Pathogen-specific antibody profiles in patients with severe systemic infections.

Infections are often caused by pathobionts, endogenous bacteria that belong to the microbiota. Trauma and surgical intervention can allow bacteria to overcome host defences, ultimately leading to sepsis if left untreated. One of the main defence strategies of the immune system is the production of highly specific antibodies. In the present proof-of-concept study, plasma antibodies against 9 major pathogens were measured in sepsis patients, as an example of severe systemic infections. The binding of plasma antibodies to bacterial extracellular proteins was quantified using a semi-automated immunoblot assay. Comparison of the pathogen-specific antibody levels before and after infection showed an increase in plasma IgG in 20 out of 37 tested patients. This host-directed approach extended the results of pathogen-oriented microbiological and PCR diagnostics: a specific antibody response to additional bacteria was frequently observed, indicating unrecognised poly-microbial invasion. This might explain some cases of failed, seemingly targeted antibiotic treatment.

Monitoring of abdominal Staphylococcus aureus infection using magnetic resonance imaging: a murine animal model for hepatic and renal abscesses.

To establish a routine workflow for in vivo magnetic resonance imaging (MRI) of mice infected with bacterial biosafety level 2 pathogens and to generate a mouse model for systemic infection with Staphylococcus aureus suitable for monitoring by MRI. A self-contained acrylic glass animal bed complying with biosafety level 2 requirements was constructed. After intravenous infection with 105 colony-forming units (CFU) (n = 3), 106 CFU (n = 11) or 107 CFU (n = 6) of S. aureus strain Newman, female Balb/c mice were whole-body scanned by 7T MRI. Abdominal infections such as abscesses were visualized using a standard T2-weighted scan. Infection monitoring was performed for each animal by measurements at 1, 3, and 7 days after infection. Intravenous pathogen application led to a dose-dependent decrease in survival probability (p = 0.03). In the group with the highest infectious dose the 7-day survival rate was 33 %. An intermediate S. aureus dose showed a survival rate of 80 %, whereas at the lowest infection dose, none of the animals died. All animals with the highest infection dose exhibited hepatic abscesses 4 days after inoculation, 80 % developed renal abscesses on the 3rd day. Mice obtaining the intermediate S. aureus load reached a plateau at day 4 with 72 % liver and 60 % renal abscess probability. No abscesses were observed in other abdominal organs at any time point. The implemented experimental setup provides a suitable and reliable in vivo MRI method to study murine abdominal infection models using BSL-2 pathogen. Systemic Staphylococcus aureus infection leads to a dose-dependent development of hepatic and renal abscesses.

High prevalence of extended-spectrum-ß-lactamase-producing Enterobacteriaceae in organic and conventional retail chicken meat, Germany.

OBJECTIVES: To determine the prevalence of extended-spectrum ß-lactamase (ESBL) production in Enterobacteriaceae in retail chicken meat in Germany. METHODS: A total of 399 chicken meat samples from nine supermarket chains, four organic food stores and one butcher's shop in two geographically distinct regions (Berlin and Greifswald) were screened for ESBL production using selective agar. Phenotypic ESBL isolates were tested for bla(TEM), bla(CTX-M) and bla(SHV) genes using PCR and DNA sequencing. Antibiotic coresistances were determined and strain typing was performed using PCR-based phylogenetic grouping and XbaI-PFGE. RESULTS: A total of 185 confirmed ESBL isolates were obtained from 175 samples (43.9%) from all tested sources. The majority of isolates were Escherichia coli producing ESBL types SHV-12 (n = 82), CTX-M-1 (n = 77) and TEM-52 (n = 16). No differences could be observed in the prevalence of ESBL producers between organic and conventional samples. 73.0% of the ESBL producers showed coresistance to tetracycline, 35.7% to co-trimoxazole and 7.6% to ciprofloxacin. Strain typing of selected E. coli isolates from Berlin revealed identical macrorestriction patterns for several isolates from samples taken from the same stores. CONCLUSIONS: This is the first comprehensive study from Germany showing a high prevalence of TEM-, CTX-M- and SHV-type ESBLs in Enterobacteriaceae isolated from retail chicken meat. The high rate of coresistance to different classes of antibiotics in the ESBL producers might reflect the common veterinary usage of these and related substances. There is an urgent need to further evaluate the role of poultry in the transmission of highly resistant ESBL-producing bacteria in humans.

