Staphylococcus epidermidis induces complement activation, tumor necrosis factor and interleukin-1, a shock-like state and tissue injury in rabbits without endotoxemia. Comparison to Escherichia coli.

Tumor necrosis factor (TNF) and IL-1 are thought to mediate many of the pathophysiologic changes of endotoxemia and Gram-negative bacteremia. In these studies, heat-killed Staphylococcus epidermidis were infused into rabbits to determine whether an endotoxin (LPS)-free microorganism also elicits cytokinemia and the physiologic abnormalities seen in Gram-negative bacteremia. S. epidermidis induced complement activation, circulating TNF and IL-1, and hypotension to the same degree as did one-twentieth the number of heat-killed Escherichia coli. Circulating IL-1 beta levels had a greater correlation coefficient (r = 0.81, P less than 0.001) with the degree of hypotension than TNF levels (r = 0.48, P less than 0.02). Leukopenia, thrombocytopenia, diffuse pulmonary capillary aggregation of neutrophils, and hepatic necrosis with neutrophil infiltration were observed to the same extent after either S. epidermidis or E. coli infusion. However, S. epidermidis infusion did not induce significant (less than 60 pg/ml) endotoxemia, whereas E. coli infusion resulted in high (11,000 pg/ml) serum endotoxin levels. S. epidermidis, E. coli, LPS, or S. epidermidis-derived lipoteichoic acid (LTA) induced TNF and IL-1 from blood mononuclear cells in vitro. E. coli organisms and LPS were at least 100-fold more potent than S. epidermidis or LTA. Thus, a shock-like state with similar levels of complement activation as well as circulating levels of IL-1 and TNF were observed following either S. epidermidis or E. coli. These data provide further evidence that host factors such as IL-1 and TNF are common mediators of the septic shock syndrome regardless of the organism.

The analysis of milk components and pathogenic bacteria isolated from bovine raw milk in Korea.

Bovine mastitis can be diagnosed by abnormalities in milk components and somatic cell count (SCC), as well as by clinical signs. We examined raw milk in Korea by analyzing SCC, milk urea nitrogen (MUN), and the percentages of milk components (milk fat, protein, and lactose). The associations between SCC or MUN and other milk components were investigated, as well as the relationships between the bacterial species isolated from milk. Somatic cell counts, MUN, and the percentages of milk fat, protein, and lactose were analyzed in 30,019 raw milk samples collected from 2003 to 2006. The regression coefficients of natural logarithmic-transformed SCC (SCCt) on milk fat (-0.0149), lactose (-0.8910), and MUN (-0.0096), and those of MUN on milk fat (-0.3125), protein (-0.8012), and SCCt (-0.0671) were negative, whereas the regression coefficient of SCCt on protein was positive (0.3023). When the data were categorized by the presence or absence of bacterial infection in raw milk, SCCt was negatively associated with milk fat (-0.0172), protein (-0.2693), and lactose (-0.4108). The SCCt values were significantly affected by bacterial species. In particular, 104 milk samples infected with Staphylococcus aureus had the highest SCCt (1.67) compared with milk containing other mastitis-causing bacteria: coagulase-negative staphylococci (n = 755, 1.50), coagulase-positive staphylococci (except Staphylococcus aureus; n = 77, 1.59), Streptococcus spp. (Streptococcus dysgalactiae, n = 37; Streptococcus uberis, n = 12, 0.83), Enterococcus spp. (n = 46, 1.04), Escherichia coli (n = 705, 1.56), Pseudomonas spp. (n = 456, 1.59), and yeast (n = 189, 1.52). These results show that high SCC and MUN negatively affect milk components and that a statistical approach associating SCC, MUN, and milk components by bacterial infection can explain the patterns among them. Bacterial species present in raw milk are an important influence on SCC in Korea.

In vitro positive controls for histochemical stains of bacteria and fungi.

Positive in vitro controls for histochemical stains of bacteria and fungi in tissue were developed from fibrin substrates of cultured microorganisms. These substrates show excellent and reproducible stain control characteristics. They are easily prepared, highly specific preparations which display greater uniformity and consistency as stain controls than do most infected tissues. These preparations may also be suitable control substrates for immunohistochemical procedures that identify microorganisms in tissue or fluid.

