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Biomedical subjects

C Cha

Publications and source records attributed to C Cha.

13 recordsLinked to original sources

The role of external beam irradiation in patients undergoing prostate brachytherapy.

6. From this cohort, a matched-pair analysis was performed to better assess the role of EBT and TIPPB (n = 215). PSA relapse-free survival was based on the American Society for Therapeutic Radiology and Oncology Consensus Panel definition. Kaplan-Meier actuarial survival curves were compared to assess various prognostic factors. The median follow-up for all 215 matched patients was 44 months (range, 24-81) with an actuarial PSA relapse-free survival (RFS) at 5 years of 81.1%. Patients treated with EBT and TIPPB had a 5-year PSA RFS of 83.5% whereas patients treated with TIPPB only had a 5-year PSA RFS of 79.4% (p = 0.715)10 ng/ml. Risk group analysis combining PSA, Gleason score, and stage failed to identify any risk group for which the addition of EBT was significant. Analysis of postimplant dosimetry using the dose to 90% of the prostate volume (D90) failed to distinguish any difference between groups. A significant advantage for combining EBT and TIPPB could not be demonstrated in this retrospective matched-pair analysis. These data indicate that the role and rationale of combined treatment in prostate brachytherapy requires better clarification, with a prospective randomized trial.

Journal Article↗

A second T-region of the soybean-supervirulent chrysopine-type Ti plasmid pTiChry5, and construction of a fully disarmed vir helper plasmid.

Agrobacterium tumefaciens Chry5, which is particularly virulent on soybeans, induces tumors that produce a family of Amadori-type opines that includes deoxyfructosyl glutamine (Dfg) and its lactone, chrysopine (Chy). Cosmid clones mapping to the right of the known oncogenic T-region of pTiChry5 conferred Amadori opine production on tumors induced by the nopaline strain C58. Sequence analysis of DNA held in common among these cosmids identified two 25-bp, direct repeats flanking an 8.5-kb segment of pTiChry5. These probable border sequences are closely related to those of other known T-regions and define a second T-region of pTiChry5, called T-right (TR), that confers production of the Amadoriopines. The oncogenic T-left region (TL) was located precisely by identifying and sequencing the likely border repeats defining this segment. The two T-regions are separated by approximately 15 kb of plasmid DNA. Based on these results, we predicted that pKYRT1, a vir helper plasmid derived from pTiChry5, still contains all of TR and the leftmost 9 kb of TL. Consistent with this hypothesis, transgenic Arabidopsis thaliana plants selected for with a marker encoded by a binary plasmid following transformation with KYRT1 co-inherited production of the Amadori opines at high frequency. All opine-positive transgenic plants also contained TR-DNA, while those plants that lacked TR-DNA failed to produce the opines. Moreover, A. thaliana infected with KYRT1 in which an nptII gene driven by the 35S promoter of Cauliflower mosaic virus was inserted directly into the vir helper plasmid yielded kanamycin-resistant transformants at a low but detectable frequency. These results demonstrate that pKYRT1 is not disarmed, and can transfer Ti plasmid DNA to plants. A new vir helper plasmid was constructed from pTiChry5 by two rounds of sacB-mediated selection for deletion events. This plasmid, called pKPSF2, lacks both of the known T-regions and their borders. pKPSF2 failed to transfer Ti plasmid DNA to plants, but mobilized the T-region of a binary plasmid at an efficiency indistinguishable from those of pKYRT1 and the nopaline-type vir helper plasmid pMP90.

Agrobacterium tumefaciens↗

Production of acyl-homoserine lactone quorum-sensing signals by gram-negative plant-associated bacteria.

