Sterility Testing Research Articles

The Flexural Strength and the Effect of the Autoclave Sterilization of Polypropylene/Natural Rubber Blended Materials

Background: Rubber dam clamps are used extensively in dentistry, especially during root canal treatment. However, existing rubber dam clamps have several drawbacks, including discomfort and potential damage to vital tissue in the oral cavity. To address these existing issues, a new rubber dam clamp should be developed. The aim of this study was to identify the optimum ratios of polypropylene and natural rubber (PP/NR) for a customized rubber dam clamp in dentistry. This study was focused on the flexural strength of PP/NR in various ratios. Moreover, the impact of autoclave sterilization was also considered. Methods: Six proportions of PP/NR blends (100/0, 90/10, 80/20, 70/30, 60/40 and 50/50) were prepared and assessed for flexural strength using a three-point bending test. After this test, the PP/NR blends with 100/0, 90/10 and 80/20 ratios were selected and underwent autoclave sterilization for 1, 5 and 10 cycles. Eventually, the flexural strength testing was repeated and investigated. An analysis of variance and Tukey’s test were used to evaluate the flexural strength of various PP/NR blends before autoclave sterilization at p < 0.05. An analysis of variance and Dunnett’s T3 test were used to evaluate the flexural strength of selected PP/NR blends before and after autoclave sterilization at p < 0.05. Results: The results revealed that the flexural strength of PP/NR blended materials showed a statistically significant difference in every group of this study. The autoclave sterilization test revealed that the flexural strength of the PP/NR 90/10 and 80/20 ratios was significantly increased after sterilization for 1, 5 and 10 cycles. In addition, the PP/NR 90/10 ratio was also comparable to the 100/0 ratio. The lower NR content in PP/NR blends resulted in significantly higher flexural strength, and autoclave sterilization had an effect on this property. Conclusions: This study suggested that the PP/NR blend with a 90/10 ratio might be considered as an alternative material for developing rubber dam clamps.

Extended-spectrum β-lactamase and carbapenemase producing Enterobacteriaceae among patients suspected with surgical site infection at Hospitals in Southern Ethiopia

BackgroundExtended-spectrum β-lactamase and carbapenemase-producing Enterobacteriaceae are an increasing problem for patients today. Data on clinical samples for ESBL and carbapenemase-producing Enterobacteriaceae for surgical site infection patients in developing countries are limited, including Ethiopia, mainly due to resource constraints. Hence, this study aimed to determine the prevalence of extended-spectrum beta-lactamase- and carbapenemase-producing Enterobacteriaceae among patients suspected to have surgical site infection at Hospital in Southern Ethiopia.Materials and methodsA hospital-based cross-sectional study was conducted on 422 suspected surgical site infections from June 1, 2022 to August 30, 2022 at Hospitals in Southern Ethiopia. Sociodemographic and clinical data were obtained by using a structured questionnaire. Clinical samples (pus, pus aspirates, and wound swabs) were collected aseptically and processed within 30 min by placing the swabs in sterile test tubes containing sterile normal saline (0.5 mL). Samples were cultured on blood and MacConkey agar plates. All positive cultures were characterized by colony morphology, Gram staining, and standard biochemical tests. Antimicrobial sensitivity tests were performed using Kirby Baur disk diffusion on Mueller–Hinton agar. ESBL production was confirmed using a double-disc synergy test (DDST) method. Carbapenemase production was assessed using the modified Hodge test. Logistic regression analysis was used to determine associated factors. A P-value < 0.05 were considered statistically significant.ResultBacteria belonging to the order Enterobacterales were cultured in 23.7% out of 422 patients with suspected surgical wound infection. Of all the isolates, Enterobacteriaceae (69 isolates) were the most frequent, with E. coli (29/69) followed by K. pneumoniae (14/69). Of 69 Enterobacteriaceae isolates, 66.6 % (46/69) were positive for ESBL production, and 21.7 (15/69) were positive for carbapenemase-producing Enterobacteriaceae. The majority of Enterobacteriaceae isolates showed sensitivity to meropenem (72.1%); however revealed 63.9% and 70.5% were resistant to gentamicin and ciprofloxacin, respectively. Similarly, a higher resistance rate to cefepime (91.8%), amoxicillin-clavulanic acid (98.4%), ceftriaxone (95.1%), and ceftazidime (91.8%). MDR rate of Enterobacteriaceae isolates was 25/61 (41%) among patients suspected for surgical site infection. The Multivariable analysis revealed that length of hospital stay in hospital [AOR = 3.81 (95% CI 2.08–6.95)] remained statistically significant factor associated with surgical site infection due to ESBL producing Enterobacteriaceae.ConclusionStudy results showed the severity of ESBL-producing Enterobacteriaceae is critical and CPE is alarming. Meropenem is the most effective antibiotic against the ESBL-producing Enterobacteriaceae. MDR rate of Enterobacteriaceae isolates was 61 (61%) among patients suspected for surgical site infection. Therefore, antibiotic selection should be based on the results of the culture and sensitivity tests.

