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Biomedical subjects

M Morrison

Publications and source records attributed to M Morrison.

At least 19 recordsLinked to original sources

Adhesion to cellulose by Ruminococcus albus: a combination of cellulosomes and Pil-proteins?

An obligatory step in cellulose degradation by anaerobic bacteria is the adhesion of the bacterium to the polysaccharide. In many anaerobic bacteria the adhesion protein, and the enzymes required for extensive polysaccharide hydrolysis, are organized into a complex and interesting structure called the cellulosome. The Gram-positive anaerobe Ruminococcus albus also produces a cellulosome-like complex, but the bacterium appears to possess other mechanism(s) for adhesion to plant surfaces and genes encoding functions relevant to growth on cellulose are conditionally expressed, as suggested by a combination of functional proteomics, differential display reverse-transcriptase PCR, and mutational analysis. A novel form of cellulose-binding protein has been identified and shown to belong to the Pil-protein family, being most similar to the type 4 fimbrial proteins of Gram-negative, pathogenic bacteria. These studies have provided new insights into the adhesion of bacteria to plant surfaces, and call attention to the likely existence of genetically analogous adhesion determinants in both pathogenic and non-pathogenic bacteria.

Adhesins, Bacterial↗

HCP-1, a protein involved in chromosome segregation, is localized to the centromere of mitotic chromosomes in Caenorhabditis elegans.

To learn more about holocentric chromosome structure and function, we generated a monoclonal antibody (mAb), 6C4, that recognizes the poleward face of mitotic chromosomes in Caenorhabditis elegans. Early in mitosis, mAb 6C4 stains dots throughout the nucleoplasm. Later in prophase, mAb 6C4 stains structures on opposing faces of chromosomes which orient towards the centrosomes at metaphase. Colocalization with an antibody against a centromeric histone H3-like protein and the MPM-2 antibody, which identifies a kinetochore-associated phosphoepitope present in a variety of organisms, shows that the mAb 6C4 staining is present adjacent to the centromere. Expression screening using mAb 6C4 identified a protein in C. elegans that we named HCP-1 (for holocentric protein 1). We also identified a second protein from the C. elegans genome sequence database, HCP-2, that is 54% similar to HCP-1. When expression of HCP-1 is reduced by RNA interference (RNAi), staining with mAb 6C4 is eliminated, indicating that hcp-1 encodes the major mAb 6C4 antigen. RNAi with hcp-1 and hcp-2 together results in aberrant anaphases and embryonic arrest at approximately 100 cells with different amounts of DNA in individual nuclei. These results suggest that HCP-1 is a centromere-associated protein that is involved in the fidelity of chromosome segregation.

Amino Acid Sequence↗

Treatment of laryngeal contact ulcers and granulomas: a 12-year retrospective analysis.

Multiple etiological factors including gastroesophageal reflux, hyperfunctional voice use, and endotracheal intubation have been implicated in the development of posterior laryngeal ulcers and granulomas. The optimal approach to treatment of these lesions remains controversial. The mainstay of treatment at Vancouver General Hospital has been aggressive medical management of gastroesophageal reflux, with complimentary voice therapy offered to patients suspected of having significant hyperfunctional phonation. The authors reserve Botulinum toxin injection or surgical excision for patients who fail initial therapy. They conducted a retrospective analysis of their voice clinic records from 1985-1997 to examine the efficacy of this approach. They identified 76 patients with the diagnosis of contact ulcer or granuloma. Fifty-two patients had follow-up data available for review. Ninety-four percent of patients were treated nonsurgically: 35 patients were treated solely by dietary and medical therapy to control gastroesophageal reflux, 10 patients were treated by a combination of medical gastroesophageal reflux control and voice therapy, 3 patients had Botox injections, 2 patients had surgical excision of granuloma, 1 patient had a Kenalog injection, and 1 patient underwent laparoscopic fundoplication. Overall, 77% of patients had complete resolution, whereas 11% had partial resolution and another 11% had no significant improvement. The data supports control of gastroesophageal reflux as a central component in treatment of posterior laryngeal ulcers and granulomas.

