Detachment of trophozoites of Acanthamoeba species from soft contact lenses with BEN22 detergent, BioSoak, and Renu multi-purpose solutions.

PURPOSE: BEN22 detergent was studied for its ability to detach Acanthamoeba from soft contact lenses without mechanical cleaning or separate cleaning agents. METHODS: Trophozoites of Acanthamoeba castellanii and A. polyphaga were adhered onto nonionic, high water content soft contact lenses. The lenses were immersed for 2 hours in contact lens care solutions and the remaining trophozoites were counted microscopically. The counts were compared to the counts on the same lens before treatment. RESULTS: BEN22 (50:50 mixture of L-alpha-L-rhamnopyranosyl-beta-hydroxydecanoyl-beta-hydroxydecanoate and 2-O-alpha-L-rhamnopyranosyl-alpha-L-rhamnopyranosyl-beta-hydroxydecanoyl-beta-hydroxydecanoate) (Kassell Industries, Inc., Wisconsin Dells, WI) in a concentration of 0.05% detached the trophozoites to a statistically significant greater extent than saline, but commercial ReNu Multi-Purpose Solution (Bausch & Lomb, Italy) and BioSoak (Finnsusp Ltd., Finland) did so as well. ReNu Multi-Purpose Solution was more effective than 0.005% BEN22 in detaching the trophozoites of both of the Acanthamoeba strains. After the 2 hour immersion period, a maximum of 97% of the initial trophozoites were detached. The variation between individual lenses was significantly greater than that within the different areas of one lens. CONCLUSIONS: BEN22 had no reliable detaching effect on Acanthamoeba. The variation between lenses was great, and the rate of detachment was low with all the agents tested indicating that immersion and rinsing in the solutions tested cannot be considered as a safe substitute for proper disinfection against Acanthamoeba in contact lens care.

Measurement of the D-arabinitol/L-arabinitol ratio in urine of neutropenic patients treated empirically with amphotericin B.

The aim of the present study was to evaluate the diagnostic significance of the D-arabinitol/L-arabinitol ratio in urine of neutropenic patients with suspected fungal infection. D-arabinitol/L-arabinitol ratios were determined in 373 serial urine samples of 104 patients with haematological malignancies receiving empirical amphotericin B treatment for suspected invasive fungal infection. Twenty-eight (8%) urine samples obtained from 17 (16%) patients were positive (ratio > or =4). Eight (47%) patients had positive urine samples at the initiation of empirical amphotericin B treatment and the rest from 7 to 30 days after empirical therapy was started. Several urine samples were positive in six patients. Only one of the five patients with candidemia had elevated D-arabinitol/L-arabinitol ratios (persistent Candida krusei fungaemia). Four patients with transient candidemia and seven patients with invasive mould infections were negative. Patients who died during the study period had significantly higher D-arabinitol/L-arabinitol ratios than patients who survived (P=0.0002). Pneumonia was the most common manifestation of infection (53% of patients with elevated D-arabinitol/L-arabinitol ratios) and was associated with an especially high mortality (67%). The present study shows that elevated urine D-arabinitol/L-arabinitol ratios are common in febrile, neutropenic patients. However, the urine arabinitol test did not detect transient candidemia at elevated levels during the course of infection. Furthermore, D-arabinitol/L-arabinitol ratios were often elevated in the late phase of infection only. This contests the use of this test in guiding the initiation of antifungal therapy. The detection of elevated arabinitol levels in neutropenic patients during empirical amphotericin B treatment is associated with poor prognosis.

Importance of intestinal colonisation in the maturation of humoral immunity in early infancy: a prospective follow up study of healthy infants aged 0-6 months.

AIM: To evaluate the role of intestinal microflora and early formula feeding in the maturation of humoral immunity in healthy newborn infants. STUDY DESIGN: Sixty four healthy infants were studied. Faecal colonisation with Bacteroides fragilis group, Bifidobacterium-like, and Lactobacillus-like bacteria was examined at 1, 2, and 6 months of age, and also the number of IgA-secreting, IgM-secreting, and IgG-secreting cells (detected by ELISPOT) at 0, 2, and 6 months of age. RESULTS: Intestinal colonisation with bacteria from the B fragilis group was more closely associated with maturation of IgA-secreting and IgM-secreting cells than colonisation with the other bacterial genera studied or diet. Infants colonised with B fragilis at 1 month of age had more IgA-secreting and IgM-secreting cells/10(6) mononuclear cells at 2 months of age (geometric mean (95% confidence interval) 1393 (962 to 2018) and 754 (427 to 1332) respectively) than infants not colonised (1015 (826 to 1247) and 394 (304 to 511) respectively); p = 0.04 and p = 0.009 respectively. CONCLUSIONS: The type of bacteria colonising the intestine of newborns and the timing may determine the immunomodulation of the naive immune system.