Determination of immunogenicity of an inactivated ND-vaccine developed experimentally with Newcastle disease virus (Genotype VII.2) local isolates of Bangladesh

BackgroundNewcastle disease virus (NDV) genotype VII severely affects poultry, causing respiratory and neurological symptoms with a high rate of morbidity and mortality. The research aimed to develop an inactivated ND vaccine using local isolates (Genotype VII.2) and assess its immunogenicity compared to other commercial live ND vaccines.MethodsAn inactivated vaccine using a candidate NDV (GenBank: OR924274.1) was developed according to WOAH guidelines following inactivation, sterility, purity, and safety tests. The birds were vaccinated through subcutaneous (SC) and intramuscular (IM) routes using three doses (0.25, 0.5, and 1.0 ml/bird). Immunogenicity and protective potentiality of the experimentally developed inactivated ND vaccine and live commercial ND vaccine (intra-ocularly/IO) were compared by challenge studies using three vaccination schedules: killed-followed-killed, live-followed-killed, and live-followed-live.ResultsThe birds vaccinated with 1.0 ml/bird SC showed higher antibody titers compared to those of IM-vaccinated groups. Birds vaccinated with the live-followed-killed commercial ND vaccines had slightly higher antibody titers compared to those vaccinated with killed-followed-killed and live-followed-live vaccines. Birds vaccinated with the killed-followed-killed ND vaccine showed a higher protection rate (100%) compared to live-followed-killed (83±5.77%) and live-followed-live (57±5.77%) vaccines. Birds vaccinated with killed-followed-killed group showed a slower decline rate of antibody titers than other groups. This regimen provided significantly better immunity, highlighting its potential in controlling ND outbreaks in Bangladesh's poultry.ConclusionThe study found that the inactivated ND vaccine, developed with the locally circulating isolate of genotype-VII.2 of NDV, might play an important role in effective control and management of ND in the commercial poultry population in Bangladesh.

A Combination of Chemotherapy and Autogenous Vaccine for Control of Bovine Papillomatosis in Cattle

The present study was conducted during the period from July, 2023 to January, 2024. Two clinical cases of cattle were reported with cutaneous papillomas, one near right umbilical region and another over the right hind teat of cattle. Both the animals were treated with a protocol designed such that it can regulate the secondary bacterial infection, speedy recovery of integument system and enhance the immune response for complete regression of the warts. The innate immune responses of infected animals were augmented with inactivated pathogen associated molecular patterns (PAMPs) as autogenous vaccine candidate. The warts were surgically excised, triturated in sterile PBS, inactivated with 0.5% formaline and prepared the autogenous vaccine. The sterility test of vaccine revealed that the vaccine was free from bacterial and fungal contaminants. The chemotherapy includes the use of drugs Thuja ointment, Inj.Enrofloxacin, Inj.Avil and Inj.Anthiomaline. The dose of the vaccine was 1 ml 20 kg-1 b.wt. subcutaneously and given at weekly intervals for a period of 4 weeks. It was observed that the treatment protocol was successfully effective and regression of warts was noticed from 2nd week onwards. By end of 4th week a complete recovery of the wart infected area was noticed with a black scar left behind. The formalin inactivated autogenous vaccine was effective, and applied for control of bovine papillomavirus infection in cattle with combination of chemotherapeutic drugs resulted in early regression of warts.

