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Biomedical subjects

M S Favero

Publications and source records attributed to M S Favero.

At least 37 records · Page 2Linked to original sources

The effect of uremia on tumor necrosis factor-alpha release after an in vitro whole-blood endotoxin challenge.

Uremia has been associated with immunologic aberrations, including anergy, increased susceptibility to infections, and reduced phagocytic activity of polymorphonuclear leukocytes. In this study, cytokine release in uremic and nonuremic blood after in vitro endotoxin stimulation was studied. Blood from nonuremic controls, chronic renal failure patients not on dialysis, and chronic hemodialysis patients predialysis and postdialysis was spiked with 10 ng/mL of Escherichia coli endotoxin and incubated for 2 and 26 h. Plasma tumor necrosis factor-alpha (TNF alpha) concentrations were determined by ELISA after each incubation period. To further study which uremic blood component may be responsible for enhanced release of TNF alpha, plasma and cellular components of chronic renal failure patients and controls were switched and then given an in vitro endotoxin stimulation (1 ng/mL). It was found that (1) TNF alpha release is enhanced by uremia and is exacerbated with progressive declines in renal function, (2) enhanced TNF alpha release is related to a blood cellular phenomenon induced by uremia, and (3) enhanced TNF alpha release in hemodialysis patients is associated with a prolonged stimulation and/or reduced plasma elimination of TNF alpha.

Blood Cells↗

The colonization of solid PVC surfaces and the acquisition of resistance to germicides by water micro-organisms.

Six common water bacteria were examined for their ability to colonize polyvinyl chloride (PVC) surfaces, survive various germicidal treatment, and re-establish themselves in sterile distilled water (SDW). For each test, two 30.4 cm PVC pipes attached to a 90 degrees PVC elbow were filled with 600 ml of distilled water inoculated with either Pseudomonas aeruginosa, Ps. cepacia, Ps. mesophilica, Acinetobacter anitratus, Mycobacterium chelonae or M. chelonae var. abscessus. After 8 weeks contaminated water was removed and the pipes were exposed to 600 ml of 1:213 iodophor disinfectant (ID), 1:128 phenolic detergent (P), 1:256 quaternary ammonium compound (QA), stock iodophor antiseptic (IA), 2% formaldehyde (F), 10-15 ppm free chlorine (C), 2% glutaraldehyde (G) and 70% ethanol (E). These germicides were periodically sampled, neutralized and examined for surviving organisms. After exposure for 7 d the germicides were removed and each pipe was refilled with SDW. This was assayed at 7 d intervals to determine microbial re-establishment. Samples were removed during microbial conditioning and examined by scanning electron microscopy (SEM). Pseudomonads were isolated directly from ID, QA, C, P and F, and mycobacteria from QA, IA, ID, P, G, C and F. Pseudomonas aeruginosa and Ps. cepacia survived in PVC pipes after 7 d of exposure to P, ID and C; Ps. mesophilica, after C and ID; and both mycobacteria, after C. SEM examination of PVC remnants revealed bacterial attachment and formation of extracellular material with embedded cells. These studies show that common water bacteria can attach and colonize the interior surface of PVC pipes and develop significant resistance to the action of certain germicides.(ABSTRACT TRUNCATED AT 250 WORDS)

Acinetobacter↗

Cytokine kinetics in an in vitro whole blood model following an endotoxin challenge.

Whole blood and peripheral blood mononuclear cell (PBMC) culture models have been used to study cytokine stimulation and release in vitro. In this study, we characterize the kinetics of the interleukins (IL-1 beta), (IL-6), (IL-8), and tumor necrosis factor-alpha (TNF-alpha) following an endotoxin (ET) challenge using our in vitro whole blood model. Whole blood samples from 10 healthy volunteers were studied. All cytokines were measured by enzyme-linked immunosorbent assay. Peak concentrations of TNF-alpha occurred 2 h after ET challenge followed by a rapid decline in free plasma TNF-alpha concentration (half-life 18.2 min). IL-1 beta was not significantly elevated until 4 h after ET challenge. IL-8 was elevated 1 h after ET challenge. IL-6 concentration exhibited a biphasic peak occurring at 6 and 74 h after ET challenge. We conclude that (1) our whole blood in vitro model produces cytokine release kinetics similar to those reported in vivo, and (2) the presence of either binding proteins or cellular metabolism of TNF-alpha in whole blood produces a similar plasma half-life to that observed in vivo.