Effect of dose and timing of prostaglandin F2α treatments during a 7-d Ovsynch protocol on progesterone concentration at the end of the protocol and pregnancy outcomes in lactating Holstein cows.

The objective of this study was to evaluate the effect of two prostaglandin F2α (PGF) treatments 24 h apart (500 µg of cloprostenol) and treatment with a double PGF dose on d 7 (1000 µg of cloprostenol) during a 7-d Ovsynch protocol on progesterone (P4) concentration and pregnancy per artificial insemination (P/AI) in lactating Holstein cows. We hypothesized that treatment leads to a decreased P4 concentration at the second GnRH treatment (G2) and an increase in P/AI compared to the traditional 7-d Ovsynch protocol. A secondary hypothesis was that the treatment effect is influenced by the presence of a corpus luteum (CL) at the first GnRH treatment (G1). Two experiments were conducted on 8 commercial dairy farms in Germany. Once a week, cows from both experiments were assigned in a consecutive manner to receive: (1) Ovsynch (control: GnRH; 7 d, PGF; 9 d, GnRH), (2) Ovsynch with a double PGF dose (GDPG: GnRH; 7 d, 2xPGF; 9 d, GnRH), or (3) Ovsynch with a second PGF treatment 24 h later (GPPG: GnRH; 7 d, PGF; 8 d, PGF; 32 h, GnRH). All cows received timed AI (TAI) approximately 16 h after G2. Pregnancy diagnosis was performed by transrectal palpation (38 ± 3 d after TAI, experiment 1) or transrectal ultrasonography (35 ± 7 d after TAI, experiment 2). Whereas farms from experiment 1 used a Presynch-Ovsynch protocol (PGF, 14 d later PGF, 12 d later GnRH, 7 d later PGF, 2 d later GnRH, and 16-18 h later TAI) to facilitate first postpartum TAI, no presynchronization protocol was used on farms from experiment 2. In experiment 1, we enrolled 1581 lactating dairy cows (60 experimental units) from 2 dairy farms. At G2, blood samples were collected from a subsample of cows (n = 491; 16 experimental units) to determine P4 concentration at G2. In experiment 2, we enrolled 1979 lactating dairy cows (252 experimental units) from 6 dairy farms. Transrectal ultrasonography was performed to determine the presence or absence of a CL at G1. In experiment 1, treatment affected P/AI (P = 0.01) and P/AI was greater for GDPG (38.2%) and GPPG (38.9%) than for control cows (29.8%). Both, GDPG and GPPG cows had decreased P4 concentration at G2 compared with control cows (P < 0.01). Whereas both treatments increased the percentage of cows with very low P4 concentration (0.00-0.09 ng/mL) at G2, only the GPPG treatment decreased the percentage of cows with high P4 concentration (≥0.6 ng/mL) at G2 compared to the control group. In experiment 2, P/AI was greater for GPPG (37.4%) than for control cows (31.0%; P = 0.03) and tended to be greater than for GDPG cows (31.8%; P = 0.05). Cows from the GDPG group had similar (P = 0.77) P/AI compared to the control group. Pregnancy per AI did not differ between cows with a CL at G1 and cows without a CL at G1 (34.1% vs. 32.6%; P = 0.50). There was no interaction between treatment and presence of a CL at G1 on P/AI (P = 0.61). Combining data from the 2 experiments but excluding cows from experiment 1 receiving presynchronization before first TAI (n = 2573; 312 experimental units), P/AI was greater for GPPG (40.3%; P < 0.01) than for control (31.8%) and GDPG cows (33.4%). Between GDPG and control cows, P/AI did not differ (P = 0.46). We conclude that overall the addition of a second PGF treatment on d 8 during a 7-d Ovsynch protocol increased P/AI compared to the traditional 7-d Ovsynch including a single PGF dose on d 7 and to a double PGF dose on d 7. Doubling the PGF dose on d 7 in a 7-d Ovsynch protocol did not affect P/AI. Use of a presynchronization protocol, however, seems to influence the effect of a dose frequency modification of PGF treatment in an Ovsynch protocol. Presynchronized cows receiving first postpartum TAI had similarly increased P/AI treated with a double PGF dose compared with treatment with a second PGF dose. Future studies need to elucidate whether the treatment effect is modified by presynchronization of the first postpartum TAI.

