type A aortic dissection with Marfan's syndrome). The aortic operations were performed with CA (58 to 104 minutes; mean +/- SD, 75 +/- 12 minutes) at a rectal temperature of 23.3 +/- 0.5 degrees C (21 degrees C to 25 degrees C). For RCP, cold (14 degrees C to 18 degrees C) oxygenated blood (300 mL/min) was pumped to the superior vena cava with internal jugular venous pressure of 15 +/- 5 mm Hg. The cardiopulmonary bypass time was 157 +/- 18 minutes. Cortical blood flow during RCP detected by subdural laser Doppler probe was 10 +/- 5% of baseline. Percent oxygen extraction and pyruvate and lactate levels (26 +/- 2% and 0.43 +/- 0.17 and 45 +/- 16 mg/dI) were insignificantly different from those before CA (28 +/- 3% and 0.71 +/- 0.08 and 62 +/- 20 mg/dL, P > .05). Creatine kinase-BB isoenzyme was undetectable. All but 1 patient survived the operation (95.5%) and woke up without neurological deficit. Follow-up (mean, 13 months) was complete in all survivors. There were no late deaths. Cerebral functional studies performed 3 months after discharge showed results insignificantly different from those of the normal control subjects. CONCLUSIONS: There is no evidence of ischemia of the brain during prolonged moderate hypothermic CA with the aid of RCP. Retrograde cerebral perfusion effectively extends the safe time of CA. Deep hypothermia during CA seems unnecessary.

Adult↗

Treatment of hypercholesterolemia in the Watanabe rabbit using allogeneic hepatocellular transplantation under a regeneration stimulus.

Numerous studies have reported successful allotransplantation of hepatocytes. However, none have shown long-term correction of a liver-related metabolic defect. In this study, we used a method of regional hepatocyte transplantation and subsequent induction of transplanted cell proliferation by regeneration response in the transplant-bearing liver lobes. New Zealand White rabbits were used as cell donors and Watanabe heritable hyperlipidemic (WHHL) rabbits were used as cell recipients (2 x 10(8) cells/rabbit). All recipient rabbits were maintained on daily cyclosporine. Two weeks after baseline serum cholesterol determination, group I WHHL rabbits (n = 7) received an infusion of cells into the right lateral liver lobe, and a loose ligature was placed around the portal venous branch supplying the anterior lobe. After 1 week, to allow engraftment, the portal venous branch was ligated, which resulted in the atrophy of the affected liver parenchyma and induction of hyperplasia in the transplant-bearing liver tissue. Group II rabbits (n = 6) were transplanted with New Zealand White hepatocytes without portal branch ligation (PBL) and group III rabbits (n = 4) were subjected to sham transplantation (saline) and PBL. The experimental period extended to 150 days after transplantation. All WHHL rabbits transplanted with normal hepatocytes showed reduction in serum cholesterol and low-density lipoprotein (LDL) levels. Group I (PBL-stimulated) recipients demonstrated a more pronounced and sustained effect than group II animals (P < 0.05). Group III controls showed only a slight, typical for aging decrease in serum cholesterol. Group I recipient livers perfused with LDL labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl indocarbocyanine perchlorate (DiI) showed much higher numbers of DiI-LDL-positive hepatocytes than those of group II recipients. In conclusion, a liver regeneration stimulus enhanced the population of transplanted hepatocytes and their functional effect in a large animal model of inborn error of liver metabolism.

Alanine Transaminase↗

Probing the active site of Tritrichomonas foetus hypoxanthine-guanine-xanthine phosphoribosyltransferase using covalent modification of cysteine residues.

The hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRTase) of Tritrichomonas foetus was inactivated by the thiol reagents iodoacetate and 5,5'-dithiobis(2-nitrobenzoic acid) (Nbs2). Iodoacetate inactivates the enzyme in a time-dependent and concentration-dependent manner that follows pseudo-first-order kinetics. However, the observation that total inactivation with iodoacetate was not achieved suggests that none of the reactive cysteine residues is directly involved in the catalytic activity of the enzyme. Nbs2 caused 50% inactivation rapidly, which was followed by gradual modifications of an additional three cysteine residues leading to complete enzyme inactivation. Analysis of the inactivation using the method developed by Tsou (1962) revealed that modification of two cysteine residues by Nbs2 is sufficient to impair the HGXPRTase activity. Tryptic digestion of HGXPRTase labeled with iodo[2-14C]acetic acid, followed by fractionation of the digest by HPLC and sequence analysis of the labeled peptides allowed the identification of Cys71, Cys129, Cys132, and Cys148 as the reactive cysteine residues. GMP and 5-phosphoribosyl-1-diphosphate provided complete protection against HGXPRTase inactivation by iodoacetate and against carboxymethylation of Cys129, Cys132, and Cys148, Cys71 was not protected by either substrate against iodoacetate, but its carboxymethylation caused no loss in enzyme activity either. There was also no substrate protection of Cys71 against Nbs2, which, however, caused 50% inactivation of the enzyme. Replacing the thionitrobenzoate (Nbs) moiety from Cys71 with cyanide resulted in a gradual recovery of the enzyme activity, which indicates that a steric hindrance at the active site was introduced by Nbs but removed by cyanide. Thus, our results demonstrate that although the reactive cysteine residues in HGXPRTase are not directly involved in the catalytic activity, modification of cysteine residues 129, 132, and 148 by iodoacetate or Nbs2 hinders substrate binding which can, in turn, protect the cysteine residues from modifications. The substrate protection of Cys129 and Cys148 is probably also indicative of a conformational change in the protein structure brought about by substrate binding.

