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1.
J Small Anim Pract ; 2024 Sep 05.
Article in English | MEDLINE | ID: mdl-39238283

ABSTRACT

OBJECTIVES: To describe the techniques for preparation and placement of peripheral intravenous catheters (PIVCs), to describe the complications associated with PIVCs, and to identify factors associated with PIVC complications in small animal practice in the United Kingdom. MATERIALS AND METHODS: A prospective multicentre observational study was undertaken between January 2022 and January 2023. Data collected included patient information, information regarding the placement and maintenance of PIVCs, and PIVC complications, from privately owned cats and dogs presenting to veterinary institutes in the United Kingdom. Patients required a PIVC to be placed as part of their care and the PIVC was anticipated to be in situ for >24 hours to be eligible for PIVC complication analysis. RESULTS: A total of 19 institutes recorded data regarding 382 PIVCs, with 325 (85.1%) placed in dogs and 57 (14.9%) in cats. The most common reasons for placement were to administer intravenous fluid therapy (74.3%) and intravenous medications (71.7%). There were 102 of 382 (26.7%) PIVCs associated with a complication, with limb swelling/suspected phlebitis in 44 of 382 (11.5%) and PIVC dislodgement/patient interference in 30 of 382 (7.9%) PIVCs. Factors associated with increased risk of complication were more than 1 attempt to place the PIVC, a second or subsequent PIVC being placed during hospitalisation, flush frequency different than every 1 to 24 hours, and flush solution with compound sodium lactate. CLINICAL SIGNIFICANCE: Veterinary professionals must be vigilant when monitoring a patient with a PIVC in situ, particularly if a PIVC is associated with one of the aforementioned factors of increased likelihood of complication.

2.
J Affect Disord ; 361: 465-471, 2024 Sep 15.
Article in English | MEDLINE | ID: mdl-38897305

ABSTRACT

BACKGROUND: Identifying patients at risk for a suicide attempt (SA) is critical in adolescents with mental disorders. The current study aimed to 1) examine whether personality dysfunction (PD) is associated with previous SA, 2) explore the incremental utility of PD over psychiatric disorders in modeling previous SA. METHODS: The sample comprised of n = 498 adolescent patients (mean age = 15.41 years, 79.12 % females, inpatient 48.8 %, outpatient 51.2 %). SA in the past year, PD according to the alternative DSM-5 model for personality disorders, and psychiatric diagnoses were assessed using semi-structured interviews. Logistic regression and principal component analysis examining the associations and specific patterns of PD and SA in the past year were conducted. Hierarchical (stepwise) logistic regression was applied to investigate the incremental utility of PD over that of psychiatric diagnoses to identify individuals with SA in the past year. RESULTS: Including all facets of PD revealed a significant model with SA in the past year as outcome (χ2(12) = 106.65, McFaddens Pseudo-R2 = 0.17, p < 0.01). Adding PD to the model explained a significant amount of variance in past SA over that of psychiatric diagnoses (Pseudo-R2 = 0.18, Wald χ2 = 43.05, p < 0.01). LIMITATIONS: As we only studied past SA and due to the cross-sectional design, no conclusion regarding the prediction of future SA can be drawn. DISCUSSION: PD should routinely be assessed in adolescent patients since individuals with PD are more likely to have attempted suicide even when controlling for comorbid psychiatric disorders. PD may represent an important target for intervention in those with suicidal thoughts and behaviors.


Subject(s)
Personality Disorders , Suicide, Attempted , Humans , Suicide, Attempted/statistics & numerical data , Suicide, Attempted/psychology , Female , Adolescent , Male , Personality Disorders/epidemiology , Personality Disorders/psychology , Personality Disorders/diagnosis , Mental Disorders/epidemiology , Mental Disorders/psychology , Risk Factors , Logistic Models
3.
Perit Dial Int ; 42(6): 640-642, 2022 11.
Article in English | MEDLINE | ID: mdl-36286404

ABSTRACT

18 F-FDG PET/CT scan is a useful diagnostic tool in patients with neoplasia or inflammatory diseases for further evaluation. Due to interference of glucose with the cellular uptake of the 18 F-FDG tracer via glucose transporter, withhold of any glucose source several hours before imaging is mandatory. This is also the case in peritoneal dialysis patients where glucose-containing peritoneal dialysis fluid cannot be used prior to 18 F-FDG PET/CT scan. Whether the same hold true for icodextrin is not known. We describe two patients with metastatic carcinomas while on peritoneal dialysis on an icodextrin-containing regimen, in whom icodextrin was not discontinued before 18 F-FDG PET/CT scan and where accurate diagnosis of metastatic lesions as well as in one case a simultaneous tunnel infection could be made. Our observation suggests that there is no significant interference of icodextrin with the metabolism of 18 F-FDG and so it is feasible to continue an established icodextrin-containing regimen in peritoneal dialysis patients in case of 18 F-FDG PET/CT scan.