Phase-variable expression of lipopolysaccharide contributes to the virulence of legionella pneumophila.

With the aid of monoclonal antibody (mAb) 2625, raised against the lipopolysaccharide (LPS) of Legionella pneumophila serogroup 1, subgroup OLDA, we isolated mutant 811 from the virulent wild-type strain RC1. This mutant was not reactive with mAb 2625 and exhibited an unstable phenotype, since we observed an in vitro and in vivo switch of mutant 811 to the mAb 2625-positive phenotype, thus restoring the wild-type LPS. Bactericidal assays revealed that mutant 811 was lysed by serum complement components, whereas the parental strain RC1 was almost serum resistant. Moreover, mutant 811 was not able to replicate intracellularly in macrophage-like cell line HL-60. In the guinea pig animal model, mutant 811 exhibited significantly reduced ability to replicate. Among recovered bacteria, mAb 2625-positive revertants were increased by fourfold. The relevance of LPS phase switch for pathogenesis of Legionella infection was further corroborated by the observation that 5% of the bacteria recovered from the lungs of guinea pigs infected with the wild-type strain RC1 were negative for mAb 2625 binding. These findings strongly indicate that under in vivo conditions switching between two LPS phenotypes occurs and may promote adaptation and replication of L. pneumophila. This is the first description of phase-variable expression of Legionella LPS.

Lack of sensitivity of an IVD/CE-labelled kit targeting the S gene for detection of SARS-CoV-2.

OBJECTIVES: New molecular tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are being rapidly launched in response to the coronavirus disease 2019 (COVID-19) pandemic. The aim of this study was to evaluate the analytical and clinical performance of the VIASURE SARS-CoV-2 S gene RT-PCR Kit on the BD Max™ system and to compare results with those obtained with the cobas® SARS-CoV-2 test on the cobas® 6800 system. METHODS: For testing the analytical performance, reference material was used. Clinical samples (n = 101) obtained from individuals with symptoms compatible with COVID-19 were studied. Oropharyngeal and nasopharyngeal swabs were collected by using either ESwab™ or UTM™ collection systems. RESULTS: When the analytical performance was evaluated, the sample containing the lowest SARS-CoV-2 concentration tested negative with the VIASURE test whereas results obtained with the cobas® test were found to be concordant with the results expected. Six out of the 101 clinical samples (5.9%) showed an inhibition with the VIASURE test. When analysing the remaining 95 clinical samples, 27 were found to be negative with both assays. Of 68 samples that were positive with the cobas® test, the VIASURE test missed 21 (30.9 %) samples. All of those 21 samples had shown Ct values ≥ 31 with the cobas® 6800 system. None of the samples tested positive with the VIASURE test and negative with the cobas® test. CONCLUSIONS: The VIASURE test was impaired by a lack of sensitivity and a relatively high number of invalid results. When using the VIASURE test for routine testing, a significant number of COVID-19-positive samples would have been missed.

Is there a risk for bacterial transmission from surgical marker pens?

