Aseptic Handling Research Articles

Improving transient gene expression and agroinfiltration‐based transformation effectiveness in Indonesian orchid Phalaenopsis amabilis (L.) Blume

Transient gene expression is an approach used to study transient genes across various species, with infiltration by Agrobacterium tumefaciens (agroinfiltration) being a commonly used method. Agroinfiltration offers a simple and effective means of delivering transgenes into the plant genome. An alternative method for enhancing the quality and productivity of orchids as ornamental plants is genetic modification through agroinfiltration. Although Agrobacterium‐mediated genetic transformation by immersion has been used on the Phalaenopsis amabilis (L.) Blume species of orchid, transformation efficiency using the immersion technique remains relatively low and the method itself is challenging due to its requirement for aseptic handling. The application of agroinfiltration in P. amabilis has not previously been reported. This study investigates the impact of the injection site, acetosyringone concentration, bacterial density (OD600), and injection volume to determine the optimum conditions for agroinfiltration on P. amabilis. The results demonstrated that injection site had a noticeably distinct impact on transformation effectiveness, with the abaxial position of the leaf being the optimal site for Agrobacterium culture suspension injection. While adjustments in acetosyringone concentration, bacterial density (OD600), and injection volume did not significantly affect transformation efficiency, they did influence the peak time of GFP fluorescence. Acetosyringone at a concentration of 200 µM, an OD600 of 1.0 for Agrobacterium culture, and an injection volume of 500 µL effectively accelerated GFP expression duration.

Bacterial contamination and greenhouse gas emissions: A randomised study of reuse versus single-use of infusion-set components for intravenous anaesthesia.

Reusing anaesthesia infusion-set components may reduce the climate impact from plastic waste and discarded medications. Infusion-set contents can be shielded from patient contact by single use of an infusion line fitted with dual antireflux valves, preventing retrograde entry of microorganisms, and eliminating the risk for patient-to-patient cross-contamination. However, infusion-set contamination from compromised aseptic handling could affect quality of care. To determine the prevalence of infusion-set bacterial contamination and compare the climate effects, we randomised operating rooms scheduled for total intravenous anaesthesia to handle procedures by infusion-set reuse or single-use. Both methods used dual single-use antireflux valves. The primary outcome was infusion-set bacterial contamination assessed by aerobic culture of infusion-set fluid collected after each procedure. The secondary outcome was CO 2 emissions (CO 2 -eq) estimated by life cycle assessment of component and medication use. To assess feasibility of detecting an inter-method difference in bacterial contamination, an interim analysis was planned after including at least 150 procedures per group. After allocating 54 operating rooms per method, 189 and 159 procedures of reuse and single use were included. Reuse permitted a median of three procedures per infusion set (range 1 to 8). Positive cultures occurred in two procedures per method [mean (95% CI)]; prevalence 1.15% (0.03 to 2.27); relative risk of reuse versus single use 0.84 (0.12 to 5.93), P = 0.861. As prespecified, inclusion was stopped due to futility. The median (95% CI) per-procedure climate emissions were 0.43 (0.41 to 0.47) and 1.39 (1.37 to 1.40) kg CO 2 -eq for reuse and single-use respectively; difference -0.96 (-0.99 to -0.93), P < 0.0005. The main sources for climate emissions were production of infusion-set components and waste handling. We conclude that the prevalence of bacterial contamination was low for both methods. A much larger study would be needed to detect an inter-method difference. Reuse of infusion-set components allowed significantly reduced intravenous anaesthesia climate emissions.

Observation of the Reconstitution Process and Factors that Influence the Final Condition of Chemotherapy Patients in Private Hospitals