Bacterial anterior chamber contamination with foldable silicone lens implantation using a forceps and an injector.

PURPOSE: To compare the rate of anterior chamber (AC) contamination during implantation of a foldable three-piece silicone lens using a forceps and an injector. SETTING: Department of Ophthalmology, University of Saarland, Homburg (Saar), Germany. METHODS: In a prospective, randomized study, we cultured AC aspirates of patients who had phacoemulsification with silicone lens implantation through a 3.5 mm temporal clear corneal incision. In Group A, the lens was implanted with a forceps and in Group B, with an injector. Immediately after lens implantation, the AC aspirate (0.05 to 0.10 ml) was collected and sent to the microbiology department. All cultures were incubated for 3 days. As a topical antibiotic, gentamicin was given preoperatively and intraoperatively. RESULTS: One hundred patients (50 in each group) were examined. In each group the bacterial contamination of the AC was 2%. CONCLUSION: Bacterial contamination of the AC was not significantly different after silicone lens implantation with a forceps and with an injector.

Application of extended single-reaction multiplex polymerase chain reaction for toxin typing of Staphylococcus aureus isolates in South Korea.

The extended single-reaction multiplex PCR (esr-mPCR) developed in this study to detect staphylococcal enterotoxins (SEs), including SEA, SEB, SEC, SED, SEE, SEH, SEI, and SEJ, requires fewer sets of primers than other conventional multiplex PCRs and can be used to detect newly identified staphylococcal enterotoxins SEs more readily. Esr-mPCR analysis of 141 isolates of Staphylococcus aureus obtained from abattoir and livestock product samples revealed that 27 of the S. aureus isolates were toxigenic, and two were 2 multitoxigenic isolates. The most prevalent SE type was SEI followed by SEA and SEH. In addition, we investigated the clonal relatedness of toxigenic S. aureus isolates by arbitrarily primed PCR (AP-PCR). AP-PCR analysis of toxigenic S. aureus isolates revealed that the discriminatory power of AP-PCR was 9 (D=0.81), 8 (D=0.77), and 10 types (D=0.83) with primers AP1, ERIC2, and AP7, respectively. The combination of three each AP-PCR result could rearrange toxigenic S. aureus isolates into 10 types and five subtypes, with the D-value of 0.92. Interestingly, our data showed that toxigenic S. aureus isolates from different sources had different fingerprinting patterns although some of them carried the same types of SE genes. These data suggest that combinations of esr-mPCR and AP-PCR can provide a powerful approach for epidemiological investigation of toxigenic S. aureus isolates.

Antimicrobial performance of alkaline ionic fluid (GC-100X) and its ability to remove Escherichia coli O157:H7 from the surface of tomatoes.

An efficacy test of GC-100X, a noncorrosive alkaline ionic fluid (pH 12) composed of free radicals and supplemented with xylitol, was carried out against six major foodborne pathogens-Staphylococcus aureus FRI 913, Salmonella enterica serovar Enteritidis ATCC 13076, S. enterica serovar Typhimurium DT104 Korean isolate, Vibrio parahaemolyticus ATCC 17803, Escherichia coli O157:H7 ATCC 43894, and Pseudomonas aeruginosa KCTC 1637-at three different temperatures (4, 25, and 36 degrees C) with or without organic load (2% yeast extract). Results revealed a more than 4-log10 (CFU/ml) reduction (1.0 x 10(4) CFU/ml reduction) against all pathogens reacted at 37 degrees C for 3 h in the absence of organic material. GC-100X solution diluted with an equal volume of distilled or standard hard water (300 ppm CaCO3) showed effective bactericidal activity, particularly against gram-negative bacteria. Washing efficacy of GC-100X solution was compared against E. coli O157:H7 on cherry tomato surfaces with those of a commercially used detergent and chlorine water (100 ppm). Viable cell counts of E. coli O157:H7 that had penetrated to the cores of tomatoes after sanitizing treatment revealed that GC-100X stock and its 5% diluted solutions had similar washing effects to 100-ppm chlorine water and were more effective than the other kitchen detergent. These results indicate that GC-100X has good bactericidal and sanitizing activities and is useful as a new sanitizer for food safety and kitchen hygiene.