Aspergillus antigen in serum, urine and bronchoalveolar lavage specimens of neutropenic patients in relation to clinical outcome.

We have used a new, commercial enzyme-linked immunosorbent assay (ELISA, Platelia Aspergillus) to detect Aspergillus antigen in serum, urine and bronchoalveolar lavage (BAL) samples of 105 haematological patients who received empirical amphotericin B treatment for suspected fungal infection. 14% (60/419) of serum and 5% (18/373) of urine samples were positive. Ten-fold concentration of urine increased the number of positive samples to 31 (8%). The antigen was detected in 5 of 20 BAL samples, but fungal culture was negative in all of them. 22 patients had positive antigen test. Serum was positive in 17, urine in 7 and concentrated urine in 12 patients. Six patients had confirmed invasive aspergillosis. In all these patients, antigen was detected in serum, but urine was positive in only 2 patients. Patients whose antigen test turned negative during the amphotericin B treatment had significantly lower mortality than patients with persistently positive antigen test (2/10 vs. 8/8, p = 0.002). We conclude that Aspergillus galactomannan can be detected by ELISA in serum, urine and BAL samples of haematological patients, but the higher sensitivity of serum testing makes it preferable for screening. Disappearance of the antigen during antifungal therapy seems to correlate with good, and persistence with poor, clinical outcome.

Fungal colonization of haematological patients receiving cytotoxic chemotherapy: emergence of azole-resistant Saccharomyces cerevisiae.

Fungal colonization during cytotoxic chemotherapy was studied in 42 patients with a recent diagnosis of a haematological malignancy. In total, 2759 surveillance cultures were taken from the nostrils, throat, urine, stool and perineal region. Seven hundred and ninety-six positive surveillance cultures (28.9%) yielded 968 fungal isolates. The rate of fungal colonization did not differ between patients with acute leukaemia, patients with other haematological malignancies and control patients in the same ward at admission (71% vs. 67% vs. 80%). Patients with acute leukaemia were colonized at a significantly lower rate in samples from the throat (32%), urine (10%), stool (45%) and perineum (29%) taken during hospitalization when compared with other haematological patients (respective values 58%, 21%, 67% and 45%; P-values 0.001). This could be attributed to differences in the use of antifungal drugs. Although 21/42 (50%) of our patients had multiple-site fungal colonization at the end of follow-up, only one systemic Candida infection was diagnosed. Extensive use of antifungal treatment may have influenced the low incidence of systemic fungal infections during the follow-up. In addition to Candida species, Malassezia furfur, Geotrichum candidum and Saccharomyces cerevisiae were frequently isolated. The rate of S. cerevisiae isolation increased significantly over time after admission (1%, vs. 18% of isolates, P<0.001), suggesting hospital-acquired transmission. These isolates were highly resistant to azole antifungals (MIC90 128 microg/mL for fluconazole and 16 microg/ml, for itraconazole), and caused persistent multiple site colonization in 12 patients. Extensive use of antifungal agents in a haematological ward may keep the incidence of invasive fungal infections low in spite of heavy fungal colonization. However, there may be a risk of emergence of resistant fungal strains.

Development of intestinal bacterial enzymes in infants--relationship to mode of delivery and type of feeding.

To evaluate the development of intestinal flora in young infants, and especially to estimate the influence of mode of delivery and type of feeding on the establishment of intestinal microflora, faecal flora was studied indirectly by measuring prospectively the faecal bacterial enzyme activities (beta-glucosidase, beta-glucuronidase and urease) in 29 full-term, healthy infants during the first 6 months of life. Mode of delivery had no influence on the faecal enzyme activities. In contrast, infants receiving formula feeds were more often urease positive at 1-2 months of age (70% vs 25%, p=0.043) and had higher median activity of beta-glucuronidase at 6 months of age (0.90 and 0.19 nmoles/mg protein x min, p= 0.0043) than exclusively breast-fed infants. Through indirect methods to measure the development of a faecal microflora our results indicate that the type of milk that infants receive during the first months of life may have an important role in the development of intestinal flora.

