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Cloning and overexpression in Escherichia coli of the gene encoding dihydroxyacetone kinase isoenzyme I from Schizosaccharomyces pombe, and its application to dihydroxyacetone phosphate production.

The gene dak1 encoding a dihydroxyacetone kinase (DHAK) isoenzyme I, one of two isoenzymes in the Schizosaccharomyces pombe IFO 0354 strain, was cloned and sequenced. The dak1 gene comprises 1743 bp and encodes a protein of 62,245 Da. The deduced amino acid sequence showed a similarity to a putative DHAK of Saccharomyces cerevisiae and DHAK of Citrobacter freundii. The dak1 gene was expressed at a high level in Escherichia coli, and the recombinant enzyme was purified to homogeneity and characterized. The acetone powder of recombinant E. coli cells was used to produce dihydroxyacetone phosphate.

Amino Acid Sequence↗

Predicting infected bile among patients undergoing percutaneous cholecystostomy.

PURPOSE: Patients may not achieve a clinical benefit after percutaneous cholecystostomy due to the inherent difficulty in identifying patients who truly have infected gallbladders. We attempted to identify imaging and biochemical parameters which would help to predict which patients have infected gallbladders. METHODS: A retrospective review was performed of 52 patients undergoing percutaneous cholecystostomy for clinical suspicion of acute cholecystitis in whom bile culture results were available. Multiple imaging and biochemical variables were examined alone and in combination as predictors of infected bile, using logistic regression. RESULTS: Of the 52 patients, 25 (48%) had infected bile. Organisms cultured included Enterococcus, Enterobacter, Klebsiella, Pseudomonas, E. coli, Citrobacter and Candida. No biochemical parameters were significantly predictive of infected bile; white blood cell count >15,000 was weakly associated with greater odds of infected bile (odds ratio 2.0, p = NS). The presence of gallstones, sludge, gallbladder wall thickening and pericholecystic fluid by ultrasound or CT were not predictive of infected bile, alone or in combination, although a trend was observed among patients with CT findings of acute cholecystitis toward a higher 30-day mortality. Radionuclide scans were performed in 31% of patients; all were positive and 66% of these patients had infected bile. Since no patient who underwent a radionuclide scan had a negative study, this variable could not be entered into the regression model due to collinearity. CONCLUSION: No single CT or ultrasound imaging variable was predictive of infected bile, and only a weak association of white blood cell count with infected bile was seen. No other biochemical parameters had any association with infected bile. The ability of radionuclide scanning to predict infected bile was higher than that of ultrasound or CT. This study illustrates the continued challenge to identify bacterial cholecystitis among patients referred for percutaneous cholecystostomy.

Adult↗

Quantification of Salmonella by 5'-nuclease real-time polymerase chain reaction targeted to fimC gene.

A new 5'-nuclease real-time polymerase chain reaction (PCR) system for the quantification of Salmonella enterica was developed, with primers and the probe oriented to a Salmonella-specific region of the fimC gene. The PCR system was specific and sensitive, its inclusivity was 100% (determined by the analysis of 53 strains of Salmonella belonging to 38 serovars) and its exclusivity was 100% (determined by the analysis of 49 non-Salmonella strains). For quantification purposes, calibration lines were constructed for three Salmonella strains belonging to three serotypes. These calibration lines were linear (r > or = 0.99) in the range from 10(3) to 10(7 )CFU/mL and practically identical in terms of very similar slopes and x-intercepts. Escherichia coli (10(6 )CFU/mL) and Citrobacter freundii (10(6) CFU/mL) had no effect on Salmonella quantification by the system.

Colony Count, Microbial↗

Diagnosis of bacteria in vitro by mass spectrometric fingerprinting:a pilot study.

The identification of bacteria by using conventional microbiological techniques can be very time-consuming and circumstantial. In contrast, the headspace screening of bacterial cultures by analyzing their emitted volatile compounds using mass spectrometry might provide a novel approach in diagnostic microbiology. In the present study different strains of Escherichia coli, Klebsiella, Citrobacter, Pseudomonas aeruginosa, Staphylococcus aureus, and Helicobacter pylori were investigated. The volatile compounds emitted by these bacteria in vitro were analyzed using proton-transfer-reaction mass spectrometry, which allows rapid and sensitive measurement. The detected patterns of volatile compounds produced by the investigated bacteria were compared and substantial differences regarding both quantity and quality were observed. In conclusion, the present study is the first to describe headspace screening of bacterial cultures as a potential diagnostic approach in medical microbiology.