Comparison of survival probabilities for living-unrelated versus cadaveric renal transplant recipients.

UNLABELLED: Any attempt to improve organ donation would be of benefit due to the growing shortage of cadaveric sources for transplantation. OBJECTIVE: We compared the graft survivals and possible predictive variables among renal transplant recipients with organs from living unrelated (LURD) versus cadaveric donors (CD). METHOD: Among 104 consecutive renal transplants performed from July 1992 to February 2003, 41 were from LURD and 24 from CD. Immunosuppressive regimens were based on cyclosporine and steroids with mycophenolate mofetil added after 1998. Patient and graft survivals were estimated using the Kaplan-Meier method and compared using log-rank tests. The significance level of predictive variables was analyzed with the Cox proportional hazard model. The follow-up period was 2 to 127 months (median 46 months). RESULTS: Eight recipients lost their grafts (six from LURD and two from CD) due to four chronic rejections, one acute rejection, one recurrence of primary disease, and one death with a functioning graft. The graft survival rates at 1, 3, 5, and 7 years were 97.6%, 91.9%, 88.5%, and 82.2% for LURD transplants versus 95.5%, 90.9%, 90.9%, and 90.9% for CD transplants, respectively (P > .05). Delayed graft function and donor age (>55 years old) were statistically significant predictors of graft survival among LURD transplants. Donor age (>55 years old) and multiple preoperative transfusion history were significant in CD transplants. CONCLUSION: LURD transplant survival was similar to that of CD transplants in our series. LURDs are an excellent source of organs to expand the donor pool.

Risk factors on graft survival of living donor kidney transplantation.

UNLABELLED: Living donors have always been the basic resources of transplantation in our country, where cadaveric harvesting is still hampered for various reasons. OBJECTIVE: The aim of this study was to compare graft survival rates between living unrelated donor (LURD) and living related donor (LRD), to assess the potential risk factors for the graft survival, and to discuss the role of LURD. METHOD: From October 1991 to February 2003, 77 living donor renal transplants were performed: 41 were LURD and 36 were LRD transplants. The analyzed variables were donor relationship, recipient age and sex, donor age and sex, HLA-DR mismatching, nonspecific blood transfusion history of donor, acute rejection episodes, repeated rejection episode (more than 3 times), delayed graft function, recurred primary disease, and immunosuppressive regimen. Graft survival rate was assessed with the Kaplan-Meier method and the significance of possible variables with the Cox proportional hazard model. RESULTS: Eleven recipients lost their grafts (6 from LURD and 5 from LRD), most of them are due to chronic rejection (n = 7). Overall 3-, 5- and 10-year graft survival in live donors were 92.8%, 86.6%, and 76.9%, respectively. Graft survival at 3, 5, and 10 years being 91.9%, 88.5%, and 74.7% for the LURD versus 94%, 84%, and 78.8% for LRD transplants (P > .05). Acute rejection episodes, especially more than 3 times (risk ratio [RR] = 11.1) and preoperative multiple transfusion history (RR = 4.2) were significant factors on graft survival in our series. CONCLUSION: Acute rejection episodes markedly decreased the long-term graft survival in live donor renal transplants. The use of LURD transplants provides graft survival comparable with LRD transplants and proper management to acute rejection is essential for long-term graft survival.

Contamination of dialysis water and dialysate. A survey of 30 centers.

The concentration of bacteria and endotoxin in dialysis water and dialysate of 30 dialysis centers in western Germany was examined. Water samples were obtained after treatment by reverse osmosis or other processing methods. Collection of dialysis samples for bacterial, fungal, and endotoxin analysis was conducted before and 2 hours after start of hemodialysis. In 17.8% of all water samples analyzed, the AAMI standard was exceeded and bacterial and fungal counts greater than 200 colony forming units/ml were found. In 11.7% of all dialysate samples, higher contamination than the recommendations for dialysate of 2000 colony forming units/ml were found. The concentration of endotoxin in water and dialysate varied between 0 and 95 endotoxin units in the water samples and 0 and 487 endotoxin units/ml in the dialysate samples. In 12.2% of all water sampled, and 27.5% of all dialysate samples, values of 5 endotoxin units/ml were found. No correlation was found between the level of contamination of either water or dialysate in a specific center and the following factors: water processing method (reverse osmosis or others), type of dialysate (acetate of bicarbonate), type of dialysate machine, or method of machine disinfection. In view of these results it is suggested that endotoxin testing, especially in the dialysate, be a part of regular quality control in dialysis.