Blood↗

Anaphylactoid reactions associated with reuse of hollow-fiber hemodialyzers and ACE inhibitors.

From July 18 through November 27, 1989, 12 anaphylactoid reactions (ARs) occurred in 10 patients at a hemodialysis center in Virginia. One patient required hospitalization; no patients died. ARs occurred within minutes of initiating dialysis and were characterized by peripheral numbness and tingling, laryngeal edema or angioedema, facial or generalized sensation of warmth, and/or nausea or vomiting. All 12 ARs occurred with dialyzers that had been reprocessed with an automated reprocessing system. A cohort study, including all patients undergoing dialysis sessions on the six days when an AR occurred, showed that the patients who experienced ARs were significantly more likely than patients who did not to be treated with angiotensin-converting enzyme (ACE) inhibitors (7/10 vs. 3/33; relative risk = 7.9; 95% confidence interval = 2.5 to 25.2) and to have been exposed to reused dialyzers rather than to new dialyzers (12/70 sessions vs. 0/31; P = 0.016). In those sessions using a reused dialyzer, the mean number of dialyzer uses in case-sessions was significantly higher than for noncase-sessions (10.3 vs. 6.2; P = 0.016). After reuse of dialyzers was discontinued at the center, no further ARs occurred, despite the continued administration of ACE inhibitors. This is the first report of an outbreak of ARs associated exclusively with reused dialyzers. We hypothesize that interactions between a dialyzer that has been repeatedly reprocessed and reused, blood, and additional factors, such as ACE inhibitors, increased the risk of developing ARs.

Ambulatory Care Facilities↗

Bacteria and endotoxin removal from bicarbonate dialysis fluids for use in conventional, high-efficiency, and high-flux hemodialysis.

The use of bicarbonate-based dialysis fluids in hemodialysis centers in the United States has increased with the advent of high-efficiency and high-flux hemodialysis. However, bicarbonate dialysis fluids can support rapid bacterial growth and high endotoxin concentrations. This study determined the efficacy of an ultrafiltration device in reducing the bacterial and endotoxin concentrations in bicarbonate dialysis fluids. A polysulfone hollow fiber dialyzer was used to ultrafilter bicarbonate concentrate before entering the central proportioner and bicarbonate dialysate after exiting the proportioner in single patient dialysis machines. Pre- and post-ultrafilter samples were collected for bacterial and endotoxin assays over 10 months. Ultrafiltration of bicarbonate concentrate reduced bacterial and endotoxin concentrations from 288,330 colony forming units (CFU)/ml and 42,804 pg/ml to 0.47 CFU/ml and 109 pg/ml, respectively. Ultrafiltration of the dialysate in single patient systems decreased bacterial and endotoxin concentrations from 15,889 CFU/ml and 1,746 pg/ml to 0.003 CFU/ml and 0.109 pg/ml, respectively. These results demonstrate that ultrafiltration of bicarbonate dialysis fluids is effective in reducing bacterial and endotoxin contamination inherently associated with the use of bicarbonate-based dialysates.

Bacteria↗

Pyrogenic reactions in patients receiving conventional, high-efficiency, or high-flux hemodialysis treatments with bicarbonate dialysate containing high concentrations of bacteria and endotoxin.