Subject(s)
Fluorodeoxyglucose F18 , Peritoneal Dialysis , Humans , Icodextrin , Peritoneal Dialysis/methods , Positron Emission Tomography Computed Tomography , Glucose/metabolism
4.
Nervenarzt ; 88(1): 3-9, 2017 Jan.
Article in German | MEDLINE | ID: mdl-27853852

ABSTRACT

The number of underage refugees arriving in Germany has rapidly increased since 2015. Many of these children and adolescents have been and still are, exposed to a large number of stressful circumstances. The group of those helping refugee minors is heterogeneous with both volunteers and professional workers from the fields of child welfare and healthcare services. Easily applicable instruments to assess both burdens and resources are needed in order to plan appropriate interventions. This paper focuses on instruments for assessing the circumstances of refugee minors and includes pilot data of an online-based screening instrument to assess burdens and resources (providing online resource and trauma assessment for refugees, PORTA). Field application was tested by the staff of a clearing and preclearing institution with 33 cases and good practical feasibility was reported. Applying a simple to use screening instrument for refugee minors and their helpers, which is available in several languages creates the possibility of a shared definition of problems and solutions and is beneficial to helpers (e.g. volunteers, youth welfare services and medical doctors) as well as refugee minors.


Subject(s)
Mass Screening/methods , Personality Assessment/statistics & numerical data , Psychometrics/methods , Refugees/psychology , Stress Disorders, Traumatic/diagnosis , Stress Disorders, Traumatic/psychology , Adolescent , Child , Child, Preschool , Female , Germany , Humans , Infant , Infant, Newborn , Male , Minors/classification , Minors/psychology , Refugees/classification , Reproducibility of Results , Sensitivity and Specificity , Translating
5.
Biochem Biophys Res Commun ; 412(3): 419-24, 2011 Sep 02.
Article in English | MEDLINE | ID: mdl-21827738

ABSTRACT

Ligand-biased receptor signaling has been proposed for several G-protein coupled receptors including the niacin receptor GPR109A. Coupling to the G(i/o) pathway has been shown to be responsible for the well described triglyceride lowering effect of nicotinic acid in mice, while activation of the ß-arrestin pathway has been suggested to be responsible for its peripheral vasodilatory effect that causes cutaneous flushing. Several ligands have been described to selectively induce triglyceride lowering without inducing flushing. Cellular impedance has been demonstrated to determine G-protein coupled receptors activation in a G-protein specific manner. Agonists, which induce triglyceride lowering, but not flushing show a profile in cellular impedance that is distinct from the one induced by niacin and those compounds that induce triglyceride lowering as well as flushing. The strength of the signal correlates with the activation of ß-arrestin.


Subject(s)
Electric Impedance , Receptors, G-Protein-Coupled/agonists , Animals , Cell Line , Humans , Ligands , Mice , Niacin/pharmacology , Receptors, G-Protein-Coupled/analysis , Receptors, Nicotinic/analysis , Signal Transduction , Triglycerides/metabolism
6.
J Appl Microbiol ; 109(5): 1521-30, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20553346

ABSTRACT

AIMS: Characterize the response of Deinococcus radiodurans R1 cells to low-pressure low-temperature nitrogen-oxygen microwave plasma and identify repair processes during recovery. METHODS AND RESULTS: Cells coated onto glass slides exhibited a biphasic plasma inactivation kinetics. Treatment with various plasmas and subsequent incubation in recovery medium prolonged the lag phase in a part of the survivors, during which the ability to grow on stress medium was recovered. This recovery strongly depended on transcriptional and translational processes and cell wall synthesis, as revealed by addition of specific inhibitors to the recovery medium. Genes involved in DNA repair, oxidative stress response, and cell wall synthesis were induced during recovery, as determined by quantitative RT-PCR. Damage to chromosomal DNA caused by plasma agents and repair during recovery was directly shown by quantitative PCR. Plasmas with less UV radiation emission were also effective in killing D. radiodurans cells but resulted in less DNA damage and lower induction of the investigated genes. CONCLUSIONS: The response of D. radiodurans to plasma indicates that DNA, proteins, and cell wall are primary targets of plasma finally leading to the cell death. Protein oxidation is more important for killing of D. radiodurans cells than of Bacillus subtilis spores. Thus, the contaminating biological material affects the plasma composition to be used for sterilization. SIGNIFICANCE AND IMPACT OF THE STUDY: The results in this study provide new insight into the interaction of plasma with bacterial cells. This knowledge contributes to the definition of useful parameters for novel plasma sterilization equipment to control process safety.