Adult↗

The irritable larynx syndrome.

Muscular tension dysphonia, episodic laryngospasm, globus, and cough may be considered to be hyperfunctional laryngeal symptoms. Suggested etiological factors for these symptoms include gastroesophageal reflux, psychological problems, and/or dystonia. We propose a unifying hypothesis that involves neural plastic change to brainstem laryngeal control networks through which each of the above etiologies, plus central nervous system viral illness, can play a role. We suggest that controlling neurons are held in a "spasm-ready" state and that symptoms may be triggered by various stimuli. Inclusion criteria for the irritable larynx syndrome are episodic laryngospasm and/or dysphonia with or without globus or chronic cough; visible or palpable evidence of tension or tenderness in laryngeal muscles; and a definite symptom-triggering stimulus. thirty-nine patients with irritable larynx syndrome were studied. Gastroesophageal reflux was felt or proven to play a major role in a large number of the group (>90%), and about one third were deemed to have psychological causative factors. Viral illness seemed quite prevalent, with one third of patients able to relate the onset of symptoms to a viral illness that we feel might lead to central nervous system changes. Our proposed hypothesis includes a mechanism whereby acquired plastic change to central brainstem nuclei may lead to this form of hyperkinetic laryngeal dysfunction. It gives structure and reason to an array of therapy measures and suggests direction for basic research.

Adult↗

Influence of different functional elements of plasmid pGT232 on maintenance of recombinant plasmids in Lactobacillus reuteri populations in vitro and in vivo.

Plasmid pGT232 (5.1 kb), an indigenous plasmid of Lactobacillus reuteri 100-23, was determined, on the basis of nucleotide and deduced protein sequence data, to belong to the pC194-pUB110 family of plasmids that replicate via the rolling-circle mechanism. The minimal replicon of pGT232 was located on a 1.7-kb sequence consisting of a double-strand origin of replication and a gene encoding the replication initiation protein, repA. An erythromycin-selectable recombinant plasmid containing this minimal replicon was stably maintained (>97% erythromycin-resistant cells) without antibiotic selection in an L. reuteri population under laboratory growth conditions but was poorly maintained (<33% resistant cells) in the L. reuteri population inhabiting the murine gastrointestinal tract. Stable maintenance (>90% resistant cells) of pGT232-derived plasmids in the lactobacillus population in vivo required an additional 1.0-kb sequence which contained a putative single-strand replication origin (SSO). The SSO of pGT232 is believed to be novel and functions in an orientation-specific manner.

Amino Acid Sequence↗

Adherence of the gram-positive bacterium Ruminococcus albus to cellulose and identification of a novel form of cellulose-binding protein which belongs to the Pil family of proteins.

The adherence of Ruminococcus albus 8 to crystalline cellulose was studied, and an affinity-based assay was also used to identify candidate cellulose-binding protein(s). Bacterial adherence in cellulose-binding assays was significantly increased by the inclusion of either ruminal fluid or micromolar concentrations of both phenylacetic and phenylpropionic acids in the growth medium, and the addition of carboxymethylcellulose (CMC) to assays decreased the adherence of the bacterium to cellulose. A cellulose-binding protein with an estimated molecular mass following sodium dodecyl sulfate-polyacrylamide gel electrophoresis of approximately 21 kDa, designated CbpC, was present in both cellobiose- and cellulose-grown cultures, and the relative abundance of this protein increased in response to growth on cellulose. Addition of 0.1% (wt/vol) CMC to the binding assays had an inhibitory effect on CbpC binding to cellulose, consistent with the notion that CbpC plays a role in bacterial attachment to cellulose. The nucleotide sequence of the cbpC gene was determined by a combination of reverse genetics and genomic walking procedures. The cbpC gene encodes a protein of 169 amino acids with a calculated molecular mass of 17,655 Da. The amino-terminal third of the CbpC protein possesses the motif characteristic of the Pil family of proteins, which are most commonly involved with the formation of type 4 fimbriae and other surface-associated protein complexes in gram-negative, pathogenic bacteria. The remainder of the predicted CbpC sequence was found to have significant identity with 72- and 75-amino-acid motifs tandemly repeated in the 190-kDa surface antigen protein of Rickettsia spp., as well as one of the major capsid glycoproteins of the Chlorella virus PBCV-1. Northern blot analysis showed that phenylpropionic acid and ruminal fluid increase cbpC mRNA abundance in cellobiose-grown cells. These results suggest that CbpC is a novel cellulose-binding protein that may be involved in adherence of R. albus to substrate and extends understanding of the distribution of the Pil family of proteins in gram-positive bacteria.