High-efficiency (HE) and high-flux (HF) hemodialysis are becoming increasingly popular methods for treating patients with chronic renal failure because they reduce the time required for dialysis treatment. HF and HE dialyzers require bicarbonate dialysate, often prepared from concentrates that can support bacterial growth with endotoxin production. There is a concern that endotoxins or bacteria may cross or interact at the membranes of these dialyzers, triggering the release of endogenous pyrogens (cytokines) by peripheral blood mononuclear cells to cause pyrogenic reactions (PR). To determine the incidence of PR and to examine the association between PR and levels of bacteria and endotoxin in dialysate, a cohort of patients receiving conventional, HE, or HF hemodialysis with bicarbonate dialysate and reprocessed dialyzers at three dialysis centers during a 12-month period was studied prospectively. All dialyzers underwent a test of membrane integrity before use. A total of 19 PR were identified among 18 patients in 26,877 hemodialysis treatments (0.7 PR/1,000 treatments). There was no significant difference in PR rates by treatment modality: conventional, 0.5 per 1,000 (7 PR/13,123 treatments) versus HE, 0.9 per 1,000 (9 PR/11,345) versus HF, 1.2 per 1,000 (3 PR/2,409) (P = 0.21; chi 2 test). Throughout the study period, bacterial counts for dialysate at each center significantly exceeded the Association for the Advancement of Medical Instrumentation's (AAMI) microbiologic standards for dialysate of less than 2,000 CFU/mL (mean, 19,000 CFU/mL), but water used in the reuse of dialyzers tested less than 200 CFU/mL.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

A prospective study of pyrogenic reactions in hemodialysis patients using bicarbonate dialysis fluids filtered to remove bacteria and endotoxin.

Pyrogenic reactions (PR) are a well-recognized complication of hemodialysis and have been associated with dialyzer reuse, high-flux dialysis, and bicarbonate dialysate. However, the roles of bacteria and endotoxin in dialysate for producing PR are not well defined. To determine the effect of removing most bacteria and endotoxin from the dialysate on the incidence of PR, a cohort of chronic hemodialysis patients receiving high-flux, high-efficiency, or conventional hemodialysis at three centers with bicarbonate dialysis fluids that had been filtered with a polysulfone high-flux hemodialyzer was prospectively studied. Unfiltered bicarbonate concentrate had median bacterial and endotoxin concentrations of 479,000 CFU/mL and 39,800 pg/mL, respectively. After filtration of the bicarbonate concentrate at the central proportioner, dialysate had a median 9.2 CFU/mL of bacteria and 17.8 pg/mL of endotoxin. Dialysate filtered at individual proportioning dialysis machines had a median 0.001 CFU/mL of bacteria and 0.19 pg/mL of endotoxin. Nine PR were identified among 303 patients after 28,007 hemodialysis treatments (0.3 PR/1,000 treatments). The rate of PR was similar for the three hemodialysis treatment modalities and for first-use compared with reused dialyzers. Although the PR rate in this study was lower (P = 0.046) than the PR rate of a previous study with unfiltered dialysis fluids (0.7 PR/1,000 treatments), it represents a difference of only 10 PR in over 28,000 treatments. It was concluded that filtration of hemodialysis fluids is efficacious in removing bacterial and endotoxin contamination and can result in a lower incidence of PR in patients receiving high-flux, high-efficiency, or conventional hemodialysis.

Adult↗

Bacteriologic and endotoxin analysis of salvaged blood used in autologous transfusions during cardiac operations.

