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PubMed2005

Preliminary investigation of bacteremia incidence after removal of the Haas palatal expander.

BACKGROUND: The aim of this study was to investigate the presence of bacteria in the bloodstream immediately after removing Haas palatal expanders; this could be considered an invasive procedure. METHODS: Eight patients (18.5 +/- 3.9 years old) wearing Haas palatal expanders had 5-mL blood samples taken immediately before and 10 minutes after removal of the apparatus. The blood was aseptically inoculated into hemoculture bottles and incubated at 37 degrees C for 7 days. Medium alterations suggesting bacterial growth were investigated by using Gram staining. RESULTS: Bottles containing blood taken before apparatus removal did not show bacterial growth. However, 4 of the 8 postremoval blood samples showed turbidity after 5 days, and bacterioscopy analysis showed Gram-positive cocci. Subsequent cultures in blood-agar medium plates allowed classifying the bacteria as viridans streptococcus. CONCLUSIONS: There is a strong possibility that trauma after removing Haas expanders might be correlated to transitory bacteremia, thereby implying the need for a more careful approach when dealing with special patients, such as those at risk of cardiopathic complications.

Adolescent↗
PubMed1995

Survey of intramammary infections in dairy heifers at breeding age and first parturition.

A survey was conducted to determine and contrast prevalence of IMI in nulligravid and primigravid dairy heifers pre- and postpartum. Contrasts were made to evaluate the risk factors of location of dairy, trimester of gestation, and season of sampling on IMI. Twenty-eight dairies in California, Louisiana, Vermont, and Washington were studied. Lacteal secretions were collected aseptically from heifers at breeding age (8 to 19 mo) from one side of the gland and again at 4 d postpartum from all quarters. Of the quarters sampled, 65.6% prepartum and 64.0% postpartum were free of IMI. The percentages of quarters with IMI from coagulase-negative staphylococci or Staphylococcus aureus IMI were 27.1 and 9% prepartum and 21.8 and 2.9% postpartum. Staphylococcus aureus IMI were most prevalent in Louisiana during the months other than summer. Location, herd, and season significantly influenced prevalence of IMI. The prevalence of IMI was greatest during the last trimester of pregnancy, ranging from 49.2% in the winter to 36.8% in the summer. The significant effects of herd location and season suggest that management variables influence prevalence of heifer IMI. Because prevalence of IMI was greatest during the last trimester of pregnancy compared with prevalence during earlier stages of pregnancy, the heifer may be most susceptible to this disease during this period of first gestation.

Aging↗
PubMed1984

Use of flow cytometry in industrial microbiology for strain improvement programs.

A flow cytometry (FCM) system was chosen to analyze and sort microbiological samples, e.g., bacteria, bacterial spores, yeasts, and fungal spores, without major changes in the commercially available state. The system was further improved by addition of a stepping motor-driven scanning table that accepts standard petri dishes or microtiter plates. The electronics of the sorting system were changed to enable the sorter to deliver only one particle at a time, working in a "handshake" mode with the scanning table. Appropriate parameters, depending on the biological material and including all fluorescent stains that do not impair growth and productivity of cells were chosen to sort distinct bioparticles under aseptic conditions and to clone colonies or cultures out of them. A mutagenized sample of spores entering the germination cycle can be followed and thus provide a means to pick only viable growing cells despite the killing effect of the mutagen. One example of a typical strain improvement is illustrated. From a spore suspension of Rhizopus arrhizus, a subpopulation of morphologically different spores comprising about 5-10% of the whole population was cloned. From approximately 8,000 clones, 10 were isolated that produced approximately five- to six-fold the amount of fungal lipase activity, compared to the original strain or to reisolated clones from the mean population of clones.

Bacteria↗
PubMed2001

[Crohn's disease complicated by multiple and recurrent aseptic splenic abscesses].

We report a case of multiple aseptic splenic abscesses occurring in a woman with Crohn's disease for three years. All microbiological samples were negative. The diagnosis was suspected on abdominal echography and CT scan and confirmed on histologic examination of the splenectomy specimen. The evolution was marked by recurrence of fever and inflammatory syndrome, associated to transitory morphological abnormalities of the accessory spleen. Aseptic splenic abscesses recurrence was suspected. This case allows us to consider aseptic splenic abscesses as an extra-intestinal manifestation and not as a splenic localisation of Crohn's disease.