Spatial and temporal characterization of Aedes albopictus oviposition activity in candidate urban settings for sterile insect technique testing in La Reunion Island

BackgroundUnderstanding of mosquito spatiotemporal dynamics is central to characterize candidate field sites for the sterile insect technique (SIT) testing, and is critical to the effective implementation and evaluation of pilot sterile male release programs. Here, we present a detailed description of Aedes albopictus (Skuse) egg-laying activity over a 6-year period in urban areas identified as potential SIT testing sites on Reunion Island.MethodWeekly entomological collections using ovitraps were carried out in residential and adjacent uninhabited habitats in two urban areas, Duparc and Bois Rouge, in the municipality of Sainte Marie, Reunion Island. Time-series data incorporating the frequency of positive ovitraps and the total number of eggs/ovitrap recorded each time at each locality during the study period from May 2013 to December 2018 were analyzed with multifaceted statistical approaches including descriptive statistics and spatiotemporal analyses incorporating the role of climatic factors on overall ovitrap productivity.ResultsDuring the ovitrap survey, the proportion of egg-positive ovitraps differed among study sites (χ2 = 50.21, df = 2, P < 0.001), being relatively lower in Duparc (89.5%) than in Bois-Rouges (95.3%) and the adjacent buffer zone (91.2%). Within each neighborhood, Ae. albopictus egg abundance varied by month in a roughly seasonal pattern marked by a single peak occurring more regularly February each year, a decline at the onset of the austral winter in July, followed by a period of lower ovitrap productivity in August and September. Fluctuation in both positivity rate and eggs densities per ovitraps were related to annual and seasonal variations in local temperature and rainfall (P < 0.001 in all cases). The spatial analysis also captured substantial between- and within-habitats heterogeneity, whereby the overall ovitrap productivity was higher in residential areas than in the buffer zone.ConclusionsCollectively, these results reveal that the distribution of Ae. albopictus oviposition activity is shaped by local habitat heterogeneity and seasonal climatic factors. Overall, this study provides baseline insights into the reproductive dynamics of Ae. albopictus, which would assist in planning locally tailored SIT interventions, while addressing concerns related to focal areas of high egg-laying intensity and potential immigration of females from natural areas.

Post-marketing quality evaluation of oxytocin injection and assessment of associated factors among selected health facilities in Addis Ababa, Ethiopia

Oxytocin is the principal drug and the primary choice for labor induction and postpartum hemorrhage prevention and treatment. The problems associated with oxytocin are not usually from its effectiveness, but they are from its compromised quality which may occur at the point of manufacturing, transportation, and storage. The main objective of this study was to assess the status of oxytocin injection quality and associated factors in the health facilities located in Addis Ababa city, Ethiopia. A laboratory based instrumental quality testing alongside a cross sectional study design was used to evaluate oxytocin injection quality in Addis Ababa city administration from January to June, 2022 according to United States Pharmacopeia (USP 20), and the World Health Organization guideline of post-market medicine quality assessment. Binary logistic regression and cross tabulation analysis were conducted using SPSS 26. Significance was considered at p ≤ 0.05. Out of 107 tested oxytocin injection samples, 92.5% met the limit for oxytocin content, 81.3% for sterility test, and 91.6% for pH value. Overall, quality failure rate was 23.4%, while sterility test failure rate was 18.7%. Lack of reliable refrigerated storage condition [(AOR = 5.62, 95% CI: (1.87, 16.88)], lack of effective cold chain system during distribution [(AOR = 5.5, 95% CI: (1.53, 19.74)], and weak national medicine regulatory system [(AOR = 3.23, 95% CI: (1.13, 9.23)] were among the significantly associated factors for the failure of some of the brands. The study demonstrated that oxytocin injection quality failure rate, failing to meet at least one quality test requirement, is demonstrated in the health facilities of Addis Ababa. The determinant factors for the reported failures are related to storage, distribution, and regulatory system. Hence, these should be addressed by strictly adhering in to the national regulation and providing a comprehensive health promotion on rational use of oxytocin injection should be implemented to assure its quality and safety.