Markers for preoperative skin marking are used several times and bear a risk of transmitting bacteria. Bacterial contamination was assessed by sonication and culture. Antimicrobial susceptibility testing (AST) was performed for facultative pathogens to assess multi-drug resistance (MDR). An accelerated failure time model was applied to assess the statistical relationship between the bacterial contamination and the filling status of markers. Of 45 markers, 13 had a colony count <10 cfu/mL and 32 had counts from 10 to 12,500 cfu/mL. Three markers were colonized by Staphylococcus aureus. No MDR bacteria were found. We recommend single use of markers to reduce transmission risk.

A simple laboratory algorithm for diagnosis of melioidosis in resource-constrained areas: a study from north-central Vietnam.

OBJECTIVES: Melioidosis may be endemic in many tropical developing countries, but diagnosis of the disease is currently unreliable in resource-limited areas. We aimed to validate a simple and cheap laboratory algorithm for the identification of Burkholderia pseudomallei from clinical specimens in parts of Vietnam where the disease has not previously been reported. METHODS: In June 2015, we conducted training courses at five general hospitals in north-central provinces in order to raise awareness of the disease and to introduce a simple and cheap laboratory identification algorithm for B. pseudomallei including the three-antibiotic disc test. RESULTS: Until the end of the year (7 months later), 94 suspected B. pseudomallei strains resistant to gentamicin and colistin but sensitive to amoxicillin/clavulanic acid were detected in clinical specimens from 70 patients. All strains were further confirmed as B. pseudomallei by using a specific TTSS1 real-time PCR assay and recA sequencing analysis. Among positive blood cultures, positive rates with B. pseudomallei ranged from 3.4% (5/147) to 10.2% (32/312) in the various clinics. A total of 82.8% (58/70) patients were bacteraemic, with a mortality of 50% (18/36) among patients with known outcome. No death occurred in nonbacteraemic patients. CONCLUSIONS: Our results demonstrate that the introduction of a simple and easy-to-perform laboratory algorithm for the identification of B. pseudomallei from clinical samples, together with clinical awareness raising, can lead to the diagnosis of a significant number of melioidosis cases in resource-limited clinical laboratories which previously did not identify the pathogen.

Bacterial colonisation of interior implant threads with and without sealing.

Premature loss of dental implants is due, apart from mechanical factors, to germrelated inflammation. Gaps and hollow spaces within the implant system, for example the gap between implant and abutment in the two-part implant system, may provide a bacterial reservoir causing or maintaining inflammation. The bacterial spectrum involved is similar to that found in periodontitis. This in vitro study aimed to scrutinise the capability of Porphyromonas gingivalis (DSM 20709), the bacterium blamed for inducing peri-implantitis, to pass the implant/abutment gap in titanium implant systems used for orthodontic anchorage and to remain vital in the interior. Additionally, the in vitro effectiveness of gutta percha for gap sealing was examined. Twelve titanium implants (Straumann, diameter: 3.3 mm, length 5.5 mm) were provided with abutments at a defined torque (20 Ncm), six of which were sealed with gutta percha before screwing in the abutment. Subsequently the implants were placed in a nutrient solution (thioglycolate boullion with haemin-menadione solution) that contained Porphyromonas gingivalis. Microbiological specimens were sampled from the implant interiors after 24 and 72 hours and analysed using culture methods. There was evidence that penetration of the periodontal pathogen Porphyromonas gingivalis to the implant interior may occur as early as after 24 hours. Microbes were also detected in the interior of implants sealed with gutta percha. The abutment/implant interface in vitro provides a microbiological leakage for the prospective peri-implantitis-inducing bacterium Porphyromonas gingivalis. Survival of the bacterium is possible in the interior, so that development of a bacterial reservoir is assumed. This in vitro trial produced no evidence that sealing with gutta percha is an effective means to prevent secondary bacterial colonisation in the implant interior.

