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PubMed1999

Short communication: emission of nitrous oxide (N2O) from transgenic tobacco expressing antisense NiR mRNA

The emission of N2 and N2O from intact transgenic tobacco (clone 271) expressing antisense nitrite reductase (NiR) mRNA, and wild-type plants grown aseptically, on NO3-, NO2- or NH4+ -containing medium was investigated. 15N contents of gas sampled from gas-sealed pots, in which the plants were grown on 15N-containing medium, were analyzed by gas chromato- graphy and mass spectrometry (GC-MS). No emission of N2 was detected in either of the gas samples from plant clone 271 or the wild-type grown on NO3--containing medium. N2O emission from clone 271 grown on NO3--containing medium was detected, but not from the wild-type plants. The N2O emission rate of clone 271 was 106 ng N2O mg-1 incorporated N week-1 and the N2O emission was inhibited by tungstate (a nitrate reductase inhibitor). No emission of N2O was found from clone 271 or wild-type plants grown on medium containing NH4+. Emission of N2O also was detected from clone 271 grown on NO2--containing medium and its emission rate increased with increasing NO2- levels in plants. We speculate that NO3- is reduced to NO2- and that a part of NO2- is metabolized to N2O in clone 271.

Journal Article↗
PubMedDate not supplied

Characterization of antibiotic-resistant bacteria in rendered animal products.

Antibiotics are used in food animal production to treat diseases and also to improve performance. Antibiotics are not used on all farms, and antibiotic resistance is occasionally found on farms that do not use antibiotics. Rendered animal protein products are often included in poultry feeds and could potentially serve as a source of antibiotic-resistant bacteria. One hundred sixty-five rendered animal protein products from cattle, poultry, and fish were aseptically collected from poultry feed mills. Fifty-five percent of the poultry meal samples had detectable levels of gram-negative bacteria ranging from 40 to 10,440 colony-forming units/g of sample. Poultry meal and meat and bone meal had the greatest number of samples with bacteria resistant to five or more antibiotics. A high percentage of feed samples (85%) contained bacteria resistant to amoxicillin, ampicillin, clavulanic acid, or cephalothin, whereas few samples contained bacteria resistant to ciprofloxacin, kanamycin, or trimethoprim/sulfamethoxazole. Acinetobacter calcoaceticus, Citrobacter freundii, and Enterobacter cloacae were the most commonly isolated antibiotic-resistant bacteria. Isolation for Salmonella was also performed, with 14% of the meat and bone meal samples containing Salmonella sp. Only one of the meat and bone meal isolates, Salmonella livingstone, was resistant to five or more antibiotics. Many of the antibiotic-resistant bacteria contained integrons, genetic elements that mediate multiple drug resistance.

Animal Feed↗
PubMedDate not supplied

Cytotoxicity of denture base resins: effect of water bath and microwave postpolymerization heat treatments.

PURPOSE: This study compared the effect of two postpolymerization heat treatments on the cytotoxicity of three denture base resins on L929 cells using 3H-thymidine incorporation and MTT assays. MATERIALS AND METHODS: Sample disks of Lucitone 550, QC 20, and Acron MC resins were fabricated under aseptic conditions and stored in distilled water at 37 degrees C for 48 hours. Specimens were then divided into three groups: (1) heat treated in microwave oven for 3 minutes at 500 W; (2) heat treated in water bath at 55 degrees C for 60 minutes; and (3) no heat treatment. Eluates were prepared by placing three disks into a sterile glass vial with 9 mL of Eagle's medium and incubating at 37 degrees C for 24 hours. The cytotoxic effect from the eluates was evaluated using the 3H-thymidine incorporation and MTT assays, which reflect DNA synthesis levels and cell metabolism, respectively. RESULTS: The components leached from the resins were cytotoxic to L929 cells when 3H-thymidine incorporation assay was employed. In contrast, eluates from all resins revealed noncytotoxic effects as measured by MTT assay. For both MTT assay and 3H-thymidine incorporation, the heat treatments did not decrease the cytotoxicity of the materials tested. CONCLUSION: Resins were graded by 3H-thymidine incorporation assay as slightly cytotoxic and by MTT assay as noncytotoxic. Cytotoxicity of the denture base materials was not influenced by microwave or water bath heat treatment.