Research on Intelligent Disinfection Robot Based on STC89C52 Microcontroller Control System

An intelligent disinfection robot based on STC89C52 microcontroller as the control system is proposed to address the current problems of high cost, scene limitations, and complex control factors. With the assistance of software development, the peripheral circuit of the chip is designed, and the robot can dynamically adjust its path according to its own and surrounding companions' position and motion status. The research results indicate that the various modules of the system can work in coordination, with high accuracy in tracking routes, precise image recognition, timely obstacle avoidance, fast response speed, and safe and reliable system operation. The disinfection robot carried out disinfection and sterilization test on the most common staphylococcus aureus and mold, and the test results showed that the pathogenic bacteria decreased significantly after 2 min, and all bacteria were removed after 3 min, and the killing rate of bacteria was up to&gt;96.0%. This method improves the obstacle avoidance ability and collaborative efficiency of robot systems in complex environments, and can effectively respond to various unexpected situations.

Improved Automated Radiosynthesis of [18F]Dolutegravir: Toward Clinical Applications.

Positron emission tomography imaging using radiolabeled dolutegravir (DTG) is an interesting approach to understand the biodistribution of this antiretroviral drug at HIV-1 sanctuary sites. In the course of clinical translation, we depict herein an improved and pharmaceutically compliant radiosynthesis of [18F]DTG from an original tin precursor. The radiosynthesis was achieved in two steps by copper-mediated radiofluorination, followed by enol ether deprotection using a kit-based AllInOne module. Ready-to-inject [18F]DTG was obtained in 20 ± 5% (n = 12) decay-corrected radiochemical yield within 90 min, representing a 4-fold increase compared to the previously published three-step radiosynthesis. Quality control was carried out with three consecutive [18F]DTG productions according to the current European Pharmacopoeia guidelines, which include pH determination, identity and purity (chemical, radiochemical, and radionuclide) assessments, residual solvent quantification, dosage of lithium, copper, and tin traces, sterility and bacterial endotoxin tests. [18F]DTG (∼2 GBq) was obtained with a molar activity of 59 ± 2 GBq/μmol at the time of injection and was suitable for human applications.

Carbon-carbon composite material as a potential basis for orthopedic implants

Aim. To determine the cytocompatibility of carbon-carbon composite materials (CCCM) and assess their ability to be impregnated with vancomycin.Materials and Methods. The study included samples of carbon-carbon composite materials (CCCM). The cytocompatibility of CCCM blocks was evaluated using a culture of eukaryotic cells (Vero cell line). Biofilms of S. aureus ATCC 29213 (MSSA), S. aureus ATCC 43300 (MRSA), S. epidermidis ATCC 12228 (MSSE), and S. epidermidis ATCC 29887 (MRSE) were formed by immersing sterile test samples of CCCM into a nutrient medium which contained bacteria. After 24-hour incubation, the samples were washed, placed in an ultrasonic bath, and sonication fluid was inoculated using the sector method. To saturate the CCCM blocks with antibiotics, they were placed into a vancomycin solution and then lyophilized under negative pressure with gradual heating. The antimicrobial activity of the resulting blocks was studied using the cup plate method against the same reference cultures of staphylococci. The dynamics of vancomycin elution from CCCM was investigated using high-performance liquid chromatography.Results. Vero cells maintained their viability in the presence of the tested material. Considering the highly porous structure of CCCM and variable diameter of the pores, we suggested a good osteointegration potential of this material. On the samples without an impregnation with an antibacterial drug, reference strains of staphylococci were able to form a biofilm with a sufficient number of bacterial cells to initiate an infectious process. The duration of antimicrobial activity of the antibioticim-pregnated samples against the reference staphylococcal strains was up to 3 days. The majority of the antibiotic eluted from the CCCM into the incubation medium during the first two days.Conclusion. The cytocompatibility and porosity of CCCM in combination with a vancomycin impregnation makes this material promising for the fabrication of implants with antimicrobial activity as well as tissue engineering constructs.