Subject(s)
Cold Temperature , Deinococcus/physiology , Microbial Viability , Pressure , Sterilization/methods , DNA Damage , DNA Ligases/genetics , DNA Repair , Deinococcus/genetics , Gene Expression Regulation, Bacterial , Microwaves , Ultraviolet Rays
7.
J Appl Microbiol ; 108(2): 521-31, 2010 Feb.
Article in English | MEDLINE | ID: mdl-19659696

ABSTRACT

AIMS: To identify structural components of Bacillus subtilis spores serving as targets for sterilization with microwave induced low-pressure, low-temperature nitrogen-oxygen plasma. METHODS AND RESULTS: The inactivation of spores followed a biphasic kinetics consisting of a log-linear phase with rapid inactivation followed by a slow inactivation phase. In the course of plasma treatment, damage to DNA, proteins and spore membranes were observed by monitoring the occurrence of auxotrophic mutants, inactivation of catalase (KatX) activity and the leakage of dipicolinic acid, respectively. Spores of the wild-type strain showed the highest resistance to plasma treatment. Spores of mutants defective in nucleotide excision repair (uvrA) and small acid-soluble proteins (Delta sspA Delta sspB) were more sensitive than those defective in the coat protein CotE or spore photoproduct repair (splB). Exclusion of reactive particles and spectral fractions of UV radiation from access to the spores revealed that UV-C radiation is the most effective inactivation agent in the plasma, whereby the splB and Delta cotE mutant spores were equally and slightly less sensitive, respectively, than the wild-type spores. Finally, the extent of damages in the spore DNA determined by quantitative PCR correlated with the spore inactivation. CONCLUSIONS: Spore inactivation was efficiently mediated by a combination of DNA damage and protein inactivation. DNA was identified to be the primary target for spore inactivation by UV radiation emitted by the plasma. Coat proteins were found to constitute a protective layer against the action of the plasma. SIGNIFICANCE AND IMPACT OF THE STUDY: The results provide new evidence to the understanding of plasma sterilization processes. This knowledge supports the identification of useful parameters for novel plasma sterilization equipment to control process safety.


Subject(s)
Bacillus subtilis/radiation effects , Spores, Bacterial/radiation effects , Sterilization/methods , Ultraviolet Rays , Bacillus subtilis/genetics , Bacillus subtilis/growth & development , Bacterial Proteins/radiation effects , Cold Temperature , Colony Count, Microbial , DNA Damage/radiation effects , Microbial Viability , Microwaves , Models, Biological , Picolinic Acids , Pressure , Spores, Bacterial/genetics , Spores, Bacterial/growth & development
8.
Microb Ecol ; 45(4): 455-63, 2003 May.
Article in English | MEDLINE | ID: mdl-12704557

ABSTRACT

Denaturing gradient gel electrophoresis (DGGE) of DNA fragments generated by PCR with primers specific for lactic acid bacteria (LAB) was applied to investigate various media and incubation conditions to recover LAB from human feces. Samples were plated on selective and nonselective media and incubated under standard condition (37 degrees C, anaerobiosis) for fecal LAB as well as alternative condition (30 degrees C, 2% O2). PCR-DGGE analyses of resuspended bacterial biomass (RBB) obtained from agar plates revealed that the species composition of the recovered LAB was affected more strongly by the incubation condition than by the used medium. It was observed that food-associated LAB, such as Lactobacillus sakei and Leuconostoc mesenteroides, hitherto not described as intestinal inhabitants, are more easily selected when the alternative incubation condition is used. Identification of randomly picked colonies grown under the alternative condition showed that L. sakei is one of the predominant food-associated LAB species, reaching counts of up to 106 CFU/g feces. Comparison of the results of bacteriological culture with those obtained by PCR-DGGE analysis of the RBB showed that investigation of RBB is a fast and reliable method to gain insight into the species composition of culturable LAB in feces.


