ABSTRACT
Cytokinins induce two specific morphological alterations in mosses: (i) the differentiation of a tip-growing cell into a three-faced apical cell (the so-called bud), and (ii) the division of chloroplasts. In a developmental mutant of the moss Physcomitrella patens (Hedw.) B.S.G. (mutant PC22) impeded in both cellular differentiation (bud production) and chloroplast division, addition of cytokinin (N6-delta 2-isopentenyladenine) led to bud production after 3 d in the wild type and after 7 d in the mutant. Hormone induced a division of the mutant macrochloroplasts starting within 24 h and ongoing for 72 h. During this period the abundances of several plastid proteins changed in both genotypes as judged by two-dimensional-protein gel electrophoresis, silver staining and subsequent quantification with novel computer software. Eight of these polypeptides were isolated independently, subjected to microsequencing and thus identified, resulting in the first protein sequence data from a moss. Three polypeptides (24 kDa, 22 kDa, 20 kDa) were found to be homologous to enhancer protein OEE2 of the oxygen-evolving complex, four to represent isoforms of phosphoglycerate kinase (EC 2.7.2.3), and one was identified as the beta-chain of chloroplast ATPase (EC 3.6.1.34). Possible involvement of these key enzymes of the chloroplast energy-conversion machinery in organelle division and in cellular differentiation is discussed. Further sequence information was obtained from both subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39). Amounts of these polypeptides were not appreciably affected by cytokinin in moss chloroplasts.
Subject(s)
Adenine/analogs & derivatives , Adenosine Triphosphatases/biosynthesis , Chloroplasts/enzymology , Cytokinins/pharmacology , Phosphoglycerate Kinase/biosynthesis , Plant Cells , Plant Proteins/biosynthesis , Plastids/drug effects , Ribulose-Bisphosphate Carboxylase/biosynthesis , Adenine/pharmacology , Adenosine Triphosphatases/chemistry , Amino Acid Sequence , Cell Nucleus/physiology , Chloroplasts/drug effects , Chloroplasts/ultrastructure , Electrophoresis, Gel, Two-Dimensional , Electrophoresis, Polyacrylamide Gel , Isopentenyladenosine , Molecular Sequence Data , Phosphoglycerate Kinase/chemistry , Plant Proteins/chemistry , Plants/drug effects , Plants/enzymology , Plastids/enzymology , Plastids/ultrastructure , Ribulose-Bisphosphate Carboxylase/chemistry , Sequence Homology, Amino AcidSubject(s)
Managed Care Programs/trends , Pathology Department, Hospital/trends , Pathology, Clinical/trends , Cost Savings , Health Care Costs/trends , Managed Care Programs/economics , Managed Care Programs/organization & administration , Pathology, Clinical/economics , Pathology, Clinical/organization & administration , Societies, Medical , United StatesSubject(s)
Electronic Data Processing/organization & administration , Hospital Information Systems/organization & administration , Patient Identification Systems/methods , Admitting Department, Hospital/organization & administration , Blood Banks/organization & administration , Computer Systems/standards , Emergency Service, Hospital/organization & administration , Laboratories, Hospital/organization & administration , Software , United StatesSubject(s)
Clinical Laboratory Information Systems/organization & administration , Electronic Data Processing/methods , Patient Identification Systems/methods , Georgia , Hospital Bed Capacity, 300 to 499 , Patient Identification Systems/standards , Planning Techniques , Quality Assurance, Health Care/organization & administrationSubject(s)
Emergency Services, Psychiatric , Ill-Housed Persons/psychology , Mental Disorders/nursing , Psychiatric Nursing , Adult , Geriatric Psychiatry/standards , HIV Seropositivity , Hospitalization , Hospitals, Psychiatric , Humans , Male , Mental Disorders/complications , Mental Disorders/psychology , Nursing Care/standards , Physical Examination , Psychiatric Nursing/standards , Substance-Related Disorders/complications , Substance-Related Disorders/diagnosisABSTRACT
Plastid DNA of the moss Physcomitrella patens has been sequenced. An open reading frame (ORF 315) was identified downstream from rbcL, between trnR-CCG and psaI. This ORF shares homology with zfpA, a putative regulatory gene in Pisum sativum. The moss ORF is preceded by a Shine-Dalgarno sequence, two plastid promoter consensus sequences, and three TATA boxes. A specific probe detected three transcripts of low abundance in the wild-type moss and a cytokinin-sensitive chloroplast mutant. Steady state levels of zfpA transcripts were different in the two genotypes. In mutant protonemata treated with cytokinin, steady state levels of the largest transcript decreased significantly.