Significant increase of Kuppfer cells associated with loss of Na+,K+-ATPase activity in rat hepatic allograft rejection.

BACKGROUND: Cholestasis is a complication that occurs during the rejection of liver transplants. The aim of this study was to investigate the association of activated Kupffer cells (KCs) and Na+,K+-ATPase activity for taurocholate cotransport and bile canalicular (BC) Mg++-ATPase activity for hepatobiliary excretion in rat liver allograft. METHODS: Quantitative analyses of KC number and size in relationship to enzyme activity of Na+,K+-ATPase and of BC Mg++-ATPase were conducted in rejected liver after allogenic transplantation and after prevention of rejection using cyclosporine. RESULTS: The animals were examined on the 10th postoperative day. In the rejection group, the number of KCs significantly increased more than fourfold in comparison with the number of KCs in the control livers. Some KCs were found in the sinusoids, but the majority were located in the space of Disse. Na+,K+-ATPase activity vanished from the basolateral plasma membrane, whereas BC Mg++-ATPase activity was restored in the apical domain. With immunosuppression, KCs showed the same behavior as in the control group, and activity of both ATPases was observed as strong electron-dense precipitates in basolateral and apical plasma membrane domains. CONCLUSIONS: In this study, we demonstrate that activated KCs migrate into the donor liver and release cytokines, which leads to the loss of Na+,K+-ATPase activity in the rejection group. BC Mg++-ATPase activity was not influenced by these mediators of activated macrophages. Since Na+,K+-ATPase is the cotransporter for hepatocyte taurocholate uptake, these data may contribute to understanding the mechanisms for cholestasis during hepatic allograft rejection.

Impairment of peroxisomal structure and function in rat liver allograft rejection: prevention by cyclosporine.

BACKGROUND: During allograft rejection, cytokines and lipid mediators contribute to cell injury and organ failure. Peroxisomes play a crucial role in lipid metabolism, including the degradation of lipid mediators by peroxisomal beta-oxidation. Therefore, we investigated the alterations of hepatic peroxisomes after allogeneic rat liver transplantation. METHODS: MHC-incompatible Dark Agouti (RT1a) donor rats and Lewis (RT1(1)) recipient rats were used for allogeneic transplantation. For immunosuppression, a group of these animals received cyclosporine (CsA) intraperitoneally (1 mg/kg body weight per day). Lewis rats were used for isogeneic transplant combination. Ten days after transplantation, livers were investigated using morphometrical methods for determination of peroxisomal diameter and volume density. The activities of peroxisomal catalase (CAT) and acyl-coenzyme A oxidase (AOX) were determined, and the corresponding proteins were evaluated by quantitative immunocytochemistry and immunoblotting. The expressions of mRNAs encoding CAT and AOX were investigated by Northern blotting. RESULTS: The volume density and diameter of peroxisomes were significantly decreased in allogeneic transplanted livers but were unchanged in CsA-treated animals. Both the activities of CAT and AOX and their protein levels were significantly reduced in liver allografts. Moreover, the corresponding mRNA levels of CAT and AOX were decreased significantly in liver allografts, whereas CsA treatment led to an increase of those mRNAs. Isogeneic transplanted livers showed only a slight reduction of the corresponding enzyme values. CONCLUSIONS: Peroxisomes are severely affected both morphologically and functionally after allogeneic liver transplantation. These results suggest that impairment of peroxisomal lipid beta-oxidation could contribute to the pathogenesis of the rejection process by decreased catabolism of lipid mediators involved in the regulation of the inflammatory response. CsA, in addition to its immunosuppressive effects, may contribute to allograft survival by maintenance of those important peroxisomal functions.

Optimal selective sentinel lymph node dissection in primary malignant melanoma.