Effect of acute burn trauma on phagocytic activity of the reticuloendothelial system in rats.

The effect of acute burn trauma on phagocytic activity of the reticuloendothelial system measured in vivo with technetium 99m sulfur colloid was examined in rats subjected to acute burn trauma. After the scald injuries (10-second, full-thickness burns) were induced, a reduction in phagocytic activity by the spleen took place with an accompanying increase in the uptake of colloid material by the lungs. Uptake of colloid material by the liver was essentially unchanged. These uptake changes, observed within hours after the inducement of acute burn trauma and apparently continuing for 7 days after burn injury, may explain, in part, the development of septicemia in patients with burns because altered phagocytic activity of the reticuloendothelial system can result in subsequent overabundance of microorganisms and bacteria in the blood.

Effects of substrates on biofilm formation observed by atomic force microscopy.

Formation of biofilm is known to be strongly dependent on substrates including topography, materials, and chemical treatment. In this study, a variety of substrates are tested for understanding biofilm formation. Sheets of aluminum, steel, rubber, and polypropylene have been used to examine their effects on formation of Pseudomonas aeruginosa biofilm. In particular, the morphological variation, transition, and adhesiveness of biofilm were investigated through local measurement by atomic force microscopy (AFM). Mechanism of removing biofilm from adhering to substrate is also analyzed, thus the understanding of the mechanism can be potentially useful to prevent the biofilm formation. The results reveal that formation of biofilm can remain on rough surface regardless of substrates in hot water, which may easily induce extra-polymeric substances detachment from bacterial surface. By probing using AFM, local force-distance characterization of extra-cellular materials extracted from the bacteria can exhibit the progress of the biofilm formation and functional complexities.

[Kinetics of resorption in the cochlear perilymph after charging with radionuclides (author's transl)]. / Zur Resorptionskinetik der cochleären Perilymphe, gemessen mit Radionucliden nach minimaler Störung

The perilymphatic space of the guinea pig's cochlea is filled within 1 to 3 min with an isotonic Ringer solution labelled with a radionuclide. The cochlea is neatly sealed immediately afterwards, thus causing only a minimal disturbance in the inner ear. The animal's cochlea is brought into the strict focus of a small detector as quickly as possible, the decay of activity is continuously controlled for 2 to 10 hrs. The radionuclides used are Cl36, Na22, and Tc99m. The decay of activity of the cochlea is strongly believed to accord to a simple exponential law; half lives range from about one to about three hours. Cl36 and Tc99m were used for the first time in studying cochlear kinetics.

Susceptibility of 50 isolates of Clostridium perfringens to cefotaxime, fosfomycin, penicillin G and vancomycin; variable tolerance for vancomycin.

50 strains of Clostridium perfringens recovered from human fecal specimens were examined for susceptibility to cefotaxime (CTX), fosfomycin (Fosfo), penicillin G (Pen G) and vancomycin (Vanco) with the aid of a microtiter procedure. The geometric mean minimal inhibitory concentrations (MICs) of CTX, Fosfo, Pen G and Vanco were 0.81, 48.3, 0.15 and 0.33 micrograms/ml, respectively; the geometric mean bactericidal concentrations (MBCs) of the four drugs were 1.25, 109.8, 0.53 and 9.05 micrograms/ml, respectively. The geometric mean MBC:MIC ratios of CTX, Fosfo, Pen G and Vanco amounted to 1.56, 1.97, 3.64 and 27.0. It was concluded that the C. perfringens strains were not tolerant for CTX, Fosfo and Pen G; however, the strains displayed variable tolerance for Vanco.