Subject(s)
DNA, Bacterial/analysis , Lactobacillus/isolation & purification , Leuconostoc/isolation & purification , Digestive System/microbiology , Electrophoresis , Feces/microbiology , Humans , Lactobacillus/genetics , Leuconostoc/genetics , Polymerase Chain Reaction , Reproducibility of Results , Specimen Handling
9.
Nature ; 417(6886): 254-9, 2002 May 16.
Article in English | MEDLINE | ID: mdl-12015594

ABSTRACT

The normal plasma protein serum amyloid P component (SAP) binds to fibrils in all types of amyloid deposits, and contributes to the pathogenesis of amyloidosis. In order to intervene in this process we have developed a drug, R-1-[6-[R-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]pyrrolidine-2-carboxylic acid, that is a competitive inhibitor of SAP binding to amyloid fibrils. This palindromic compound also crosslinks and dimerizes SAP molecules, leading to their very rapid clearance by the liver, and thus produces a marked depletion of circulating human SAP. This mechanism of drug action potently removes SAP from human amyloid deposits in the tissues and may provide a new therapeutic approach to both systemic amyloidosis and diseases associated with local amyloid, including Alzheimer's disease and type 2 diabetes.


Subject(s)
Amyloidosis/drug therapy , Amyloidosis/metabolism , Serum Amyloid P-Component/metabolism , Amyloidosis/blood , Animals , Calcium/metabolism , Carboxylic Acids/chemistry , Carboxylic Acids/metabolism , Carboxylic Acids/pharmacology , Carboxylic Acids/therapeutic use , Cross-Linking Reagents/chemistry , Cross-Linking Reagents/metabolism , Cross-Linking Reagents/pharmacology , Cross-Linking Reagents/therapeutic use , Crystallography, X-Ray , Dimerization , Humans , Inhibitory Concentration 50 , Liver/metabolism , Mice , Models, Molecular , Protein Binding , Protein Structure, Quaternary/drug effects , Pyrrolidines/chemistry , Pyrrolidines/metabolism , Pyrrolidines/pharmacology , Pyrrolidines/therapeutic use , Serum Amyloid P-Component/antagonists & inhibitors , Serum Amyloid P-Component/chemistry
10.
Syst Appl Microbiol ; 24(2): 232-7, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11518326

ABSTRACT

Batch cultures inoculated with human faeces were used to study the prebiotic properties of levan-type exopolysaccharides (EPS) from Lactobacillus sanfranciscensis as well as levan, inulin, and fructooligosaccharide (FOS). Denaturing gradient gel electrophoresis of 16S rDNA fragments generated by PCR with universal primers was used to analyse the cultures. Characteristic changes were revealed in the composition of the gut bacteria during fermentation of the carbohydrates. An enrichment of Bifidobacterium spp. was found for the EPS and inulin but not for levan and FOS. The bifidogenic effect of the EPS was confirmed by culturing on selective medium. In addition, the use of EPS and FOS resulted in enhanced growth of Eubacterium biforme and Clostridium perfringens, respectively.


Subject(s)
Fructans/metabolism , Intestines/microbiology , Lactobacillus/metabolism , Polysaccharides/metabolism , Probiotics , Colony Count, Microbial , Culture Media , Electrophoresis, Agar Gel , Fermentation , Humans , Inulin/metabolism , Oligosaccharides/metabolism , Polymerase Chain Reaction , Polysaccharides/chemistry , RNA, Ribosomal, 16S
11.
Appl Environ Microbiol ; 67(6): 2578-85, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11375166

ABSTRACT

Denaturing gradient gel electrophoresis (DGGE) of DNA fragments generated by PCR with 16S ribosomal DNA-targeted group-specific primers was used to detect lactic acid bacteria (LAB) of the genera Lactobacillus, Pediococcus, Leuconostoc, and Weissella in human feces. Analysis of fecal samples of four subjects revealed individual profiles of DNA fragments originating not only from species that have been described as intestinal inhabitants but also from characteristically food-associated bacteria such as Lactobacillus sakei, Lactobacillus curvatus, Leuconostoc mesenteroides, and Pediococcus pentosaceus. Comparison of PCR-DGGE results with those of bacteriological culture showed that the food-associated species could not be cultured from the fecal samples by plating on Rogosa agar. On the other hand, all of the LAB species cultured from feces were detected in the DGGE profile. We also detected changes in the types of LAB present in human feces during consumption of a milk product containing the probiotic strain Lactobacillus rhamnosus DR20. The analysis of fecal samples from two subjects taken before, during, and after administration of the probiotic revealed that L. rhamnosus was detectable by PCR-DGGE during the test period in the feces of both subjects, whereas it was detectable by culture in only one of the subjects.


