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PubMed2002

Concentrations of cartilage oligomeric matrix protein in dogs with naturally developing and experimentally induced arthropathy.

OBJECTIVE: To assay concentrations of cartilage oligomeric matrix protein (COMP) in canine sera and synovial fluid (SF), to compare COMP concentrations in clinically normal dogs and dogs with joint disease, and to analyze changes in COMP concentrations in dogs with experimentally induced acute synovitis. ANIMALS: 69 control dogs without joint disease, 23 dogs with naturally occurring aseptic arthropathy, and 6 dogs with experimentally induced synovitis. PROCEDURE: Serum (n = 69) and SF (36) were obtained from control dogs. Samples of serum (n = 23) and SF (13) were obtained from dogs with naturally occurring aseptic arthropathy with or without radiographic features of osteoarthritis (OA). Serum and SF were obtained before and 1, 2, 3, and 7 days after induction of synovitis. The COMP concentrations were determined by use of an inhibition ELISA that had canine cartilage COMP and monoclonal antibody against human COMP. RESULTS: Concentrations of COMP in serum and SF of control dogs were 31.3+/-15.3 and 298.7+/-124.7 microg/ml, respectively. In naturally occurring OA, COMP concentrations in serum (44.9+/-177 microg/ml) and SF (401.7+/-74.3 microg/ml) were significantly higher than corresponding concentrations in control dogs. The COMP concentration in SF peaked 24 and 48 hours after induction of synovitis, whereas concentration in serum peaked on day 3. CONCLUSIONS AND CLINICAL RELEVANCE: These results supported the hypothesis that COMP concentration in serum and SF of dogs may be altered after cartilage degradation or synovitis. Measurement of COMP concentrations can be useful when differentiating arthropathies in dogs.

Animals↗
PubMed2001

Integrin expression at the bone/biomaterial interface.

The aim of this study was to visualize integrin expression by cells in interface tissue in relation to their ligands. Tissue samples were obtained from 25 patients undergoing revision of aseptically loose total joint replacements. Serial sections were immunolabeled for the integrins alpha(2)beta(1), alpha(v)beta(3), alpha(4)beta(1), alpha(L)beta(2) (CD11a), alpha(M)beta(2) (CD11b), and alpha(X)beta(2) (CD11c), and the ligands fibronectin, laminin, vitronectin, intercellular adhesion molecule-1, and vascular adhesion molecule-1. Most cells were found to express alpha(2)beta(1), most macrophages and giant cells expressed CD11b, and the majority of CD11a was found on perivascular T lymphocytes. From the small amount of alpha(4)beta(1) and vascular adhesion molecule-1 expression in the interface tissue and the combination of CD11a, CD11b, and intercellular adhesion molecule-1 expression, it would seem that macrophages use beta(2) integrins to transmigrate.

Aged↗
PubMed1989

Definition of high-proficiency serological markers for diagnosis of varicella-zoster virus infections by enzyme immunoassay.

The levels of Varicella-zoster virus (VZV)-specific IgG, IgM, and IgA antibodies produced in 22 cases of varicella, 22 cases of cutaneous zoster, and 12 cases of acute aseptic meningitis due to VZV in the absence of cutaneous lesions, were measured by indirect enzyme immunoassay (EIA) and compared with those observed in a group of 34 age-matched controls. The definition of cutoff titres for each serological marker and combinations of them allowed early diagnosis of infection in 82% of varicella patients and 91% of patients with acute aseptic meningitis lacking cutaneous lesions on a single serum sample, the specificity being over 90%. The system was as sensitive as the demonstration of intrathecally produced IgG antibodies for the early diagnosis of the infection in 22 cases of neurological disease due to VZV. A working protocol for the serological diagnosis of VZV infections, using currently available EIA reagents, is described.

Adolescent↗
PubMed2004

Effectiveness of topical chlorhexidine powder as an alternative to hexachlorophane for the control of Staphylococcus aureus in neonates.

