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[Sterility of non-preserved autologous serum drops for treatment of persistent corneal epithelial defects].

PURPOSE: To investigate the sterility of non-preserved autologous serum drops in hospitalized patients with persistent corneal epithelial defects. METHODS: Thirty patients with persistent corneal epithelial defects (aged between 28 and 85, mean 67+/-14 years) were treated with autologous serum drops in five different wards of a university eye hospital between October 2001 and March 2002. After centrifugation of freshly collected autologous blood, the serum was stored in sterile drop bottles at refrigerator temperature (7 degrees C) and applied undiluted every 1 or 2 h up to 7 days. Using blood, chocolate, endo, and Sabouraud agar media, we assessed the sterility of autologous serum drops on the day before application and on the 4th and 7th days. Examinations performed on the 1st and 4th days included 40 autologous serum drops each and on the 7th day 120 samples (40 serum drops, 40 bottle tops, and 40 bottle bottoms). The agar media for bacterial examination were read after 48 h of incubation time, and those for fungal examination were read after 21 days. All patients had received topical autologous serum therapy and prophylactic antibiotic drops for at least 7 days. RESULTS: The time period of autologous serum treatment ranged from 7 to 28 (10+/-5) days. On the 1st and 4th days autologous serum drops were sterile. On the 7th day, 3 of 40 (7.5%) samples were contaminated with Staphylococcus epidermidis. Two of these contaminations were due to a combined serum drop and bottle top contamination (5%). A third case resulted from additional bottle bottom contamination (2.5%). None of the samples examined showed signs of fungal infection. During application of autologous serum as well as during the follow-up period no patient showed any symptoms of ocular infection. CONCLUSIONS: The application of autologous serum has proven to be a practicable therapy in patients with persistent corneal epithelial defects. When the drops are applied by trained personnel, the absence of contamination can be ensured up to the 4th day. By additional application of prophylactic antibiotic drops, infections may be avoided even if refrigerated non-preserved autologous serum is used up to 7 days.

Adult↗

A next generation, pilot-scale continuous sterilization system for fermentation media.

A new continuous sterilization system was designed, constructed, started up, and qualified for media sterilization for secondary metabolite cultivations, bioconversions, and enzyme production. An existing Honeywell Total Distributed Control 3000-based control system was extended using redundant High performance Process Manager controllers for 98 I/O (input/output) points. This new equipment was retrofitted into an industrial research fermentation pilot plant, designed and constructed in the early 1980s. Design strategies of this new continuous sterilizer system and the expanded control system are described and compared with the literature (including dairy and bio-waste inactivation applications) and the weaknesses of the prior installation for expected effectiveness. In addition, the reasoning behind selection of some of these improved features has been incorporated. Examples of enhancements adopted include sanitary heat exchanger (HEX) design, incorporation of a "flash" cooling HEX, on-line calculation of F(o) and R(o), and use of field I/O modules located near the vessel to permit low-cost addition of new instrumentation. Sterilizer performance also was characterized over the expected range of operating conditions. Differences between design and observed temperature, pressure, and other profiles were quantified and investigated.

Bioreactors↗

Sterile and economic instrumentation in laparoscopic surgery. Experiences with 6,000 surgical laparoscopies, 1990-1996.

BACKGROUND: Because so many common surgical problems can now be addressed by the laparoscopic approach, the issue of sterile processing has to be reconsidered. METHODS: Selected laparoscopic instrumentation was analyzed regarding wear and tear and decontamination after sterile processing following 6,000 surgical laparoscopies carried out between 1990 and 1996 at the Academic Hospital Moabit, Berlin. RESULTS: Fewer than 7.9 (parts of) instruments failed per 100 laparoscopies. Most of the repairs involved scissors. The main problems were blunting, burnt or disconnected electromechanical components, defective insulation, and damaged or lost parts of dismantable instruments. Residues of human blood proteins were detected on a few instruments. The effect of intraluminal rinsing was documented by measuring the iron content (as an indicator for blood contamination). A comparison of costs showed that it was >10 times cheaper to use instrumentation with reusable components. CONCLUSIONS: The sterile processing of economic reusable instrumentation for laparoscopies needs staff well trained in sterile supply. Instrument design should allow easy dismantling and rinsing of internal parts. Insulating compounds present a problem for decontamination. Disinfection with aldehydes before cleaning the lumina of instruments must be avoided because protein coagulation will occur. A tube-in-tube concept for tubular instruments offering compatibility should be favored.

Data Collection↗

Changing distribution of Candida species isolated from sterile and nonsterile sites in Israel.

