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Biomedical subjects

W Campbell

Publications and source records attributed to W Campbell.

At least 37 records · Page 2Linked to original sources

Diagnosis of Jamestown Canyon encephalitis by polymerase chain reaction.

In recent years, polymerase chain reaction (PCR) has been under study as a potential technique to improve the accuracy of diagnosis of suspected central nervous system viral infections. We describe a case of severe encephalitis in a previously healthy 20-year-old woman from New York who presented with headache, fever, and photophobia. Her illness was characterized by progressive worsening of her neurological status, leading to confusion, delirium, and status epilepticus. The diagnosis of Jamestown Canyon encephalitis was established by positive reverse transcriptase (RT)-PCR and nucleic acid sequencing of the band from both cerebrospinal fluid and brain tissue. The nucleotide sequence and the deduced amino acid sequence of the Jamestown Canyon virus from this patient were very similar to Jamestown Canyon virus isolates from mosquito pools in New York. This report suggests that RT-PCR assays could be important tools in the diagnostic workup of cases of encephalitis.

Adult↗

Head-injured patients who are nasal carriers of Staphylococcus aureus are at high risk for Staphylococcus aureus pneumonia.

OBJECTIVE: To determine if head-injured patients with premorbid nasal colonization with Staphylococcus aureus are at increased risk for S. aureus infection. DESIGN: Patients admitted over a 2-yr period were enrolled if they met the following criteria: Injury Severity Score > or = 9, intensive care unit (ICU) admission, hospitalization in another hospital < 24 hrs, no recent use of antibiotics. SETTING: Acute care trauma facility. PATIENTS: Any patient sustaining acute, blunt, or penetrating injury and meeting the enrollement criteria were eligible. INTERVENTIONS: Swab cultures of both internal nares were performed within 72 hrs of readmission and cultured for S. Aureus. Patients were prospectively monitored for S. Aureus infections until discharge. MEASUREMENTS AND MAIN RESULTS: Admission nasal cultures were positive (NC+) for S. aureus in 144 of the 776 patients cultured. Forty of the 144 NC+ patients had isolated head (37) or high cervical spine (3) injury, and 11 of that group (27.5%) developed S. aureus infections. The remaining 104 patients positive for S. aureus on admission had no head injury (74) or head combined with torso and extremity injuries (30). S. aureus infection was diagnosed in 11 of the 104 patients (10.6%). The difference in incidence of infections is significant (p <.01), as is the difference in incidence of pneumonia (20% vs. 3.8%, respectively [p <.01]). Organisms causing pneumonia were often the same organisms isolated from the nares on admission. CONCLUSIONS: Nasal colonization with S. aureus at the time of severe head injury increases the risk of S. aureus pneumonia during hospitalization. Prophylactic measures against S. aureus pneumonia may help reduce the length and cost of hospitalization.

Adult↗

Arginine carboxypeptidase (CPR) in human plasma determined with sandwich ELISA.

There are two types of carboxypeptidases present in human blood, carboxypeptidase N (CPN) and arginine carboxypeptidase (CPR). CPR is generated during coagulation from a precursor (proCPR) which can be converted to the active form by trypsin in vitro. Since it is difficult to distinguish the two types of carboxypeptidases in human blood by the measurement of enzyme activity, we established a quantitative sandwich ELISA by which CPR can be quantitated. The amount of CPR in plasma, fresh serum and heated serum were essentially the same. Therefore the ELISA assay does not distinguish proCPR, activated CPR and inactivated CPR. With the ELISA method, CPR was quantitated in plasma from fifty patients with rheumatoid arthritis and eleven patients with severe hepatitis as well as healthy individuals. The amount of CPR in plasma obtained from patients with rheumatoid arthritis was not found to be lower than that of normal subjects. Furthermore, the patients who suffered severe hepatitis and had very low levels of CPR-total were fatal. This suggests that a decrease of CPR level might be a good indication of a patient's prognosis to death by hepatitis.

Antibodies, Monoclonal↗

Transverse arrest: a review of outcomes of rotational forceps and cesarean delivery at a single center.

