Biomedical subjects

G Burns

Publications and source records attributed to G Burns.

At least 19 recordsLinked to original sources

PubMed2001

A simple turbidimetric assay designed for the routine screening as well as therapeutic monitoring of native LDL particles.

We describe the development and performance of a homogeneous assay for the direct turbidimetric determination of LDL particles in human serum. The assay is based upon the specific agglutination of LDL particles by the polyanion PAMPS. The co-agglutination of VLDL is avoided by the addition of a zwitterionic detergent. Yielding results within 10 min, the assay requires only a small sample volume taken directly from primary serum tubes, i.e., no pretreatment of the sample is necessary. It can be easily applied to routine clinical chemistry analyzers. The results are highly correlated with LDL cholesterol determinations by ultracentrifugation (r>0.95) and dextran sulfate precipitation (r>0.95), but an increased recovery of small, high density LDL particles is observed, which more adequately reflects the atherogenic risk of LDL. The assay provides excellent intra- and inter-assay CVs in the range of 0.6--1.6 and 1.7--2.4%, respectively, on Roche Diagnostics/Hitachi analyzers. The method is well suited to the high-throughput screening of LDL cholesterol levels.

Artifacts↗

PubMed2000

Glucose absorption in acute peritoneal dialysis.

OBJECTIVE: During acute peritoneal dialysis (APD), it is known that glucose found in the dialysate solution contributes to the provision of significant calories. It has been well documented in continuous ambulatory peritoneal dialysis (CAPD) that glucose absorption occurs. In APD, however, it remains unclear how much glucose absorption actually does occur. Therefore, the purpose of this study was to determine whether it is appropriate to use the formula used to calculate glucose absorption in CAPD (Grodstein et al) among patients undergoing APD. METHODS: Actual measurements of glucose absorption (Method I) were calculated in 9 patients undergoing APD treatment for >24 hours who were admitted to the intensive care unit. Glucose absorption using the Grodstein et al formula (Method II) was also determined and compared with the results of actual measurements. The data was then further analyzed based on the factors that influence glucose absorption, specifically dwell time and concentration. RESULTS: The mean total amount of glucose absorbed was 43% +/- 15%. However, when dwell time and concentration were further examined, significant differences were noted. Method I showed a cumulative increase over time. Method II showed that absorption was fixed. This suggests that with the variation in dwell time commonly seen in the acute care setting, the use of Method II may not be accurate. In each of the 2 methods, a significant difference in glucose absorption was noted when comparing the use of 1.5% and 4.25% dialysate concentrations. CONCLUSION: The established formula designed for CAPD should not be used for calculating glucose absorption in patients receiving APD because variation in dwell time and concentration should be taken into account. Because of the time constraints and staffing required to calculate each exchange individually, combined with the results of the study, we recommend the use of the percentage estimate of 40% to 50%.

Absorption↗

PubMed1999

Chronic survival of calves implanted with the DeBakey ventricular assist device.

The DeBakey ventricular assist device (VAD) is a miniaturized, electromagnetically driven axial flow pump capable of generating in excess of 10 L/min output. The VAD was evaluated in 19 calves during experiments designed to test iterative modifications in the system and to determine the safety of the DeBakey VAD for intermediate to long-term implant. Five of the animals died or were euthanized during the perioperative period (i.e., Days 1-5) due to complications associated with bleeding (n = 3), sudden cardiac arrest (n = 1), or pump occlusion due to a muscle remnant associated with coring (n = 1). The remaining 14 animals survived from 7-145 days. Ten of the 14 animals survived 30 or more days, and 2 animals survived 93 and 145 days before elective euthanasia. Pump function was evaluated in the 14 calves that survived beyond the perioperative period. Pump output at implantation averaged 3 L/min while output at 100 days (n = 2) averaged 4.22 L/min. The electrical current did not change across time during the study, indicating normal operation of the bearings. Pumps consumed less than 10.5 W of power for all support durations. Hemolysis did not occur; the average daily plasma free hemoglobin varied from 2.0 to 8.0 mg/dl. Evaluation of serum biochemical data showed that implantation of the DeBakey VAD in calves with normal hearts did not impair end organ function; BUN, creatinine, and total bilirubin varied minimally within the normal range. The white blood cell count of implanted animals remained within the normal range throughout the study.