 Handling of cytostatic preparations is the aseptic handling of cancer drugs in ready-to-use packaging according to patient needs by trained pharmacists with control of environmental safety, officers, and drug preparations from toxic effects and contamination, using personal protective equipment. Resistant to handling cytostatics preparations, including preparation, mixing, handling of spills, and cytostatics waste management. The purpose of this study was to determine the suitability of the handling of cytostatic preparations and the description of the use of chemotherapy drugs in cancer patients. This research was conducted by descriptive analytic observation, data on the handling of cytostatics preparations was obtained by observing directly in the chemotherapy drug mixing room and data on the use of chemotherapy drugs were obtained retrospectively by looking at medical records and service sheets of mixing cytostatics preparations. The results obtained in the process of handling cytostatic preparations in the preparation, mixing and labeling stages are 100% to the guidelines, while the labeling stage is 86% appropriate. Evaluation of the type of use of chemotherapy drugs is known that the taxane drug class is the most widely used by 91,7%. The appropriate evaluation of chemotherapy drugs in breast cancer patients is 92%, lung cancer is 100% and uterine cancer is 0%. Evaluation of the correct dose of chemotherapy drugs obtained results of 0%. The results of the Chi Square statistical test found that there was a relationship between the last cycle of chemotherapy and the patient's final condition p = &lt;0.001

Printing Process Quality Control of Bioprinting Medical Devices

This study analyzes the risk points in the quality control of bioink and the main processes of bioprinting, clarifies and explores the quality control and supervision model for bioprinting medical devices, and provides theoretical and practical guidance to ensure the safety and effectiveness of bioprinting medical devices. The quality control risk points throughout the bioprinting process were comprehensively analyzed, with a particular focus on bioprinting materials and key processes. The regulatory model and methods for bioprinting medical devices were examined. This research concentrated on critical technologies such as extrusion, laser-assisted, and in situ bioprinting, assessing their potential for clinical applications and regulatory challenges. Bioink from different sources should meet regulatory requirements. It is essential to ensure aseptic handling of raw materials and to validate sterilization under "worst-case" conditions. As bioprinting technology advances rapidly, corresponding research into materials, processes, and quality risk control should be conducted to ensure the concurrent development of the regulatory system. This will continuously contribute to the orderly progression of the entire industry and human health.

Contamination of liposomal bupivacaine during multi-dose usage in a clinical setting evaluated by culture and PCR.

Liposomal bupivacaine, a long-acting local anesthetic, is sold in single-dose vials at a cost of approximately $200/20 mL vial. As many veterinary patients are not dosed an entire vial, the vials have been used for multiple doses at our institution to provide cost savings. Multiple punctures of a vial can lead to increased opportunity for contamination of the contents. This study aims to describe our institutional procedure for multi-dose use of single-dose liposomal bupivacaine vials and to evaluate clinically utilized liposomal bupivacaine for bacterial and fungal contamination using molecular and bacteriological methods. The first (Control) and last (Sample B) 0.5 mL from each vial were collected and submitted for bacterial and fungal PCR, anaerobic and aerobic bacterial culture, and opportunistic fungal culture. All 40 bacterial cultures yielded no growth; Bacterial or fungal DNA was identified in 19 samples (50%). Of the 19 samples in which bacterial or fungal DNA was identified, 10 (52.6%) were from Control, and 9 (47.4%) were from Sample B. PCR does not appear to be useful in detecting bacterial or fungal contamination from liposomal bupivacaine. Results support the aseptic handling protocol described in this article is successful in preventing detectable bacterial and fungal contamination of liposomal bupivacaine vials for up to 7 individual punctures and vials open for up to 5 days.

Aseptic handling of ultrasound probe during pain interventions: A low-cost and easily available solution

Aseptic handling of ultrasound probe during pain interventions: A low-cost and easily available solution

Human nasal olfactory stem cells, purified as advanced therapy medicinal products, improve neuronal differentiation.