To determine if the prevalence of non- albicans Candida spp. has increased at two institutions in Israel, the distribution of 6,954 Candida isolates obtained from sterile and nonsterile sites during the periods 1995-1996 and 1999-2000 were compared. In the latter period, a slight decrease was observed in the prevalence of non- albicans Candida spp. isolated from sterile sites (from 39% to 37%) and nonsterile sites (from 38% to 35%). Specifically, the prevalence of Candida glabrata increased significantly in sterile sites, from 26% to 35% (P=0.0095), and in nonsterile sites, from 18% to 27% (P<0.0001), and the prevalence of Candida krusei increased significantly in sterile sites, from 2% to 7% (P=0.0072). The prevalence of Candida parapsilosis decreased significantly in nonsterile sites, from 31% to 23% (P=0.0002). Continuous surveys of the distribution of Candida spp. and analysis of the clinical significance of changes are warranted.

Candida↗

Validation and substantiation of 25 kGy as sterilization dose for lyophilized human amnion membrane.

The validation and substantiation of sterilization dose for lyophilized human amnion membrane by gamma irradiation delivered by Co60 source were investigated. The validation experiments were conducted according to ISO 13409 method B. A total of 120 human amnion membranes were collected. Of these, 10 membranes were used for estimation of bioburden and 20 membranes were used for the individual sterility test at verification dose. The average bioburden per product unit with sample item portion (SIP = 1) for lyophilized human amnion membrane was 572 cfu. The verification dose experiments were done at dose of 8.1 kGy and the results of sterility tests showed that human amnion membrane got one positive. Consequently, the sterilization dose of 25 kGy was confirmed and substantiated.

Amnion↗

New technology for sterilization and disinfection.

Sterilization with low temperature steam and formaldehyde is a well-known process in many European countries, but little known in the United States. It sterilizes reliably and reproducibly at temperatures greater than or equal to 65 degrees C. With a well-designed cycle, it leaves residues of formaldehyde on sterilized items below 5 micrograms/cm2, measured on a standard filter paper. Formaldehyde levels in air near the autoclave are well below official exposure limits, if at all measurable. Occurrence of late growers in bioindicators, and penetration of the sterilizing media into long narrow lumina, should be validated for new processes. Automated cleaning and disinfection in closed washer-disinfectors and flushing disinfectors are likewise processes relatively little known in the United States. Disinfection is achieved by a final rinse with hot water or steam. Washer-disinfectors are used for surgical instruments, nondisposable anesthesia and other equipment, flushing disinfectors for nondisposable bedpans, wash-bowls, urinals, and similar equipment. They clean well, washer-disinfectors excellently so, and disinfect reliably. With the use of such equipment in wards, surgical departments, and other areas, reliance on chemical germicides can be dramatically reduced and disposables can be replaced by disinfectable nondisposables.

Disinfection↗

Female sterilization. III. vaginal hysterectomy.

One hundred and eleven cases of vaginal hysterectomy for sterilization, performed over a period of five and a half years, were analyzed and compared with alternative methods of female sterilization. The mean operating time for vaginal hysterectomy was 98 minutes, and the mean length of hospitalization was 9.5 days. Immediate morbidity of one sort or another occurred in 89 cases (90.0 per cent), while the incidence of standard morbidity was 40.9 percent (35 cases). These data compare most unfavorably with findings for interval tubal ligation by either the vaginal or laparoscopic approach. Our data indicate that vaginal hysterectomy for sterilization should be used only in selected patients where there is a clear indication for hysterectomy above and beyond the desire for sterilization.

Adolescent↗

Comparison of culdoscopic and laparoscopic tubal sterilization.

The growing number of women requesting interval sterilization (apart from term delivery or abortion procedures) necessitates the re-evaluation of traditional surgical procedures. A total of 800 women--the first 400 women undergoing culdoscopic and laparoscopic tubal sterilization procedures at Siriraj Hospital in Bangkok, Thailand--evaluate the appropriateness of the two sterilization procedures for use in interval sterilization programs. No serious operative complications occurred in either group. However, the rate of postoperative complications was considerably higher for the culdoscopic group than for the laparoscopic group. Hospitalization was required for only a small percentage of the patients and most had resumed their normal activities within two days of the operation. This study indicates that both procedures are effective and may be safely performed on an outpatient basis without general anesthetics and with minimal discomfort for the patients.

Culdoscopy↗

A lifetable analysis of sterilization failure: data from the Profamilia Clinic, Bogota, Colombia.

Sterilization failure rates were computed using lifetable methods for sterilizations occurring between 1973 and 1982 in the Profamilia family planning clinic in Bogota, Colombia. Out of a total of nearly 45,000 sterilizations occurring during this period 503 sterilization failures were detected. Comparing methods, laparoscopy with the silastic band method had a significantly higher rate of failure, 1.5% after 5 years compared to 0.9% for laparoscopy with cautery and 0.6% for minilaparotomy.