Abscess↗
PubMedDate not supplied

Single-stage revision using an uncemented, porous-coated, anatomic endoprosthesis in two dogs: case report.

OBJECTIVE: To describe the clinical and radiographic features of septic and aseptic failure of two femoral endoprostheses and their successful revision. STUDY DESIGN: Case report. ANIMALS OR SAMPLE POPULATION: Two skeletally mature male research dogs. METHODS: An uncemented porous-coated anatomic (PCA) endoprosthesis was implanted in a single-stage revision procedure after thorough debridement and lavage of the femoral canal. An autogenous cancellous bone graft was used in dog 2 (aseptic loosening). Serial clinical and radiographic examinations were performed postoperatively. The dogs were euthanatized 1 year (dog 1) and 2 years (dog 2) after revision surgery, and necropsy was performed. High-resolution contact radiographs and histopathologic evaluation of femoral sections were obtained. RESULTS: The cause of implant failure was septic loosening in dog 1 and aseptic loosening in dog 2. In both dogs, clinical function returned to normal after revision. Serial radiographic assessment after revision documented disappearance of the bone pedestal and the periprosthetic lucency. Cancellous hypertrophy seen adjacent to the proximal porous-coated region of the implants provided radiographic evidence of bony fixation. Histological evaluation of femoral sections documented successful implant integration with bone and fibrous tissue. CONCLUSION: Revision with an uncemented implant in a single-stage procedure was successful in the two dogs described in this report. CLINICAL RELEVANCE: This report provides a detailed description of the clinical course and serial radiographic assessment of septic and aseptic loosening of two femoral endoprostheses. Single-stage revision is a potential treatment for either condition as demonstrated by the successful outcome in these two dogs.

Animals↗
PubMed1991

Oligoclonal free kappa and lambda bands in the cerebrospinal fluid of patients with multiple sclerosis and other neurological diseases. An immunoaffinity-mediated capillary blot study.

We describe an affinity-mediated capillary blotting technique for the detection of free kappa or lambda light chains in native cerebrospinal fluid (CSF) after isoelectric focusing in agarose gel. Interferences by light chains bound to immunoglobulins were carefully excluded. An absolute amount of 20-50 ng of free kappa or lambda Bence-Jones proteins were detectable by this method, under the form of several discrete bands with isoelectric points between 5 and 8.5. No free light chains were observed in CSF and sera from patients without neurological disorders (n = 26). Such bands were present in most CSF samples in the case of central nervous system (CNS) infections, except in aseptic meningitis. In a group of 48 multiple sclerosis (MS) patients, 44 (92%) displayed oligoclonal free kappa bands restricted to the CSF; oligoclonal IgG bands were observed in 40 cases, and oligoclonal free lambda bands in 33. In this group, the presence of CSF free light chain bands was highly correlated with their absolute levels (p less than 0.001). In other neurological diseases (n = 44), oligoclonal free kappa and free lambda bands were detected much more rarely, in seven (16%) and four (9%) cases respectively. Surprisingly, the CSF from three unrelated patients with Huntington's disease (out of five tested) contained both oligoclonal IgG and free kappa bands.

Humans↗
PubMedDate not supplied

In vitro comparison of equine cancellous bone graft donor sites and tibial periosteum as sources of viable osteoprogenitors.