Bacteria↗

Isolation and identification of bacteria associated with adult laboratory Mexican fruit flies, Anastrepha ludens (Diptera: Tephritidae).

From the guts of new and old colonies (female and male) of Mexican fruit flies, Anastrepha ludens (Diptera: Tephritidae), we identified a total of 18 different bacterial species belonging to the family Enterobacteriaceae, Pseudomonadaceae, Vibrionaceae, Micrococcaceae, Deinococcacea, Bacillaceae, and the genus Listeria. Enterobacter, Providencia, Serratia, and Staphylococcus spp. were the most frequently isolated genera, with Citrobacter, Streptococcus, Aerococcus, and Listeria found less frequently. We found Bacillus cereus, Enterobacter sakazakii, Providencia stuartii, and Pseudomonas aeruginosa only in the new colony, Aeromonas hydrophila and Klebsiella pneumoniae spp. pneumoniae only in the old colony. We also studied resistance/sensitivity to 12 antibiotics for six bacterial isolates such as Enterobacter cloacae, E. sakazakii, K. pneumoniae spp., Providencia rettgeri, P. aeruginosa, and Bacillus cereus. Isolates on the whole were resistant to penicillin and ampicillin (five of six isolates) and sensitive to rifampin and streptomycin (six of six isolates). Antibiotic resistance profiles might be useful characteristics for distinguishing among species and strains of these bacteria, probably having ecological significance with respect to intra- and inter-specific competition within host cadavers, and could have implications for the utility of these organisms for biological control, including the alternative control strategy, paratransgenesis.

Animals↗

Liver hematoma following endoscopic retrograde cholangiopancreatography (ERCP).

We report the case of an 81-year-old man who presented with abdominal pain following endoscopic retrograde cholangiopancreatography (ERCP) for choledocholithiasis. A diagnosis of infected hematoma was made. A CT-guided puncture produced bloody matter that grew Citrobacter freundii. A catheter was left in place for 3 weeks before the patient could be discharged from hospital. We hypothesize that the hepatic parenchyma had been torn by the guide used during the ERCP. This case represents the first report of this type of iatrogenic injury.

Abdominal Pain↗

Comparative evaluation of two commercial chromogenic media for detection and presumptive identification of urinary tract pathogens.

The performance of two commercial chromogenic media for the isolation and presumptive identification of urinary tract pathogens, the CPS ID2 (bioMérieux, France) and the CHROMagar Orientation (BBL Becton Dickinson, USA), was evaluated and compared with that of cystine-lactose-electrolyte-deficient agar and tryptic soy agar with 5% sheep blood. The detection, determination of bacterial counts, and presumptive identification of bacteria causing urinary tract infections were evaluated in 3,000 urine specimens. The two chromogenic media showed excellent correlation with the standard media for the detection and the bacterial count of urinary pathogens. The Escherichia coli strains produced the expected colour on the CHROMagar Orientation and the CPS ID2 media in 99% and 90% of the cases, respectively. The Klebsiella-Enterobacter-Citrobacter and the Proteus-Morganella-Providencia groups were easily identified on both chromogenic media, but further biochemical tests were needed to differentiate them to a species level. Both media enabled the differentiation, with varying degrees of difficulty, of Pseudomonas spp. strains from members of the family Enterobacteriaceae. All isolates of Enterococcus spp. were correctly identified and were easily distinguished from the Streptococcus agalactiae isolates. Staphylococcus saprophyticus isolates were easy to identify only on the CHROMagar Orientation medium. No substantial difference was observed when comparing the results of the susceptibility tests, which were performed according to the standardized disk diffusion method as described by the National Committee for Clinical Laboratory Standards, for colonies recovered from the blood agar versus those recovered from the chromogenic media. In conclusion, the CPS ID2 and CHROMagar Orientation media enabled excellent detection, count determination, and presumptive identification of urinary pathogens, both in pure and mixed cultures, and reliable and accurate antimicrobial susceptibility testing directly from primary isolates. Moreover, these media allowed a remarkable reduction in the workload and a significant savings of time. On the basis of their performance, these media can replace the standard primary plating media used in the routine diagnosis of urinary tract infections.