We routinely phage-type Staphylococcus aureus isolates from high-risk inpatients each week. This surveillance approach previously identified a five-year outbreak of a methicillin-susceptible S. aureus strain (MSSA, PT 53,85), which affected 202 babies on a regional neonatal unit. We previously reported this outbreak and the multiple staged infection control measures that were required to end it. These included strict emphasis on hand hygiene, environmental and staff surveillance sampling, application of topical triclosan solution and hexachlorophane powder, aseptic handling of a skin protectant material, and use of topical mupirocin for staff nasal carriers of the endemic MSSA strain and for babies colonized or infected with S. aureus. In summer 2000 topical hexachlorophane powder became unavailable and we therefore substituted topical 1% chlorhexidine powder as part of routine umbilical decontamination. We have continued prospective S. aureus surveillance for the past five years to monitor the effect of this practice change. We observed a continued decline in the numbers of monthly MSSA isolates from neonatal unit babies. Since the substitution of chlorhexidine for hexachlorophane, the median monthly number of MRSA isolates has been 0.5 (range 0-4). Only sporadic S. aureus PT 53,85 isolates were recovered. Control of S. aureus in our regional neonatal unit, in particular an endemic MSSA strain, was maintained when topical umbilical hexachlorophane powder was substituted with 1% chlorhexidine powder.

Administration, Topical↗
PubMed1997

Detection of enteroviral RNA and specific DNA of herpesviruses by multiplex genome amplification.

A reverse transcription (RT) multiplex polymerase chain reaction (PCR) assay was developed to allow rapid, sensitive and simultaneous detection of enteroviral RNA and herpesviral DNA specific sequences in a single tube. The method involves a reverse transcription step followed by a multiplex nested PCR in which the combination of primers amplifies cDNA from enteroviruses and specific herpesviruses DNA. Nested amplification utilises primers designed to anneal into the amplification product from the first reaction. Individual viruses were then detected and differentiated by the size of their PCR products determined using ethidium bromide stained agarose gels. To exclude false negatives due to sample inhibitors an internal amplification control, a cloned fragment of DNA from Pseudorabies virus (PRV DNA) was included in the reaction mixture. Detection levels between 0.01 and 0.001 TCID50 of prototype strains of Polio and Coxsackie type B viruses and between 1 and 100 molecules of cloned-DNA of herpesviruses prototype strains were achieved. The RT multiplex PCR method proved capable of detecting enteroviral RNA or herpesviral DNA in cerebro spinal fluid (CSF) samples from patients with aetiologically well characterized encephalitis or aseptic meningitis.

Child↗
PubMed1976

The plastic surgical adhesive drape: an evaluation of its efficacy as a microbial barrier.

A microbial evaluation was made of adhesive plastic surgical drapes and cloth surgical drapes. These studies were done both during surgery and in the laboratory. The plastic drape does not allow bacterial penetration, lateral migration does not occur, skin bacteria do not multiply under the drape within the time periods studied and the patient drapes are held in place with their use. When wet, cloth drapes showed profuse bacterial penetration. Dry cloth showed less bacterial penetration as compared to wet cloth. Lateral migration under cloth drapes was not possible to assess due to a high level of penetration. The surface of cloth showed a higher level of bacterial contamination during the surgical procedures. Deep wound cultures collected just prior to closing showed 60% contamination when cloth was used compared to 6% when plastic was employed. The micro-organisms recovered from the various sites sampled were identified. Finally, in addition to the positive aseptic benefits afforded by plastic adhesive drapes, aesthetic features such as a more delineated operative field and elimination of towel clips make this product a useful adjunct to the surgeon's armamentarium.

Air Microbiology↗
PubMed1995

Preventing posttraumatic pneumonia.

Between 20% and 40% of patients admitted to the hospital with multiple trauma develop bacterial pneumonias. The most immediate causes are aspiration, blunt chest trauma, and prolonged immobilization. Traumatic shock and the need for immediate head and chest surgery are contributing factors. Signs of systemic infection, infiltrates on the chest film, and microbiological evidence obtained from sputum samples are the principal diagnostic criteria. Prophylactic measures include aseptic management of the airway, tracheostomy when indicated, adequate pain relief, and early mobilization. When the incipient signs of pneumonia appear, prompt aggressive antimicrobial therapy is indicated.