Animals↗

PubMed1999

Management of blunt splenic trauma in patients older than 55 years. Southern Connecticut Regional Trauma Quality Assurance Committee.

BACKGROUND: Many experts have suggested that blunt splenic trauma in patients older than 55 years should not be managed by observation because of supposed increased fragility of the spleen and decreased physiologic reserve in elderly patients. We sought to determine the outcome of nonoperative management of blunt splenic trauma in patients older than 55 years. METHODS: For the years 1994 through 1996, data for patients with splenic injury older than 55 years from seven trauma centers in a single state were reviewed. RESULTS: Blunt splenic trauma occurred in 41 patients older than 55 years. Eight patients were excluded from further analysis because of death from massive associated injuries within 24 hours of admission. The remaining 33 patients (mean age, 72+/-10 years) were divided into two groups: immediate exploration (10 patients) and observation (23 patients). Observation of blunt splenic injury failed in 4 of 23 patients (17%). No patient deaths were related to the method of management of the splenic injury. CONCLUSIONS: Observation of the elderly patient with blunt splenic trauma has an acceptable failure rate of 17%.

Age Factors↗

PubMed1999

Dilatational percutaneous tracheostomy: modification of technique.

BACKGROUND: Major inherent risks associated with percutaneous dilatational tracheostomy include loss of airway during endotracheal tube manipulation, inability to cannulate the trachea below the endotracheal tube, and difficulties related to neck anatomy. METHOD: Percutaneous dilatational tracheostomy technique was modified to make the incision in the suprasternal area, and the use of air leak technique confirmed tracheal penetration below the endotracheal cuff. Bronchoscopy was not used. RESULTS: One hundred patients underwent percutaneous dilatational tracheostomy using the modification mentioned above. Although three patients had minor bleeding complications, there was no loss of airway; nor were there other complications. CONCLUSION: This technique provides improved safety from loss of airway and illuminates the need for concomitant bronchoscopy.

Dilatation↗

PubMed1998

Computed tomography in the initial evaluation of hemodynamically stable patients with blunt abdominal trauma: impact of severity of injury scale and technical factors on efficacy.

BACKGROUND: Although computed tomography is used widely in evaluating injuries from blunt abdominal trauma, grading of injuries does not reliably predict the need for intervention. Objective reporting is essential to evaluate accuracy and facilitate patient triage. We established and tested a five-point grading system for overall severity of injury. METHODS: A total of 392 computed tomographic trauma cases were objectively classified according to the type and severity of abdominal injuries, by two experienced radiologists. Interobserver variability between the original interpretation and the consensus of the film reviewers was evaluated. The computed tomographic grading system was measured against rate of admission, exploratory laparotomy, and further imaging. RESULTS: Patients with higher grades of injury on computed tomography were increasingly likely to have surgical management (odds ratio, 3.99; 95% confidence interval, 1.86-8.58; p < 0.0006), with sensitivity 100%, specificity 89.5% for level 2 injuries and higher. Although there was almost perfect agreement between the official interpretation and the reviewers' blinded consensus interpretation (raw agreement 84%, weighted kappa 0.86), indeterminate studies were reduced from 23% (16 of 67) to 12% (8 of 67) on review: these were more likely to have metallic or motion artifacts (5 of 16 vs. 1 of 51 p < 0.002). CONCLUSIONS: Standardizing reporting of injuries enhances accuracy, and grading eliminates equivocation. Diagnostic certainty in computed tomography of blunt abdominal trauma is reduced by motion and metallic artifacts.

Abdominal Injuries↗

PubMed1998

Organ failure, infection, and the systemic inflammatory response syndrome are associated with elevated levels of urinary intestinal fatty acid binding protein: study of 100 consecutive patients in a surgical intensive care unit.