Candida↗

Evaluation of a new chromogenic medium for isolation and identification of common urinary tract pathogens.

In the study presented here, a new chromogenic medium (CPS ID 3; bioMérieux, Marcy l'Etoile, France) was compared to routine media for the isolation, identification and antimicrobial susceptibility testing of bacteria recovered from urine specimens, and a cost analysis was performed. Escherichia coli, Proteeae tribe, Pseudomonas aeruginosa, Enterococcus spp. and Streptococcus agalactiae grew on the chromogenic medium as typical differentiated colonies and were accurately identified even in mixed cultures. Although the similarity of colors produced by isolates belonging to the Klebsiella, Enterobacter, Serratia and Citrobacter (KESC) group prevents differentiation among them, members of KESC were easily identified as coliforms. No substantial difference was observed when comparing the results of antimicrobial susceptibility testing performed on colonies selected from reference media versus CPS ID 3. Use of the new medium was associated with a savings of 75% over the conventional methods and the API system. Furthermore, this medium facilitated a remarkable reduction in the laboratory workload and consequently resulted in additional time and cost savings.

Anti-Bacterial Agents↗

Cefcapene inactivates chromosome-encoded class C beta-lactamases.

The stability of cefcapene and cefpodoxime, oral antibacterial cephalosporins, toward different classes of beta-lactamases was evaluated. For the class A beta-lactamases, TEM-1, SHV-1, and NMC-A, only the steady-state kinetic parameter ( k(cat)/ Km) values were calculated (3100 - 1.1 x 10(7) M(-1) x s(-1)), because these enzymes have very high Km values for cefpodoxime and cefotaxime. As for class B beta-lactamases L1, IMP-1, and CcrA, in general, similar k(cat)/ Km values were obtained. However, regarding class C beta-lactamases from Enterobacter cloacae, Escherichia coli, Pseudomonas aeruginosa, and Citrobacter freundii, we found major differences in stability between the two compounds. Cefpodoxime acted as a good substrate for the class C beta-lactamases, except for the enzyme from E. cloacae; its k(cat) and Km values were successfully calculated ( k(cat)/ Km, 1.8 x 10(5) - 1.2 x 10(7) M(-1) x s(-1)). On the other hand, cefcapene acted as a poor substrate or an inactivator for class C beta-lactamases; its k(2)/ K value was successfully calculated (8.7 x 10(5) - 7.0 x 10(6) M(-1) x s(-1)). In addition, k(3) values were determined for beta-lactamases from P. aeruginosa (2.3 x 10(-2) x s(-1)) and C. freundii (2.1 x 10(-1) x s(-1)). Even though these values could be calculated, transient inactivation as an enzyme reactivation reaction for all these enzymes was observed. These findings suggest the potential of cephem compounds as inhibitors of class C beta-lactamases.

Ceftizoxime↗

Evaluation of antibacterial efficacy of drugs for urinary tract infections by genotyping based on pulsed-field gel electrophoresis (PFGE).

We attempted to determine whether the same bacterium isolated from patients with urinary tract infections (UTI) before and after treatment with antimicrobial agents was of the same strain ("persisted") or of a different strain ("changed"). Furthermore, to verify the effectiveness of antimicrobial agents for UTI, we investigated whether bacterial strains could be classified as persisted or changed based on their electrophoretic patterns in pulsed-field gel electrophoresis (PFGE). We examined eight species of bacteria ( Enterococcus avium, Staphylococcus epidermidis, Pseudomonas fluorescens, Serratia marcescens, Citrobacter freundii, Enterobacter cloacae, Enterobacter aerogenes, and Escherichia coli), consisting of 41 strains (19 pairs isolated before and after treatment with antimicrobial agents) isolated from patients with complicated UTI. It was concluded that all bacteria were unchanged, based on the clinical effect on bacteriuria. The chromosomal DNA of these bacteria was digested with restriction enzymes and classified based on their electrophoretic pattern in PFGE. A comparison of the patterns of the fragments revealed that 14 pairs were indistinguishable before and after treatment, 3 pairs were closely related, 1 pair was possibly related, and 1 pair was different. These results demonstrated that 18 pairs of isolates (indistinguishable, closely related, and possibly related) were highly likely to be continued infections by the same strain, and that 1 pair (different) had been replaced by another strain. Based on these results, we believe that bacterium genotyping by PFGE is a more effective method for evaluating the antibacterial efficacy of antimicrobial agents.