Olfactory ecto-mesenchymal stem cells (OE-MSC) are mesenchymal stem cells derived from the lamina propria of the nasal mucosa. They display neurogenic and immunomodulatory properties and were shown to induce recovery in animal models of spinal cord trauma, hearing loss, Parkinsons's disease, amnesia, and peripheral nerve injury. As a step toward clinical practice, we sought to (i) devise a culture protocol that meets the requirements set by human health agencies and (ii) assess the efficacy of stem cells on neuron differentiation. Nasal olfactory mucosa biopsies from three donors were used to design and validate the good manufacturing process for purifying stem cells. All processes and procedures were performed by expert staff from the cell therapy laboratory of the public hospital of Marseille (AP-HM), according to aseptic handling manipulations. Premises, materials and air were kept clean at all times to avoid cross-contamination, accidents, or even fatalities. Purified stem cells were cultivated for 24 or 48 h and conditioned media were collected before being added to the culture medium of the neuroblastoma cell line Neuro2a. Compared to the explant culture-based protocol, enzymatic digestion provides higher cell numbers more rapidly and is less prone to contamination. The use of platelet lysate in place of fetal calf serum is effective in promoting higher cell proliferation (the percentage of CFU-F progenitors is 15.5%), with the optimal percentage of platelet lysate being 10%. Cultured OE-MSCs do not show chromosomal rearrangement and, as expected, express the usual phenotypic markers of mesenchymal stem cells. When incorporated in standard culture medium, the conditioned medium of purified OE-MSCs promotes cell differentiation of Neuro2a neuroblastoma cells. We developed a safer and more efficient manufacturing process for clinical grade olfactory stem cells. With this protocol, human OE-MSCs will soon be used in a Phase I clinical based on their autologous transplantation in digital nerves with a neglected injury. However, further studies are required to unveil the underlying mechanisms of action.

Monkeypox outbreak in a piercing and tattoo establishment in Spain

Monkeypox outbreak in a piercing and tattoo establishment in Spain

Design and Evaluation of Solid Lipid Nanoparticles of an Antibiotic-mannitol as Cryoprotectant

Mannitol was used as a cryoprotactant to freeze-dry Doxycycline nanoparticles for colloidal drug administration in this work. in this research, the benefits include Medicines like vaccines and other injectables, which have a short shelf life and are commonly freeze-dried to increase their shelf life. A liquid may be treated more easily with aseptic handling because of this. The stability of a dry powder has been increased. In addition, water may be removed from the device without causing it to overheat. Reconstituted product dissolves quickly and readily. All of these concerns are avoided so that the product may be transported or stored at room temperature without suffering deterioration or degradation due to aggregation. Preparation of doxycycline SLNs included homogenization and sonication. Antimicrobial activity and in vitro anticancer activity are among the techniques employed in this investigation. Freeze drying was also a component of the process, as was particle size, drug loading, and drug release. The lipid solid’s particle size distribution At various magnifications of the investigation, nanoparticles are determined to be 3.66, 6.58, 9.96, 4.97, 17.36, and 25.48mm in diameter. A freeze-dryer was used to freeze-dry the particles at -20°C and 0.4 pressure. Mannitol at various concentrations (5, 10, and 15 percent) was utilised as a cryoprotectant. Drug loading was determined to be 78 percent, while drug release was found to be 98 percent after 24 hr. Maximum inhibition of cell growth (69.9422) was seen at an IC50 concentration of 100g/mL of leaf crude extract (48.69g/mL). Doxycycline SLNs might be inferred to be more stable, less aggregate, and have fewer adverse effects when freeze-dried.

The Development and Implementation of Airflow Visualization Studies ("Smoke" Studies) as a Training Tool in Aseptic Hospital Compounding Facilities.

In the compounding facilities of hospital pharmacies, extemporaneous preparations for parenteral administration are produced using aseptic handling. The designated environment for this practice is a clean area, such as a laminar airflow (LAF) cabinet placed in a classified cleanroom complying with good manufacturing practices (GMP) and International Organization for Standardization (ISO) 14644-1 guidelines. The European GMP Annex 1 (Revision 2020) and United States Pharmacopeia (USP) <797> monograph state that airflow visualization studies (“smoke” studies) should be performed to substantiate the cleanroom and LAF cabinet performance and their qualification status. Even though smoke studies are required by these guidelines, current literature does not describe detailed practical protocols and acceptance criteria. The objective of this study was to develop and implement a practical smoke study protocol to ensure compliance with aseptic handling guidelines in hospital pharmacies. First, a literature search was performed to collect information about smoke study protocols and acceptance criteria. Subsequently, a smoke study protocol was developed for a downflow and crossflow LAF cabinet as well as for grade C/B cleanroom areas. As a proof of concept, the smoke study protocol for the downflow LAF cabinet was executed in the at-rest and in-operation states. Video recordings of the smoke studies were analyzed to assess the performance of the cabinet. Finally, the video recordings obtained from the smoke studies were used in a training program for hospital pharmacy operators, which showed that smoke studies might aid in operators’ aseptic handling awareness. To the best of our knowledge, the present study provides for the first time a practical approach for the development of smoke study protocols in a hospital pharmacy setting and shows potential for training operators, process optimization, and continuous quality improvement.