Quality assessment of common anti-malarial medicines marketed in Gambella, National Regional State, South Western-Ethiopia

BackgroundOver the past years, there has been a growing concern that a considerable amount of anti-malarial supply in the underdeveloped world particularly in the private sector, is of poor quality. The World Health Organization (WHO) has received about 1500 reports that mentions instances of substandard and falsified products since 2013. The majority of the reports concerned antibiotics and anti-malarials. The majority of reports (42%) originate from the WHO African region.ObjectiveThis study intends to assess the quality of the most widely used anti-malarial medications [artemether-lumefantrine tablets, chloroquine phosphate tablets, primaquine phosphate tablets, artesunate, and artemether injections] in Gambella, South-West, Ethiopia.MethodsA total of 52 samples were collected on June 2022 from Gambella National Regional State, Ethiopia. Half of the districts (six) located in the four zones of the region were chosen using simple random sampling technique. All drug retail outlets available in the selected districts (locally known as woredas) were included. The samples were subjected to visual inspection with a tool adopted from the joint WHO/FIP/ USP checklist. The pharmacopeial tests for identification, uniformity of dosage forms, assay, thickness, diameter, hardness, friability, disintegration test, dissolution, and sterility tests were carried out according to the USP 44-NF 39 and International Pharmacopoeia 11th edition, 2022 monographs.Results and DiscussionOnly 25% of the samples were registered on the Ethiopian Food and Drug Authority (EFDA’s) electronic regulatory/ registration system (ERIS). Besides, 88.8% of artemether injection products were presented in clear glass ampoules. This might expose the products to photochemical degradation that leads to in loss of anti-plasmodial activity. In addition, 50% of the artemether products assessed were not bioequivalent with the comparator product in the in vitro dissolution comparison tests. Overall, the study findings reveal a high prevalence (58.3%) of substandard anti-malarial drugs in the region. The stated percent of the samples had failed in one or more of the quality test parameters assessed in this study.ConclusionThe study findings reveal a high prevalence (58.3%) of substandard anti-malarial drugs in the region. Only a quarter were registered and 38% of the unregistered products failed the quality tests. Hence, the national, regional medicine regulatory bodies and other stake holders should perform the required roles to circumvent presence of Substandard and Falsified (SF) anti-malarial drugs in the study sites.

Evaluation of comparative effect between aluminum hydroxide gel and montanide (ISA 70) in potency and protection of locally prepared rabbit hemorrhagic disease virus 2 (RHDV2) vaccines in rabbits

BackgroundRabbit hemorrhagic disease (RHD) is an acute infectious disease that damages the rabbit industry by producing significant mortality rates in young and adult rabbits. RHD is better controlled by vaccination.ObjectiveThe current study's goal was to prepare and evaluate the immuno-enhancing effect of montanide ISA70 and aluminum hydroxide (Al(OH)3) gel incorporated within the inactivated RHDV2 vaccine and assess the vaccine's protective efficacy against the homologous and heterologous local RHDV2 strains in rabbits.MethodsInactivated RHDV vaccines were prepared using Montanide ISA70 oil or Al(OH)3 gel adjuvants and submitted to sterility, safety, and potency tests. 200 rabbits were equally divided into 4 groups: G1 (control), G2 (vaccinated with gel-incorporated vaccine), G3 (vaccinated with montanide-incorporated vaccine), and G4 (vaccinated with gel- and montanide-incorporated vaccines). Individual blood samples were collected from one week to six months from all groups. The vaccine's potency was measured by the HI test and protection percentage post challenge.ResultsData revealed slightly increasing HI titer means reaching the 1st peak at 4 weeks post-vaccination (7.33, 7.67, and 7.33 log2 in the 2nd, 3rd, and 4th groups, respectively), then slightly decreasing and peaked again, giving 9.33 log2 for the2nd group at 3 months post-vaccination (MPV), 10.67 log2 for 3rd the group, and 10.33 log2 for the 4th group at 5 months post-vaccination. Titer gradually decreased but remained protective. The protection rate ranged from 80–100% and 80–90% for homologous and heterologous local RHDV2 vaccines, respectively, within 3 weeks and 6 months post-challenge. The montanide oil RHDV2 vaccine induced better protection than the aluminum gel RHDV2 vaccine.ConclusionThe results demonstrated evidence of cross-protection between RHDV2 strains. The oil emulsion vaccine induced higher and longer-lasting antibody titers than those obtained with the RHDV2 aluminum gel vaccine.