Subject(s)
Feces/microbiology , Lactobacillaceae/isolation & purification , Polymerase Chain Reaction/methods , Streptococcaceae/isolation & purification , Adult , Clinical Trials as Topic , DNA Primers , DNA, Ribosomal/genetics , DNA, Ribosomal/isolation & purification , Electrophoresis/methods , Female , Humans , Lactobacillaceae/genetics , Lactobacillus/isolation & purification , Leuconostoc/isolation & purification , Male , Nucleic Acid Denaturation , Pediococcus/isolation & purification , Probiotics/pharmacology , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 16S/isolation & purification , Streptococcaceae/genetics
12.
J Neurosci Res ; 61(3): 263-72, 2000 Aug 01.
Article in English | MEDLINE | ID: mdl-10900073

ABSTRACT

Nerve growth factor (NGF) binds to two neurotrophin receptors: p75(NTR) and p140(Trk) (TrkA). Both receptors dimerize in response to NGF binding. TrkA homodimers and heteromers of TrkA and p75(NTR) promote cell survival whereas homodimers of p75(NTR) mediate apoptosis upon binding of NGF. The interaction between receptor and NGF can be inhibited either on the level of the ligand by altering NGF conformation so that NGF is no longer recognized by the receptor or on the level of the receptor by blocking the binding site of p75(NTR) or TrkA. The effect of altering NGF conformation on NGF signaling was investigated in two neuron-like cell lines: in human SK-N-MC cells that express only p75(NTR) and in rat PC12 cells that express both p75(NTR) and TrkA. In the present study we demonstrate that Ro 08-2750 binds to the NGF dimer thereby probably inducing a change in its conformation such that NGF cannot bind to p75(NTR) anymore. In SK-N-MC cells this leads to inhibition of NGF-induced programmed cell death. In PC12 cells enhanced signaling through TrkA was observed.


Subject(s)
Nerve Growth Factor/physiology , Pteridines/pharmacology , Receptor, Nerve Growth Factor/physiology , Receptor, trkA/physiology , Animals , Apoptosis , Cell Survival , Choline O-Acetyltransferase/metabolism , Flavins , Humans , Ligands , Nerve Growth Factor/chemistry , Nerve Growth Factor/metabolism , Neurites/physiology , PC12 Cells , Phosphorylation , Protein Conformation , Pteridines/metabolism , Rats , Receptor, Nerve Growth Factor/metabolism , Receptor, trkA/metabolism , Tumor Cells, Cultured , Tyrosine/metabolism
13.
Mol Microbiol ; 35(1): 173-9, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10632887

ABSTRACT

An alpha-L-rhamnosidase clone was isolated from a genomic library of the thermophilic anaerobic bacterium Clostridium stercorarium and its primary structure was determined. The recombinant gene product, RamA, was expressed in Escherichia coli, purified to homogeneity and characterized. It is a dimer of two identical subunits with a monomeric molecular mass of 95 kDa in SDS polyacrylamide gel electrophoresis. At pH 7.5 it is optimally active at 60 degrees C and insensitive to moderate concentrations of Triton X100, ethanol and EDTA. It hydrolysed p-nitrophenyl-alpha-L-rhamnopyranoside, naringin and hesperidin with a specific activity of 82, 1.5 and 0.46 U mg-1 respectively. Hydrolysis occurs by inversion of the anomeric configuration as detected using 1H-NMR, indicating a single displacement mechanism. Naringin was hydrolysed to rhamnose and prunin, which could further be degraded by incubation with a thermostable beta-glucosidase. The secondary structure of RamA consists of 27% alpha-helices and 50% beta-sheets, as detected by circular dichroism. The primary structure of the ramA gene has no similarity to other glycoside hydrolase sequences and possibly is the first member of a new enzyme family.