BACKGROUND: Intestinal mucosal ischemia and subsequent barrier dysfunction have been related to the development of organ dysfunction and death in the critically ill. We hypothesized that urine concentrations of intestinal fatty acid binding protein (IFABP), a sensitive marker of intestinal ischemia, might predict the development of the systemic inflammatory response syndrome (SIRS) and organ dysfunction. METHODS: One hundred consecutive critically ill patients were prospectively studied for the development of infectious complications, organ dysfunction, and SIRS. Urine was collected daily for measurement of IFABP. RESULTS: A total of 58 males and 42 females (mean age, 56 years; range,16-85 years) were studied. Of these 100 patients, 40 patients developed complications and 5 patients developed SIRS. IFABP was significantly elevated in all patients with SIRS, and IFABP levels peaked an average of 1.4 days (range, 0-7 days) before the diagnosis of SIRS. CONCLUSION: Elevated concentrations of urine IFABP correlated with the clinical development of SIRS. Studies to assess the utility of IFABP as a predictor of organ dysfunction and SIRS in the critically ill are warranted.

Adolescent↗

PubMed1995

Purification of active E1 alpha 2 beta 2 of Pseudomonas putida branched-chain-oxoacid dehydrogenase.

Active E1 component of Pseudomonas putida branched-chain-oxoacid dehydrogenase was purified from P. putida strains carrying pJRS84 which contains bkdR (encoding the transcriptional activator) and bkdA1 and bkdA2 (encoding the alpha and beta subunits). Expression was inducible, however, 45-, 39- and 37-kDa proteins were produced instead of the expected 45-kDa and 37-kDa proteins. The 45-kDa protein was identified as E1 alpha and the 37-kDa and 39-kDa proteins were identified as separate translational products of bkdA2 by their N-terminal sequences. The N-terminal amino acid of the 39-kDa protein was leucine instead of methionine. The 45-, 39- and 37-kDa proteins were also produced in wild-type P.putida. Translation of bkdA1 and bkdA2 from an Escherichia coli expression plasmid produced only 45-kDa and 39-kDa proteins, with N-terminal methionine on the 39-kDa protein. The insertion of guanine residues 5' to the first ATG of bkdA2 did not affect expression of E1 beta in P. putida including the N-terminal leucine which appears to eliminate the possibility of ribosome jumping. The Z-average molecular mass of the E1 component was determined by sedimentation equilibrium to be 172 +/- 9 kDa compared to a calculated value of 166 kDa for the heterotetramer and a Stokes radius of 5.1 nm. E1 alpha Ser313, which is homologous to the phosphorylated residue of rat liver E1 alpha, was converted to alanine resulting in about a twofold increase in Km, but no change in Kcat. S315A and S319A mutations had no effect on Km or Kcat indicating that these residues do not play a major part in catalysis of E1 alpha beta 2.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗

PubMed1995

Non-metric multidimensional scaling in the analysis of neuroanatomical connection data and the organization of the primate cortical visual system.

Neuroanatomists have established that the various gross structures of the brain are divided into a large number of different processing regions and have catalogued a large number of connections between these regions. The connectional data derived from neuroanatomical studies are complex, and reliable conclusions about the organization of brain systems cannot be drawn from considering them without some supporting analysis. Recognition of this problem has recently led to the application of a variety of techniques to the analysis of connection data. One of the techniques that we previously employed, non-metric multidimensional scaling (NMDS), appears to have revealed important aspects of the organization of the central nervous system, such as the gross organization of the whole cortical network in two species. We present here a detailed treatment of methodological aspects of the application of NMDS to connection data. We first examine in detail the particular properties of neuroanatomical connection data. Second, we consider the details of NMDS and discuss the propriety of different possible NMDS approaches. Third, we present results of the analyses of connection data from the primate visual system, and discuss their interpretation. Fourth, we study independent analyses of the organization of the visual system, and examine the relation between the results of these analyses and those from NMDS. Fifth, we investigate quantitatively the performance of a number of data transformation and conditioning procedures, as well as tied and untied NMDS analysis of untransformed low-level data, to determine how well NMDS can recover known metric parameters from artificial data. We then re-analyse real connectivity data with the most successful methods at removing the effects of sparsity, to ensure that this aspect of data structure does not obscure others. Finally, we summarize the evidence on the connectional organization of the primate visual system, and discuss the reliability of NMDS analyses of neuroanatomical connection data.