Extended aseptic handling and anaesthetic efficacy of propofol preserved under different conditions

Twenty-seven vials of propofol and 20 Albino rats were used in this study to determine the effect of refrigeration and aseptic handling on contamination and anaesthesia of propofol. The vials were assigned to three groups of nine (9) vials per group in 3 replicates. They were opened aseptically and the contents transferred to sterile bottles. VR group were stored in a refrigerator at 40C, VS in an infrared hood chamber, and VT stored on a bench. Propofol was aseptically aspirated from each vial immediately after opening and at 8, 10, 12, 24, 48 and 168 hours for culture on nutrient and McConkey agar. Qualitative and quantitative analysis of the isolates were carried out. Twenty adult Albino Wistar rats were assigned to 4 groups of 5 rats per group. One group (VF) of rats was anaesthetized with fresh propofol, while the remaining 3 groups were anaesthetized with the preserved propofol from each drug group, stored at the longest duration at which contamination was not evident at culture. Durations of anaesthesia, corneal reflex loss, and abdominal twitch and tail sensation loss, were measured. Data was analyzed using ANOVA and Duncan Multiple Range Test using SPSS version 22. Values of p &lt; 0.05 were considered significant. Contamination occurred in the VS, VR and VT groups at 168, 48 and 12 hours respectively. This contravenes the opinion that opened propofol vials become contaminated after 8 hours. Bacillus subtilis, Streptococcus pneumonia, Lactobacillus spp, Klebsiella pneumonia and Staphylococcus epidermidis were isolated from the various groups. The indices of anaesthesia did not differ significantly among the groups. Therefore, following strict adherence to aseptic techniques, propofol may be safely used for up to 24 hours if stored either in a refrigerator or in a sterile environment, without any significant alterations in its anaesthetic characteristics.

Microbiological controls of aseptic handling in Dutch hospital pharmacies: Results, limits, and methods for assessing.

Introduction‘Aseptic handling is the procedure to enable sterile products to be made ready to administer using closed systems’ (EU Resolution CM/Res(2016)2). Microbiological controls are an important part of the overall assurance of process and product quality of aseptic handling. They consist of the End-of-Session Broth Test (ESBT) using Tryptone Soya Broth, Microbiological Monitoring (MM) using Ø 55- and 90-mm agar plates and a periodical Operator Broth Transfer Validation Test (OBTVT) using Tryptone Soya Broth. This study describes the results of these controls over a 7-year period, involving between 44 and 49 pharmacies (mostly hospital pharmacies). All pharmacies use a web-based programme for processing, evaluation and assessing microbiological controls (‘Microbio’). Aggregated results in Microbio are used for benchmarking and feedback information. ObjectiveThe objective of this study is to analyse the results of the 7-year period and to develop methods for assessing of, and determining realistic limits for, microbiological controls during aseptic handling. As secondary objective the role of Microbio is highlighted. Results and discussionResults of ESBT are expressed as Contamination Rate (CR), which is the percentage of units filled in a process simulation that are positive for microbial growth after incubation. Compared with the first 3 years of the study, the results in the last 4 years were significantly better: mean CRs are 0.20 and 0.11, respectively (p-value <0.01). For assessing CRs of ESBT, the approach ‘the more frequent samples with growth, the stronger the corrective actions’ was adopted. Levels of investigative and corrective actions, based on the 95% Upper Confidence Limit, are suggested. Microbiological Monitoring (MM) during aseptic handling into a laminar airflow cabinet or safety cabinet consists of settle plates, glove prints, contact plates of the worktop and surface bioburden determination of disinfected ampoules and vials. The results are expressed as the Contamination Recovery Rate (CRR), which is the rate at which MM samples contain any level of contamination. During the study period, the results of glove prints and contact plates improved substantially. The most probable explanation of this finding is improved disinfection procedures of the gloved hands, the worktop inside LAF/SC and the ampoules and vials. Results of settle plates did not change. There were too few results available to evaluate the surface bioburden of disinfected ampoules and vials. Benchmarking and feedback information from Microbio may have contributed to the improved ESBT and MM results. Results of the Operator Broth Transfer Validation Test (OBTVT) are expressed as Contamination Rate (CR). The target is zero samples with growth (CR = 0). The overall CR result over the study period is 0.50%. This is worse than ESBT (overall CR is 0.14%). This is probably due to the high number of critical steps in OBTVT compared to ESBT. ConclusionResults of microbiological controls improved during the study period. Realistic limits as well as methods for assessing ESBT and MM results are given and discussed.