Adhesins, Bacterial↗

Comparison of nine commercially available enzyme-linked immunosorbent assays for detection of Giardia lamblia in fecal specimens.

Overall performance, including ease of use, total hands-on time, incubation and processing times, sensitivity, and specificity, of each of nine enzyme-linked immunosorbent assays (ELISAs) were compared by using 222 individual fecal samples submitted for the detection of Giardia lamblia. The assays evaluated were manufactured by Alexon, Inc., Cambridge Biotech Corp., Meridian, Inc., and Trend Scientific, Inc. All assays used polyclonal antibodies except the "new and improved" Microplate (direct and diluted methods) by Alexon, which is a monoclonal antibody assay. Seventy specimens were positive for G. lamblia by ELISA, ova and parasite test, and/or direct fluorescent-antibody assay. One hundred fifty two were negative by all three methods. Sensitivities and specificities ranged from 88.6 to 100% and 99.3 to 100%, respectively. The total hands-on time needed to run one specimen ranged from 1 min to 2 min 17 s per specimen. All except one commercially available ELISA were found to be rapid, sensitive, and specific for the detection of G. lamblia in fecal specimens.

Animals↗

Evaluation of the Abbott LCx ligase chain reaction assay for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in urine and genital swab specimens from a sexually transmitted disease clinic population.

The Abbott LCx ligase chain reaction (LCR) assay for the simultaneous detection of Chlamydia trachomatis and Neisseria gonorrhoeae was evaluated by using swab and urine specimens from 562 patients. C. trachomatis results by LCR were compared to those by the Gen-Probe PACE 2 assay, whereas N. gonorrhoeae results by LCR were compared to those by culture. The Gen-Probe and LCR assays were performed according to the manufacturers' instructions. Gram-negative diplococci growing on modified Thayer-Martin medium were confirmed as N. gonorrhoeae by the GonoGen II assay. Supplemental data analysis was performed by major outer membrane protein PCR for C. trachomatis and probes for pilin gene detection for N. gonorrhoeae. A true-positive result for each pathogen was defined as a positive result for all three or two of three assays. Overall agreement among the six assays was 94.8%. C. trachomatis prevalence was 16.2%; N. gonorrhoeae prevalence was 5.5%. The overall sensitivity and specificity, respectively, for each test (after supplemental data analysis) were as follows: for C. trachomatis, Gen-Probe, 65.9 and 100%; LCR on urine, 90.1 and 100%; LCR on swab specimens, 96.7 and 100%; and for N. gonorrhoeae, culture, 80.6 and 100%; LCR on urine, 93.5 and 99.8%; and LCR on swab specimens, 96.8 and 100%. For women, the N. gonorrhoeae culture was very insensitive compared to its performance in men (58.3 versus 94.7%, respectively). For C. trachomatis, the Gen-Probe assay's sensitivity was lower for men than for women (62.3 versus 71.1%, respectively). The sensitivity for C. trachomatis detection by LCR on urethral and cervical swab specimens was 96.2 and 97.4% for men and women, respectively. For men, swab results were slightly better than urine results for both pathogens (sensitivity for C. trachomatis in swab and urine specimens, 96.2 and 92.5%, respectively; sensitivity for N. gonorrhoeae in swab and urine specimens, 100 and 94.7%, respectively), while for women, cervical swabs were superior in sensitivity to urine samples for detecting C. trachomatis (swab, 97.4%; urine, 81.6%) and equivalent for N. gonorrhoeae (swab, 92.3%; urine, 91.6%). The LCx LCR appears to be both sensitive and specific for the detection of C. trachomatis and N. gonorrhoeae when performed on urine or genital swab samples. Swab samples had better sensitivity than urine samples for the detection of both pathogens.