Acrylic Resins↗
PubMed2004

[Microbiological findings in children with an indwelling central venous catheter].

INTRODUCTION: Besides their obvious advantages for the patient, central venous catheters (CVC) also carry the risk of possible infectious complications. The purpose of our investigation was to carry out a microbiological evaluation of a 5-year set of paediatric patients with indwelling CVCs. PATIENTS AND METHODS: In the group were 218 CVCs inserted to 165 children over a period of 5 years. There were 26 multi-lumen catheters (11.927 %) and 192 single-lumen catheters (88.073 %). The mean indwelling period was 10.1 days per 1 CVC. Blood for microbiology was removed by a physician from the CVC after disinfecting its opening under standard sterile conditions into a commercial sampling vessel HEMOD (Imuna, Sarisske Michalany, Slovak Republic) or into a vessel of an automated haemoculture system BactecPeds PLUS/F (Becton Dickinson and Comp., Spark,MA, USA). When removing the tip of the CVC we disinfected, before removing the CVC, the area around the insertion with isopropyl or ethyl alcohol. We released the fixed CVC and 1 minute after disinfection we pulled out the CVC and cut off the end or rather the tip of the catheter (approx. 1-3 cm of the tip) into a sterile test tube. To establish the diagnosis of infectious complications we used the 1995 Sirges-Serra classification and the CDC criteria. RESULTS: In 5 years (1995-1999) we had 71 infectious complications. There were 31 contaminated catheters, 27 cases of catheter sepsis and 11 cases of catheter bacteraemia. With 147 catheters (67.43%) there were no infectious complications. Dominant microbes were Staphylococcus epidermidis (32 cases - 11 from haemocultures and 21 from CVCs) and Candida spp. (30 cases, 17 from haemocultures and 13 from CVCs). Among the microbiological agents of catheter sepsis predominated Gram-negative bacteria. Out of the whole analysed group 41 children (24.8 %) died. CVC as the cause of death was demonstrated in 6 children (3.636 % of patients with CVC). CONCLUSIONS: Microbiological findings in our group are in line with literary data. To reduce the incidence of infectious complications it is important to limit sampling from CVC to a minimum. Insertion of CVCs under strict sterile conditions and aseptic handling of all entries into the central bloodstream reduces to a minimum the risk of infectious complications.

Bacteremia↗
PubMedDate not supplied

[Contamination of enteral diet solutions in nosocomial environment].

The gastrointestinal tract is comparatively resistant to food-born germs, but recent studies suggest that nosocomial infections may be triggered by this route. In a study with industrialized diets, prepared with aseptic technique and stored up to 24 hours, aerobic and anaerobic contaminants were searched. Samples were taken after 0.8 and 24 hours, whereas half of these last two analyses were carried out in material left at room temperature, and the other half in refrigerated diets. Initial examination revealed 50% of positive cultures, but part of this was due to non-pathogenic Bacillus germs. After 8 and 24 hours 90% of the samples grew organisms, again with a large proportion of Bacillus, but also with several Gram-negative bacteria, as well as rare Gram-positives. Diarrhea and fever were not registered in patients submitted to enteral nutrition during the study period, nor could any episodes of bacteremia or septic shock be attributed to contaminated feeding material. This lack of clinical consequences of the reported bacterial isolations is not unexpected, and suggests that low concentrations of microorganisms were probably present in the preparations, below a critical level. Nevertheless, attention will be required in the future for better quality control of enteral nutrition mixtures, specially when resistant strains of Gram-negative species are identified, and also in the management of debilitated or immunologically compromised hosts.

Bacteria, Aerobic↗
PubMed1988

Cross reactivity among human enterovirus serotypes as revealed by microneutralization assay technique.