Many gram-negative bacteria regulate expression of specialized gene sets in response to population density. This regulatory mechanism, called autoinduction or quorum-sensing, is based on the production by the bacteria of a small, diffusible signal molecule called the autoinducer. In the most well-studied systems the autoinducers are N-acylated derivatives of L-homoserine lactone (acyl-HSL). Signal specificity is conferred by the length, and the nature of the substitution at C-3, of the acyl side-chain. We evaluated four acyl-HSL bioreporters, based on tra of Agrobacterium tumefaciens, lux of Vibrio fischeri, las of Pseudomonas aeruginosa, and pigment production by Chromobacterium violaceum, for their ability to detect sets of 3-oxo acyl-HSLs, 3-hydroxy acyl-HSLs, and alkanoyl-HSLs with chain lengths ranging from C4 to C12. The traG::lacZ fusion reporter from the A. tumefaciens Ti plasmid was the single most sensitive and versatile detector of the four. Using this reporter, we screened 106 isolates representing seven genera of bacteria that associate with plants. Most of the Agrobacterium, Rhizobium, and Pantoea isolates and about half of the Erwinia and Pseudomonas isolates gave positive reactions. Only a few isolates of Xanthomonas produced a detectable signal. We characterized the acyl-HSLs produced by a subset of the isolates by thin-layer chromatography. Among the pseudomonads and erwinias, most produced a single dominant activity chromatographing with the properties of N-(3-oxo-hexanoyl)-L-HSL. However, a few of the erwinias, and the P. fluorescens and Ralstonia solanacearum isolates, produced quite different signals, including 3-hydroxy forms, as well as active compounds that chromatographed with properties unlike any of our standards. The few positive xanthomonas, and almost all of the agrobacteria, produced small amounts of a compound with the chromatographic properties of N-(3-oxo-octanoyl)-L-HSL. Members of the genus Rhizobium showed the greatest diversity, with some producing as few as one and others producing as many as seven detectable signals. Several isolates produced extremely nonpolar compounds indicative of very long acyl side-chains. Production of these compounds suggests that quorum-sensing is common as a gene regulatory mechanism among gram-negative plant-associated bacteria.

4-Butyrolactone↗

Detecting and characterizing N-acyl-homoserine lactone signal molecules by thin-layer chromatography.

Many Gram-negative bacteria regulate gene expression in response to their population size by sensing the level of acyl-homoserine lactone signal molecules which they produce and liberate to the environment. We have developed an assay for these signals that couples separation by thin-layer chromatography with detection using Agrobacterium tumefaciens harboring lacZ fused to a gene that is regulated by autoinduction. With the exception of N-butanoyl-L-homoserine lactone, the reporter detected acyl-homoserine lactones with 3-oxo-, 3-hydroxy-, and 3-unsubstituted side chains of all lengths tested. The intensity of the response was proportional to the amount of the signal molecule chromatographed. Each of the 3-oxo- and the 3-unsubstituted derivatives migrated with a unique mobility. Using the assay, we showed that some bacteria produce as many as five detectable signal molecules. Structures could be assigned tentatively on the basis of mobility and spot shape. The dominant species produced by Pseudomonas syringae pv. tabaci chromatographed with the properties of N-(3-oxohexanoyl)-L-homoserine lactone, a structure that was confirmed by mass spectrometry. An isolate of Pseudomonas fluorescens produced five detectable species, three of which had novel chromatographic properties. These were identified as the 3-hydroxy- forms of N-hexanoyl-, N-octanoyl-, and N-decanoyl-L-homoserine lactone. The assay can be used to screen cultures of bacteria for acyl-homoserine lactones, for quantifying the amounts of these molecules produced, and as an analytical and preparative aid in determining the structures of these signal molecules.

4-Butyrolactone↗

Hyaluronan polymer size modulates intraocular pressure.

To investigate whether the occasional increase in intraocular pressure that may arise following injection of sodium hyaluronan into the anterior segment during intraocular surgery is related to the polymer size of hyaluronan, controlled fragmentation of hyaluronan chains in vitro was obtained using progressive incubation with testicular hyaluronidase. The profile of molecular sizes of the hyaluronan polymers in various preparations was determined using molecular sieve column chromatography. Individual preparations were injected into six rabbit eyes and intraocular pressures were measured every one-half hour for 12 hours. Longer incubations of hyaluronan with hyaluronidase resulted in more extensive degradation with accumulation of shorter chain lengths. In the rabbit, mean intraocular pressure for 12 hours following intracameral injection of hyaluronic acid (HA) is proportional to the polymer size of HA. The occasional elevation of intraocular pressure that occurs following injection of hyaluronan during ophthalmic surgery can be avoided in part by assuring the rapid fragmentation of the large molecular size hyaluronan polymer.