Bacteria↗

Surveillance of bacterial resistance among isolates in Shanghai in 1999.

We report here surveillance data on the bacterial resistance of clinical isolates from 11 Shanghai hospitals in 1999, for guidance in the clinical use of antibacterial agents. Of the 14,855 strains collected, 5130 (34.5%) were Gram-positive cocci and 9725 (65.5%) were Gram-negative bacilli. The most common organisms in descending order of frequency, were: Escherichia coli (16%), coagulase-negative staphylococci (CNS; 14.3%), Klebsiella spp. (12.3%), Staphylococcus aureus (11.5%), Pseudomonas aeruginosa (9.2%), Acinetobacter spp. (8.1%), and Enterococcus spp. (6.6%). Methicillin-resistant strains accounted for 64% and 77% of S. aureus and CNS, respectively. The methicillin-sensitive strains were susceptible to most agents tested, while most methicillin-resistant strains were resistant to these agents. No vancomycin-resistant staphylococci were identified. Vancomycin-resistant strains accounted for 3.6% of Enterococcus fecalis and 1.7% of E. fecium. E. coli strains resistant to piperacillin, gentamicin, and fluoroquinolones accounted for 50% or more of the strains, and the resistance rates of Klebsiella spp., Enterobacter spp., Citrobacter spp., and Acinetobacter spp. to third-generation cephalosporins had increased markedly compared with rates in recent years. Resistance rates of P. aeruginosa to ceftazidime and imipenem (27% and 20%, respectively) had also increased compared with rates in recent years. A national strategy on the limited and prudent use of antibiotics is urgently needed.

Anti-Bacterial Agents↗

Biofilm formation in an ice cream plant.

The sites of biofilm formation in an ice cream plant were investigated by sampling both the production line and the environment. Experiments were carried out twice within a 20-day period. First, stainless steel coupons were fixed to surfaces adjacent to food contact surfaces, the floor drains and the doormat. They were taken for the analysis of biofilm at three different production stages. Then, biofilm forming bacteria were enumerated and also presence of Listeria monocytogenes was monitored. Biofilm forming isolates were selected on the basis of colony morphology and Gram's reaction; Gram negative cocci and rod, Gram positive cocci and spore forming isolates were identified. Most of the biofilm formations were seen on the conveyor belt of a packaging machine 8 h after the beginning of the production, 6.5 x 10(3) cfu cm(-2). Most of the Gram negative bacteria identified belong to Enterobacteriaceae family such as Proteus, Enterobacter, Citrobacter, Shigella, Escherichia, Edwardsiella. The other Gram negative microflora included Aeromonas, Plesiomonas, Moraxella, Pseudomonas or Alcaligenes spp. were also isolated. Gram positive microflora of the ice cream plant included Staphyloccus, Bacillus, Listeria and lactic acid bacteria such as Streptococcus, Leuconostoc or Pediococcus spp. The results from this study highlighted the problems of spread of pathogens like Listeria and Shigella and spoilage bacteria. In the development of cleaning and disinfection procedures in ice cream plants, an awareness of these biofilm-forming bacteria is essential for the ice cream plants.

Biofilms↗

Efficiency of modified H2S test for detection of faecal contamination in water.

Faecal contamination is a major causative factor for incidence of water borne infectious diseases. Certain hydrogen sulfide (H(2)S) producing enteric bacteria such as Salmonella sp. and Citrobacter sp., associated with coliforms, have been considered for rapid detection of recent faecal contamination in water. The basic H(2)S test medium, modified by adding 0.25 gm/L of L-cystine HCl, was examined for its efficiency with 90 water samples collected from 40 pipe supplies, 20 open wells, 15 hand pumps and 15 different surface water bodies (river, streams and ponds). Sterilized modified culture medium in glass vials was inoculated with 100 mL of each sample and incubated at 20, 25, 30, 35 and 44 degrees C for 18, 24, 42, 48, 66 and 72 h. Blackening of content in incubated vials was considered positive. For comparison, most probable number (MPN) of coliform and faecal coliform per 100 mL was also estimated in each sample by multiple tube fermentation (MTF) method. H(2)S positive result was exhibited by 78% of samples. Coliform (> 10) and faecal coliform/100 mL were also detected in 59% of samples. Maximum H(2)S positive results (100%) were found with well and surface water samples incubated at 30, 35 and 44 degrees C for 18 h. Coliform (> 10) and faecal coliform/100 mL were also detected in most of these samples. Pipe supplies (60%) and hand pumps (73%) also exhibited considerable H(2)S production. Coliforms and faecal coliforms were also found in significant number of these samples. Thus, the modified H(2)S test may prove a useful alternative indicator of faecal contamination for water quality surveillance and screening of large number of water samples in short duration, particularly during any outbreak of water borne disease among rural population.