Subject(s)
Clostridium/enzymology , Flavanones , Glycoside Hydrolases/metabolism , Amino Acid Sequence , Circular Dichroism , Cloning, Molecular , DNA, Recombinant , Enzyme Stability , Flavonoids/metabolism , Glycoside Hydrolases/chemistry , Glycoside Hydrolases/genetics , Glycosides/metabolism , Hesperidin/metabolism , Hydrogen-Ion Concentration , Magnetic Resonance Spectroscopy , Molecular Sequence Data , Temperature
14.
Int J Syst Evol Microbiol ; 50 Pt 6: 2013-2020, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11155975

ABSTRACT

Acetic acid bacteria have been isolated from submerged high-acid spirit vinegar fermentations in the Southern part of Germany. Four strains (LTH 4560T, LTH 4341, LTH 4551 and LTH 4637) were characterized in more detail and it was revealed that they have in common certain properties such as requirement of acetic acid, ethanol and glucose for growth, and no over-oxidation of acetate. Growth occurs only at total concentrations (sum of acetic acid and ethanol) exceeding 6.0%. A method for their preservation was developed. Comparative analysis of the 16S rRNA revealed sequence similarities of >99% between strain LTH 4560T and the type strains of the related species Gluconacetobacter hansenii. However, low levels of DNA relatedness (<41 %) were determined in DNA-DNA similarity studies. In addition, specific physiological characteristics permitted a clear identification of the strains within established species of acetic acid bacteria. The strains could also be differentiated on the basis of the distribution of IS element 1031 C within the chromosome. Based on these results, the new species Gluconacetobacter entanii sp. nov. is proposed for strain LTH 4560T ( = DSM 13536T). A 16S-rRNA-targeted oligonucleotide probe was constructed that was specific for G. entanii, and the phylogenetic position of the new species was derived from a 16S-rRNA-based tree.


Subject(s)
Acetic Acid/metabolism , Acetobacter/classification , Acetobacter/genetics , Acetobacter/isolation & purification , Acetobacter/physiology , Bacterial Typing Techniques , DNA, Ribosomal/analysis , Fermentation/physiology , Genotype , Hydrogen-Ion Concentration , Industrial Microbiology , Molecular Sequence Data , Nucleic Acid Hybridization , RNA, Ribosomal, 16S
15.
Syst Appl Microbiol ; 22(3): 321-8, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10553284

ABSTRACT

The use of lactobacilli as starter organisms in food fermentation processes requires thorough knowledge of their reaction to the multitude of ecological factors including their response to stress. We have characterised the dnaK gene region of Lactobacillus sakei LTH681. Two chromosomal EcoRI fragments of 2.5 and 4.0 kb were identified using a homologous dnaK probe generated by PCR. The sequence analysis of the cloned fragments showed that the dnaK gene region consists of four heat shock genes with the organisation hrcA-grpE-dnaK-dnaJ. Comparison of the deduced amino acid sequences revealed high similarity to the corresponding heat shock proteins of Gram-positive bacteria. An upstream located orfY was found which exhibited substantial similarity (41.5%) to the chloramphenicol acetyltransferase of Enterobacter aerogenes. Northern hybridisation analysis revealed that the transcription of the genes is induced by heat shock (42 degrees C) as well as salt (6%) or ethanol (10%) stress. Several transcripts were detected including a polycistronic mRNA of 4.9 kb which represents the transcript of the complete dnaK gene region indicating a tetracistronic organisation of the dnaK operon. The other RNA fragments were identified as shorter transcripts (3.7 and 1.3 kb) or cleavage products of the polycistronic mRNAs. The transcription start sites of the dnaK operon were determined under inducing and non-inducing conditions. The site varied with the applied stress condition. A regulatory CIRCE element was identified located between the transcription and translation start site. The promoter region including CIRCE was transcriptionally fused to the beta-glucuronidase reporter gene gusA and expressed in L. sakei LTH681. The kinetics of transcriptional induction of gusA by heat shocking were identical to those of the dnaK operon confirming the involvement of the CIRCE element in regulation of gene expression.