Bacterial Infections↗
PubMedDate not supplied

Evaluation of skin bacterial flora before and after aseptic preparation of clipped and nonclipped arthrocentesis sites in horses.

OBJECTIVE: This study evaluates skin bacterial flora before and after aseptic preparation of clipped and nonclipped arthrocentesis sites in horses. STUDY DESIGN: The hair over one midcarpal joint and one distal interphalangeal joint on each horse was clipped. The contralateral joint served as the nonclipped comparison. ANIMALS OR SAMPLE POPULATION: Twelve adult horses. METHODS: A prescrub sample for microbial culture was taken from the dorsal surface of all four joints for each horse. Each site was aseptically prepared with povidone iodine and 70% alcohol, followed by postscrub sampling for microbial culture. Colony forming units (CFUs) were determined for each sample, 24 hours after inoculation of blood agar plates. RESULTS: There was no significant difference (P > .05) in number of postscrub CFUs between clipped and nonclipped skin over the midcarpal or distal interphalangeal joints. Percent bacterial reduction (mean +/- SD%) after aseptic preparation differed significantly (P = .02) between clipped (99.8 +/- .003%) and nonclipped (96.2 +/- .05%) skin at the midcarpal joint, but not at the distal interphalangeal joint (clipped, 98.5 +/- .03% and nonclipped, 97.8 +/- 0.21%). There was a significant difference (P = .009) in number of prescrub CFUs obtained from clipped and nonclipped skin for the midcarpal joint. There was no significant difference in number of prescrub CFUs between clipped and nonclipped skin at the distal interphalangeal joint. Bacteria isolated from both clipped and nonclipped skin sampled postscrub included Bacillus sp, nonhemolytic Staphylococcus sp, and Micrococcus sp. CONCLUSIONS: The presence of hair over the midcarpal and distal interphalangeal joints does not appear to inhibit the ability of antiseptics to effectively reduce bacterial flora to an acceptable level for arthrocentesis. CLINICAL RELEVANCE: Aseptic preparation of the skin over the midcarpal and distal interphalangeal joints can be accomplished without hair removal in horses.

Animals↗
PubMed1993

Role of protein and lactose interactions in the age gelation of ultra-high temperature processed concentrated skim milk.

Skim milk was pasteurized, diafiltered, and concentrated three times by UF. Lactose or sucrose was then added at 3 or 6%. The five samples containing < .05% lactose, 3 and 6% lactose, and 3 and 6% sucrose were UHT processed at 140 degrees C for 4 s using indirect heating, collected aseptically in presterilized containers, and stored at 4, 20, and 35 degrees C. All samples stored at 4 and 20 degrees C gelled after 21 wk of storage. Samples stored at 35 degrees C did not gel. Browning occurred only in samples containing lactose stored at 35 degrees C. Proteolysis in gelled samples was shown by SDS-PAGE. Bands were due to proteolysis, protein crosslinking, and a streaking pattern in ungelled samples. Electron micrographs of gelled samples showed that various casein particles were connected by hairlike protrusions, but the micelles in ungelled samples were not connected and had few protrusions. The Maillard reaction neither promoted nor deferred age gelation. Protein modifications prevented gelation in samples stored at 35 degrees C. Age gelation was probably a two-step process in which dissociated proteins from the casein micelles reformed on micelles as hairlike protrusions. This process was followed by aggregation of the protein particles.

Animals↗
PubMed2000

Cerebrospinal fluid tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, and interleukin-8 as diagnostic markers of cerebrospinal fluid infection in neurosurgical patients.