Autologous blood transfusion is a common method of reducing the need for heterologous blood transfusion during cardiac operations. Recently we investigated an outbreak of severe, nonsurgical postoperative bleeding among patients undergoing heart operations and receiving intraoperative transfusion of blood from a cell conservation device (Cell Saver System, Haemonetics Corp., Braintree, Mass.). As a result of this investigation, we conducted a prospective study to determine if bacterial or endotoxin contamination of the blood collected in the Cell Saver System and used for reinfusion during heart operations contributes to postoperative bleeding complications. Patients' blood samples were collected immediately before operation, at the end of cardiopulmonary bypass, 1 hour postoperatively, and from the Cell Saver System. All blood samples were cultured for bacteria, and all plasma samples were assayed for endotoxin. Preoperatively all patients having heart operations were without signs of infection, 33 of 37 blood cultures taken were negative, and none of the plasma samples had detectable endotoxin. After discontinuance of cardiopulmonary bypass but before delivery of blood from the Cell Saver System, bacteria and endotoxin were detected in 11 of 36 (30.6%) and five of 35 (14.3%) of the patients' blood samples, respectively. The blood aspirated from the open chest and collected by the Cell Saver System was culture positive in 30 of 31 (96.8%) samples, and seven of 29 (24.1%) contained endotoxin. One of 28 blood samples collected 1 hour postoperatively was culture positive, and five of 25 samples contained endotoxin. Of 61 total microorganisms isolated, 50 (82%) were coagulase-negative staphylococci, four (6.6%) aerobic diphtheroids, five (8.2%) anaerobic "diphtheroids" (Propionibacterium acnes), and two (3.2%) gram-negative bacilli. Plasma endotoxin concentrations ranged from 10 to 765 pg/ml. No signs of endotoxemia or unusual bleeding were observed intraoperatively or postoperatively in any of the 38 patients. Although blood collected in the Cell Saver System and used for reinfusion during heart operations often was contaminated with gram-positive bacterial commensals of the skin and low concentrations of endotoxin, no adverse effects were noted in the patients.

Bacteria↗

Application of multilocus enzyme electrophoresis to epidemiologic investigations of Xanthomonas maltophilia.

OBJECTIVE: To test the utility of a newly developed multilocus enzyme electrophoresis typing method for Xanthomonas maltophilia. DESIGN: Isolates were first screened by slide agglutination, which served as the standard to characterize the outbreak strains. All isolates were then subjected to multilocus enzyme electrophoresis and the results analyzed based on epidemiological data. SETTING: This outbreak occurred in a shock-trauma intensive care unit of a large general community hospital. PATIENTS: Patients admitted to the shock-trauma intensive care unit who had X maltophilia isolated from any site greater than or equal to 24 hours after admission met the case definition. Specimens from patients who fit the case definition were characterized, as were specimens from other patients that were used as controls for nonoutbreak isolates. Environmental samples were also evaluated for X maltophilia. RESULTS: Most of the 64 isolates received during this outbreak were serotype 10, and when they were subjected to multilocus enzyme electrophoresis, one electrophoretic type predominated and correlated to most outbreak isolates. Unrelated isolates of serotype 10 from other institutions all exhibited unique electrophoretic types. CONCLUSION: Application of multilocus enzyme electrophoresis to X maltophilia outbreaks is a valuable addition to the characterization of suspected outbreak strains.

Bacterial Infections↗

Investigations of intrinsic Pseudomonas cepacia contamination in commercially manufactured povidone-iodine.

OBJECTIVE: Laboratory investigations were initiated with a povidone-iodine antiseptic solution that was intrinsically contaminated with Pseudomonas cepacia. These investigations were helpful in understanding the microbicidal and chemical properties of iodophor solutions and the mechanism by which P cepacia can survive in iodine-containing antiseptics. DESIGN: Included in these studies were: prolonged survival of P cepacia; available and free iodine determinations; microbial challenge studies; and scanning electron microscopic examination of contaminated antiseptic. RESULTS: P cepacia survived in this iodophor antiseptic up to 68 weeks from the date of manufacture. A uniform concentration of 1% available iodine was found in all lots of povidone-iodine tested as specified on the product label, but free iodine (I2) values varied greatly. Low free iodine levels of 0.23 to 0.46 ppm were associated with the contaminated lot of povidone-iodine. Solutions of povidone-iodine with varying levels of free iodine were rapidly microbicidal when challenged with cells of P cepacia derived from culture broth and washed or adapted to growth in water. P cepacia cells taken directly from contaminated povidone-iodine survived for significantly longer periods of time. Large numbers of P cepacia were found embedded in extracellular material and among strands of glycocalyx between cells as shown by scanning electron microscopy. CONCLUSIONS: The physical thickness of cellular and extracellular material that forms on surfaces could protect embedded organisms from the microbicidal action of disinfectants and antiseptics and subsequently allow for extended microbial survival times. Manufacturers should be aware that distribution piping surfaces colonized with bacteria may be a source of product contamination and resistant organisms.