Animals↗

Evaluation of neuromuscular symptoms in veterans of the Persian Gulf War.

OBJECTIVE: To comprehensively evaluate complaints of muscle fatigue, weakness, and myalgias in Persian Gulf veterans (PGV). BACKGROUND: Approximately 700,000 American troops were deployed to the Persian Gulf during Desert Shield and Desert Storm. Upon return from the Gulf, some PGV developed unexplained illnesses, and special referral centers were established for the evaluation of these patients. Among the most common symptoms of these PGV are fatigue, weakness, and myalgias. An Institute of Medicine committee recommended further exploration into the possible etiologies of these complaints. METHODS: Twenty PGV with severe muscle fatigue, weakness, or myalgias that interfered with their daily activities were referred for an extensive prospective neuromuscular evaluation. Routine laboratory studies included serum creatine kinase (CK), erythrocyte sedimentation rate, thyroid function tests, and exercise forearm tests. All patients received nerve conduction studies (NCS), repetitive nerve stimulation, quantitative and single-fiber electromyography (EMG), and muscle biopsies. RESULTS: Manual muscle strength examinations were normal in all patients. Six patients had mildly elevated CKs (range 223 to 768 IU/l); otherwise, laboratory tests were unremarkable. NCS were normal except in 2 patients with carpal tunnel syndrome. Quantitative EMGs were normal. One patient had mildly increased jitter on single-fiber EMG. Muscle biopsies demonstrated minor nonspecific abnormalities in 5 patients (i.e., increased central nuclei, rare necrotic fibers, tubular aggregates). CONCLUSIONS: Despite severe subjective symptoms, most of our patients had no objective evidence of neuromuscular disease. Mildly increased CKs or nonspecific histologic abnormalities on muscle biopsy were evident in 8 patients but were not believed to be clinically significant in most. We found no evidence of a specific neuromuscular disorder in any patient. Exposures to toxins during the Persian Gulf War were not likely responsible for our patients' symptoms.

Action Potentials↗

Pelvic radiation therapy combined with hepatic artery chemotherapy for resected rectal carcinoma with liver metastases.

PURPOSE: Patients with hepatic metastases from rectal cancer treated with hepatic artery (HA) chemotherapy have a life expectancy great enough to be at risk for pelvic failure. Therefore, a treatment plan was developed for patients with resected rectal cancer and unresectable hepatic metastases, when the pathologic features of transmural invasion and perirectal lymph node metastases were present. Treatment consisted of concurrent pelvic radiation therapy (RT) and HA 5-fluorouracil (FUra), as systemic levels of FUra are achievable with HA administration, followed by HA fluorodeoxyuridine (FdUrd). METHODS AND MATERIALS: Fifteen patients were offered combined pelvic RT and HA FUra. Radiation was given to an initial dose of 45 Gy to the pelvis, followed by boost treatment for an additional 5.4-10.8 Gy. Concurrent HA chemotherapy was given using FUra or FUra/leucovorin administered in two cycles of 14 days for each cycle. If HA chemotherapy could not be done, then intravenous FUra was given during RT. Following completion of RT and HA FUra, patients were evaluated for treatment with HA FdUrd. RESULTS: Eleven patients received concurrent HA FUra or FUra/leucovorin and pelvic RT. Of these, six continued to receive HA FdUrd after completion of RT, as five patients were found to have progressive hepatic disease. Four patients could not have therapy as outlined, but did receive pelvic RT with concurrent intravenous FUra (two patients), FUra/leucovorin (one patient), or sequential HA FUra (one patient). There were four pelvic recurrences at 1, 4, 14, and 17 months after RT. One was the first site of progression, two occurred simultaneously with other failure, and one occurred after hepatic progression. The liver was the most frequent site of first progression (alone in seven patients; as a component of progression in four patients). Treatment was well tolerated with three Grade > or = 3 toxicities. The median survival was 14 months. CONCLUSIONS: These data support the hypothesis that patients with metastatic rectal cancer are also at risk for pelvic recurrence. The frequency of hepatic progression supports continued aggressive therapy directed to this site. As systemic and regional therapy of metastatic rectal cancer improves, we anticipate that more patients will be at risk for a pelvic recurrence, making it increasingly important to explore the role of pelvic radiation therapy despite the presence of metastatic disease.