Objective: To assess whether Kielland forceps rotational delivery causes a significant increase in maternal or perinatal morbidity in comparison with cesarean delivery in the clinical situation of transverse arrest. Rotational forceps delivery has been reported to have unacceptably high maternal and neonatal complication rates when compared with either spontaneous vaginal delivery or non-rotational forceps delivery. Clinically, however, we considered the more relevant comparison between Kielland rotational forceps delivery and cesarean delivery as these are generally the two modes of delivery for the fetus in transverse arrest.Study Design: The charts of all the mothers delivered by Kielland forceps or cesarean delivery for the main indication of transverse arrest between July 1995 and June 1996 were reviewed. The following were evaluated: Apgar scores at 1 and 5 minutes, cord pH, birth weight, neonatal superficial facial laceration or bruising, bone or nerve injuries, maternal injuries, 3rd and 4th degree perineal or vaginal lacerations, post-partum fever. Statistical analysis was by chi(2) (for Apgar scores), two-tailed Fisher Exact test (for post-partum fever), and t test (for birth weight and cord pH). A P value <.05 was considered statistically significant.Results: Fifty-two Kielland forceps and 30 cesarean deliveries were performed. One-minute Apgar scores of less than or equal to 7 were noted in 11.5% and 6.7% of the forceps and cesarean section groups, respectively (P = NS); mean arterial cord pH was 7.26 for both groups; no cases of post-partum fever were noted in the forceps versus 16% in the cesarean group (P <.05); the mean weight was 3,316 and 3,569 g for the forceps and cesarean group, respectively (P <.05). The Kielland group was also characterized by the following: 19 facial bruising and 2 temporary facial nerve injury (resolved by the time of discharge), 3 perineal lacerations, and 3 vaginal tears. No blood transfusions were required for either group.Conclusions: To our knowledge, this is the first study that compares rotational forceps delivery for transverse arrest with its true clinical alternative, cesarean delivery.No significant difference in cord pH and Apgar scores were noted between the two groups. The birth weight was greater and post-partum fever was more common in the cesarean section group. Neonatal facial bruising, temporary facial nerve palsy, and maternal perineal and vaginal lacerations were more common in the forceps groups. This study suggests that rotational forceps may be a reasonable alternative to cesarean delivery in the management of transverse arrest.

Journal Article↗

Lung volume histograms after computed tomography of the chest with three-dimensional imaging as a method to substantiate successful surgical expansion of the rib cage in achondroplasia.

The authors describe a radiographic method to quantify a surgical procedure of thoracic expansion in a 2-year-old patient with achondroplasia, small chest cage, and severe restrictive lung disease. The patient had undergone three surgical procedures of thoracic expansion since birth. The authors utilized computer-generated lung volume histograms after spiral computed tomographic scanning and three-dimensional imaging of the lungs to calculate his lung volumes before and after the third surgical thoracic expansion. The lung volumes, calculated by the histograms, were 363 mL and 406 mL before and after surgery, respectively. This 40-mL difference in the patient's lung volumes (4 mL/kg) accounted for a significant clinical improvement. Lung volume histograms obtained by this radiographic method are very helpful in substantiating a successful surgical chest expansion or provide an explanation for an unsuccessful repair.

Achondroplasia↗

Antisense homology box-derived peptides represent a new class of endothelin receptor inhibitors.

Several peptides encoded by the sense and corresponding antisense DNA have been found to recognize and bind to each other. We developed software to search for sense-antisense regions within proteins taking into account the degeneracy of the genetic code, i.e., one amino acid can have several "antisense" counterparts. Using this approach, we searched endothelin receptor type A for intramolecular regions related in sense-antisense fashion. After locating these regions (termed "antisense homology boxes"), several corresponding peptides were synthesized. The four new ET(A) receptor fragment peptides ETR-P1 ("CALSVDRYRAVASW"), ETR-P3 ("QGIGPLITAIEI"), ETR-P4 ("IADNAERYSANLSSHV") and ETR-P6 ("LNRRNGSLRIALSEHLKNRREVA") reported here can inhibit ET-1 activity.

3T3 Cells↗

Low-dose spiral computed tomography of the thorax: comparison with the standard-dose technique.