Colony Count, Microbial↗

Microbial growth and endotoxin production in the intravenous anesthetic propofol.

OBJECTIVE: In this study, we measured microbial growth and endotoxin production in the intravenous anesthetic propofol using 10 different microbial strains; 6 isolated from outbreak cases and 4 from laboratory stock cultures. DESIGN: In each trial, endotoxin-free glass tubes containing 10 ml propofol were inoculated with 10(0)-10(3) CFU/ml of the test organism and incubated at 30 degrees C for 72 hours. SETTING: In May and June 1990, the Centers for Disease Control received reports of 5 outbreaks in 5 states of postsurgical patient infections and/or pyrogenic reactions. Epidemiologic and laboratory investigations implicated extrinsic contamination of an intravenous anesthetic, propofol, as the probable source of these outbreaks. RESULTS: After 24 hours, 9 of the 10 cultures increased in viable counts by 3 to 6 logs. At least 1 ng/ml of endotoxin was produced within 24 hours by Escherichia coli, Enterobacter cloacae, and Acinetobacter calcoaceticus subspecies anitratus. CONCLUSIONS: Propofol can support rapid microbial growth and endotoxin production. To avoid infectious complications, scrupulous aseptic technique should be used when preparing or administering this anesthetic.

Anesthesia, Intravenous↗

Comparison of microbiologic assay methods for hemodialysis fluids.

To help prevent pyrogenic reactions and bacteremia in hemodialysis patients, the Association for the Advancement of Medical Instrumentation and the Centers for Disease Control recommend microbiologic assay of hemodialysis fluids at least monthly. Five commercially available assay systems were evaluated by using the membrane filtration technique with standard methods agar and trypticase soy agar as the standards for comparison. Each assay system was challenged with dialysate and reverse-osmosis water from local dialysis centers, aqueous suspensions of eight laboratory strains of gram-negative bacilli and nontuberculous mycobacteria, and a mixed microbial flora inoculated into reverse-osmosis water and laboratory-prepared dialysate. Mean viable counts from triplicate samples were obtained after incubation at 37 degrees C for up to 72 h. The efficiency of recovery varied with the specific type of microbial challenge. The SPC water sampler (Millipore Corp., Bedford, Mass.) was the most consistent in obtaining the highest viable counts. Other commercial systems were comparable to each other in overall performance. All assay systems tested provided an acceptable balance between microbial recovery and required sampling time, equipment, and expertise.

Bacteria↗

Effects of incubation time and temperature on microbiologic sampling procedures for hemodialysis fluids.

To prevent pyrogenic reactions and bacteremia in hemodialysis patients, the Association for the Advancement of Medical Instrumentation and the Centers for Disease Control recommend culturing of hemodialysis fluids (water and dialysate) at least once a month. The recommendations for total microbial counts are (i) less than or equal to 200 CFU/ml in water used to prepare dialysate or reprocess hemodialyzers and (ii) less than or equal to 2,000 CFU/ml for the dialysate. In accordance with the Association for the Advancement of Medical Instrumentation recommendations all cultures should be incubated at 37 degrees C for 48 h on suitable culture media, such as Trypticase soy agar, standard methods agar, or one of several commercially available assay systems. There have been suggestions that lower temperatures and longer incubation might improve the recovery of bacteria from water and dialysate. In this study bacterial recovery from various dialysis fluids (water, bicarbonate dialysate, and bicarbonate concentrate) at 30 and 37 degrees C was compared. Duplicate sets of samples were membrane filtered (pore size, 0.45 microns); one set was incubated at 30 degrees C and the other was incubated at 37 degrees C for 72 h. The number of visible colonies was counted every 24 h by using a dissecting microscope. No significant difference was observed in specimens incubated at 37 degrees C for 48 h compared with those incubated at 30 degrees C for 72 h. Also, bacterial recovery was significantly better when samples of bicarbonate dialysate or bicarbonate concentrate were plated on Trypticase soy agar as opposed to standard methods agar.

Bacteria↗