A Shipping Container-Based Sterile Processing Unit for Low Resources Settings

Deficiencies in the sterile processing of medical instruments contribute to poor outcomes for patients, such as surgical site infections, longer hospital stays, and deaths. In low resources settings, such as some rural and semi-rural areas and secondary and tertiary cities of developing countries, deficiencies in sterile processing are accentuated due to the lack of access to sterilization equipment, improperly maintained and malfunctioning equipment, lack of power to operate equipment, poor protocols, and inadequate quality control over inventory. Inspired by our sterile processing fieldwork at a district hospital in Sierra Leone in 2013, we built an autonomous, shipping-container-based sterile processing unit to address these deficiencies. The sterile processing unit, dubbed “the sterile box,” is a full suite capable of handling instruments from the moment they leave the operating room to the point they are sterile and ready to be reused for the next surgery. The sterile processing unit is self-sufficient in power and water and features an intake for contaminated instruments, decontamination, sterilization via non-electric steam sterilizers, and secure inventory storage. To validate efficacy, we ran tests of decontamination and sterilization performance. Results of 61 trials validate convincingly that our sterile processing unit achieves satisfactory outcomes for decontamination and sterilization and as such holds promise to support healthcare facilities in low resources settings.     

Radiation and Ethylene Oxide Terminal Sterilization Experiences with Drug Eluting Stent Products

Radiation and ethylene oxide terminal sterilization are the two most frequently used processes in the medical device industry to render product within the final sterile barrier package free from viable microorganisms. They are efficacious, safe, and efficient approaches to the manufacture of sterile product. Terminal sterilization is routinely applied to a wide variety of commodity healthcare products (drapes, gowns, etc.) and implantable medical devices (bare metal stents, heart valves, vessel closure devices, etc.) along with products used during implantation procedures (catheters, guidewires, etc.). Terminal sterilization is also routinely used for processing combination products where devices, drugs, and/or biologics are combined on a single product. High patient safety, robust standards, routine process controls, and low-cost manufacturing are appealing aspects of terminal sterilization. As the field of combination products continues to expand and evolve, opportunity exists to expand the application of terminal sterilization to new combination products. Material compatibility challenges must be overcome to realize these opportunities. This article introduces the reader to terminal sterilization concepts, technologies, and the related standards that span different industries (pharmaceutical, medical device, biopharmaceuticals, etc.) and provides guidance on the application of these technologies. Guidance and examples of the application of terminal sterilization are discussed using experiences with drug eluting stents and bioresorbable vascular restoration devices. The examples provide insight into selecting the sterilization method, developing the process around it, and finally qualifying/validating the product in preparation for regulatory approval and commercialization. Future activities, including new sterilization technologies, are briefly discussed.     

苹果蠹蛾不育昆虫释放技术研究进展

不育昆虫释放技术(sterile insect technique,SIT)是一种环境友好、可作为大面积害虫综合治理(AW-IPM)的防治技术,是以压倒性比例释放不育昆虫来减少田间同种害虫繁殖量的害虫治理方法。苹果蠹蛾Cydia pomonella(L.)是世界重要的梨果类害虫,现已入侵世界5洲71国。本文综述了苹果蠹蛾大规模饲养技术、辐射不育技术、释放技术3个关键环节的研究与技术进展,主要包括:苹果蠹蛾人工饲料、实验种群建立、饲养设备与条件、收集和质量评估、长距离运输、辐射源与设备、辐射剂量与敏感性、释放方法、释放标记和释放量等,并介绍了各国采用SIT技术的应用效果。苹果蠹蛾在我国新疆、甘肃、宁夏、内蒙、黑龙江、吉林6个省区发现,对我国苹果产业安全生产构成严重威胁,我国很有必要引进并建立苹果蠹蛾SIT防治技术体系。     

The effect of ultrasonication on the immunomodulatory activity of low‐quality ginseng