Sterility of Sustained-release Buprenorphine.

Sustained-release formulations of controlled substances are commonly used to provide analgesia in research animals. These formulations represent refinements that offer the advantage of prolonged, multiday pain relief with a single injection, thereby decreasing handling stress in animals and saving time for scientists. Compounding pharmacies produce sustainedrelease buprenorphine for veterinary use (i. e., buprenorphine SR-LAB); one of these pharmacies has shortened the original 6-mo shelf-life to 28 d to comply with United States Pharmacopeia standards for ensuring sterility. This limitation risks increasing the waste of controlled substances, which require an expensive destruction process that is legally enforced in our state. To assess whether the sterility of buprenorphine SR-LAB is preserved for at least 6 mo in a general laboratory setting, we tested 5 bottles for the presence of endotoxin and bacterial and fungal contamination monthly for 6 mo. Overall, results of the study showed that the bottles remained sterile over the 6-mo duration as no endotoxin was detected and the bottles did not become contaminated with bacteria or fungi. In conclusion, when stored securely and used with aseptic handling techniques, buprenorphine SR-LAB can be maintained in a sterile state for 6 mo in a general laboratory setting.

Complications associated with central venous lines for paediatric oncology patients at Universitas Academic Hospital, Bloemfontein, from 1992 to 2018

Background. Central venous access devices are associated with complications such as central-line infections and systemic sepsis. Objective. To determine the complication rates associated with central venous lines used to administer chemotherapy at the Paediatric Haematology Oncology Unit, Universitas Academic Hospital, Bloemfontein, from January 1992 to March 2018. Methods. A retrospective descriptive analysis of paediatric oncology patients who received intravenous catheterisation and were treated at the unit. Cases with incomplete data, age &gt;16 years and treatment after 31 March 2018 were excluded. Results. Records of 293 Hickman lines were analysed. The median patient age was 64.7 months. Sepsis was noted in 13.3% of the cases; no in situ local complications were found in 62.5% of the lines. Of the 39 cases that presented with line sepsis, 23.1% showed no signs of systemic sepsis, whereas 61.5% were neutropenic and septic. In total, 190 patients had symptoms of systemic sepsis and accompanying neutropenia. Of these, 67.4% did not develop in situ line complications. Lines were removed mostly because the end of treatment had been reached (44.7%); line sepsis was noted as the reason for removal in only 16.3% of cases. Conclusion. Despite the study population having a high risk for infections because of a young age, a large proportion of haematological malignancies and surgical placement of Hickman lines, there was a low incidence of line sepsis complications. The special care taken intraoperatively and meticulous aseptic postoperative handling and maintenance are suggested as contributing to limited infective complications.

Study of Microbial Contamination of Some Commercial Contact Lens Solutions

Sixty samples of commercially available contact lens solutions were collected from students at the Pharmacy College/Baghdad University. The types of lenses used varied from medical to cosmetic. They were cultured to diagnose any microbial contamination within the solutions. Both used and unused solutions were subject for culturing. Thirty six (60%) used samples showed bacterial growth, fungal growth was absent. Pseudomonas aeruginosa accounts for the highest number of isolates (25%) followed by E. coli (21%), Staphylococcus epidermidis (6.6%), Pseudomonas fluorescence (5%) and Proteus mirabilis (1.6%) respectively. Only one (1) unused (sealed) sample showed growth of P. fluorescence. These bacterial contamination likely came from bad personal hygiene and improper or misuse of the solutions where these bacteria especially P. aeruginosa are frequently found in various environments from skin to solid materials and surfaces and are known to thrive in harsh environments. No relation was found between eye associated diseases and solution contamination among contact lens users. Special care should be paid in maintaining aseptic solutions and proper handling to avoid transmitting harmful bacteria to the eye where it may lead to serious eye infections