Subject(s)
Bacterial Proteins/genetics , Escherichia coli Proteins , HSP70 Heat-Shock Proteins/genetics , Lactobacillus/genetics , Operon/genetics , Amino Acid Sequence , Base Sequence , Blotting, Northern , DNA-Binding Proteins , Ethanol/pharmacology , HSP40 Heat-Shock Proteins , Heat-Shock Proteins/genetics , Heat-Shock Response/genetics , Lactobacillus/drug effects , Lactobacillus/growth & development , Molecular Sequence Data , Promoter Regions, Genetic/genetics , RNA, Bacterial/analysis , RNA, Messenger/analysis , Repressor Proteins/genetics , Sodium Chloride/pharmacology
16.
J Pediatr Hematol Oncol ; 21(6): 486-93, 1999.
Article in English | MEDLINE | ID: mdl-10598659

ABSTRACT

The outcome for 82 pediatric patients with Ewing sarcoma (ES) and primitive neuroectodermal tumor (PNET) of bone is reported; the patients were treated at the Dana-Farber Cancer Institute (DFCI) and Children's Hospital (CH) in Boston, MA (USA) from 1971-1988. The charts of all patients with ES/PNET of bone treated during this period were reviewed for disease status, therapy, sites of relapse, information on second malignancies, and survival status. Eighty-two patients with ES/PNET of bone treated at DFCI/CH were identified. The 10-year event-free survival (EFS) rates were 12% (95% confidence interval [CI] 0, 27%) and 38% (95% CI 26, 51%) for patients with and without metastases, respectively (P = 0.002); the overall survival (OS) rates were 17% (95% CI 1, 33%) and 48% (95% CI 35, 61%) for patients with and without metastases (P = 0.001). Median follow-up for surviving patients is 10.2 years. Primary site in the pelvis also was associated with a poor outcome for patients with no metastatic disease (P = 0.006 OS, P = 0.03 EFS). Thirty-one patients survived in first remission at least 5 years from diagnosis, and of these, five experienced relapse of original disease, and five experienced secondary malignancies. Pediatric patients treated for ES/PNET of bone remain at risk for life-threatening events into the second decade of follow-up. After 5 years, the risk of second malignant neoplasm is at least as high as the risk of late relapse. Prolonged follow-up of patients with ES and PNET of bone is indicated.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Bone Neoplasms/drug therapy , Neuroectodermal Tumors, Primitive/drug therapy , Sarcoma, Ewing/drug therapy , Adolescent , Adult , Bone Neoplasms/mortality , Bone Neoplasms/pathology , Child , Child, Preschool , Cyclophosphamide/administration & dosage , Dactinomycin/administration & dosage , Female , Follow-Up Studies , Humans , Male , Neoplasm Metastasis , Neuroectodermal Tumors, Primitive/mortality , Neuroectodermal Tumors, Primitive/pathology , Retrospective Studies , Sarcoma, Ewing/mortality , Sarcoma, Ewing/pathology , Survival Rate , Time Factors , Treatment Outcome , Vincristine/administration & dosage
17.
J Food Prot ; 62(10): 1150-6, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10528718

ABSTRACT

A polymerase chain reaction-based system for detection of Staphylococcus aureus was developed. The system consisted of the following components: (i) selective enrichment, (ii) DNA isolation, (iii) amplification of DNA with primers targeted against the 23S rRNA gene, and (iv) evaluation of the specificity of the polymerase chain reaction by Southern hybridization and nested polymerase chain reaction. The method achieved a high degree of sensitivity and unambiguity as required for the detection of contaminants in food starter preparations. The method permitted detection of Staphylococcus aureus in preparations of meat starter cultures containing Staphylococcus carnosus either alone or in combination with lactobacilli, pediococci, and/or Kocuria varians. Detection limits were sufficiently low to show within 12 h the presence of 10(0) CFU of S. aureus in starter preparations containing 10(10) CFU of S. carnosus. The system was also applied to dried skim milk and cream. For detection without selective enrichment, a protocol was developed and permitted detection of 120 CFU of S. aureus in 1 ml of cream within 6 h. With nested polymerase chain reaction, the detection limit was decreased by one order of magnitude.


Subject(s)
DNA, Ribosomal , Dairy Products/microbiology , Meat/microbiology , Polymerase Chain Reaction/methods , Staphylococcus aureus/isolation & purification , Animals , Milk/microbiology , Oligonucleotides/chemistry
18.
J AOAC Int ; 82(4): 923-8, 1999.
Article in English | MEDLINE | ID: mdl-10490320