This study investigated the effects of ultrasonication extraction (UE) on the immunomodulatory activity of low‐quality ginseng. The results indicate that the optimal conditions for extracting low‐quality ginseng are ultrasonication at 60 kHz and 85°C for 60 min. The extraction yield from the UE was 20% higher than that of the water extraction (WE) at 100°C. The low quality ginseng obtained from the UE exhibited relatively low cytotoxicity toward normal human cells, with an observed toxicity of 15–18% at a concentration of 1.0 mg/mL. The ginseng product obtained following UE induced human B and T cells growth and resulted in concentrations of up to 9.33 × 10⁴ cells/mL and 15.33 × 10⁴ cells/mL, respectively. The ginseng extract also increased the secretion of interleukin‐6 and tumor necrosis factor‐α from these cells by up to 35%, and natural killer/ cell growth was also improved by up to 30%. The UE effectively released 2‐ to 3‐fold higher levels of ginsenosides than the WE. Specifically, the obtained levels of Rb₁, Re, and Rg₁, which are likely immunomodulatory factors, were approximately three times higher after ultrasonication than after WE. These results were further supported by the finding that UE product‐treated macrophages produced higher levels of nitric oxide (21 μM) than macrophages treated with the WE product or with standard ginsenosides. These results demonstrate that this optimized ultrasonication process effectively destroyed the more rigid cell walls of low‐quality ginseng and released high levels of ginsenosides. This work is the first to correlate extraction parameters with both extraction yields and biological activity. The use of low‐quality ginseng can thus be expanded by utilizing a low‐temperature ultrasonic extraction process. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2013     

Modification of a commercial cell sorter to support efficient and reliable preparation of ALDH-bright cells for clinical use

BACKGROUND: Cell populations manufactured by conventional commercial cell sorters have been safely infused into patients, but reliably sterilizing these instruments remains challenging. We are developing clinical protocols involving use of ALDH bright cells manufactured by cell sorting in patients. However, we encountered problems when we attempted to reliably sterilize the FACSAria cell sorter using standard methods. RESULTS: We have identified and modified potential sources of microbial contamination in several FACSAria systems. We added new filter systems to the sheath and sample air lines, to the wet cart fluid supply, and to the sample line. Sheath was provided from an external sterile, disposable bag through sterile disposable tubing sets. The plenum reservoirs were modified in several ways to allow efficient decontamination of internal surfaces. A new bubble filter assembly was added and one valve was eliminated from the sample pathway to improve flow cell sterilization. A new cleaning and sterilization protocol was developed and validated. All cell products manufactured using the modified instrument and validated cleaning protocol have met lot release criteria for prevention of microbial contamination and safe clinical use. DISCUSSION: The instrument modification and cleaning protocol described enable reliable manufacture of ALDH bright cell populations that are suitable for clinical trials. We have manufactured nineteen consecutive samples that meet all clinical release criteria in an on-going Phase 1 human trial.     

A review: taking the sterile out of sterility

Users of in-dwelling medical devices, prostheses and surgical dressings rely implicitly on their sterility. Rarely do consumers give any thought to what sterility really means. The general assumption is that manufacturers have adopted the most efficient and cost-effective methods of achieving sterility. Currently, terminal sterilization processes appropriate for the manufacture of medical devices are those that are deemed to give less than one chance in a million of a single, finished product item containing a viable organism. Such a definition of sterility is embodied in the European standard EN556 as a Sterility Assurance Level of 10(-6), based on the properties of heat-resistant endospores. However, is this level of sterility assurance appropriate for all categories of medical device? Moreover, do all medical devices which are labelled as sterile require the same level of treatment? This paper will demonstrate that in some instances, the high standards set for sterilization processing are unreasonable, not cost-effective and exclude new sterilization technologies from being accepted by the regulatory authorities.     

Potential occurrence of adhering living Bacillus spores in milk product processing lines

AIMS: The hygienic risk associated with microbial soil on surfaces of milk processing lines was evaluated, based on experimental results. METHODS AND RESULTS: From a panel of Bacillus spores isolated from milk products, B. cereus CUETM 98/4, was found to be highly resistant to heat (D100=3.32 min in whole milk) and oxidant disinfectant (70% lethality of adherent spores with Ikalin 2%). From adhesion trials, up to 1.1 x 10(7) spores cm(-2) were found to be adherent to solid surfaces when suspended in saline or in custard (10(5) and 10(7) cfu ml(-1)), and over 10% of these adherent spores would resist the cleaning procedure. CONCLUSION: A highly contaminated milk (10(5) cfu ml(-1)) subjected to a current sterilization process (8 log reduction) led to a residual contamination of less than 1 cfu in the representative processing line after a complete production run. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlighted the fact that under appropriate processing conditions (efficient sterilization and cleaning procedures), even disinfection would be sufficient to eliminate any contamination risk. Conversely, the disinfection procedure becomes an essential step under inappropriate processing conditions.     