Mode of delivery directs the phagocyte functions of infants for the first 6 months of life.

Factors that direct the immune responsiveness of the newborn beyond the immediate post-natal period are not known. We investigated the influence of mode of delivery and type of feeding on the phagocyte activity during the first 6 months of life. Sixty-four healthy infants (34 delivered vaginally and 30 by elective Caesarean section) were studied at birth and at the ages of 2 and 6 months. Phagocyte functions were studied by measuring the chemiluminescence (CL) activity of whole blood and isolated leucocytes and by investigating the expression of phagocyte receptors (FcgammaRI (CD64), FcgammaRII (CD32), FcgammaRIII (CD16), CR1 (CD35), CR3 (CD11b) and FcalphaR (CD89)) on neutrophils, monocytes and eosinophils by using receptor-specific MoAbs and immunofluorescence flow cytometry. Infants born by elective Caesarean section had significantly higher CL activity than those delivered vaginally during the entire 6-month follow up. In addition, infants who received formula feeds had significantly higher CL activity at 6 months of age and higher expression of FcgammaRI-, Fcalpha- and CR3-receptors on neutrophils than infants exclusively breast-fed. We suggest that stress reaction associated with labour influences the phagocytic activity measured in the cord blood but later during infancy the intraluminal antigens, gut microflora and diet, become important determinants in immune programming of human individuals.

Papillary hypertrophy of the upper tarsal conjunctiva during contact lens wear: a 4-month study with ethyl-6-O-decanoyl-glucoside.

PURPOSE: We studied the potential effect of ethyl-6-O-decanoyl-glucoside (EDG) on papillary hypertrophy in contact lens wearers who were recruited on the basis of papillary hypertrophy and a long history of contact lens wear. The contact lens care solutions were 0.00025% chlorhexidine acetate (CHX) with or without 0.005% EDG. METHODS: Nineteen subjects wearing both ionic and non-ionic contact lenses for 6-18 hours used either CHX or CHX+EDG as a cleaning and disinfecting agent. CHX and CHX+EDG was used simultaneously by each subject but in different eyes during two consecutive periods of 8 weeks. Symptoms and signs were recorded at three examinations during the study. The protein content of contact lenses and tryptase activity of tear fluids were measured. RESULTS: The degree of papillary hypertrophy did not decrease in either the CHX or CHX+EDG groups. Also, there were no differences in protein content of lenses nor tryptase activity of tear fluids in either group. There was a significant correlation between papillary hypertrophy and tryptase activity during the study. CONCLUSIONS: Despite the earlier finding that EDG prevents development of papillary hypertrophy in contact lens wearers, EDG still cannot reverse established signs of papillary hypertrophy.

Fecal microflora in healthy infants born by different methods of delivery: permanent changes in intestinal flora after cesarean delivery.

BACKGROUND: Newborn infants in modern maternity hospitals are subject to numerous factors that affect normal intestinal colonization--for example, cesarean delivery and antimicrobial agents. To study the duration of the effect of external factors on intestinal colonization, two groups of infants with different delivery methods were investigated. METHODS: The fecal flora of 64 healthy infants was studied prospectively. Thirty-four infants were delivered vaginally, and 30 by cesarean birth with antibiotic prophylaxis administered to their mothers before the delivery. The fecal flora was cultured on nonselective and selective media in infants 3 to 5, 10, 30, 60, and 180 days of age. Gastrointestinal signs were recorded daily by the mothers for 2 months. RESULTS: The fecal colonization of infants born by cesarean delivery was delayed. Bifidobacterium-like bacteria and Lactobacillus-like bacteria colonization rates reached the rates of vaginally delivered infants at 1 month and 10 days, respectively. Infants born by cesarean delivery were significantly less often colonized with bacteria of the Bacteroides fragilis group than were vaginally delivered infants: At 6 months the rates were 36% and 76%, respectively (p=0.009). The occurrence of gastrointestinal signs did not differ between the study groups. CONCLUSIONS: This study shows for the first time that the primary gut flora in infants born by cesarean delivery may be disturbed for up to 6 months after the birth. The clinical relevance of these changes is unknown, and even longer follow-up is needed to establish how long-lasting these alterations of the primary gut flora can be.