Reducing the risk of non-sterility of aseptic handling in hospital pharmacies, part C: applying risk assessment and risk control in practice

ObjectivesTo describe the application of the model described in part A and part B of this series of articles for risk assessment (RA) and risk control (RC) of non-sterility during...

The quality assessment of the University hospital bone bank in Central Serbia: the second audit after fourteen years.

We analyzed the prevalence and predisposing factors for the overall rejection rate after retrieval of 267 fresh femoral head allografts over the past 7 years. The present study aimed to assess the quality system of institutional bone banking that can provide high-standard allografts with a low infection rate. Retrospective analysis of bone banking from June 2013 to December 2019 was conducted on 267 donors and 153 recipients. Of the 267 donated femoral heads, 74 were rejected, giving an overall rejection rate of 27.71%. The leading cause of allograft rejection was the inability to perform serology tests due to donor death; the absence of serological tests itself, and the donor refusal to perform the serology 6-month retest in 42 donors (15.72%). At retrieval, 12 allografts were positive, giving an overall contamination rate of 4.49%. Seven (2.62%) of the 267 allografts failed the blood screening tests. Thirteen allografts (4.86%) were discarded because of suspected damage to the packaging or disuse during surgery. An infection rate of 1.30% was found following transplantation. Over the past 7 years of bone banking, our results show that the overall rejection rate and the allograft-related infection rate correlate with international standards. The leading cause of allograft rejection was the inability to perform serology tests due to donor death and their refusal to perform the serology retests. Besides stringent aseptic allograft handling, donor motivation to participate in bone banking is extremely important for its efficient functioning.

BioSamplr: An open source, low cost automated sampling system for bioreactors

Autosampling from bioreactors reduces error, increases reproducibility and offers improved aseptic handling when compared to manual sampling. Additionally, autosampling greatly decreases the hands-on time required for a bioreactor experiment and enables sampling 24 h a day. We have designed, built and tested a low cost, open source, automated bioreactor sampling system, the BioSamplr. The BioSamplr can take up to ten samples from a bioreactor at a desired sample interval and cools them to a desired temperature. The device, assembled from low cost and 3D printed components, is controlled wirelessly by a Raspberry Pi, and records all sampling data to a log file. The cost and accessibility of the BioSamplr make it useful for laboratories without access to more expensive and complex autosampling systems.

Microbiological monitoring during aseptic handling: Methods, limits and interpretation of results

Aseptic handling is the procedure to enable sterile products to be made ready to administer using closed systems (EU Resolution CM/Res(2016)2). Microbiological monitoring (MM) and media fills are used for environmental and process control.In this study, the application of MM methods during aseptic handling inside, or related to working in, a laminar airflow cabinet or safety cabinet in hospital pharmacies is described and evaluated. Results are expressed as colony forming units (cfu) and Contamination Recovery Rate (CRR; the rate at which MM samples contain any level of contamination -USP<1116>-). For trend analysis, a rolling CRR is developed (a rolling CRR calculates a CRR using a predetermined number of most recent samples).Of all MM methods, glove print is the most informative. The added value of air sampling is doubtful. Because of microbiological as well as statistical considerations, the use of CRR for assessing MM results is advised. Glove prints, in general, give the highest CRR. A CRR < 10% is a realistic limit for MM during aseptic handling in hospital pharmacies. A rolling CRR, calculated using the last 100 samples, is a good compromise between reliability of the CRR value and timely prediction of process changes.

Reducing the risk of non-sterility of aseptic handling in hospital pharmacies, part B: risk control

ObjectivesTo determine prospectively the risk reducing measures of non-sterility during aseptic handling and to develop a method for prioritising these measures.MethodsIn the first part of this series of articles, we...