OBJECTIVE: To evaluate whether cerebrospinal fluid concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, or IL-8 may be used as diagnostic markers for the differential diagnosis of aseptic vs. bacterial meningitis and/or ventriculitis in neurosurgical patients. DESIGN: Prospective, observational study. SETTING: University teaching hospital. SUBJECTS: A total of 112 cerebrospinal fluid samples from 14 asymptomatic patients with normal cerebrospinal fluid after neurosurgery, 27 asymptomatic and 19 symptomatic patients with postneurosurgical aseptic meningitis, 32 patients with postneurosurgical cerebrospinal fluid infection, and 20 with severe subarachnoid and/or cerebral hemorrhage. MEASUREMENTS AND MAIN RESULTS: Specific ELISA kits were used to analyze TNF-alpha, IL-1beta, IL-6, and IL-8 concentrations on cerebrospinal fluid samples. Elevations in cerebrospinal fluid concentrations of TNF-alpha, IL-1beta, IL-6, and IL-8 were induced by different diseases or neurosurgical procedures, but cerebrospinal fluid bacterial infection induced the highest concentrations. To discriminate between aseptic cerebrospinal fluid pleocytosis and cerebrospinal fluid infection with a specificity of 95%, cerebrospinal fluid leukocyte count >1700/mL, TNF-alpha >150 pg/mL, and IL-1beta >90 pg/mL showed sensitivities of 51%, 74%, and 90%, respectively. Sufficiently sensitive and specific cutoff points could not be found for cerebrospinal fluid IL-6 or IL-8. CONCLUSION: Cerebrospinal fluid IL-1beta appears to be the best biochemical marker of cerebrospinal fluid infection in neurosurgical patients.

Adolescent↗
PubMed1999

Expression of laminins and their integrin receptors in different conditions of synovial membrane and synovial membrane-like interface tissue.

OBJECTIVE: To demonstrate the expression of laminins (Lns) and their integrin (Int) receptors in different synovial samples and synovial membrane-like interface tissues from well fixed and aseptically loosened total hip replacement (THR), and the potential role of Ln-Int interaction in the production of collagenases and cytokines. METHODS: Immunohistochemical staining was done to detect the distribution of EHS Ln, Ln alpha2, alpha3, alpha5, beta1, beta2 chains and Int alpha1, alpha2, alpha3, alpha6, beta1, beta4 subunits in different samples. Double immunofluorescence labelling was used to find colocalisation of Int alpha6 subunit and collagenase-1/collagenase-3/TNFalpha/IL6. RESULTS: General Ln immunoreactivity was detected in all specimens. Ln alpha5, beta1 and beta2, but not alpha2 and alpha3 chains were seen in the synovial lining and the basement membrane of blood vessels with the intensity/extent of labelling in the following rank order: rheumatoid arthritis (RA) loosened prostheses, osteoarthritis, well fixed prostheses, traumatic knees. Among Int subunits, staining for beta1 was usually the strongest, followed by staining for Int alpha6, alpha1, alpha3, and alpha2 subunits, with the same rank order for overall expression of Lns. Int beta4 subunit was not detectable in most of the specimens. Double labelling focused on Int alpha6 subunit disclosed its frequent colocalisation with collagenases 1 and 3 and with tumour necrosis factor alpha and interleukin 6 in synovial lining. CONCLUSION: Synovial lining contains Ln-10, Ln-11, and Int alpha6beta1 and alpha1beta1 receptors. In aseptic loosening of THR, interface tissue has a similar Ln subtype and Int receptor composition as RA synovium, which confirms its "lining-like" phenotype. Synovial lining does not contain Ln-5 (alpha3beta3gamma2) or Int alpha6beta4, which are components of epithelial hemidesmosomes. The expression of Lns and their Int receptors is upregulated in inflammation. The close spatial relation between Ln and its Int receptors in synovial lining cells containing proteinases and cytokines suggests a potential role in joint destruction and prosthetic loosening.

Adult↗
PubMedDate not supplied

Establishment of critical contamination risk locations ("hot spots") in environmental monitoring by means of three-dimensional airflow analysis and particulate evaluation.