Animals↗

PubMed1995

Cytokines in Crohn's colitis.

BACKGROUND: Increasing evidence points to a pathologic role for cytokines in Crohn's colitis. Levels of cytokines are increased in diseased segments of colon in Crohn's colitis, but no one has studied the concentration of cytokines in clinically and histologically nondiseased segments. METHODS: Mucosal biopsies were obtained from 7 patients with active segmental Crohn's colitis and from 7 controls without inflammatory bowel disease. The concentration of Interleukin (IL)-1 beta, IL-2, IL-6, and IL-8 in patients and controls were determined using enzyme linked immunosorbent assay and compared. Histologic sections were also performed to confirm diseased and nondiseased segments of colon. RESULTS: The concentrations of IL-1 beta, IL-6, and IL-8 were significantly higher in the involved segments of colon (10.3 +/- 4.1, 3.7 +/- 1.0, 34.4 +/- 6.9 picograms [pg] per mg) when compared to controls (1.8 +/- 0.5, 1.1 +/- 0.5, 5.3 +/- 1.0 pg/mg). The concentrations of IL-1 beta, IL-2, and IL-8 (8.5 +/- 2.9, 5.3 +/- 1.2, 26.3 +/- 8.8 pg/mg) in normal appearing segments of colon of patients with Crohn's colitis were also significantly higher than in controls, whose IL-2 level was 2.0 +/- 0.5 pg/mg. IL-1 beta and IL-8 were significantly more concentrated in both the involved and uninvolved colonic segments of patients with Crohn's colitis compared to controls. IL-2 and IL-6 were also more concentrated in Crohn's patients than in controls, but not significantly. The differences in interleukin concentrations between involved and uninvolved segments of colon in patients with segmental Crohn's colitis were not significant. CONCLUSIONS: Although Crohn's colitis is often a segmental disease, concentrations of IL-1 beta and IL-8 are increased throughout the entire colon. These observations reinforce the hypothesis that Crohn's colitis involves the whole colon even when this is not apparent clinically or histologically.

Adult↗

PubMed1995

Kringle 4 of human apolipoprotein[a] shares a linear antigenic site with human catalase.

Monoclonal antibody (mab) 1A2, directed against human apolipoprotein[a] (apo[a]), revealed a strong reaction with peroxisomes as shown by immuno-gold labeled cryosections of human liver biopsies. This reactivity was not due to the presence of apo[a] in peroxisomes but to a cross-reactivity of mab 1A2. Immunoblot analysis of peroxisomal fractions and purified human catalase demonstrated that mab 1A2 reacts with catalase. Conversely, an anti-catalase antibody also recognized apo[a]. By sequence comparison we identified a 4-amino acid motif (Y-Y-P-N) that is shared between the highly repetitive kringle 4 motif of apo[a] and the carboxy-terminal third of the peroxisomal marker enzyme catalase. No other identical sequences were identified in these proteins. Results from the following experiments indicated that 1A2 recognizes this short linear epitope. i) Mab 1A2 reacted only with the 4 amino acid peptide sequence in a pin-ELISA using immobilized overlapping peptides. ii) A synthetic peptide including this sequence completely inhibited the 1A2 immunoreactivity to apo[a] and catalase. iii) A recombinant fusion protein tagged with the putative epitope was recognized by mab 1A2. Our findings demonstrate that unknown linear epitopes in native proteins can be identified by sequence comparison between known proteins. The practical implication is that antibodies against apo[a] must be controlled for this cross-reactivity before using them for immunohistochemical studies of intracellular apo[a] in tissues or cells.