OBJECTIVE: To compare the osteogenic potential of cancellous bone of conventional graft sites with that of one nonconventional site (fourth coccygeal vertebra) and to investigate the tibial periosteum as a donor site with respect to osteogenic potential. STUDY DESIGN: In vitro osteogenic cell culture system. SAMPLE POPULATION: Eight adult horses. METHODS: Cancellous bone or tibial periosteum was aseptically collected and cut into bone chips or periosteal strips of 1 to 2 mm(3) for primary explant cultures. After 2 weeks, primary tissue cultures that yielded a population of osteogenic cells were counted and subcultured at 1 x 10(5) cells/35-mm dish in osteogenic media. After 7 to 10 days, subcultures were stained with Von Kossa (VK) to assess mineralized bone nodule formation. VK-positive bone nodules were counted as osteoprogenitors and compared among 3 donor sites, which provided consistent primary osteogenic cells (tuber coxae, fourth coccygeal vertebra, periosteum) using ANOVA (P <.05). RESULTS: Sternal and tibial bone yielded viable osteogenic cells from 25% and 50% of horses, respectively, whereas yields from tuber coxae, coccygeal vertebra, and periosteum were 75%, 100%, and 100%, respectively. Tuber coxae and periosteum had significantly greater numbers of osteoprogenitors compared with fourth coccygeal vertebra. CONCLUSIONS: Among the conventional donor sites, tuber coxae most consistently yielded viable osteogenic cells with an acceptable percentage of osteoprogenitors. Sternal and tibial sites were unreliable in providing osteogenic cells. Two new donor sites, the fourth coccygeal vertebra and tibial periosteum, were tissues with good osteogenic potential. CLINICAL RELEVANCE: When a source of transplantable viable osteoprogenitor cells is desired, use of the tuber coxae as a conventional donor site is warranted. Use of tibial periosteum or fourth coccygeal vertebra as reliable sources of transplantable osteoprogenitors should be considered.

Animals↗
PubMed1991

Serum C-reactive protein in childhood meningitis in countries with limited laboratory resources: a Chilean experience.

Quantitative C-reactive protein (CRP) was determined sequentially by nephelometry and photometry from a finger prick serum sample in 67 children with bacterial meningitis (BM) and 16 children with aseptic meningitis (AM). The initial mean CRP value of 180 mg/liter in children with BM differed significantly from the 12 mg/liter found in those with AM (P less than 0.001). In BM a slow descent instead of rapid normalization or a secondary increase in sequential CRP values were early indicators of complications during recovery, such as resistance to the antibiotic. A significant difference in the mean CRP values between uneventful and complicated courses of BM was observed from the fourth day on (P less than 0.001). The measurements obtained with nephelometry correlated reliably with the more widely available photometry (r = 0.99). Easily performed rapid CRP determinations can considerably improve the quality of care in meningitis patients, especially in those situations where facilities for performing bacterial cultures or antibiotic susceptibility testing are not available.

C-Reactive Protein↗
PubMed1970

An automated method for the measurement of folate activity.

A new approach to the estimation of folate activity is described. The system may be used as a semi-automated method or as a fully automated method using the AutoAnalyzer or Mecolab equipment. The methods use a chloramphenicol-resistant strain of L. casei as the test organism, and this eliminates the need for sterilization or aseptic addition. The systems are able to handle extremely large numbers of samples in a relatively short time. Results are similar to those obtained by standard techniques.

Autoanalysis↗
PubMed2001

Disinfection/sterilization of extracted teeth for dental student use.

Extracted human teeth are used in many preclinical courses. While there has been no report of disease transmission with extracted teeth, sterilization of teeth used in the teaching laboratory should be a concern. The purpose of this study was to determine the effectiveness of different sterilization/disinfection methods of extracted human teeth using Bacillus stearothermophilus, a bacteria resistant to heat and frequently used to test sterilizers. In this study, 110 extracted molars with no carious lesions were collected and stored in buffered saline. An endodontic occlusal access preparation was cut into the pulp chamber of each tooth. Pulp tissue in the chamber was removed with a broach. Approximately 1 x 10(5) B. stearothermophilus endospores in culture medium were injected into the pulp chamber, sealed with Cavit G, and then placed in sterile saline for twelve hours. Ten teeth were placed into each of eleven groups. Seven groups were immersed for one week in one of the following solutions: a) sterile saline (control group), b) 5.25% NaOCl, c) 2.6% NaOCl, d) 1% NaOCl, e) 10% buffered formalin, f) 2% gluteraldehyde, g) 0.28% quaternary ammonium. Four additional groups were treated by h) 10% formalin for two days, i) 10% formalin for four days, j) autoclaving at 240 degrees F and 20 psi for twenty minutes, and k) autoclaving at 240 degrees F and twenty psi for forty minutes. Each tooth was then aseptically split and placed in an individual test tube with growth medium. Samples were examined for evidence of growth (turbidity) at forty-eight hours. Only autoclaving for forty minutes at 240 degrees F and 20 psi or soaking in 10 percent formalin for one week were 100 percent effective in preventing growth. A chi-square analysis of the data indicates these two methods were significantly better than all other methods (p<0.001).