Identification of Particles in Raw Materials

Raw materials need to be of a certain quality with respect to physical and chemical composition. They also need to have no contaminants in the form of particles because these could get into the product or indicate the raw materials are not pure enough to make a good quality product. When particles are found, it is important to identify their chemical and elemental composition to correct any process errors that can cause them and to have acceptable quality of the final product. Sources of materials can be the environment, process equipment and processing, and packaging. Microscope versions of Raman spectroscopy, laser-induced breakdown spectroscopy (LIBS), and IR spectroscopy are excellent tools for identifying particles in materials because they are fast and accurate techniques needing minimal sample preparation that can provide chemical composition as well as images that can be used for identification. The micro analysis capabilities allow for easy analysis of different portions of samples so that multiple components can be identified and sample preparation can be reduced or eliminated. The complementarity of the techniques provides the advantage of identifying various chemical components, as well as elemental and image analyses. The sources of materials were seen to be the environment, process equipment and processing, and packaging.     

膜分离技术在蜂蜜饮品中的应用

蜂蜜饮品在蜂蜜用量为4%时感观指标最好;经0.22、0.45、0.65μm孔径滤膜过滤后样品澄清透明,可滤掉蜂蜜中较大分子的蛋白质等影响样品澄清的物质,同时一定程度起到膜除菌的作用;蜂蜜饮品不应通过加热的方式杀菌,以减少营养物质的损失;通过保质期试验,经0.65μm孔径滤膜可以保证样品的澄清度,结合无菌灌装及非加热杀菌...     

Quality assessment for processed and sterilized bone using Raman spectroscopy

To eliminate the potential for infection, many tissue banks routinely process and terminally sterilize allografts prior to transplantation. A number of techniques, including the use of scanning electron microscopy, bone graft models, and mechanical property tests, are used to evaluate the properties of allograft bone. However, as these methods are time consuming and often destroy the bone sample, the quality assessment of allograft bones are not routinely performed after processing and sterilization procedures. Raman spectroscopy is a non-destructive, rapid analysis technique that requires only small sample volumes and has recently been used to evaluate the mineral content, mineral crystallinity, acid phosphate and carbonate contents, and collagen maturity in human and animal bones. Here, to establish a quality assessment method of allograft bones using Raman spectroscopy, the effect of several common sterilization and preservation procedures on rat femoral bones were investigated. We found that freeze-thawing had no detectable effects on the composition of bone minerals or matrix, although heat treatment and gamma irradiation resulted in altered Raman spectra. Our findings suggest Raman spectroscopy may facilitate the quality control of allograft bone after processing and sterilization procedures.     

Preliminary evaluation of several disinfection/sterilization techniques for use with microdialysis probes

This study evaluated the suitability of some disinfection and sterilization methods for use with microdialysis probes. Disinfection or sterilization should minimize the tissue inflammatory reaction and improve the long-term health of rats on study and ensure the quality of data obtained by microdialysis sampling. Furthermore, the treatment should not negatively impact probe integrity or sampling performance. The techniques chosen for evaluation included two disinfection methods (70% ethanol and a commercial contact lens solution) and two sterilization methods (hydrogen peroxide plasma, and e-beam radiation). Linear microdialysis probes treated by these processes were compared to untreated probes removed from the manufacturer's packaging as if sterile (the control group). The probes were aseptically implanted in the livers of rats and monitored for 72 hours. The parameters chosen to evaluate probe performance were relative sample mass recovery and the relative in vivo extraction efficiency of the probe for caffeine. Post mortem bacterial counts and histopathology examination of liver tissue were also conducted. The probes remained intact and functional for the entire study period. The methods tested did not acutely alter the probes although hydrogen peroxide plasma and contact lens solution groups showed reduced extraction efficiencies. Minimal tissue damage was observed surrounding the probes and acute inflammatory reaction was mild to moderate. Low numbers of bacterial colonies from the implantation sites indicates that the health of animals in this study was not impaired. This was also true for the control group (untreated probe).     