Amino Acid Sequence↗

PubMed1994

O6-methylguanine-DNA methyltransferase in normal and malignant tissue of the breast.

An important component of high-dose chemotherapy/autologous bone marrow support regimens for adjuvant treatment of breast cancer is carmustine. Preclinical studies have shown that the level of the DNA repair protein O6-methylguanine-DNA methyltransferase is correlated with the resistance of cultured human tumor cells to this drug, but little is known about transferase levels of breast tissue in vivo. We measured the DNA repair activity in 80 tissue samples from 65 patients, including normal, abnormal, benign, and malignant specimens. Wide interindividual variations was observed and average transferase levels were similar in normal and benign tissue. However, transferase levels were significantly elevated in stage I-IV disease. In addition, the frequency of samples with no detectable transferase was greatly reduced in this malignant group, and transferase was positively correlated with the presence of positive nodes, a marker for disease progression. In contrast, transferase levels were not correlated with age or estrogen receptor status, and the levels in normal tissue did not vary between patients with benign or malignant disease. These results suggest that this DNA repair activity may be increased in breast cancer relative to normal tissue and encourage further study of the predictive value of transferase measurements in high-dose chemotherapy/autologous bone marrow transplant for breast cancer.

Adult↗

PubMed1993

Construction of chromosomal recA mutants of Pseudomonas putida PpG2.

The recA gene of Pseudomonas putida PpG2 was cloned by complementation of the recA mutations of Escherichia coli strains DH5 alpha and HB101. The nucleotide sequence of the DNA fragment was determined and shown to contain recA and a downstream partial open reading frame. Two mutants of P. putida PpG2, strains JS387 and JS388, were constructed by insertional inactivation of recA with a tetracycline-resistance gene in both orientations. Both mutants acquired sensitivity to methyl methanesulfonate (MMS) and both failed to undergo homologous recombination. While the recA mutation of P. putida JS388 was complemented in trans by recA of P. putida, the JS387 mutant was difficult to transform and transformants exhibited varying degrees of sensitivity to MMS. Therefore, P. putida JS388 can be used as a carrier of recombinant plasmids, but JS387 is not a suitable host for this purpose.

Amino Acid Sequence↗

PubMed1993

Transforming growth factor beta 1-like immunoreactivity in the pituitary gland of the rat: effect of estrogen.

We have recently shown that transforming growth factor-beta 1 (TGF beta 1) is produced in the rat pituitary gland and inhibits the secretion of PRL and the estrogen-induced growth of lactotropes. In this study, we sought to ascertain whether TGF beta 1 inhibits lactotropic functions by an autocrine or paracrine mechanism. Our techniques consisted of localizing the pituitary distribution of the growth factor immunoreactivity and measuring changes in pituitary TGF beta 1 and PRL levels in the presence and absence of estrogen in ovariectomized animals. With the use of standard immunohistochemical techniques, we observed that in cyclic females and ovariectomized rats, 60 +/- 7% of the cells exhibiting TGF beta 1-like immunoreactivity in the anterior pituitary were lactotropes. In addition, the melanotropes in the intermediate lobe appeared to be TGF beta 1 immunopositive. Treatment with estrogen for 7 days reduced the number of TGF beta 1-immunoreactive lactotropes. Immunoreactive TGF beta 1 was also detectable in anterior pituitary extracts using a specific RIA. Estrogen treatment decreased the level of TGF beta 1 in the anterior pituitary extracts from ovariectomized rats. TGF beta 1 immunoreactivity was inversely proportional to the overall size of the anterior pituitary and the concentrations of PRL, as measured in both the anterior lobe extracts and plasma. These results suggest that lactotropes may serve as a site of TGF beta 1 synthesis and that the production of TGF beta 1 in these cells can be negatively influenced by PRL stimulation through the action of the lactotrope-proliferating hormone, estrogen. Furthermore, these data support the notion that TGF beta 1 controls lactotropic function by an autocrine mechanism.

Animals↗

PubMed1991