Chi-Square Distribution↗
PubMed1986

Fragment D-dimer levels: an objective marker of vaso-occlusive crisis and other complications of sickle cell disease.

Although abnormalities in coagulation tests have been reported during vaso-occlusive crises in patients with sickle cell disease, objective, readily performed laboratory tests that document the occurrence of this complication have not been available. We examined the relationship between fibrin D-dimer levels and the occurrence of complications in patients with sickle cell disease, using a commercially available latex bead agglutination assay. The patients were either asymptomatic, hospitalized for vaso-occlusive crisis, or had other complications of sickle cell disease including leg ulcers, chronic cholecystitis, aseptic necrosis, joint pain and infection. Fifty-seven percent of 187 samples on 96 patients had elevated levels of fibrin D-dimer. Ninety percent of 75 samples from asymptomatic patients were negative for fibrin D-dimer (less than 1 microgram/ml) but 97% of 29 samples from patients with vaso-occlusive crisis and 85% of 83 samples from patients with other complications of sickle cell disease were positive. In serial studies, worsening or amelioration in clinical complications were reflected in increasing or decreasing levels of fibrin D-dimer, respectively. The molecular species of fibrin identified by the latex agglutination test was shown to be fragment D-dimer by successive immunoprecipitation and protein blot analysis. We conclude that the complications of sickle cell disease, including vaso-occlusive crisis, result in the production of fibrin D-dimer, and its detection may be used as a marker for the presence of the complication.

Anemia, Sickle Cell↗
PubMed2006

Simple differentiation method of mumps Hoshino vaccine strain from wild strains by reverse transcription loop-mediated isothermal amplification (RT-LAMP).

Mumps virus is still circulating and annual mumps outbreaks occur with fluctuating magnitudes in Japan. Aseptic meningitis has been reported after vaccination and it would be of importance to determine whether this was related to the vaccination. The objective of this study was to develop a sensitive, specific and rapid diagnostic method for the differentiation of the Hoshino vaccine strain from circulating wild types. We developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method of the hemagglutinin neuraminidase (HN) region for the detection of mumps virus genome from clinical samples. The typical ladder pattern disappeared after the LAMP products of the Hoshino vaccine strain were digested with ScaI, but those of wild types were not cut by ScaI. We obtained 19 cerebro spinal fluids (CSF) from the patients with aseptic meningitis and 17 salivary swab samples from the patients with acute parotitis after mumps vaccination, in which one case was complicated with orchitis. Mumps virus genome was detected in 18 CSF samples and in all NPS by RT-LAMP. The Hoshino vaccine strain was identified in 16 out of 18 CSF RT-LAMP positives and in 11 out of 17 NPS samples and the remaining samples were identified as wild types. RT-LAMP followed by ScaI digestion is a sensitive, simple and rapid differential method and useful for laboratory surveillance for vaccine-adverse events.

Base Sequence↗
PubMed1992

Ultrasonographic and quantitative histologic assessment of sequelae to testicular biopsy in stallions.