A practical approach for the qualification of the surrounding environment of the critical area in aseptic processing has been developed. This method uses three-dimensional air velocity measurements combined with airborne particle monitoring. The analysis of the results obtained using the methods described in this article are beneficial in the selection of sample sites and frequencies and in refining personnel procedures and materials flow in aseptic processing. We propose that this improved qualification method can be widely applicable for both existing and new aseptic processing areas. This paper shows the results of one case study utilizing this method. The particle distribution map of a Grade B environment based upon extensive analysis was found to correspond to room airflow, as visualized by air vector mapping. The actual annual environmental monitoring data, which include airborne particles and microbes, as well as other microbial monitoring data, are also presented with respect to their relationship to the airflow pattern.

Air Microbiology↗
PubMed1995

Relationship between somatic cell count and intramammary infection of the half udder in dairy ewes.

A total of 1382 milk samples from half udders of three breeds of ewes (686 Assaf, 422 Churra, and 274 Castellana) were collected aseptically at midlactation from 18 flocks in the Castile-León region of Spain. Bacteriological tests were carried out on the samples to ascertain the status and type of IMI. Somatic cell counts were determined for all samples after bacteriological analysis. Analysis of variance showed significant effects of breed, IMI status, flock nested within breed by milking type, organism within IMI status, and interactions of breed by milking type and of breed by organism. The most frequent bacterial groups were staphylococci, especially coagulase-negative staphylococci, followed by streptococci, micrococci, and yeasts. The SCC response was related to the bacterial group involved in the IMI; coagulase-negative staphylococci showed significantly lower SCC values than those for coagulase-positive staphylococci and streptococci. Highest percentages of IMI by major pathogens and highest SCC, both in the absence of IMI and in staphylococcal IMI, were in Assaf ewes. The threshold of 300,000 cells/ml allowed 81% of the half udders to be correctly classified according to IMI status. However, the existence of breed differences in SCC response and IMI type could make the use of specific SCC thresholds for each breed advisable.

Animals↗
PubMed1987

Comparison of various criteria for determining the health status of the bovine udder.

The International Dairy Federation's (IDF) criteria for classification of subclinical bovine health conditions does not provide a true picture of the health status of the udder. When combining these criteria with the serum albumin levels of milk samples, the IDF's classification of sub-clinical mastitis could be extended to include 8 possible conditions such as normal quarters, septic mastitis, aseptic mastitis, relevant and irrelevant teat canal infection, specific and unspecific hyperalbumingalactia and unspecific cellular reaction. When the bacteriological results of teat canal swab samples were combined with the IDF's standards, these conditions could be scaled down to 4 diagnoses which included normal quarters, aseptic mastitis, teat canal infections, subclinical mastitis and a combination of the latter 2 conditions. When applying the teat canal swabbing technique, up to 16% of quarters classified as normal and 10% as cases of aseptic mastitis according to the IDF's criteria in fact, have TCI. Since classification of the health status of quarters according to the IDF's criteria and with due regard to the bovine serum albumin values of milk, did not include the bacteriological results of the teat canal swab samples, further research must be done to clarify the existence of conditions in the IDF classification. Teat canal infections may result in damage to the udder parenchyma as manifested by the elevated bovine serum albumin content of milk.

Animals↗
PubMed1994

Automated monitoring of cell concentration and viability using an image analysis system.

In order to automate measurements of cell concentration and viability in a suspended animal cell culture, we have developed an in situ microscopic image analysis system with an effective cell recognition algorithm. With a small amount of sample, this system can measure the cell density rapidly and aseptically. In addition, it can measure a cell size histogram including cell debris small particle distribution. These small particles have been found to be related to the viability of the mouse-mouse hybridoma STK1 cell line. By using cell debris small particle density as an indicator of cell viability, the developed system provides non-destructive viability monitoring without trypan blue staining.

Animals↗
PubMed2000

Evaluation of probiotic treatment in a neonatal animal model.