Adolescent↗

Management of unilateral vocal cord paralysis by injection medialization with teflon paste. Quantitative results.

This paper prospectively analyzes eight patients who were selected for injection medialization of a unilateral vocal cord paralysis by a transcutaneous technique. Preoperative and postoperative videolaryngoscopic recordings of the position of the paralyzed cord were made with a scoring system correlated to the laryngeal airflow measurements. Quantitative acoustic recordings of the patient's voice were also taken and compared with the patient's own subjective analysis of his or her voice. Results indicate that Teflon injection by the transcutaneous method gives a statistically and clinically improved voice in the short term.

Adult↗

Effect of soybean hulls, soy lecithin, and soapstock mixtures on ruminal fermentation and milk composition in dairy cows.

Two experiments were conducted to determine effects of soybean hulls, soy lecithin, and soapstock mixtures on ruminal fermentation, milk composition, and ruminal microbial populations. In Experiment 1, 20 Holstein dairy cows were assigned to one of five total mixed rations (TMR) in replicated 5 x 5 Latin squares to measure the effect of TMR on intake and milk composition. Four ruminally fistulated cows were used in a 4 x 5 Youden square to measure the effect of TMR on ruminal fermentation. The TMR consisted of 40 or 50% alfalfa and corn silages, 13% soybean hulls, and 47 or 37% of a concentrate containing either 2.25% soybean oil; 2.25% lipid from a mixture of soybean hulls, soy lecithin, and soapstock; or no added lipid. The ratios of soy lecithin to soapstock evaluated were 1:1, 2.5:1, and 4:1 (dry basis). The TMR containing soy lecithin and soapstock at ratios of 1:1 and 2.5:1 resulted in higher milk C18:2 than did the soybean oil TMR. Rate of ruminal NDF digestion of soybean hulls was reduced for the soybean oil TMR. In Experiment 2, a semi-continuous culture system was used to determine effects of soybean oil or an optimal mixture of soy lecithin and soapstock (1:1, wt/wt) on fermentation of soybean hulls and microbial populations. The TMR containing the lipid mixture increased the ratio of acetate to propionate compared with the TMR containing soybean oil and resulted in higher total protozoal counts than did the control TMR. The TMR containing mixtures of soy lecithin and soapstock at ratios of 1:1 and 2.5:1 elevated milk C18:2, and no negative effect on NDF digestion was detected, indicating some degree of ruminal protection.

Animal Feed↗

smg mutants affect the expression of alternatively spliced SR protein mRNAs in Caenorhabditis elegans.

The expression of alternatively spliced mRNAs from genes is an ubiquitous phenomenon in metazoa. A screen for trans-acting factors that alter the expression of alternatively spliced mRNAs reveals that the smg genes of Caenorhabditis elegans participate in this process. smg genes have been proposed to function in degradation of nonsense mutant mRNAs. Here we show that smg genes affect normal gene expression by modulating the levels of alternatively spliced SRp20 and SRp30b mRNAs. These SR genes contain alternatively spliced exons that introduce upstream stop codons. The effect of smg genes on SR transcripts is specific, because the gene encoding the catalytic subunit of the cAMP-dependent protein kinase, which also contains an alternatively spliced exon that introduces upstream stop codon, is not effected in a smg background. These results suggest that the levels of alternatively spliced mRNAs may, in part, be regulated by alternative mRNA stability.

Alternative Splicing↗

Growth inhibition of Prevotella ruminicola by protamine.

Growth of Prevotella ruminicola strains B(1)4 (subsp. brevis) and D31d (subsp. ruminicola), was inhibited by protamine, a polycationic, low molecular mass protein. Results showed that protamine has a bacteriocidal effect when present in concentrations exceeding 30 micrograms ml-1. Protamine exerted its toxic effects by disrupting the outer membrane, which was demonstrated by: (i) an increased sensitivity to hydrophobic antibiotics (novobiocin and monensin) and (ii) release of the periplasmic enzyme alkaline phosphatase following short-term exposure to protamine. Although the concentrations of protamine inhibitory to P. ruminicola are relatively low, the effects of such a compound are probably too broad to permit its successful use in terms of manipulating ruminal proteolysis.