A sample of testicular parenchymal tissue, approximately 2 x 7 x 7 mm, was aseptically removed from 1 testis in each of 9 stallions on day 0. Slight to moderate hemorrhage from the tunica albuginea was observed in 8 stallions, but bleeding from the parenchyma was detected in only 2 stallions. Stallions were castrated 27 days later. Normal development of granulation tissue was evident at the biopsy site, but hematomas were not observed. In situ measurement of the widths of the right and left testes, total scrotal width, and evaluation of testicular echogenicity during ultrasonography were variables used to monitor changes in the testicular parenchyma from 14 days before biopsy through 27 days after biopsy. The control testis was consistently larger than the biopsied testis, except for day 3. Ultrasonography revealed signs of a localized change in the parenchyma of the biopsied testis in 4 stallions, but each lesion decreased in size by day 27. Tissues removed during biopsy enabled an excellent appraisal of spermatogenesis at that time. Detailed examinations of seminiferous tubules in the testes were performed to assess for damage to testicular function. At castration, samples were taken from 6 sites in each testis. Quantitative histologic evaluations of testicular tissues revealed low numbers of spherical spermatids and pachytene spermatocytes in biopsied testes, compared with control testes. It was concluded that there was a transitory increase in degeneration of preleptotene spermatocytes and B spermatogonia at the time of biopsy. A mild inflammatory response at the biopsy site in some testes was evidenced by an increased number of leukocytes at the biopsy site and at a dorsal site.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Legionella pneumophila grows adherent to surfaces in vitro and in situ.

Legionella pneumophila continues to play a role in both community- and nosocomially-acquired pneumonia. We investigated the ability of L pneumophila to adhere to various types of materials such as those found in the hospital air-cooling and portable water distribution systems. Through the use of a unique sampling apparatus, we were able to regularly acquire planktonic and sessile samples and determine the numbers of bacteria present in both populations, in vitro and in situ. Portions of these apparatuses could be aseptically removed for examination by scanning electron microscopy, or for the determination of the number of viable adherent L pneumophila. The number of bacteria present in each sample was determined by direct plate count, with presumptive L pneumophila colonies being positively identified by direct fluorescent antibody staining techniques. The results demonstrated that not only are legionellae capable of colonizing various metallic and nonmetallic surfaces but that they are preferentially found on surfaces. Surface-adherent bacteria may play a profound role as a reservoir of these potential pathogens in aquatic environments. Furthermore, these results suggest that any comprehensive legionella monitoring program must include not only water samples but also an examination of the adherent populations.

Air Conditioning↗
PubMed2006

Blood culture contamination in Tanzania, Malawi, and the United States: a microbiological tale of three cities.

We conducted retrospective, comparative analyses of contamination rates for cultures of blood obtained in the emergency rooms of Muhimbili National Hospital (MNH) in Dar es Salaam, Tanzania; Lilongwe Central Hospital (LCH) in central Malawi; and the Duke University Medical Center (DUMC) in the United States. None of the emergency room patients had indwelling intravascular devices at the time that the blood samples for cultures were obtained. In addition, we reviewed the contamination rates for a cohort of patients already hospitalized in the DUMC inpatient medical service, most of whom had indwelling intravascular devices. The bloodstream infection rates among the patients at MNH (n=513) and LCH (n=486) were similar (approximately 28%); the contamination rates at the two hospitals were 1.3% (7/513) and 0.8% (4/486), respectively. Of 54 microorganisms isolated from cultures of blood collected in the DUMC emergency room, 26 (48%) were identified as skin contaminants. Cultures of blood collected in the DUMC emergency room were significantly more likely to yield growth of contaminants than the cultures of blood collected in the emergency rooms at MNH and LCH combined (26/332 versus 11/1,003; P<0.0001) or collected in the DUMC inpatient medical service (26/332 versus 7/283; P<0.01). For the MNH and LCH blood cultures, lower contamination rates were observed when skin was disinfected with isopropyl alcohol plus tincture of iodine rather than isopropyl alcohol plus povidone-iodine. In conclusion, blood culture contamination was minimized in sub-Saharan African hospitals with substantially limited resources through scrupulous attention to aseptic skin cleansing and improved venipuncture techniques. Application of these principles when blood samples for culture are obtained in U.S. hospital emergency rooms should help mitigate blood culture contamination rates and the unnecessary microbiology workup of skin contaminants.

2-Propanol↗
PubMedDate not supplied

Evaluation of arthrocentesis site bacterial flora before and after 4 methods of preparation in horses with and without evidence of skin contamination.