The clinical use of probiotic agents such as enteral Lactobacillus to enhance intestinal defense against potential luminal pathogens has been tested in vivo; however, an understanding of the mechanisms responsible for the observed protection is lacking. The purpose of this study was to evaluate the effects of Lactobacillus on bacterial translocation (BT) in a neonatal animal model. Newborn New Zealand white rabbit pups were enterally fed a 10% Formulac solution inoculated with or without a 10(8) suspension of ampicillin-resistant Escherichia coli K1 (E. coli K1A) and/or Lactobacillus casei GG (Lacto GG). Pups received either no bacteria (n = 10), Lacto GG (n = 8), E. coli K1A (n = 26), or a combination of Lacto GG and E. coli K1A (n = 33). On day 3, representative tissue specimens from the mesenteric lymph nodes (MLN), spleen (SPL), and liver (LIV) were aseptically harvested in addition to a small-bowel (SB) sample that was rinsed to remove luminal contents. The specimens were then cultured in organism-specific media. Statistical analysis was by one-way ANOVA with P values less than 0.05 considered significant. Neonatal rabbits receiving Lacto GG-supplemented formula exhibited a 25% decrease (P < 0.05) in small-bowel colonization by E. coli K1A. In addition, Lacto GG decreased the frequency of extraintestinal BT by 46% (P < 0.05), 61% (P < 0.05), and 23%, respectively, in the MLN, SPL, and LIV. We have shown that enterally-administered Lacto GG decreases the frequency of E. coli K1A translocation in a neonatal rabbit model. These results may have significant implications for the treatment of BT and sepsis in the human neonate and provide a model for further studies.

Animals↗
PubMed1999

Laboratory diagnosis of common viral infections of the central nervous system by using a single multiplex PCR screening assay.

A multiplex PCR assay that detects the four commonest causes of viral meningitis and encephalitis in the United Kingdom (herpes simplex virus [HSV] type 1 [HSV-1], HSV type 2 [HSV-2], varicella-zoster virus [VZV], and enteroviruses) was developed, and its sensitivity was compared with those of similar assays described previously for this application. Compared to the previous assays, this single multiplex PCR assay had higher molecular sensitivities for the detection for each of the viruses and improved utility for routine use in a diagnostic laboratory. The assay was used to test a series of 1,683 consecutive cerebrospinal fluid (CSF) samples between June 1997 and March 1998 inclusively. Viral nucleic acid was detected in 138 (8.2%) of the CSF samples, including enteroviruses in 51 samples, HSV-2 in 33 samples, VZV in 28 samples, and HSV-1 in 25 samples. Compared to the accepted relative incidence of viral etiologies, aseptic meningitis due to HSV-2 infection was high, and in adult female patients with symptoms of aseptic meningitis, HSV-2 was the virus most commonly detected in the CSF.

Adult↗
PubMed1997

Interleukin 6 in cerebrospinal fluid of patients with meningitis is not a useful diagnostic marker in the differential diagnosis of meningitis.

We assayed interleukin 6 (IL-6) concentrations in cerebrospinal fluid (CSF) from patients affected by meningitis of different aetiologies, and verified whether IL-6 can be used as a diagnostic marker in the differential diagnosis of meningitis. We used a monoclonal antibody enzyme immunoassay to test 98 CSF samples classified as pyogenic (15), viral (15), self-resolving aseptic meningitis (20), other infectious meningitis (9), neoplastic (4) and normal CSF from patients with (20) and without (15) non-infectious neurological diseases. CSF IL-6 concentrations were increased in pyogenic meningitis (100%) and in more than 50% of viral and other subarachnoid space infections, and rarely in patients without central nervous system infections. Though patients affected by pyogenic meningitis showed the highest levels of CSF IL-6, only a cut-off point > or = 10000 pg/mL was able to discriminate pyogenic meningitis from those of other aetiologies with a specificity > or = 94% and a positive predictive value of > or = 0.75 but the sensitivity was < or = 60%. Therefore, CSF IL-6 concentration is not a good diagnostic marker in the differential diagnosis of meningitis.

Adolescent↗