RATIONALE AND OBJECTIVES: This study was conducted to determine whether significant reduction in radiation dose (250 mAs-->50 mAs), in chest computed tomography (CT) using volume acquisition affects image quality or the detectability of pathologic findings in the lung and mediastinum. METHODS: Phantom studies were conducted to evaluate resolution and noise level, then a patient study was then done. Fifty consecutive patients (10 normal and 40 abnormal) cases were examined. After a scan (250 mAs, 120OkVp) of the entire thorax, five additional slices (50 mAs, 120kVp) at the level of the abnormality were obtained. Three independent observers evaluated the visibility of normal lung and mediastinal structures, as well as image noise. The mean score was compared between the standard and low doses. In a second study, an independent evaluation of the presence or absence of pathologic findings was made by four observers. RESULTS: Lucite phantom studies demonstrated diminished low-contrast resolution and increased noise level for the low-dose technique. Observers assessed more noise in the low-dose images (P < 0.001). The normal structures were judged to be more visible with standard dose (P < 0.01), but the magnitude of the judged differences was small especially in the lung. No differences were found in the detection of lung and mediastinal abnormalities (P > 0.10). CONCLUSIONS: The low-dose technique was effective in demonstrating pathologic findings for the lung and mediastinum. Low-dose spiral CT should be considered as a viable alternative to standard-dose spiral CT.

Adult↗

Measurement of arginine carboxypeptidase-generating activity of adult plasma.

Arginine carboxypeptidase (CPR) is a novel carboxypeptidase which was first described by Campbell and Okada. CPR is generated from a stable precursor of CPR (proCPR) during coagulation or under other circumstances and is promptly inactivated at 37 C. Therefore, it is not easy to determine CPR in blood samples. Since proCPR can be separated from the other basic carboxypeptidase (carboxypeptidase N; CPN) by passing plasma through DEAE gel, we have established a method to determine the amount of proCPR after converting it to active CPR by trypsin treatment. We first separated the proCPR from CPN using a filter cup tube (FC tube) packed with DEAE Sephadex, and measured activity after conversion of the enzyme to its active form using trypsin. With this method, no significant decrease in proCPR was noted in the plasma of patients including those with rheumatoid arthritis (RA), although CPR activity in fresh sera has been reported to be decreased. This discrepancy suggests that proCPR is not depleted in most patient sera, but that the level of activity of the enzyme which converts proCPR into active CPR may be compromised in RA patients.

Adult↗

Phenotypic and genotypic characterization of nosocomial Staphylococcus aureus isolates from trauma patients.

Staphylococcus aureus is a major cause of nosocomial infections. During the period from March 1992 to March 1994, the patients admitted to the intensive care unit of the University of Maryland Shock Trauma Center were monitored for the development of S. aureus infections. Among the 776 patients eligible for the study, 60 (7.7%) patients developed 65 incidents of nosocomial S. aureus infections. Of the clinical isolates, 43.1% possessed a polysaccharide type 5 capsule, 44.6% possessed a type 8 capsule, and the remaining 12.3% had capsules that were not typed by the type 5 or type 8 antibodies. Six antibiogram types were noted among the infection-related isolates, with the majority of the types being resistant only to penicillin and ampicillin. It was noted that the majority of cases of pneumonia were caused by relatively susceptible strains, while resistant strains were isolated from patients with bacteremia and other infections. Only 16 (6.3%) of the isolates were found to be methicillin-resistant S. aureus (MRSA). DNA fingerprinting by pulsed-field gel electrophoresis showed 36 different patterns, with characteristic patterns being found for MRSA strains and the strains with different capsular types. Clonal relationships were established, and the origins of the infection-related isolates in each patient were determined. We conclude that (i) nosocomial infection-related isolates from the shock trauma patients did not belong to a single clone, although the predominance of a methicillin-resistant genotype was noted, (ii) most infection-related S. aureus isolates were relatively susceptible to antibiotics, but a MRSA strain was endemic, and (iii) for practical purposes, the combination of the results of capsular and antibiogram typing can be used as a useful epidemiological marker.

Adolescent↗

Smoking cessation treatment for patients with schizophrenia.