Actuarial Analysis↗

Changes in the circulating levels of hemoglobin, hematocrit, serum iron and total iron binding capacity after four methods of surgical tubal sterilization.

One-hundred women requesting voluntary surgical sterilization were included and enrolled into four groups (each n = 25). Four sterilization modalities were used namely: Falope ring, electrocoagulation, clip and Pomeroy ligation. The circulating levels of hemoglobin, hematocrit, serum iron and total iron binding capacity were studied before sterilization and at 3, 6 and 12 months thereafter. No significant changes were observed at 3 and 6 months post-sterilization. At 12 months a significant increase in the hemoglobin and serum iron was observed with a significant decrease in total iron binding capacity. Falope ring, clip and Pomeroy ligation were of a higher rank in this context versus electrocoagulation.

Adult↗

Safe storage times for sterile dental packs.

Safe storage times were evaluated for three sterile packaging materials that are commonly used in dentistry. No significant differences were found among packaging materials. Only three packs of 300 produced positive cultures during the 1-year test period. There were no time-related trends. The contamination levels found were within the range that has been reported to occur as a result of inadvertent contamination during unpackaging and transfer of sterile instruments. The findings of this study support the proposal that contamination is event related, not time related; that is, sterile instrument packs remain sterile for at least 1 year unless a specific event causes contamination.

Dental Instruments↗

Microwave: practical cost-effective method for sterilizing urinary catheters in the home.

We used a standard microwave oven to sterilize red rubber catheters used for intermittent self-catheterization. Catheters were incubated for sixty minutes in a suspension of microorganisms isolated from the urine of patients with urinary tract infections. For each trial, 6 catheters were removed from their respective suspensions, placed in separate plastic freezer bags, distributed evenly in a microwave oven (avoiding cold spots), and microwaved simultaneously for twelve minutes. A control catheter was not microwaved. Two strains of each microorganism were tested. The urinary isolates were Escherichia coli, Klebsiella sp., Proteus sp., Enterobacter sp., Pseudomonas sp., Streptococcus sp., Staphylococcus sp., and Candida sp. In each experiment, all 6 catheters were sterilized. Repeat sterilization in the microwave oven did not affect the integrity of the catheters or the plastic bags. A water heat sink of constant volume was employed. A home microwave oven may be used as a method to sterilize red rubber catheters for reuse with a recommended time of twelve minutes at full power. This technique makes aseptic intermittent self-catheterization a practical possibility.

Bacteriuria↗

Failure of ethylene oxide to sterilize extracted human teeth.

OBJECTIVES: The purpose of this study was to determine if ethylene oxide could sterilize extracted human teeth to be used in research. METHODS: An occlusal preparation was cut in freshly extracted molars and a small hole was drilled into the pulp chamber. A suspension of Bacillus subtilis (globigii) endospores, the standard biological monitor used for ethylene oxide sterilization, was injected into the pulp cavity, and the pulp cavity access was filled with composite material and sealed with a light-cured sealant. The teeth were exposed to either a 30 degrees C or 63 degrees C ethylene oxide sterilization process. Following exposure, the teeth were aseptically split and cultured to reveal viable spores. RESULTS: Sixty-four percent of the teeth exposed to "cold" ethylene oxide treatment and 80% of the teeth exposed to the "warm" treatment still contained viable spores. SIGNIFICANCE: Ethylene oxide cannot be relied on to sterilize extracted human teeth. Therefore, before they are used in research, other methods should be used to ensure killing of bloodborne pathogens that may be present within the teeth.

Bacillus subtilis↗

Degradative effects of conventional steam sterilization on biomaterial surfaces.

Prior to implantation trials in animals, the effect of steam sterilization on the surface properties of metallic and coated biomaterials was studied. Pure germanium plates and cast surgical Vitallium discs and subperiosteal implants were treated to present three standard types of biomaterials surfaces prior to steam sterilization, ranging from scrupulously clean, high-energy metals to uniformly low-energy organic layers. Both before and after sterilization, the sample surfaces were characterized by a variety of nondestructive physiochemical techniques. The results indicate that steam sterilization is likely to compromise the properties of otherwise carefully prepared biomedical implants by depositing hydrophobic organic and hygroscopic salt contaminants over the implant surfaces.

Biocompatible Materials↗

Effect of ethylene oxide sterilization on human demineralized bone.

Demineralized human bone used as an autologous bone graft substitute is ethylene oxide sterilized, which is reported to reduce the bone's osteoinductive potential. To determine the minimum sterilization time required and the effect of ethylene oxide on bone's osteoinductive capacity and chemical structure, samples were exposed to ethylene oxide for different times (5-240 min). Results showed that 1 h exposure sterilized the bone (assessed by Attests). This exposure time reduced the bone's osteoinductive potential and caused surface ultrastructural changes. These findings raise questions concerning the suitability of ethylene oxide for sterilizing demineralized human bone.