Optimization of batch fermentor sterilization

This article presents a calculation procedure useful for the optimization and scale up of batch sterilization cycles in large-scale fermentors. This technique determines the sterilization temperature and hold-time necessary to minimize nutrient damage in a specific fermentor. The method can also be used for "scaledown" experiments to eliminate sterilization conditions as a scale up parameter. A method for the systematic evaluation of different sterilization conditions on product yield is also presented. This procedure is useful in determining if scale up of sterilization conditions is important for a given process. The validity of the techniques presented are supported by data showing significant yield improvements in a 1.2 x 10(5) L antibiotic fermentation.     

Reducing the Risk of Contamination of Sterile Parenteral Products via Ready-to-Use Closure Components

The preparation of sterile parenteral products requires careful control of all ingredients, materials, and processes to ensure the final product has the identity and strength, and meets the quality and purity characteristics that it purports to possess. Contamination affecting these critical properties of parenteral products can occur in many ways and from many sources. The use of closures supplied by manufacturers in a ready-to-use state can be an effective method for reducing the risk of contamination and improving the quality of the drug product. This article will address contamination attributable to elastomeric container closure components and the regulatory requirements associated with container closure systems. Possible contaminants, including microorganisms, endotoxins, and chemicals, along with the methods by which these contaminants can enter the product will be reviewed. Such methods include inappropriate material selection, improper closure preparation processes, compromised container closure integrity, degradation of closures, and leaching of compounds from the closures.     

高温杀菌对预浸泡豆杆的杀菌效果及品质影响

本研究以预浸泡豆杆为研究对象,研究高温杀菌对预浸泡豆杆中微生物的杀灭效果及其品质的影响,以期获得具有良好感官品质和理化品质的产品。结果表明:当温度≥105℃和时间≥10 min时,样品的菌落总数由初始的8.72×108cfu/g减少到50 cfu/g以内,样品的大肠菌群从1.6×105MPN/g减少到3 MPN/g以内,致病菌均未检出。杀菌温度和杀菌时间对预浸泡豆杆中脂肪含量、pH、弹性影响差异不显著(p>0.05),对水分含量、蛋白质、色差、硬度和咀嚼性影响显著(p<0.05),综合微生物、理化指标、色泽、质构的变化结果,经105℃、15 min和110℃、10 min处理后的预浸泡豆杆既能满足产品卫生要求也可以较好的保留产品原有的品质。     

Use of focused ultrasonication in activity-based profiling of deubiquitinating enzymes in tissue

To develop a reproducible tissue lysis method that retains enzyme function for activity-based protein profiling, we compared four different methods to obtain protein extracts from bovine lung tissue: focused ultrasonication, standard sonication, mortar & pestle method, and homogenization combined with standard sonication. Focused ultrasonication and mortar & pestle methods were sufficiently effective for activity-based profiling of deubiquitinases in tissue, and focused ultrasonication also had the fastest processing time. We used focused-ultrasonicator for subsequent activity-based proteomic analysis of deubiquitinases to test the compatibility of this method in sample preparation for activity-based chemical proteomics.     

Mechanical strength of bone allografts subjected to chemical sterilization and other terminal processing methods

Infectious disease transmission through the use of human donor allografts can be a catastrophic complication in an otherwise straightforward surgical procedure. The use of bone allograft in reconstructive orthopedic surgeries is increasing, yet severe complications, including death, can result if the transplanted tissues transmit a communicable disease to the tissue recipient. The BioCleanse((R)) tissue sterilization process is a fully automated, low-temperature chemical sterilization process that renders allograft tissue sterile. The purpose of this study was to evaluate the effect of a chemical tissue sterilization process on the mechanical strength of cortical bone allografts prior to implantation. Cylindrical cortical bone specimens were harvested from seven human cadaver donors and treated either by: chemical sterilization alone; chemical sterilization and terminal sterilization by gamma irradiation; chemical sterilization, lyophilization, terminal sterilization by STERRAD and rehydration; or untreated. The specimens were tested to failure in axial compression, diametral compression, shear, or bending. There were no significant differences in ultimate stress, strain, or fracture energy between the chemically sterilized and control groups in any of the testing modes.     