Bacteriological Techniques↗

Mixed infection in adult bacterial meningitis.

12 adult patients suffering from bacterial meningitis caused by mixed infection were identified at Kaohsiung Chang Gung Memorial Hospital over a period of 13 years (1986-1998), and they accounted for 6.5% (12/184) of our culture-proven adult bacterial meningitis. The 12 cases included seven males and five females, aged 17-74 years. Six of the 12 cases had community-acquired infections and the other six had nosocomially-acquired infections. Ten of the 12 cases had associated underlying diseases, with head trauma and/or neurosurgical procedure being the most frequent. Both gram-negative and gram-positive pathogens were identified in these 12 cases with gram-negative pathogens outnumbering the gram-positive ones. The implicated pathogens, starting with the most frequent, included Enterobacter species (Enterobacter cloacae, Enterobacter aerogenes), Klebsiella species (Klebsiella pneumoniae, Klebsiella oxytoca), Escherichia coli, Staphylococcus species (Staphylococcus aureus, Staphylococcus haemolyticus), Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus, Serratia marcescens, Citrobacter diversus, Proteus mirabilis, Streptococcus viridans and Neisseria meningitidis. Six of the 12 cases were found to have multi-antibiotic-resistant strains, which included E. cloacae in one, A. baumannii in one, K. pneumoniae in one and S. aureus in three. The management of these 12 cases included appropriate antibiotics and neurosurgical procedures including shunt revision. Despite the complexity of implicated pathogens and the high incidence of emergence of resistant strains, the overall mortality rate (8.3%, 1/12) was not higher than that in adult bacterial meningitis. However, complete recuperation was difficult in adult patients with mixed bacterial meningitis.

Adolescent↗

Contribution of beta-lactamases to bacterial resistance and mechanisms to inhibit beta-lactamases.

Resistance of bacteria to beta-lactam antibiotics has become a serious problem in the past several decades. Virtually all Staphylococcus aureus, and many Hemophilus influenzae, Branhamella catarrhalis, Neisseria gonorrhoeae, Enterobacteriaceae, and Bacteroides species possess beta-lactamases that hydrolyze penicillins and cephalosporins. The most common plasmid-mediated beta-lactamase is the TEM enzyme (Richmond-Sykes type IIIa), which is present in Hemophilus, Neisseria, and Enterobacteriaceae. One technique to overcome bacterial resistance has been the development of beta-lactamase inhibitors. Clavulanic acid is a beta-lactamase inhibitor that inhibits the beta-lactamases of S. aureus, Hemophilus, Neisseria, Branhamella, Eschericia coli, Klebsiella, and Bacteroides. Clavulanate acts as a "suicide" inhibitor, forming a stable enzyme complex that binds to serine at the active site of the enzyme. Clavulanate readily crosses the outer cell wall of most Enterobacteriaceae to interact with beta-lactamases in the periplasmic space. Clavulanate does not inhibit beta-lactamases such as the Richmond-Sykes type I enzymes found in Pseudomonas aeruginosa, Enterobacter, and Citrobacter species, which are inducible enzymes that function primarily as cephalosporinases.

Anti-Bacterial Agents↗

Timentin in the treatment of symptomatic complicated urinary tract infections in adult patients.