Urinary bacterial flora of women with urethral syndrome and interstitial cystitis.

The aerobic and anaerobic bacterial flora in the first voided and in the midstream urine of healthy females (n = 5) and female patients with either urethral syndrome (US) (n = 5) or interstitial cystitis (IC) (n = 14) were studied. Bacteria were grown on media enabling isolation of fastidious and aerobic as well as obligatory anaerobic species. In healthy females only gram-positive rods were found whereas US patients also harbored streptococci. Patients with IC presented also with Enterobacteriae and anaerobic bacteria. Five IC patients with severe symptoms were treated with metronidazole; 2 out of 3 patients with anaerobic bacteria in the pretreatment specimens had no anaerobes after metronidazole therapy and in 1 patient streptococci disappeared after the therapy. One patient with severe symptoms and Bacteroides fragilis in the midstream urine became symptom-free after 2 weeks of metronidazole treatment. Although there is uncertainty whether the US and IC are not related to an infectious etiology, the bacterial flora in urethral and in midstream urine in these conditions differs considerably from that of healthy females.

Diagnosis of meningococcal meningitis by broad-range bacterial PCR with cerebrospinal fluid.

We used broad-range bacterial PCR combined with DNA sequencing to examine prospectively cerebrospinal fluid (CSF) samples from patients with suspected meningitis. Fifty-six CSF samples from 46 patients were studied during the year 1995. Genes coding for bacterial 16S and/or 23S rRNA genes could be amplified from the CSF samples from five patients with a clinical picture consistent with acute bacterial meningitis. For these patients, the sequenced PCR product shared 98.3 to 100% homology with the Neisseria meningitidis sequence. For one patient, the diagnosis was initially made by PCR alone. Of the remaining 51 CSF samples, for 50 (98.0%) samples the negative PCR findings were in accordance with the negative findings by bacterial culture and Gram staining, as well as with the eventual clinical diagnosis for the patient. However, the PCR test failed to detect the bacterial rRNA gene in one CSF sample, the culture of which yielded Listeria monocytogenes. These results invite new research efforts to be focused on the application of PCR with broad-range bacterial primers to improve the etiologic diagnosis of bacterial meningitis. In a clinical setting, Gram staining and bacterial culture still remain the cornerstones of diagnosis.

Information system for clinical resource management in a regional setting.

The Hospital District of Varsinais-Suomi consists of five hospitals for non-psychiatric diseases to deliver specialised health care for the population of 440,000 inhabitants in South-western Finland. We have applied an open regional information system to aid clinical resource management in such a complex expert organisation. The main areas of application have been assessment of inpatient care usage in different communities and in different diagnosis-related-groups (DRG), measurement of length of hospital stay in different DRGs and comparison of DRG unit weight costs of different hospitals. The open structure of the data base and its distribution to all parties of health care professionals has resulted in mutual decisions on structural changes and increased cost containment activities.

Alcohol abuse: a risk factor for surgical wound infections?

BACKGROUND: The incidence of postoperative surgical site infections (SSIs) is difficult to estimate because of the current trend of early discharge after surgery. Both operation-related and host factors should be taken into consideration in the prevention of SSIs. We wanted to determine the actual incidence of SSIs and evaluate the risk factors in our clinic, using an extended follow-up period of 30 days after operations. METHODS: We performed a prospective follow-up survey of SSIs over a 3.5-month period including a 1-month follow-up after discharge with written instructions and a telephone survey. The SSIs were defined according to Centers for Disease Control and Prevention criteria. Forty-three patient parameters were recorded, and risk factors for SSI were sought and tested by using multiple logistic regression analysis. RESULTS: The follow-up was completed in 772 of 807 patients. The SSI rates in these patients were 5.3% in clean, 7.1% in clean-contaminated, 6.2% in contaminated, and 28.1% in dirty operations. Seventy-one percent of infections were not diagnosed until after discharge from the hospital. According to multiple logistic regression analysis, alcohol abuse (p < 0.0001), wound contamination class (p < 0.05), and operation duration of over 2 hours (p < 0.05) were independently significant risk factors for SSI. CONCLUSIONS: A major portion of SSIs are found only after follow-up is extended during the postdischarge period. Alcohol abuse is a significant risk factor for SSI and should be taken into account when determining the susceptibility of an individual patient.