Effect of methods of sterilization on thermoplastics with special reference to modified surfaces]

For materials intended for use in the medical setting their sterilizability is an indispensable prerequisite. In the case of most polymers the usual sterilization methods result in changes that even extend to cleavage of the polymer chains. A particular problem in this respect are the surfaces modified for improved biocompatibility investigated in the present study, which are characterised by enlarged contact areas. For this reason, possible changes to three different thermoplastics commonly used for medical applications (polyethylene, thermoplastic polyurethane, polycarbonate) were investigated. Steam, gas and radiation were used for sterilization. Tensile tests were employed to identify changes in mucosal characteristics caused by different sterilization techniques irrespective of the surface modification. Sterilization-related changes to the structure of the modified surfaces were investigated with the scanning electron microscope (SEM). Differential thermo analysis (DTA) was used to determine changes in the thermal characteristics of the plastics. Clear tendencies with regard to the behaviour of the plastics after sterilization with various techniques were found. A general statement about the compatibility of plastic materials with a specific sterilization method is not possible on the basis of this study. For every new polymeric product used for medical purposes, the characteristics required must first be defined and compliance with the permissible variations of these characteristics investigated for each of the various sterilization techniques available.     

Solvent-dehydrated calvarial allografts in craniofacial surgery

Craniofacial surgery almost always requires the use of bone grafting. Although autografts are the standard procedure for bone grafting, it is sometimes not possible to harvest bone, and autografts have particular risks. The use of allograft bone provides a reasonable alternative to meet the need for graft material. Solvent dehydration is a multistage procedure in which human cadaveric bone is processed by osmotic exchange baths and gamma sterilization. This processing avoids the risk of infection transmission, decreases antigenicity, and does not weaken the mechanical properties of the bone. Solvent-dehydrated, gamma-irradiated human calvarial bone allografts were used for reconstruction of craniofacial deformities in 24 patients between 1988 and 2002. Resorption of the allografts and results of the surgical intervention were evaluated with plain radiographs and three-dimensional computed tomography 12 months after surgery, in 21 patients. Serologic tests for human immunodeficiency virus-1 antibody, hepatitis B surface antigen, and hepatitis C antigen were also performed. Biopsy specimens were taken from the allografts. Average follow-up in this group was 30 months (range, 8 to 60 months), and results of serologic tests were negative in all patients. Seventy-one percent of the patients (15 of 21) showed no resorption, with partial and complete allograft fusion. One patient had nearly total graft loss and the remaining five patients had 10 to 25 percent graft resorption. Rigid fixation of the allograft, contact with the dura and periosteum, and prevention of dead spaces around the allograft are the most important factors in achieving a satisfactory result. In solvent-dehydrated bone allografts, sterilization and antigenic tissue cleaning are achieved after several steps with a minimal dose of radiation. The result is a nonantigenic, sterile mechanical scaffold that can tolerate external forces. Although autografts are the standard in craniofacial surgery, solvent-dehydrated calvarial bone allografts produced successful results in selected cases.     

Effects of Ethylene Oxide on the Tissue and Flora of Fresh and Freeze Dried Cod Fillets

S ummary . Attempts to sterilize fresh cod fillets with ethylene oxide resulted in protein denaturation with the commensurate exudation of soluble tissue materials. The bacterial colony forming units were reduced to a level of between zero and 100/g of tissue, but sterilization was never achieved. Freeze dried cod fillet sections treated with ethylene oxide and reconstituted with sterile Ringer's solution gave a sterile product which when inoculated with a natural bacterial flora produced normal spoilage odours.     

植物内生放线菌的选择性分离

[目的]更好地发掘内生菌资源,建立有效的植物内生放线菌分离方法.[方法]比较不同消毒剂和消毒程序、样品预处理、选择性分离培养基等分离内生放线菌的效果,通过形态及16S rRNA基因序列分析进行菌种鉴定.[结果]用5％的次氯酸钠处理样品4-7 min消毒效果最好;100℃处理样品15 min能较好地减少真菌和细菌的干扰.丙酸钠、琥珀酸钠等培养基分离放线菌出菌率较高且类群多样性丰富.[结论]植物样品表面消毒干燥后,100℃处理15 min,用无菌搅拌杯打碎,直接撒植物于分离培养基中的分离方法效果较好.