Animals↗

Effect of surface sterilization, fumigation and gamma irradiation on the microflora and germination of barley seeds.

Sodium hypochlorite (NaOCl) and mercuric chloride (HgCl2) surface sterilization, methyl bromide and propylene oxide fumigation and gamma irradiation treatments were compared for their effectiveness in killing microorganisms on or within barley seeds. Surface sterilization with 12.5, 25 or 50% (v/v) NaOCl for 5, 15 or 30 min, decreased Fusarium spp., Epicoccum purpurascens, and Bacillus spp. but did not kill Alternaria alternata. However, surface sterilization with 0.1 or 0.2% (w/v) HgCl2 for 3 min significantly decreased A. alternata, Fusarium spp. and E. purpurascens but Bacillus spp. were only killed by 0.3% (w/v) HgCl2 used for 10 min, which also decreased seed germination. Aspergillus flavus inoculated onto barley seeds as spores, was completely killed by surface sterilization with NaOCl but not with HgCl2, while Fusarium culmorum was killed by both NaOCl and HgCl2 treatments. Fumigation with methyl bromide yielding a concentration-time product of 3000 mg h l-1 or with propylene oxide giving a concentration-time product of 2400 mg h l-1 eliminated all filamentous fungi but Bacillus spp. and yeasts survived, and both treatments adversely affected seed germination. Gamma irradiation at a dose of 4 kGy eliminated most Alternaria, Fusarium and Epicoccum spp. but a dose of 12 kGy was required to kill Bacillus spp., yeasts and Aureobasidium pullulans. Germination was improved slightly up to a dose of 8 kGy but gradually decreased with increase in dosage to 15 kGy of gamma irradiation.

Alternaria↗

Comet assay analysis of multigenerational genomic instability (F0-F2) in Aedes aegypti exposed to gamma radiation in Sterile Insect Technique.

The use of irradiation in the Sterile Insect Technique (SIT) is a sustainable and environmentally friendly strategy for controlling Aedes aegypti populations by the release of sterile males. However, the potential toxic effects of radiation on mosquito genetic material, as well as the heritability of such damage, remain insufficiently understood. In this study, we evaluated gamma radiation-induced DNA damage (20, 30, 40, and 50 Gy) in male pupae (F0 generation) and assessed the persistence of these effects in subsequent generations (F1 and F2) using the comet assay in hemocytes. In the parental generation, a significant dose-response relationship was observed, with increasing radiation doses associated with higher damage index and damage frequency (p < 0.05). In the F1 generation, both larvae and adults exhibited significantly greater DNA damage than the control group, particularly at doses of 30 and 40 Gy, supporting the inheritance of radiation-induced genomic instability. In the F2 generation, genotoxic effects were attenuated, although residual damage remained detectable in adults, suggesting partial recovery of genomic stability, possibly influenced by DNA repair mechanisms and/or selective pressures. No viable offspring were obtained at 50 Gy, confirming the sterilizing efficacy of higher doses. Integration of comet assay results with micronucleus data and reproductive parameters reinforces the association between DNA damage, mutagenic effects, and reduced fertility. These findings indicate that radiation-induced genotoxic effects may persist beyond the irradiated generation but tend to decline across generations. Overall, this study provides insights into the balance between achieving sterility and preserving biological quality in SIT programs, contributing to optimizing radiation doses and enhancing the safety and efficacy of vector control strategies.

Comet assay↗

Finalizing the properties of porous scaffolds of aliphatic polyesters through radiation sterilization.

Porous scaffolds made of various L,L-lactide (LLA), 1,5-dioxepane-2-one (DXO) and epsilon-caprolactone (CL) copolymers were sterilized by EB- and gamma-irradiation. Differences in the comonomers, composition and the microstructure of the starting materials were used to influence the degradation mechanism and susceptibility towards irradiation and by this means to achieve sterilized scaffolds with predicted end-properties. The chemical changes and the formation of low-molecular-weight products were determined by SEC, 1H nuclear magnetic resonance (NMR), 13C NMR and gas chromatography-mass spectrometry (GC-MS). The degradation mechanism changed from random chain scission to cross-linking depending on the choice of monomers, the copolymer composition and the monomer sequences. Copolymerization of LLA with small amounts of CL or DXO increased the stability compared to that of the LLA homopolymer. Changing DXO to CL in a LLA copolymer also increased the stability. The type of radiation and the microstructure of the copolymer chains determined which of the monomer sequences were more prone to degrade. The most abundant low-molecular-weight product identified after sterilization was DXO monomer. Traces of LLA and CL monomers were also identified. Modification of the copolyester microstructure changed the degradation mechanism and the susceptibility towards irradiation. This allows the use of radiation sterilization to finalize the scaffold properties.

Biocompatible Materials↗