Ligase chain reaction in detection of Chlamydia DNA in synovial fluid cells.

Synovial fluid cells from 12 patients with reactive arthritis (ReA) triggered by Chlamydia trachomatis were studied for the presence of Chlamydia DNA using the ligase chain reaction (LCR) LCx (Abbott) and the polymerase chain reaction (PCR) Amplicor (Roche). In addition, peripheral blood leucocytes from 11 of these patients were analysed by LCR. As controls, seven patients with newly diagnosed rheumatoid arthritis (RA) were included. Chlamydia trachomatis DNA was detectable by LCR in samples of synovial fluid cells from 4/12 patients with C. trachomatis-triggered ReA, and in none by PCR. Chlamydia trachomatis DNA was not detectable in the synovial fluid cells of the seven RA patients by either method, neither was C. trachomatis DNA detectable in the peripheral blood leucocytes of the ReA patients (0/11) or controls (0/6) by LCR. The LCR technique may be useful in the demonstration of Chlamydia DNA in synovial fluid cells.

Faecal fatty acids and gastrointestinal upset in newborn infants.

Patterns of long-chain faecal fatty acids were studied by gas-liquid chromatography in 55 newborn infants in a neonatal intensive care unit. Decreased fractions of fatty acid C16:1 and increased fractions of C16:0 and C17:0 were associated with the occurrence of abdominal distension. Decreased fractions of C16:1 and C18:2 were associated with diarrhoea. Flatulence was found in infants who had relatively smaller amounts of fatty acids C17:0D and C15:0 in their faecal samples. The differences in the patterns of faecal fatty acids are due to differences in bacterial flora. The results support the hypothesis that the initial intestinal colonization plays a role in the later gastrointestinal signs of newborn infants.

Bacteriological evaluation of 194 adult patients with acute frontal sinusitis and findings of simultaneous maxillary sinusitis.

In a prospective study bacteriological findings from frontal and maxillary sinus secretes were analyzed. Frontal sinus trephination was performed to 103 and maxillary irrigation to 192 patients. Main pathogens in the frontal as well as in the maxillary sinus secretes were Haemophilus influenzae and Streptococcus pneumoniae. beta-lactamase production of H. influenzae was found in 17% of frontal and in 16% of maxillary sinus samples. Bacteriological findings were supported by the staining results. The portion of anaerobic pathogens in cultures were poor, though Bacteroides fragilis antigen was detected in 8 frontal and 6 maxillary sinus samples. The acute frontal sinusitis should be treated with respective antibiotics. Complications seem not to develop in the acute phase and the disease can primarily be treated conservatively. It should however be controlled properly to avoid prolonged disease and late complications.

Bacteriology of lacrimal duct obstruction in adults.

AIMS: To determine the current bacteriology of lacrimal duct obstruction (LDO) and to relate the bacteriological findings to the type of symptoms. METHODS: 127 samples were obtained from the lacrimal sac in 118 consecutive adult patients with LDO, including nine bilateral cases. RESULTS: Altogether, 156 isolates were recovered from the 127 samples cultured. Cultures were positive from 84% of the samples. Gram positive bacteria were isolated in 79 (62%) samples. The most frequently cultured bacterial species was Staphylococcus epidermidis, representing 27% of the isolates. Gram negative bacteria were recovered from 26 (20%) samples, and these bacteria were statistically significantly more common in cases with copious discharge than in cases with minor discharge (p = 0.000). Cases with simple stenosis of the lacrimal duct (SSLD) showed significantly less Streptococcus sp (p = 0.004) and Gram negative organisms (p = 0.004) than those with chronic dacryocystitis. CONCLUSION: The bacteriology of SSLD resembles that of normal conjunctival flora. Chronic dacryocystitis in adults is associated with an increased proportion of Gram negative bacteria which may be a reservoir for postoperative intraocular infection. They should also be taken into account in selecting antimicrobial prophylaxis in lacrimal drainage surgery.