OBJECTIVE: This study was an uncontrolled trial to assess the efficacy of a smoking cessation group program modified for individuals with schizophrenia. METHOD: Fifty outpatients with schizophrenia were divided into five groups who met separately for seven weekly sessions of a smoking cessation program. The subjects' schizophrenic and extrapyramidal symptoms were assessed before the group sessions began and after they had been completed. Assessments of smoking were made at those times and at 3-month and 6-month follow-ups. RESULTS: Forty-two percent of the subjects had stopped smoking at the end of the group sessions; 16% remained abstinent at 3 months, and 12% at 6 months. These changes were statistically significant. There was no change in the positive or negative symptoms of schizophrenia. CONCLUSIONS: The results suggest that it is possible for individuals with schizophrenia to stop smoking.

Adult↗

Antisense homology boxes in C5a receptor and C5a anaphylatoxin: a new method for identification of potentially active peptides.

A computer program was designed to locate regions termed antisense homology boxes (AHB), i.e., 8-15 amino acid regions corresponding to complementary DNA strands. Two AHBs were found in C5a and eight within the C5a receptor. The majority of intermolecular AHBs were found to overlap those found in the intramolecular AHBs. Several AHB peptides were synthesized and tested for their ability to interfere with C5a receptor functions. A peptide fragment of the C5a receptor corresponding to the loop between the fifth and sixth hypothetical transmembrane regions (amino acids 226-245) antisense to C5a and an intramolecular AHB in C5a receptor proved to be an antagonist of C5a when preincubated with C5a at high concentrations (>0.5 microM). However, when U937 cells bearing the C5a receptor were preincubated with this peptide at a much lower concentration (even as little as 40 pmol), the AHB peptide behaved as an agonist. Another AHB peptide corresponding to region 10-27 in the C5a receptor bound to two of its corresponding antisense peptides derived from C5a anaphylatoxin, corresponding to amino acids 37-43 and 61-74. This observation raises the possibility that the C5a receptor may bind C5a with two distinct orientations. Two other AHB peptides derived from C5a, PL12 (amino acids 12-27), and PL61 (amino acids 61-74), were also shown to inhibit activity. Incubation of dibutyryl cyclic AMP-stimulated U937 cells with PL37 (amino acids 37-51) resulted in increased intracellular Ca2+ levels and an anergy to subsequent challenge with C5a. Locating regions with sense-antisense relationships in proteins might help in identification of peptides that can interfere with the function of target proteins.

Amino Acid Sequence↗

Detection of Pasteurella pneumotropica in laboratory mice and rats by polymerase chain reaction.

A 16S rDNA-based polymerase chain reaction (PCR) method specific for Pasteurella pneumotropica was developed. The PCR product, a 395-base pair DNA fragment, was amplified from P. pneumotropica and not from 42 other bacterial species tested, including four other Pasteurella species and Actinobacillus ureae. The PCR method was used to identify 13 previously isolated strains that had been identified as P. pneumotropica by conventional methods: 12 were confirmed by PCR; one that was PCR-negative was re-examined by biochemical methods and determined to be A. ureae. The PCR detection of P. pneumotropica in nasopharyngeal swab specimens from 121 surveillance animals (15 inbred mice and 5 inbred rats from 20 animal rooms) had a high carrier state in healthy laboratory animals; for example, rat swab specimens were 89.6% (43/48) positive by PCR, 8.3% were positive by the direct culture-biochemical method, and 16.7% were positive by the enrichment culture-biochemical method. The positive rate for mice (21.9% [16/73]) was lower than that for rats.

Animals↗

Structural studies on corn nitrate reductase: refined structure of the cytochrome b reductase fragment at 2.5 A, its ADP complex and an active-site mutant and modeling of the cytochrome b domain.