Paired or tripled serum specimens examined for cross reactions between poliomyelitis viruses and other non-polio enteroviruses were obtained from 356 patients (217 males and 139 females) with various clinical diagnoses (aseptic meningitis, meningoencephalitis, paretic neuro-infections, upper respiratory tract infections, bronchopneumonia) who were sampled in 1984 and 1985. The sera were examined by the microneutralization assay technique in flat-bottomed 96-well polyvinyl chloride plates against 100-300 TCD50 of these enterovirus serotypes: CA9, CB1-CB5, E1, E2, E4-E9, E17, E20, E24, E30, EV71 and polioviruses P1-P3. The tests were carried out on BGM or RD cells. Confirmed significant seroconversions to at least one enterovirus serotype were observed in 87 (24.2%) patients. Of these, 27 (31%) reacted by parallel significant or insignificant increases in the heterotypic antibody titre to poliovirus, which accounted for 7.6% of all patients examined. Heterotypic antibody responses may become source of errors in individuals examined for poliovaccine efficacy or the state of specific immunity response to polio, but are unlikely to discredit the overall outcomes of the seroepidemiological surveys conducted in the general population.

Antibodies, Viral↗
PubMed1993

Biocompatibility of polyethylene and host response to loosening of cementless total hip replacement.

A human lymphocyte culture protocol was used to identify the biocompatibility pattern of fine particulate ultra-high molecular weight polyethylene. Polyethylene did not cause an increase in lymphocyte DNA synthesis as assessed by the 3H-thymidine incorporation method on culture Days 1, 3, nor 5. As analyzed with monoclonal activation markers, the polyethylene dependent expression of major histocompatibility complex (MHC) class II antigen as well as interleukin-2 receptor (CD25) was virtually nonexistent. An apparent increase in the amount of CD11b positive monocytes/macrophages from 7% +/- 2% to 22% +/- 6% was recorded. Samples of pseudocapsules of the totally replaced hips (THR) obtained at revision operations for aseptic loosening of cementless prostheses with polyethylene lining of the acetabular component, were obtained from ten patients for immunopathologic studies. In seven cases the prostheses consisted of chromium-cobalt-molybdenum steel alloy and in three cases of titanium. Revisions were performed on average 4.6 years (range, 2.5-7) after insertion of the prostheses. The predominant cell in the lining cell layer of the periprosthetic cavity was in each case the CD11b, CD68, and nonspecific esterase positive but endogenous peroxidase-negative macrophage. Proline 4-hydroxylase positive fibroblasts dominated the stroma that was also inhabited by usually perivascular mononuclear cell infiltrations of mainly CD11b/CD68 phenotype with occasional CD4-positive cells. Only few mononuclear cells were activated CD25 positive T cells or CD19-positive B-lymphocytes. In titanium-based THRs, the cytoplasm of the macrophages contained a large number of small metallic particles, although this phenomenon was not seen in chromium-cobalt-molybdenum steel-based THRs. Fine particulate ultra-high molecular weight polyethylene is immunologically relatively insert. Nevertheless, it causes a clear foreign body type of phenomenon in vitro. The loosening of cementless acetabular components was associated with CD11b- and CD68-positive macrophage reaction in the pseudocapsular tissue. In any case, there is no clinical or experimental evidence to suggest that the use of cementless THR prostheses with polyethylene sockets would prevent an adverse biologic host response.

Adult↗
PubMed2004

Isolation and molecular identification of echovirus 13 isolated from patients of aseptic meningitis in Korea, 2002.

During 2002, several epidemics of aseptic meningitis were attributed to echovirus 13 in Korea. The causative agents of these outbreaks were isolated and identified using rhabdosarcoma cells, HEp-2 and Buffalo green monkey kidney cells, and a neutralization test using monospecific antiserum. Fifty-four echovirus 13 isolates were isolated from patients with aseptic meningitis in the provinces, Seoul, Kyonggi, Gwangju, Jeonju, Busan, and Ulsan. Symptoms associated with aseptic meningitis infection in patients included the occurrence of headaches and mild fever. Molecular characterization of echovirus 13 samples was achieved by sequence and phylogenetic analyses on partial VP1 sequences from 20 Korean isolates and 10 foreign isolates listed in Genbank. Minor variation was observed among the Korean isolates, which formed a unique cluster with isolates of German and Japanese origin. The marked similarities between isolates could be attributed to a relatively recent arrival of the virus in Korea. This is the first such investigation of aseptic meningitis caused by echovirus 13 on the Korean peninsula.

Animals↗
PubMed2002

Detection of infectious enteroviruses and adenoviruses in tap water in urban areas in Korea.