OBJECTIVE: To determine the optimal approach of selective sentinel lymph node (SLN) dissection in primary malignant melanoma. DESIGN: Consecutive patient study. Prior to selective SLN dissection and wide local excision of the primary melanoma biopsy site, technetium Tc 99m sulfur colloid was injected intradermally around the primary melanoma or biopsy site to mark the SLN. Isosulfan blue (Lymphazurin, Hirsch Industries Inc, Richmond, Va) was injected at the primary biopsy site immediately before the surgical procedure. SETTING: Teaching hospital tertiary care referral center. MAIN OUTCOME MEASURES: Successful identification of SLNs being defined as positive for microscopic metastatic melanoma by blue dye staining, radioisotope uptake, or both. RESULTS: Selective intraoperative mapping by gamma probe and visualization of blue dye-stained SLN(s) resulted in a 98% (160/163) successful identification rate. Thirty patients (18.4%) had microscopic metastatic melanoma of the SLN(s), 22 of whom had subsequently completed lymphadenectomy. In 4 (18.2%) of these 22 patients, further microscopic metastatic disease was found in 1 of 8 nodes, 1 of 8 nodes, 1 of 28 nodes, and 1 of 9 nodes. No notable complications were encountered. Five recurrent cases from patients with SLNs without microscopic metastatic melanoma (3.8%) and 2 from patients with SLNs with microscopic metastatic melanoma (6%) were found during a median follow-up period of 463 days. A second primary melanoma developed in 2 patients; neither had no local recurrence. CONCLUSIONS: Sequential combination of preoperative lymphoscintigraphy and intraoperative mapping is a reliable way to identify regional SLN. The frequency of microscopic metastatic melanoma of the SLN(s) is 18.4%. Gamma-probe--guided resection minimizes the extent of lymph node dissection. Further follow-up is needed to assess the outcome of this group of patients for regional and systemic recurrences.

Monoclonal antibody against species-specific epitope of Pseudomonas aeruginosa Hsp60 protein cross-reacts with Pseudomonas stutzeri and other Pseudomonas species.

In a previous study, we determined the epitope of the Pseudomonas aeruginosa Hsp60 heat shock protein which is recognized by the specific monoclonal antibody (MAb) 2528. Subsequent investigations revealed a weak cross-reactivity of MAb 2528 with P. stutzeri, P. alcaligenes, P. mendocina and P. pseudoalcaligenes. To elucidate the molecular structure for these cross-reactions, we cloned the P. stutzeri hsp60 gene in Escherichia coli and determined the nucleotide sequence of the gene. In addition, the hsp60 gene of further Pseudomonas species was amplified and sequenced and amino acid substitutions within the epitope recognized by MAb 2528 were determined. The decapeptide QADIEARVLQ is unique to the P. aeruginosa Hsp60 protein, and cross-reaction of MAb 2528 reflects the phylogenetic relationship of Pseudomonas species as P. aeruginosa and all four cross-reacting species constitute a DNA homology group within the rRNA group I of the family Pseudomonadaceae, which belong to the gamma-subclass of the Proteobacteria.

Secretory monoclonal IgA class-switch variants against bacterial enteric pathogens in bile and intestinal secretions.

In a previous study we analyzed the molecular forms of monoclonal IgA class-switch variants (moIgA variants) and their transport into murine respiratory secretions. The aim of the present study is to characterize the transport of moIgA variants into bile and intestinal secretions so that their applicability in a passive immunization model of the gut can be evaluated. Different moIgA variants were directly isolated from IgG1 and IgG2a producing hybridoma clones specific for the same surface determinants of bacterial enteric pathogens (Salmonella typhimurium and Campylobacter jejuni) as their respective parent IgG clones. Hepatobiliary transport experiments clearly revealed the selective transport of biologically active polymeric forms of the IgA variants into the murine and rat bile after intravenous injection. Biotinylation of polymeric IgA variants prior to intravenous injection resulted in the recovery of functional, labeled SIgA. Moreover biotin-labeled polymeric IgA variant was recovered in bile with an increased molecular weight, suggesting that the secretory component had been added during passage through the liver. When IgA variant and IgG parent clones were both used in a murine backpack tumor model for passive immunization, IgA variant was selectively transported into intestinal secretions in comparison to IgG. The experimental model described here is suitable for use in comparative studies on the role of IgA and IgG with identical specificity in invasive infections of the intestinal tract.