Amino Acid Sequence↗

Expression of Epstein-Barr virus-encoded RNAs as a marker for metastatic undifferentiated nasopharyngeal carcinoma.

BACKGROUND: Epstein-Barr virus (EBV) is associated with undifferentiated nasopharyngeal carcinoma (NPC). EBV-encoded nonpolyadenlyated RNAs (EBERs) are often used as a marker to detect EBV-infected NPC cells. This study was conducted to document the expression and determine the significance of EBERs in NPC cells at various metastatic sites. METHODS: An in situ hybridization (ISH) technique was used to identify the presence of EBERs in paraffin embedded tissues of primary and metastatic sites obtained from 21 patients with NPC. Nineteen of these patients had undifferentiated lesions, and 2 had squamous cell carcinoma. One hundred and fifty specimens of normal tissues and tissues from patients with a variety of benign and malignant diseases other than NPC served as controls. In the NPC specimens, the expression of latent membrane protein (LMP) and a lytic protein, BZLF-1, were also examined by immunohistochemistry. RESULTS: Tissues from all patients with undifferentiated NPC and one patient with squamous cell carcinoma contained EBERs in the malignant cells; the other case of squamous cell carcinoma was negative. In metastatic NPCs, LMP was expressed in 18% (4 of 22) of tissues whereas BZLF-1 was not expressed in any tissues. EBERs were not detected in the 43 patients with normal tissues and benign lesions. In malignant diseases other than NPC, EBERs were detected in only 2 of 12 cases of non-Hodgkin's lymphoma, 1 of 2 cases of Hodgkin's lymphoma, and 1 of 6 cases of gastric cancer. CONCLUSIONS: By virtue of the direct correlation between latent EBV infection and NPC, the authors conclude that EBERs can be used as a sensitive marker to identify NPC cells at various metastatic sites by techniques of in situ hybridization, and that demonstration of EBERs in lesions of undifferentiated histology may be useful as a diagnostic adjunct for NPC presenting as metastatic cancer of unknown origin.

Biomarkers, Tumor↗

Crystal structure of the hypoxanthine-guanine-xanthine phosphoribosyltransferase from the protozoan parasite Tritrichomonas foetus.

The crystal structure of the hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRTase) from Tritrichomonas foetus has been determined and refined against X-ray data to 1.9 A resolution. T. foetus HGXPRTase crystallizes as an asymmetric dimer, with GMP bound to only one of the two molecules that form the asymmetric unit. Each molecule of HGXPRTase is formed by two lobes joined by a short "hinge" region, and the GMP binds in a cavity between the two lobes. A comparison of the two molecules in the asymmetric unit shows that the hinge region is flexible and that ligand binding affects the relative positions of the two lobes. The binding of GMP brings the two lobes closer together, rotating one lobe by about 5 degrees relative to the other. T. foetus appears to depend on HGXPRTase for its supply of GMP, making this enzyme a target for antiparasite drug design. A comparison of the structures of T. foetus HGXPRTase and human HGPRTase reveals that, while these enzymes retain a similar polypeptide fold, there are substantial differences between the active sites of these two homologs. These differences suggest that it will be possible to find compounds that selectively inhibit the parasite enzyme.

Amino Acid Sequence↗

Local control of T3 carcinomas after accelerated fractionation: a look at the "gap".

PURPOSE: To study the effects of midcourse treatment break or gaps related to the local control of T3 carcinoma of the oropharynx and larynx following accelerated hyperfractionated radiation therapy. METHODS AND MATERIALS: All patients were treated at the Massachusetts General Hospital from 1979 through 1994 with treatment consisting of 1.6 Gy per traction, two fractions a day for the treatment of T3 carcinoma of the oropharynx and larynx. They were entered in the head and neck data base. Their treatment dates, treatment breaks, and doses vs. local control were analyzed and compared. A p-value of 0.05 was considered statistically significant. RESULTS: A total of 162 patients were available for review. Due to the acute severe mucosal effects, most of the patients required a midcourse pause or "break" after a dose of 38.4-48 Gy before treatment could be resumed and completed. The data indicate that (a) prolongation of the treatment gap for more than 14 days, (b) total treatment course longer than 45 days, (c) total dose less than 67 Gy, and (d) male gender adversely affected local control. In spite of the gaps, the female patients with advanced carcinomas enjoyed the benefits of improved local control after the accelerated hyperfractionated radiation therapy. CONCLUSIONS: Accelerated hyperfractionation radiation therapy using 1.6 Gy per fraction/twice-a-day (b.i.d.) for a total dose of 70.4 Gy in 6 weeks is effective in achieving high local control of T3 squamous cell carcinoma of the oropharynx and larynx. The midcourse treatment gap should be as short as possible with the projected total dose and time. Should the gaps be unduly prolonged due to various circumstances, further increase in the total dose, for example, 72-75 Gy, and/or increase of the fraction sizes, for example, 1.8-2.0 Gy/f b.i.d. after the gap may be necessary to compensate for the adverse effects of the tumor regeneration from the prolonged gap.