The safety and effectiveness of Timentin were evaluated in 34 adult patients with symptomatic complicated urinary tract infections, principally due to multiply-drug-resistant bacteria. Although a wide variety of organisms, particularly gram-negative bacilli, were found, Escherichia coli was the most frequent, accounting for 14 of 45 (31 percent) pathogens isolated. Ten (22 percent) isolates were Pseudomonas aeruginosa; 11 (24 percent) were Proteus or Morganella species; three (7 percent) were Citrobacter; one (2 percent) was Klebsiella pneumoniae; two (4 percent) were Staphylococcus aureus; and two (4 percent) were enterococci. Ninety-three percent of all pathogens isolated produced a beta-lactamase. Eight (24 percent) infections were polymicrobial; seven (21 percent) were associated with bacteremia. Clinical improvement occurred in 30 of 34 (86 percent) patients. All bacteremias were cured. Although bacteriologic cure occurred in only 32 percent of patients, control of sepsis and temporary eradication of bacteria (bacteriologic improvement) occurred in 96 percent. Not surprisingly, the rates of relapses and reinfections were high. It was concluded that Timentin is a useful agent in the management of complicated urinary tract infection and offers clinicians an alternative to more toxic antibiotics, such as aminoglycosides.

Adult↗

Treatment of pediatric infections with amikacin as first-line aminoglycoside.

Because of increased aminoglycoside resistance of hospital bacterial isolates, aminoglycoside sensitivity patterns of isolates in a large children's hospital were assessed before and during a 33-month period of almost exclusive amikacin use. There was no significant change in overall resistance rates of gram-negative enteric bacteria to gentamicin (4.8 percent and 4.6 percent), tobramycin (2.5 percent and 3.6 percent), and amikacin (1.2 percent and 1.8 percent) from the pre-amikacin period to the amikacin usage period, respectively. No significant differences were observed for isolates of Escherichia coli, Klebsiella, Serratia, Acinetobacter, and Pseudomonas species. In contrast, significant decreases in gentamicin and tobramycin resistance rates for Enterobacter, Citrobacter, and Pseudomonas aeruginosa and in gentamicin resistance of Proteus were found. Very little change in resistance of staphylococcal isolates was seen during a shorter evaluation period. Pediatric aminoglycoside usage includes therapy of neonatal infections, cystic fibrosis, febrile neutropenic episodes in patients with cancer, abdominal surgery, bacterial endocarditis, and gram-negative central nervous system infections. Amikacin has also been used successfully as single-dose therapy of urinary tract infections, and acceptable cerebrospinal fluid levels of amikacin have been documented in hydrocephalic patients with ventriculitis. In vitro studies of 22 bacterial isolates demonstrated synergy between amikacin and penicillin or newer cephalosporins in 13, an additive effect in seven and indifference in two. No antagonism was found. In addition, in vivo synergy between imipenem and amikacin was found in neutropenic infant rats with P. aeruginosa sepsis using a strain with which no synergy was demonstrable in vitro. Amikacin is effective in pediatric infections and is well tolerated by children. Because excessive or inadequate levels are frequent with usually recommended doses, particularly in neonates and patients with compromised renal function or cystic fibrosis, serum levels should be monitored to minimize risk and to ensure therapeutic levels.

Amikacin↗

Treatment of gram-negative infections with aztreonam.

Twenty-one patients with serious gram-negative infections were treated with aztreonam. Twenty of these were clinical and microbiologic cures; there was one clinical improvement with microbiologic persistence. No bacteria became resistant. Cure rates were: bone and joint (11 of 11); skin and soft tissue (six of six); pneumonia (two of two); perinephric abscess (one of one); and intra-abdominal abscess (zero of one). The bacteria responsible for these infections included Pseudomonas aeruginosa (12), Serratia marcescens (two), Enterobacter gergoviae (three), Enterobacter aerogenes (two), Escherichia coli (one), Citrobacter diversus (one), and Hemophilus influenzae (one). Aztreonam was well tolerated. Significant serum glutamic-oxaloacetic transaminase/serum glutamic-pyruvic transaminase elevations developed in three patients, but none was symptomatic and all resolved after therapy was stopped. Two patients in whom a rash developed were receiving other antibiotics (vancomycin and metronidazole), making the cause of the rash unclear. Diarrhea developed in a single patient with Pseudomonas osteomyelitis, who also was receiving cefazolin for Staphylococcus aureus superinfection of his decubitus ulcer. Aztreonam was highly effective against gram-negative bacilli, including P. aeruginosa. The only clear-cut side effect was an asymptomatic rise in serum glutamic-oxaloacetic transaminase/serum glutamic-pyruvic transaminase levels in three patients.

Abscess↗