We investigated the viral contamination of tap water at 11 urban sites in Korea between 1997 and 1998 over a period of 11 months. A total of 23 tap water samples were examined for infectious enteroviruses and adenoviruses by a cell culture technique followed by polymerase chain reaction (PCR) amplification. To identify the recovered viruses, sequence analysis of PCR products was performed. Infectious enteroviruses and adenoviruses were detected in 11 (47.8%) and 9 (39.1%) of the samples, respectively. Both enteroviruses and adenoviruses were detected in five samples (21.7%). The level of viral contamination was quite high, ranging from 2 x 10(-3) to 2.9 x 10(-2) Most Probable Number of Infectious Unit L(-1), far above the recommended virus level in drinking water set by the US EPA. Poliovirus type I derived from vaccine was frequently detected and the remainder comprised coxsackievirus B type or echovirus type 6, which were causative agents of aseptic meningitis in Korea in 1997 and 1998, respectively. Several types of adenovirus were detected in tap water samples and some water samples were found to contain adenoviruses which were closely related to enteric adenovirus types 40 and 41.

Adenoviridae↗
PubMed2001

A study of U.S. orchards to identify potential sources of Escherichia coli O157:H7.

The association of unpasteurized apple cider with Escherichia coli O157:H7 foodborne illness has led to increased interest in potential reservoirs of this pathogen in the orchard. Fourteen U.S. orchards were surveyed in autumn 1999 to determine the incidence and prevalence of E. coli O157:H7, E. coli, total aerobic microflora, and yeasts and molds. Fruit samples (n = 63) (eight apple and two pear varieties) and soil, water, and fecal samples were collected. Samples were plated on (i) tryptic soy agar for total mesophilic aerobic count, (ii) E. coli and coliform Petrifilm for total coliforms and E. coli, and (iii) yeast and mold Petrifilm. Samples positive for coliforms and E. coli were enriched and tested for E. coli O157:H7. Fruit was also tested for internalization of microflora by aseptically removing the core, stem, and calyx areas, and the individual sections were assessed for the categories of microflora listed above. E. coli was detected in soil and water and in 6% of fruit samples (three pear samples and one apple sample), generally collected from areas previously designated as high risk in this study. However, no E. coli O157:H7 was found. Coliforms were found in 74% of fruit samples and were internalized in the cores of 40% of fruit tested. Yeasts and molds were internalized in 96.7% of samples and aerobic bacteria in 89.6%. E. coli was not found to be internalized. Total aerobic counts and total coliforms were higher in dropped and damaged fruit (P < 0.05). Findings suggest that dropped or damaged fruit should not be included in fruit designated for the production of unpasteurized juice or for the fresh or fresh-cut market. In addition, orchards should be located away from potential sources of contamination, such as pastures.

Bacteria, Aerobic↗
PubMed1994

Use of subcutaneous vascular access ports in rhesus monkeys.

To investigate new approaches for treatment of drug abuse, subcutaneous vascular ports were implanted in rhesus monkeys (Macaca mulatta) for use in drug self-administration studies. Internal or external jugular veins were cannulated, and the catheter was attached to a subcutaneous port that was positioned on the back between the shoulder blades. This port/catheter system allowed easy serial blood sampling and intravenous drug administration. Daily use of the ports in awake monkeys was simple, effective, and caused no apparent stress over a 2-h experimental session. At the time of submission, the mean functional lifetime of the port/catheter system in 20 monkeys implanted longer than 6 months was 243.61 days, with the upper range being 540 days. Several complications developed with this system, and solutions to these problems are described. With proper aseptic use, the subcutaneous vascular port system provided a safe and enduring method for daily access for intravenous sampling and/or drug delivery. The duration of the preparation is a considerable improvement compared with exteriorized catheterization.

Animals↗
PubMed2004

Ductal lavage using medically aseptic technique in women at high risk for breast cancer.