Alkaline Phosphatase↗

Medical training in psychiatry residency. A proposed curriculum.

During the coming decades, psychiatrists will be asked to participate to a greater extent in the physical evaluation and treatment of patients with behavioral or emotional problems. Despite the high frequency with which psychiatric symptoms are caused or exacerbated by organic disease, psychiatrists have been reluctant, and in some ways, even discouraged to include physical assessments. Psychoanalysis and concerns about boundary issues have influenced psychiatrists to cede physical assessment and physical illness to other physicians. To help overcome these barriers to improved care of psychiatric patients, a curriculum is proposed for psychiatry residents. It will allow them to better use their medical backgrounds while increasing their contributions as mental health specialists.

Clinical Competence↗

Pattern recognition in muscle misuse voice disorders: how I do it.

The diagnosis of the patient with a muscle misuse voice disorder can be a complex process. The dysphonia is usually caused by problems stemming from a number of interacting factors that may include habitual vocal technique and postural use, vocal behavior, emotion, manifestations of gastroesophageal reflux, neuromuscular abnormalities, and associated organic mucosal disease. Sorting out the relative importance of the various factors is the first step towards planning an effective treatment program. This article addresses the difficulties caused by trying to classify dysphonia too rigidly, and presents a strategy for collecting the necessary information in a manner that facilitates the development of effective tools for clinical decision making. For each dysphonic patient, a pattern of causation will emerge in a way that helps the clinician to disentangle the interrelated factors.

Affect↗

Biochemical and mutational analysis of a gingipain-like peptidase activity from Prevotella ruminicola B(1)4 and its role in ammonia production by ruminal bacteria.

A chemical mutagenesis protocol was used with the ruminal bacterium Prevotella ruminicola strain B(1)4 to generate mutant strains defective in peptidase activity. Compared with the wild-type parent strain, the isolated mutants possessed 1/10 of the enzyme activity responsible for cleavage of glycine-arginine-4-methoxy-beta-naphthylamide (Gly-Arg-MNA). A concomitant loss in activity against arginine-arginine-4-methoxy-beta-naphthylamide (Arg-Arg-MNA) was also observed. Both activities were similarly affected by various proteinase inhibitors, suggesting that the same enzyme is responsible for the Arg-Arg-MNA peptidase and Gly-Arg-MNA peptidase activities. Growth rates of wild-type and mutant strains grown in batch culture with various nitrogen sources did not differ. However, a role for the Gly-Arg-MNA peptidase activity was demonstrated in coculture experiments with gram-positive, ammonia-producing ruminal bacteria. The rate and extent of ammonia production were reduced by approximately 25% in cocultures containing the mutants when compared with that of wild-type-containing cultures. These reductions could not be accounted for simply by the decrease in ammonia production by the mutant strain alone. To our knowledge, this paper reports the first successful use of chemical mutagenesis with ruminal microorganisms.

2-Naphthylamine↗

Glutamate dehydrogenase activity profiles for type strains of ruminal Prevotella spp.

The glutamate dehydrogenase (GDH) activities for the type strains of Prevotella ruminicola (strain 23), Prevotella brevis (strain GA33), and Prevotella bryantii (strain B(1)4) were assessed by a combination of enzyme assays and analysis of migration patterns of GDH proteins following nondenaturing polyacrylamide gel electrophoresis. Unlike results with most other prokaryotes, but similar to results with other members of the family Bacteroidaceae, NADPH-utilizing specific activity was greatest in all species following ammonia-limited growth. Similar also to previous findings with P. bryantii, the NAD(P)H-utilizing GDH activity of P. ruminicola can be attributed to a single protein. However, P. brevis produces an additional GDH protein(s) in response to growth with peptides. These results conclusively demonstrate that all type strains of the ruminal Prevotella sp. grouping possess GDH activity.

Ammonia↗