Subject(s)
Cytokinins/pharmacology , DNA-Binding Proteins/genetics , Genes, Plant , Plants/genetics , Promoter Regions, Genetic , Transcription, Genetic/drug effects , Amino Acid Sequence , Base Sequence , Blotting, Northern , Chloroplasts , Cloning, Molecular , DNA, Circular , Gene Expression Regulation , Molecular Sequence Data , Open Reading Frames , Phylogeny , RNA, Ribosomal , Zinc FingersSubject(s)
Decision Making , Food Service, Hospital , Interprofessional Relations , Diet , Goals , Humans , Male , Patient Care TeamABSTRACT
The morphological and biochemical changes that occur after inoculation of sterile blood into a blood culture medium (tryptic soy broth) with sodium polyanetholesulfonate and CO(2) were investigated. Cellular changes, pH, PCO(2), and PO(2) were monitored and evaluated. Erythrocytes became crenated and developed precipitated hemoglobin inclusions within 4 h. The lymphocytes appeared morphologically intact at 24 h, and by 48 h a few cells had undergone transformation. Many neutrophils were vacuolated at 24 h. Neutrophils capable of phagocytizing Staphylococcus aureus were observed after 18 h of incubation. Identifiable eosinophiles were present on day 6 of the study. A decrease in PO(2) in the unvented bottles from 44.4 to 8 mm of Hg occurred by 24 h. PO(2) remained low for 6 days, after which a slight increase occurred. An increase in PO(2) in the vented bottle from 51 to 58 mm of Hg occurred by 24 h of incubation. In both the vented and unvented bottles the PCO(2) increased. This increase was markedly more rapid in the unvented bottle. From a pH of 7.06 a decrease occurred for the first 24 h after inoculation, with the pH stabilizing at 6.8 in the vented bottles and at 6.6 in the unvented bottles. The biochemical changes that occurred in the vented culture bottles stabilized more rapidly than those of the unvented bottles. Changes caused by the addition of sterile blood to a blood culture medium resulted in conditions which departed considerably from accepted optima for the isolation of clinically important microorganisms. The phagocytosis of organisms that occurred may also have reduced the yield.
Subject(s)
Blood , Culture Media , Bacteria/isolation & purification , Blood/microbiology , Blood Cells/ultrastructure , Carbon Dioxide , Humans , Hydrogen-Ion Concentration , Neutrophils/immunology , Oxygen , Partial Pressure , Phagocytosis , Staphylococcus aureus/immunologyABSTRACT
Laboratory procedures used to establish the diagnosis of Pneumocystis carinii pneumonia were evaluated using an experimental murine model. Touch preparations and suspension smears were prepared from lung tissue know to contain Pneumocystis cysts. These preparations were stained by a variety of methods known to demonstrate either cyst forms or sporozoites and trophozoites. Suspension smears proved to be superior to touch preparations in terms of cyst content and homogeneity of staining. Also, methods that stain cyst forms were superior to those that stain sporozoites and trophozoites for location and identification of organisms. The authors believe that suspension smears prepared from lung tissue and stained with toluidine blue O should be examined initially as a rapid screening method for Pneumocystis cysts. When the results of this initial screen are negative or inconclusive, additional suspension smears stainded by the modified Gomori methenamine silver nitrate method should be examined, pending availability of histologic sections.
Subject(s)
Clinical Laboratory Techniques , Disease Models, Animal , Pneumonia, Pneumocystis/diagnosis , Animals , Azure Stains , Methenamine , Methylene Blue , Rats , Silver Nitrate , Staining and Labeling , Tolonium ChlorideABSTRACT
Increasing concentrations of malonaldehyde and beta-propiolactone were increasingly mutagenic with 7 mutants of Salmonella typhimurium, 5 of which mutated bya frameshift mechanism and 2 of which mutated through base-pair substitution. The antioxidants vitamin C, vitamin E, selenium and butylated hydroxytoluene (BHT) at 3 logarithmic concentrations markedly reduced mutagenesis in those strains which mutated by frameshift mechanism.