ABSTRACT

This paper presents results of a collaborative trial study (IUPAC project No. 650/93/97) involving 29 laboratories in 13 countries applying a method for detecting genetically modified organisms (GMOs) in food. The method is based on using the polymerase chain reaction to determine the 35S promotor and the NOS terminator for detection of GMOs. reference materials were produced that were derived from genetically modified soy beans and maize. Correct identification of samples containing 2% GMOs is achievable for both soy beans and maize. For samples containing 0.5% genetically modified soy beans, analysis of the 35S promotor resulted also in a 100% correct classification. However, 3 false-negative results (out of 105 samples analyzed) were reported for analysis of the NOS terminator, which is due to the lower sensitivity of this method. Because of the bigger genomic DNA of maize, the probability of encountering false-negative results for samples containing 0.5% GMOs is greater for maize than for soy beans. For blank samples (0% GMO), only 2 false-positive results for soy beans and one for maize were reported. These results appeared as very weak signals and were most probably due to contamination of laboratory equipment.


Subject(s)
Food Analysis , Genetic Engineering , Glycine max/genetics , Plants, Genetically Modified , Polymerase Chain Reaction , Zea mays/genetics , Agrobacterium tumefaciens/genetics , Caulimovirus/genetics , Electrophoresis, Polyacrylamide Gel , False Negative Reactions , False Positive Reactions , Promoter Regions, Genetic , Terminator Regions, Genetic
19.
Syst Appl Microbiol ; 22(2): 161-8, 1999 May.
Article in English | MEDLINE | ID: mdl-10390866

ABSTRACT

The plasmid transfer by transformation of Escherichia coli in 12 foods was investigated under conditions commonly found in processing and storage of food. Transformation occurred in all foods with frequencies of at least 10(-8) when a simplified standard transformation protocol with non-growing cells was applied. Higher rates (ca. 10(-7)) were found in milk, soy drink, tomato and orange juice. Furthermore, E. coli became transformed at temperatures below 5 degrees C, i.e. under conditions highly relevant in storage of perishable foods. In soy drink this condition resulted in frequencies which were even higher than those determined after application of a temperature shift to 37 degrees C. The transformation of cells growing in milk and carrot juice at a constantly kept temperature of 37 degrees C provides evidence for the potential of E. coli to become transformed naturally. With purified DNA frequencies were determined in these substrates of ca. 2.5 x 10(-7) and 2.5 x 10(-8), respectively. Similar frequencies were also obtained in milk containing the crude nucleic acids of homogenised cell suspensions of E. coli (pUC18). Moreover, the release of plasmid DNA from E. coli during food processing and the subsequent uptake of this DNA by growing E. coli cells was shown to take place after homogenisation in milk indicating a horizontal plasmid transfer by transformation of E. coli.


Subject(s)
Escherichia coli/genetics , Transfection/genetics , Transformation, Bacterial/genetics , Animals , Beverages/microbiology , Culture Media/chemistry , Escherichia coli/growth & development , Food Handling , Milk/microbiology , Plasmids/genetics , Soybean Proteins , Temperature , Time Factors
20.
J Neurosci Res ; 57(3): 399-404, 1999 Aug 01.
Article in English | MEDLINE | ID: mdl-10412031

ABSTRACT

In clinical studies, it has been shown that estrogen replacement therapy in menopause is strongly correlated with a reduced risk of the development of Alzheimer's disease (AD). In in vitro experiments, it was demonstrated that estradiol protects cells against the toxic effects of beta-amyloid, the major component of plaques in brains of AD patients. Therefore, estrogens have become interesting candidates for a possible treatment of neurodegeneration. In plants, a class of compounds has been identified that bind to human estrogen receptor, so-called phytoestrogens, which are part of our daily diet. Here, we compared the effects of alpha- and beta-estradiol with plant-derived kaempferol on beta-amyloid peptide-induced toxicity in PC12 neuroblastoma and T47D human breast cancer cells. The present results demonstrate a protective effect of kaempferol comparable to that observed with estradiol. The effects of the weak estrogen receptor agonists alpha-estradiol and kaempferol were found to be similar to the effects of the strong estrogen receptor agonist beta-estradiol, suggesting a mode of action independent from the nuclear estrogen receptor.


Subject(s)
Amyloid beta-Peptides/toxicity , Flavonoids , Kaempferols , Neuroprotective Agents/pharmacology , Quercetin/analogs & derivatives , Alzheimer Disease/prevention & control , Animals , Drug Evaluation, Preclinical , Estradiol/pharmacology , Humans , L-Lactate Dehydrogenase/metabolism , PC12 Cells , Quercetin/pharmacology , Rats , Receptors, Estrogen/drug effects , Tumor Cells, Cultured
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