Discordancy between clinical predictions vs lymphoscintigraphic and intraoperative mapping of sentinel lymph node drainage of primary melanoma.

OBJECTIVE: To evaluate discordancy between clinical predictions and lymphatic drainage patterns of primary cutaneous melanoma as determined by preoperative lymphoscintigraphy and intraoperative lymphatic mapping of sentinel lymph nodes (SLNs). DESIGN: Before selective SLN dissection, 226 consecutive patients with melanoma underwent preoperative lymphoscintigraphy. SETTING: Teaching hospital tertiary care center. MAIN OUTCOME MEASURE: Correlation of lymphatic drainage patterns from the following 3 data sources: clinical predictions preoperatively based on anatomical location of primary melanoma, lymphatic drainage patterns as determined by preoperative lymphoscintigraphy, and identification of SLNs during surgery. RESULTS: Preoperative lymphoscintigraphy was successful in identifying at least 1 SLN in all 226 patients. In head and neck melanomas, at least 1 SLN was identified in an area outside what would have been clinically predicted in 11 (36.7%) of 30 cases. Discordancy for trunk melanomas was seen in 24 (25.3%) of 95 cases. Extremity melanomas showed drainage to unexpected SLNs in 6 (13.6%) of 44 and 3 (5.3%) of 57 patients for the upper and lower extremities, respectively. The overall rate of discordancy was 44 (19.5%) of 226. The SLNs were identified in surgery in all but 4 cases. CONCLUSIONS: Discordancy is most frequent in melanomas of the head and neck region, followed by that of the trunk. Preoperative lymphoscintigraphy identifies the occasional cases in the upper and lower extremities where drainage occurs to a basin that is not clinically predictable. Preoperative lymphoscintigraphy is a prerequisite for characterizing the lymphatic drainage pattern in patients with primary melanoma, especially for sites such as head and neck as well as trunk, before selective SLN dissection.

Economic and clinical outcomes of microlaparoscopic and standard laparoscopic sterilization. A comparison.

OBJECTIVE: To compare micro-laparoscopic surgical sterilization and standard laparoscopic sterilization with respect to cost effectiveness and patient preferences. STUDY DESIGN: A retrospective study of all laparoscopic surgical sterilizations performed under general anesthesia at Johns Hopkins Bayview Medical Center--16 micro-laparoscopies and 34 standard laparoscopies. Cases selected for review were limited to patients undergoing surgical contraception and not requiring additional, concurrent procedures. Laparoscopic surgical sterilization was performed using a double-puncture technique with silicone band application. In each case either a standard, 10-mm laparoscope or a 2-mm micro-laparoscope was used, and the procedure was performed under general anesthesia. Postoperative pain management was achieved by nonsteroidal antiinflammatory drugs and/or narcotic analgesia. All cases were performed by residents under faculty supervision. Medical records and hospital billing records were reviewed, and a standardized telephone interview was conducted to assess postoperative quality of life and patient satisfaction. RESULTS: Both techniques were comparable in cost effectiveness. There was no significant difference in operating room time, average operating room costs, average ancillary department costs, instrument and supply costs, or length of stay. Postoperative discomfort was significantly less with microlaparoscopy (P = .05), and patient satisfaction was higher in the microlaparoscopy group. CONCLUSION: Microlaparoscopy and the standard laparoscopic approach for surgical sterilization are associated with similar hospital charges. Postoperative pain and overall patient satisfaction were significantly better with microlaparoscopy than standard laparoscopy.