The arterial switch operation in transposition of the great arteries: anatomic indications and contraindications.

1. The arterial switch procedure is now the operation of choice for typical D-transposition of the great arteries at The Children's Hospital, Boston, USA, the operative mortality rate since 1985 being 3%. 2. There are many anatomic considerations suggesting the morphologically left ventricle (LV) may well be a better systemic pump than the morphologically right ventricle (RV) which, if true, would favor the arterial switch procedure as opposed to an atrial switch operation: (1) The LV consists almost entirely of the sinus or pumping portion, and has only a minimal distal infundibular (conal) component. The RV, by contrast, has a relatively much larger infundibular component, the primary function of which is to prevent regurgitation rather than to pump. (2) Phylogenetically, the LV is the ancient "professional" pump. By contrast, the RV is a comparatively recent modification of the bulbus cordis. (3) The LV is a two-coronary ventricle, whereas the RV is a one-coronary ventricle. (4) The LV has relatively much more compact myocardium (stratum compactum) than does the RV. (5) The mitral valve leaflets are better designed to occlude a circular systemic atrioventricular orifice than are the tricuspid valve leaflets. (6) The papillary muscles of the LV are large, paired, well balanced, and both arise from the same ventricular wall--the LV free wall. By contrast, the papillary muscles of the RV are comparatively small, numerous, unbalanced, and arise from both ventricular septal and free walls. Hence, dilatation of the LV does not pull the LV papillary muscles apart, whereas dilatation of the RV does pull the RV muscles apart, favoring the development or exacerbation of tricuspid regurgitation. (7) The LV has two conduction system radiations, whereas the RV has only one. 3. The current anatomic contraindications to the arterial switch operation in typical D-TGA include the following: (1) an unprepared LV; (2) an aortic intramural left coronary artery arising from the right coronary sinus of Valsalva; (3) pulmonary outflow tract stenosis (with small annulus and subvalvar obstruction) or atresia; (4) aortic outflow tract stenosis (with small annulus and subvalvar obstruction) with tubular hypoplasia of the aortic arch and preductal coarctation; (5) tricuspid or mitral atresia; (6) marked underdevelopment or absence of either the RV sinus or the LV sinus; and; (7) 2 major anomaly of the systemic and/or pulmonary veins, as in the heterotaxy syndrome with asplenia.(ABSTRACT TRUNCATED AT 400 WORDS)

Osmotic water permeability through liposomes in the presence of alpha 1-acid glycoprotein.

Alpha 1-acid glycoprotein (orosomucoid) from human blood serum was isolated in pure form and then reconstituted into large multilamellar liposomes, consisting of a binary mixture of hen-egg phosphatidylcholine and cholesterol. These liposomes were found to be osmotically sensitive. The osmotic water permeability of proteoliposomes was determined by light-scattering measurements of the osmotic volume changes after mixing with hyperosmotic solutions of potassium salts and aminoglycoside antibiotics. The initial rate of water outflow was measured as a function of glycoprotein concentration in the mixture for the preparation of proteoliposomes. This can serve as an indication for membrane permeability to the solutes used in these experiments. It was shown that aminoglycoside antibiotics passed much faster across the membrane than potassium salts, in the presence of glycoprotein in the liposomes. A recognition pattern in the osmotic behavior of these proteoliposomes was assumed.

Manganese oxidation by an intracellular protein of a Pseudomonas species.

Cultures of a Pseudomonas sp. strain MnB 1 produce an intracellular, manganese oxidizing protein (abbrev. as Mn ox. protein) during the stationary phase of growth. This protein is heat labile, can be inactivated by protease and has a pH-optimum for manganese oxidation at pH 7.0. Mn2+ is oxidized only at concentrations below 3-10(-5) M. The occurrence of the protein is not dependent on the presence of Mn2+, but is clearly related to the cessation of growth after the end of the exponential growth phase. Oxygen, coenzymes, and low molecular weight components of the cell extract seem not to be involved in the reaction as electron acceptors for the oxidation of Mn2+. Continued manganese oxidation by Mn ox. protein results in a progressive decrease in activity which corresponds to the amount of formed manganese oxide.