Adult↗

MDM2 gene amplification in metastatic osteosarcoma.

The human homologue of the murine double minute 2 gene (MDM2), a p53-binding protein which may act as a regulator of p53 protein function, has recently been cloned. Initial studies of this gene in a variety of human tumors have shown frequent gene amplification in most types of sarcomas, including osteosarcomas. Amplification of the MDM2 gene may produce a functional inactivation of the p53 protein. To examine possible clinical or pathological correlates of MDM2 gene amplification in osteosarcoma, we studied 28 specimens on 26 patients with high grade osteosarcoma (16 primary, 11 metastatic, and 1 local recurrence) for MDM2 gene amplification by Southern blot analysis, using two MDM2 complementary DNA probes isolated by polymerase chain reaction. Four specimens (14%) showed amplification, including 3 metastases and 1 local recurrence. None of the primary osteosarcoma specimens had detectable MDM2 gene amplification. None of the specimens tested showed MDM2 gene rearrangement. In the present series, MDM2 gene amplification was detected significantly more frequently in metastatic or recurrent osteosarcomas than it was in primary osteosarcomas (P = 0.02). Our data suggest that MDM2 gene amplification may be associated with tumor progression and metastasis in osteosarcoma. Further investigation is warranted on the potential clinicopathological correlates of MDM2 gene amplification in osteosarcoma.

Adolescent↗

Glucose transport carrier of human erythrocytes. Radiation-target size of glucose-sensitive cytochalasin B binding protein.

Apparent molecular sizes of D-glucose-sensitive cytochalasin B binding protein of human erythrocyte membranes are assessed by applying classical target theory to irradiation-inactivation data. Molecular weights of this protein as it occurs in untreated ghosts, EDTA-treated ghosts, and reconstituted vesicles of Triton extract of ghosts are 220,000, 180,000, and 220,000, respectively. These results, in conjunction with other findings in the literature, suggest that the native form of the glucose transport carrier of human erythrocytes is a tetrameric assembly of a 50,000-dalton monomer or is a dimer of 100,000 daltons.

Biological Transport, Active↗

Cranial transillumination norms of the premature infant.

Ninety-five infants from 26 to 42 weeks of gestational age were studied with a simple standardized cerebral transillumination technique utilizing a Chun gun fitted with a clear flexible disc to establish normal measurements and to test the hypothesis that transillumination is useful in differentiating "catch-up" growth from hydrocephalus. One group of infants were measured at birth; the other group were infants grown to similar gestational age and serially measured. Data from these groups were found to be similar. Transillumination was not found to be a useful technique for separating "catch-up" growth infants from the normal population, but was found to be potentially useful in separating the normal and "catch-up" population from children with hydrocephalus or serious anatomic abnormalities of the central nervous system.

Brain↗

Rationale for the combination of cryoablation with surgical resection of hepatic tumors.