OBJECTIVE: To evaluate the effectiveness of four methods of povidone-iodine preparation on skin bacterial flora of arthrocentesis sites, in horses, with and without evidence of skin contamination. STUDY DESIGN: Prospective randomized study. ANIMALS: Twenty-four adult horses. METHODS: Horses were assigned to either the clean or contaminated group based on housing environment and visual evidence of contamination. Using a moist sterile swab, microbial culture samples were obtained from the skin over the distal interphalangeal joints immediately before and after preparation. Each site was aseptically prepared with 1 of 4 povidone-iodine techniques: 10-minutes scrub, 5-minutes scrub, three 30-second scrubs, or commercial one-step iodophor surgical solution. Colony forming units (CFUs) were determined for each sample, 24 hours after inoculation, on blood agar plates. RESULTS: Mean (+/-SD) pre-scrub CFUs/mL was significantly higher in the contaminated group (9588.33+/-1223.65) compared with the clean group (4489.00+/-3842.03) (P<.01). After preparation of the arthrocentesis sites, there were no significant differences in post-scrub CFUs/mL among the 10 minutes (mean clean, 46.00+/-64.36; mean contaminated, 28.67+/-18.04), 5 minutes (mean clean, 84.17+/-109.80; mean contaminated, 40.33+/-44.52), three 30 seconds povidone-iodine scrubs (mean clean, 95.50+/-172.29; mean contaminated, 46.67+/-56.94), or application of a commercial one-step iodophor surgical solution (mean clean, 102.17+/-161.78; mean contaminated 117.67+/-143.78); or between the clean (81.96+/-131.69) and contaminated groups (58.33+/-85.90) (P<.01). CONCLUSIONS: Preparation of the distal interphalangeal joint arthrocentesis site with each of these techniques significantly reduces the bacterial flora to a similar level for arthrocentesis in horses with and without evidence of skin contamination. Clinical Relevance- Aseptic preparation of the skin over the distal interphalangeal joint may be accomplished with any of these techniques.

Administration, Cutaneous↗
PubMedDate not supplied

Expression of parathyroid hormone related protein in the tissue around loosened hip prostheses.

OBJECTIVE: To investigate the eventual presence and cellular source ofparathyroid hormone related protein (PTHrP) in the synovial-like interface membrane from aseptic loosening of total hip replacement (THR). METHODS: A polyclonal rabbit antiserum to the amino-terminal peptide of human PTHrP was used to stain 10 interface membrane samples from loose THR and 10 synovial tissue samples from hip osteoarthritis (OA). Quantitative microscopic assessment was done with a computer-assisted image analysis system. Western blotting was applied to verify the presence of PTHrP in both tissue samples. Double immunofluorescence labelling aimed to reveal the cellular sources of PTHrP. RESULTS: Immunoreactive PTHrP was found in all interface membrane and OA synovial tissue samples. The number of PTHrP positive cells in interface membrane was much higher than in OA synovial tissue. Positive cells were most commonly seen in the lining-like layers and sublining area of interface membrane. Double immunofluorescence labelling showed that most macrophages and fibroblasts in interface membrane were PTHrP positive. Western blotting revealed the 24-25 KD bands in both tissue samples. CONCLUSIONS: PTHrP expression is upregulated in interface membrane around loosened hip prostheses. Locally accumulated PTHrP may contribute to periprosthetic osteolysis and aseptic loosening of THR through its direct effects on bone, or indirectly via the induction of inflammatory mediators.

Aged↗
PubMed1983

Evaluating aseptic technique of pharmacy personnel.

A procedure for validating the effectiveness of a training program in aseptic technique for admixture personnel and for monitoring the aseptic technique of those trained is described. Double-strength soybean casein digest broth was prepared as the culture media. Technicians were instructed to prepare 50 sample i.v. admixtures after the structured one-week orientation and training period. The sample admixtures were cultured for sterility by a total-culturing method. An ongoing random-sampling plan was implemented to monitor the technique of the trained technicians. Six technicians completed the orientation program, and five compounded the validation admixtures aseptically. The ongoing monitoring revealed that the technicians who were validated have continued to perform at acceptable levels. This program validated that the personnel had adequate training in aseptic technique and the necessary skills for performing the aseptic manipulations required in i.v. admixture compounding.

Antisepsis↗