Ductal lavage of the breast is a minimally invasive procedure used to collect epithelial cells from the lumen of the ductolobular system for cytologic analysis. The purpose of this study was to determine the safety of using an aseptic technique in performing ductal lavage. The study included 114 consecutive patients at high risk for breast cancer with clinically negative mammograms and no palpable masses. Ductal lavage could not be performed on 32 of the 114 patients (28%): 26 because nipple aspirate fluid could not be obtained, and 6 because a fluid-yielding duct could not be cannulated. Ductal lavage was successfully performed on 82 patients (72%). Local anesthetic cream was used on the nipple area and local anesthetic was infused into each duct. The nipple, the cup of the nipple aspirator device, and any dilator tools were cleansed with alcohol before use. Separate sterile microcatheters were used for each duct. Sterile saline solution was used for the infusions. No patient reported any symptoms indicating infection after lavage. Cytologic analysis from the 82 lavage cases showed 45 benign cells (55%), 17 cases of mild atypia (21%), 1 case of marked atypia, and 1 case with malignant cells. Eighteen samples (22%) had inadequate cellularity for medical diagnosis. The results of the study indicate that an aseptic technique is safe and appropriate for ductal lavage of the breast.

Asepsis↗
PubMed2001

Hyaluronan synthases, hyaluronan, and its CD44 receptor in tissue around loosened total hip prostheses.

Aseptic loosening of prosthetic components, the most common long-term complication after total hip replacement (THR), is characterized by the formation of a synovial membrane-like interface tissue (SMLIT). It was hypothesized that the hyaluronan synthase (HAS)/hyaluronan (HA)/HA receptor CD44 signalling system is responsible for the synovial-like differentiation of the interface membrane. SMLIT was therefore compared with osteoarthritis (OA) synovial membrane by using reverse transcriptase polymerase chain reaction (RT-PCR) of HAS 1, 2 and 3, histochemical HA assay, and immunohistochemistry of CD44 and its non-HA ligands. All three isoforms of HAS were found in these samples. HA and CD44 were most abundant in the lining, but the signal was actually stronger in aseptic loosening than in OA (p<0.01). The non-HA CD44 ligands, collagen type VI, fibronectin, osteopontin, and MCP-1, had a similar distribution pattern in both tissues. These results confirm the synovial-like structure of the interface tissue lining. The pressure waves and movement of the HA-rich pseudosynovial fluid seem to drive HA into the implant-to-host interface, which itself also produces HA. HA may be responsible for the induction of a synovial-like lining at the interface through HA-CD44 signalling.

Adult↗
PubMed2006

Hydrogen peroxide as an irrigation solution in arthroplasty - a potential contributing factor to the development of aseptic loosening.

The reason for revision of primary total hip arthroplasty is quoted as aseptic loosening in 60.6% of cases between 1979 and 2003 in the Swedish National Hip Arthroplasty Registry. Much research effort has been directed toward enhancing the bone-cement interface of total joint arthroplasties, in an attempt to reduce this complication. Haemostatic agents have been popularized as effective means of retarding the development of potentially harmful debris interposition adjacent to, and blood lamination patterns within, the methylmethacrylate. Such agents include hydrogen peroxide (H(2)O(2)), local freezing saline, saline at room temperature and adrenaline solution. One main concern with the use of hydrogen peroxide is whether it affects the material properties of bone cement such that in the long term it contributes to aseptic loosening. This would have enormous clinical consequences. Preliminary studies indicate that porosity increases and that the tensile strength and yield stresses are reduced by up to a factor of 10 by contaminating samples with increasing concentrations of hydrogen peroxide. We postulate that the current use of hydrogen peroxide as an irrigation solution in arthroplasty contributes to the development of aseptic loosening.

Adhesiveness↗
PubMed1992

High concentrations of intrathecal interleukin-6 in human bacterial and nonbacterial meningitis.

Interleukin-6 (IL-6) is multipotent cytokine that acts in a network of factors directing the inflammatory reaction of purulent bacterial meningitis (PBM). However, little is known about the role of IL-6 in aseptic or "viral" meningitis (AM). IL-6 was assayed by RIA in cerebrospinal fluid (CSF) and serum samples obtained from patients with AM (n = 65), PBM (n = 8), and lymphocytic bacterial meningitis (LBM, n = 11). Of patients with AM, 89% had detectable IL-6 in CSF, with high IL-6 titers (median, 2160 pg/mL; 95% confidence interval [CI], 1320-2540 pg/mL) compared with 100% in patients with PBM (median, 6575 pg/mL; 95% CI, 450-32,000 pg/mL) and 90.9% in patients with LBM (median, 875 pg/mL; 95% CI, 150-2180 pg/mL). There was a highly symmetrical correlation between IL-6 and the percentage of polymorphonuclear cells in CSF of patients with PBM (r = .97, P = .01) and AM (r = .49, P = .002). In conclusion, this study shows evidence that IL-6 is released into the meningeal space in aseptic meningitis and is correlated with the local acute inflammatory response.