Only 5% to 10% of metastatic and primary liver tumors are amenable to surgical resection. Hepatic cryoablation has increased the number of patients who are suitable for curative treatment. The aim of this study was to evaluate survival and intrahepatic recurrence in patients treated with cryoablation and resection. From June 1994 to July 1999, thirty-eight surgically unresectable patients underwent a total of 42 cryoablative procedures for 65 malignant hepatic lesions. Twenty patients underwent cryoablation alone, and 18 patients were treated with a combination of resection and cryoablation, with a minimum of 18 months' follow-up. The 38 patients had the following malignancies: primary hepatocellular carcinoma (n = 8) and metastases from colorectal cancer (n = 21), neuroendocrine tumors (n = 3), ovarian cancer (n = 3), leiomyosarcoma (n = 1), testicular cancer (n = 1), and endometrial cancer (n = 1). Patients were evaluated preoperatively with spiral CT scans and intraoperatively with ultrasound examinations for lesion location and cryoprobe guidance. Local recurrence was detected by CT. Major complications included bleeding in three patients and acute renal failure, transient liver insufficiency, and postoperative pneumonia in one patient each. Two patients (5%) died during the early postoperative interval; mean hospital stay was 7.1 days. Median follow-up was 28 months (range 18 to 51 months). Overall survival according to Kaplan-Meier analysis was 82%, 65%, and 54% at 12, 24, and 48 months, respectively. Forty-eight-month survival was not significantly different between those patients undergoing cryoablation alone (64%) and those treated with a combination of resection and cryoablation (42%). Disease-free survival at 45 months was 36% for patients undergoing cryoablation plus resection compared to 25% for those undergoing cryoablation alone. Local recurrences were detected at five cryosurgical sites, for a rate of 12% overall (5 of 42), 11% (2 of 18) for patients in the cryoablation plus resection group, and 12% (3 of 24) for those in the cryoablation alone group. For patients with colorectal metastases, survival was 70% at 30 months compared to 33% for hepatocellular cancer and 66% for other types of tumors. Patients with tumors larger than 5 cm or numbering more than three did not have significantly decreased survival. Cryoablation of hepatic tumors is a safe and effective treatment for some patients not amenable to resection. The combination of cryoablation and resection results in survival comparable to that achieved with cryoablation alone.

Adult↗

Risk profiles to predict PSA relapse-free survival for patients undergoing permanent prostate brachytherapy.

PURPOSE: Combined risk profiles are helpful in assessing the likelihood of prostate-specific antigen relapse-free survival (PSA-RFS) for patients with localized prostate cancer. This retrospective analysis reports the 5-year biochemical outcome of a large cohort of patients according to prognostic factors and risk profiles for patients undergoing ultrasound-guided transperineal interstitial permanent prostate brachytherapy (TIPPB). PATIENTS AND METHODS: Seven hundred seventeen consecutive patients with clinically localized prostate cancer treated between June 1992 and November 1997 underwent TIPPB. Palladium-103 (103Pd) (n = 539) or iodine-125 (125I) (n = 178) sources were utilized to a prescribed dose of 120 Gy and 144 Gy, respectively. One hundred eleven patients received combined external-beam irradiation (41.4 or 45 Gy) and TIPPB (103Pd, 90 Gy, and 125I, 108 Gy). One hundred twelve patients received androgen ablation therapy in advance of TIPPB. Patients were grouped into risk profiles based on pretreatment factors that consisted of PSA < or = 10 and Gleason score < or = 6. Patients meeting both criteria were classified as favorable (n = 334). Those not fulfilling one criteria were classified as intermediate risk (n = 261), and those not fulfilling two criteria were classified as unfavorable risk (n = 261). PSA-RFS was calculated based on the American Society of Therapeutic Radiology and Oncology consensus conference definition of PSA failure. The median follow-up was 41 months (range, 14-82 months). RESULTS: The actuarial PSA-RFS survival at 5 years was 82%. Multivariate Cox regression analysis identified PSA and Gleason score as highly significant factors predicting for biochemical outcome. Hazard risk ratios using a nadir definition of the PSA as < or = 1.0 ng/mL and < or = 0.5 ng/mL were each significant predictors of outcome with the 0.5 ng/mL nadir level only slightly better (risk ratio for nadir < or = 1.0 = 2.78; risk ratio for nadir < or = 0.5 = 3.57). Favorable-risk patients had a 5-year actuarial PSA-RFS of 93%, whereas intermediate-risk and unfavorable-risk groups had 5-year PSA-RFSs of 77% and 62%, respectively. Four hundred ninety-three patients underwent brachytherapy without external-beam irradiation or androgen ablation. From this group, the 5-year PSA-RFSs for the favorable-risk, intermediate-risk, and unfavorable-risk patients were 92%, 74%, and 55%, respectively. DISCUSSION: This retrospective study examining TIPPB demonstrated excellent 5-year actuarial PSA-RFS rates. The assignment of risk profiles based on the results of multivariate analysis of prognostic factors identifies the expected PSA-RFS for patients undergoing TIPPB.

Adenocarcinoma↗