Adolescent↗

Localization of a putative second membrane association site in penicillin-binding protein 1B of Escherichia coli.

We have shown previously that the periplasmic domain of penicillin-binding protein 1B (PBP 1Bper; residues 90-844) from Escherichia coli is insoluble in the absence of detergents, and can be reconstituted into liposomes [Nicholas, Lamson and Schultz (1993) J. Biol. Chem. 268, 5632-5641]. These data suggested that native PBP 1B contains a membrane association site in addition to its N-terminal transmembrane anchor. We have studied the membrane topology of PBP 1B in greater detail by assessing detergent binding and solubility in the absence of detergents for PBP 1Bper and a set of proteolytic fragments of PBP 1B. PBP 1Bper was shown by three independent methods to bind to detergent micelles, which strongly suggests that the periplasmic domain interacts with the hydrophobic milieu of membrane bilayers. Digestion with high weight ratios of thrombin of purified PBP 1B containing an engineered thrombin cleavage site on the periplasmic side of the transmembrane anchor generated four fragments in addition to PBP 1Bper that varied in size from 71 to 48 kDa. In contrast to PBP 1Bper, all fragments of 67 kDa and smaller were eluted from a gel-filtration column in the absence of detergents and did not bind to detergent micelles. The N-terminal sequences of the four fragments were determined, allowing the cleavage sites to be located in the primary sequence of PBP 1B. These data localize the membrane association site of PBP 1B to a region comprising the first 163 amino acids of the periplasmic domain, which falls within the putative transglycosylase domain. Lipid modification does not appear to be the mechanism by which PBP 1Bper associates with membranes.

Amino Acid Sequence↗

Radiofrequency ablation for supraventricular and ventricular tachycardia in young patients.

Radiofrequency ablation therapy was conducted in 86 consecutive children and young patients with a mean age of 14 +/- 3 years (range = 3-18). Fifty-two patients had Wolff-Parkinson-White syndrome, one had re-entry tachycardia incorporating a nodoventricular fiber, 22 had atrioventricular node re-entry tachycardia, two had atrial tachycardia and nine had idiopathic ventricular tachycardia. Radiofrequency ablation was successful in 50 of the 52 patients (96%) with Wolff-Parkinson-White syndrome and the one with nodoventricular fiber. Radiofrequency modification of the atrioventricular node using the inferior approach was successful in eliminating atrioventricular node re-entry tachycardia in 20 of the 22 patients (91%). Radiofrequency ablation in the two patients with atrial tachycardia was unsuccessful. Of the nine patients with idiopathic ventricular tachycardia, eight from the left ventricle and one from the right ventricular outflow tract, eight were successfully ablated (88%). Follow-up over a period ranging from 1 to 46 months (21 +/- 13) revealed a recurrence of tachycardia in seven patients; a late electrophysiological study in 38 patients revealed the induction of tachycardia in 11 patients (seven with accessory pathway-mediated tachycardia, three with atrioventricular node re-entry tachycardia and one with idiopathic ventricular tachycardia). All 11 patients were successfully ablated by a second trial. In conclusion, radiofrequency ablation therapy is effective and safe in pediatric patients with supraventricular and ventricular tachycardia and should be considered as the therapy of choice in this group of patients.

Adolescent↗

Measurement of adenosine by capillary zone electrophoresis with on-column isotachophoretic preconcentration.

An on-column isotachophoretic (ITP)-capillary electrophoresis (CE) system capable of preconcentrating polyhydroxyl species is reported. The ITP-CE system utilizes borate complexation of the neutral diol species to form anionic compounds that can be directly separated by CE. Borate buffer functions as both the terminating electrolyte for the ITP preconcentration and the operating buffer for the subsequent CE separation. Isotachophoretic preconcentration allows injection volumes as large as 50% of the column volume, without compromising separation integrity, to yield detection limits about 70-fold lower than direct CE separation (with borate operating buffer). In this paper we also present an application of the ITP-CE system, with laser-induced fluorescence (LIF) detection, to the quantitative analysis of adenosine from urine. Nanomolar concentration levels of adenosine are successfully derivatized with chloroacetaldehyde (CAA) to form a fluorescent derivative whose spectral characteristics match the He-Cd laser. The technique is shown to be capable of quantitative measurement of adenosine as low as 10(-9) M, the levels expected in plasma and urine.

Acetaldehyde↗