Humans↗
PubMed1998

Herpes simplex virus type 2 DNA detected in cerebrospinal fluid of 9 patients with Mollaret's meningitis.

We present clinical and virological data on 9 patients, 7 women and 2 men aged 31-56 years, with recurrent aseptic meningitis (Mollaret's meningitis). Polymerase chain reaction detected Herpes simplex virus type 2 DNA in cerebrospinal fluid samples from all patients collected during their latest attacks of meningitis. Six patients had no history of genital herpes. Only 1 patient was offered prophylactic antiviral treatment during the study period (45 months).

Adult↗
PubMed1996

Elevations of amniotic fluid macrophage inflammatory protein-1 alpha concentrations in women during term and preterm labor.

OBJECTIVE: To determine whether elevated concentrations of macrophage inflammatory protein-1 alpha (MIP-1 alpha) in amniotic fluid (AF) are related to term and preterm labor. METHODS: Amniotic fluid was obtained from women from five different clinical situations: 1) term cesarean delivery, no labor (n = 29); 2) normal term labor, no infection (n = 36); 3) preterm labor, delivery more than 1 week from sampling, no infection (n = 19); 4) preterm labor, delivery within 1 week from sampling, no infection (n = 18); and 5) preterm chorioamnionitis (n = 8). Amniotic fluid was collected aseptically at the time of amniocentesis, amniotomy, or hysterotomy. Concentrations of MIP-1 alpha were determined by enzyme-linked immunosorbent assay. Statistical analysis was by Wilcoxon rank-sum test, Kruskal-Wallis test, and unpaired t test. RESULTS: Women in normal term labor had significant elevations of AF MIP-1 alpha concentrations when compared with women at term undergoing repeat cesarean delivery (P < .001). In women with term gestation, AF MIP-1 alpha correlated well with cervical dilation (r2 = 0.479, P < .001). In women with preterm labor who later delivered within 1 week of presentation, AF MIP-1 alpha concentrations were higher than those from women who did not deliver within 1 week. Women who presented with clinically evident chorioamnionitis had the highest concentrations of AF MIP-1 alpha (P = .001). CONCLUSION: Women in labor have significantly elevated AF concentrations of MIP-1 alpha, particularly if labor is associated with intrauterine infection. We suggest that MIP-1 alpha is involved in the physiology of normal labor and in the pathogenesis of infection-associated preterm labor.

Amniotic Fluid↗
PubMed2000

Changes in mRNA abundance within Heterodera schachtii-infected roots of Arabidopsis thaliana.

Gene expression changes in plant roots infected by plant-parasitic cyst nematodes are involved in the formation of nematode feeding sites. We analyzed mRNA abundance changes within roots of Arabidopsis thaliana during the early compatible interaction with Heterodera schachtii, the sugarbeet cyst nematode. Approximately 1,600 root sections, each containing a single parasitic nematode and its feeding site, and 1,600 adjacent, nematode-free root sections were excised from aseptic A. thaliana cultures 3 to 4 days after inoculation with H. schachtii. These tissue samples were termed infected and uninfected, respectively. Preparasitic nematodes were added to the uninfected tissue sample to maintain the nematode to plant tissue proportion. mRNA extracted from these two tissue samples was subjected to differential display analysis. Thirty-six cDNA clones corresponding to mRNA species with different abundance between both tissue samples were isolated. Of these clones, 24 were of A. thaliana origin and 12 were from H. schachtii. Differential display data predicted that the A. thaliana cDNA clones corresponded to 13 transcripts that were more abundant in the infected root sections and 11 transcripts that were more abundant in the uninfected root sections. H. schachtii cDNA clones were predicted to correspond to four transcripts that were more abundant in parasitic nematodes and to eight transcripts that were more abundant in preparasitic nematodes. In situ hybridization experiments confirmed the mRNA abundance changes in A. thaliana roots predicted by the differential display analyses for two A. thaliana clones.

Animals↗