The refined crystal structures of the recombinant cytochrome b reductase fragment of corn (Zea mays) nitrate reductase, its ADP complex and the active-site mutant Cys242Ser are reported here. The native structure has been refined at 2.5 A resolution to a crystallographic R-factor of 18.7% with root-mean-square (r.m.s) deviations from standard bond lengths and angles of 0.013 A and 2.0 degrees. The diffraction pattern of the crystals is highly anisotropic and correction of this effect lowered the crystallographic R-factor by 5% during the refinement. The structure of the enzyme co-crystallized with ADP has been solved at 2.7 A resolution and refined to an R-factor of 18.6% with r.m.s. deviations from standard bond lengths and angles of 0.014 A and 2.1 degrees. It revealed the binding site of the ADP moiety of the NADH cofactor, which is the electron donor for nitrate reduction. Based on this structure, a model of NADH at the active site of the enzyme was built and the implications for electron transfer from NADH to the flavin cofactor are discussed. The crystal structure of an active-site mutant enzyme, Cys242Ser, has been solved by difference Fourier synthesis and refined to an R-factor of 19.0% to 3.0 A resolution with standard deviations of bond lengths and angles of 0.017 A and 2.5 degrees. This structure analysis suggests that the observed decrease in catalytic activity of this mutant might be due to misalignment of the nicotinamide ring in its binding site. A model of the heme-containing domain of nitrate reductase has been built based on the X-ray structure of bovine cytochrome b5 and has been docked with the cytochrome b reductase fragment of nitrate reductase. The model of the complex contains six salt-bridges at the domain-domain interface and a hydrophobic core. In this model, His48, an invariant residue in the cytochrome b reductase family, forms an interaction with the propionic acid group of the D-ring of the heme cofactor. This group is in contact with the C-8 methyl group of the flavin ring. Residues that might influence the redox potential of the flavin cofactor are proposed and their possible role in electron transfer is discussed.

Adenosine Diphosphate↗

The antisense homology box: a new motif within proteins that encodes biologically active peptides.

Amphiphilic peptides approximately fifteen amino acids in length and their corresponding antisense peptides exist within protein molecules. These regions (termed antisense homology boxes) are separated by approximately fifty amino acids. Because many sense-antisense peptide pairs have been reported to recognize and bind to each other, antisense homology boxes may be involved in folding, chaperoning and oligomer formation of proteins. The antisense homology box-derived peptide CALSVDRYRAVASW, a fragment of human endothelin A receptor, proved to be a specific inhibitor of endothelin peptide (ET-1) in a smooth muscle relaxation assay. The peptide was able to block endotoxin-induced shock in rats as well. Our finding of endothelin receptor inhibitor among antisense homology box-derived peptides indicates that searching proteins for this new motif may be useful in finding biologically active peptides.

Amino Acid Sequence↗

Determining priorities for family physician education in substance abuse by the use of a survey.

As the initial stage in developing a curriculum to assist family physicians to diagnose and manage alcohol abuse in their practices, questionnaires were mailed to a selected group of family physicians. A total of 117 physicians (34%) completed the questionnaire. The majority of physicians (70.1%) reported that fewer than 10% of their caseload experienced alcohol-related problems. Most physicians (59.3%) did not use any of the standard diagnostic instruments but reported that screening and detection was the most challenging alcohol-related problem along with patient management. The questionnaire identified a number of areas that could be used in the development of educational strategies to increase the expertise of primary care physicians in the diagnosis and management of alcohol-related problems.

Canada↗

A 16S rDNA-based PCR method for rapid and specific detection of Clostridium perfringens in food.

A 16S rDNA-based polymerase chain reaction (PCR) method was developed for the rapid and specific detection of Clostridium perfringens in food. The PCR primers were designed by a GenBank computer search and they are complementary only with the 16S rRNA gene of C. perfringens by sequence alignment. The PCR product is a 279 BP DNA fragment. All C. perfringens strains tested were positive in the PCR assay and all other species tested were negative, including 11 other species of Clostridium and 38 species of other common bacteria. As few as two cells of C. perfringens in pure culture were detectable. High numbers of other bacterial species did not interfere with the detection of C. perfringens. The PCR amplification required only 30 min to complete. The method can be used for detection of C. perfringens in contaminated food. Samples from 100 g of chicken drumsticks which were inoculated with 20, 200 or 2000 cells of C. perfringens were subjected to the PCR assay and all produced positive results.

Animals↗

Pro-carboxypeptidase R cleaves bradykinin following activation.

Arginine carboxypeptidase (CPR) is a labile enzyme present in human serum which is unrelated to carboxypeptidase N. In this study we demonstrate that CPR exists in a precursor form in plasma and can be converted to the active form by trypsin and presumable trypsin-like enzymes. The trypsin-generated active form can not only cleave a small synthetic substrate, hippuryl-L-arginine, but can remove terminal arginine from bradykinin.

Bradykinin↗