Immunohistological characterization of leukocytes in the lungs of healthy mice and after bacterial intratracheal infection.

Leukocytes in the peripheral lung parenchyma of mice have not been characterized histologically during bacterial infection. The aim of this study was to investigate (a) the immunohistological characteristics of healthy murine lungs and (b) the cell kinetics during acute inflammation. BALB/c and MF1 mice were examined; as well as transgenic mice with the gene defect of cystic fibrosis (CF) in the airways as an animal model for this disease. MF1 mice served as controls for the transgenic animals. Lavaged and perfused lungs were snap frozen. B and T lymphocytes, CD4+ and CD8+ cells, dendritic cells, neutrophils and a subset of macrophages were enumerated on cryostat lung sections. The lung tissue and bronchoalveolar lavage (BAL) of BALB/c mice, infected intratracheally with Haemophilus influenzae type b (Hib), were studied at different time points after infection. In the lungs of healthy mice, including CF mice, the largest population was that of T cells, CD4+ cells being always more frequent than CD8+ cells. During acute inflammation the number of neutrophils in the lung parenchyma and BAL increased strongly within the first hours after bacterial instillation and reached baseline levels within one week. This study provides a semi-quantitative analysis of immunocompetent cells in normal and infected murine lung tissue. Differences in cell numbers are found between different strains. Moreover, the cellular reaction during Hib infection in mouse lungs is dominated by neutrophils, as expected in a primary immune response. In uninfected CF mice the numbers and distribution of immune cells in the lung tissue are normal, indicating that the cellular defense is adequate.

[Surveillance reports on pathogens and their antibiotic resistance patterns - a recommendation for standardization]. / Surveillanceberichte zu Erregern und Antibiotikaresistenzen - eine Empfehlung zur Standardisierung.

Surveillance reports on infectious agents and their antibiotic resistance patterns as well as on the usage of antibiotics are now enforced by law for many medical institutions in Germany. However, specific practice-oriented recommendations concerning the appropriate extent and informative mode of presentation are lacking. This consensus statement resulted from the experience from five German university hospitals in handling data from infection epidemiology and in the various possibilities for the presentation of surveillance reports. The consensus statement provides recommendations for the preparation of the legally demanded surveillance reports, extending the existing regulations. The relevance of statements on frequency and quality of microbiological tests is included. Furthermore, modes for the standardization of the data analysis are suggested in order to achieve a regional and national comparability of the results on a high quality level, similarly to the established standardized surveillance of nosocomial infections. This consensus statement describes the form in which the legally enforced reports can be presented in an informative and standardized way in order to facilitate the deduction and realization of preventive measurements.

Clonal distribution of superantigen genes in clinical Staphylococcus aureus isolates.

Staphylococcus aureus is both a successful human commensal and a major pathogen. The elucidation of the molecular determinants of virulence, in particular assessment of the contributions of the genetic background versus those of mobile genetic elements (MGEs), has proved difficult in this variable species. To address this, we simultaneously determined the genetic backgrounds (spa typing) and the distributions of all 19 known superantigens and the exfoliative toxins A and D (multiplex PCR) as markers for MGEs. Methicillin- sensitive S. aureus strains from Pomerania, 107 nasal and 88 blood culture isolates, were investigated. All superantigen-encoding MGEs were linked more or less tightly to the genetic background. Thus, each S. aureus clonal complex was characterized by a typical repertoire of superantigen and exfoliative toxin genes. However, within each S. aureus clonal complex and even within the same spa type, virulence gene profiles varied remarkably. Therefore, virulence genes of nasal and blood culture isolates were separately compared in each clonal complex. The results indicated a role in infection for the MGE harboring the exfoliative toxin D gene. In contrast, there was no association of superantigen genes with bloodstream invasion. In summary, we show here that the simultaneous assessment of virulence gene profiles and the genetic background increases the discriminatory power of genetic investigations into the mechanisms of S. aureus pathogenesis.