Pharmacopeial parametric release strategy in microbiological quality control of radiopharmaceuticals

Nuclear medicine is used to assess body functionality at various levels of biological organization and is increasingly employed for both diagnostic and therapeutic purposes. Nearly 5% of radiopharmaceuticals are used therapeutically, whereas approximately 95% are used for diagnosis. Diagnostic radiopharmaceuticals are generally considered safe because they are administered in subpharmacological doses, and the radiotracer components are employed in minimal doses. Consequently, the radiation exposure to the body during nuclear medicine investigations is typically low. However, when radiopharmaceuticals are used for therapeutic applications such as tumor ablation and bone pain palliation, the emitted radiation is highly localized and potent. Radiotracers and radiation doses are more carefully selected for young patients, as the radiation-induced risk of adverse health effects is greater in children than in adults. Radiopharmaceutical preparations should adhere to the established standards of pharmacopoeia worldwide to ensure safety and efficacy. Adherence to current good manufacturing practices, sterility, and good radiation practices are prerequisites for ensuring effective quality assurance and quality control systems during radiopharmaceutical preparation. The quality control test parameter for radiopharmaceuticals primarily involves physicochemical and biological tests. However, significant concerns exist regarding the biological testing of radiopharmaceuticals, necessitating their parametric release. The time-intensive traditional sterility test, bacterial endotoxin test, and rabbit pyrogen tests have been critical quality control issues due to the short half-lives of most radiopharmaceuticals, as they should be dispensed at the earliest to ensure optimal diagnostic/therapeutic benefit. Therefore, manufacturers of radiopharmaceuticals, dispensers, and nuclear medicine specialists should be cognizant of the issues posed by biological quality control as a primary obstacle to ensuring consistent quality through parametric release from a compendial test method standpoint.

Long-Term Stability, Sterility, And Cost-Effectiveness of 0.05% Chlorhexidine Gluconate as Antisepsis for Intravitreal Injection.

Purpose: Commercially available chlorhexidine gluconate (CHG) has a beyond-use date of 24 h. This study evaluated the stability and sterility of 0.05% CHG for 30 days after opening and compared its cost to povidone iodine (PI) for intravitreal injection antisepsis. Methods: 0.05% CHG was aliquoted into 1-mL syringes and stored at room temperature or refrigerated. Turbidity, pH, high-performance liquid chromatography (HPLC), and sterility testing were performed. A cost analysis was conducted. Results: 0.05% CHG remained stable for at least 30 days. All samples had measured turbidity <0.5 nephelometric turbidity units. The pH of all samples remained between 5.0 and 7.0. HPLC demonstrated CHG concentration at day 30 relative to day 0 of 98.52% ± 4.16% at room temperature and 99.99% ± 3.38% at 2°C -6°C. The cost per week to perform 150 injections using 0.05% CHG was $463.25 when opening a new bottle daily compared with $16.73 for 5% PI. This cost decreased to $23.16 when utilizing a bottle of CHG for 30 days. Conclusion: 0.05% CHG remains stable and sterile for at least 30 days after opening. The ability to use CHG for at least 30 days after its opening significantly decreases its utilization expense.

In vitro corrosion-fatigue behaviour of rare-earth containing magnesium WE43 in sterile complex cell culture medium

Rare-earth containing magnesium alloys are promising biomedical materials for a new generation of biodegradable orthopaedic implant systems due to their excellent biocompatibility, mechanical and biodegradation properties. However, chemo-mechanical interactions in aggressive physiological corrosion environments result in rapid degradation and early loss of mechanical integrity, limiting its broader application for orthopaedic implants. To date, only few studies have assessed the corrosion-fatigue behaviour of medical-grade magnesium alloys in an organic physiological corrosion environment, especially under sterile test conditions. In the present work, the corrosion-fatigue behaviour of fine-grained medical-grade magnesium alloy WE43MEO was systematically analysed under in vitro conditions using an organic physiological fluid DMEM. The experimental results showed that the fatigue strength of the alloy is nearly unaffected by a 1-day precorrosion, while a 7-day precorrosion resulted in a significant deterioration of mechanical integrity. In corrosion-fatigue experiments, the fatigue life was considerably reduced by interactions between corrosion and fatigue damages. The SEM analysis revealed that the mixed mode of intergranular and transgranular fracture in the crack propagation zone transits to intergranular cracking dominant mode under the corrosion-fatigue conditions due to hydrogen embrittlement.

Allogenic and autologous nondiluted serum eye drops-validation strategy compliant with good manufacturing practice.

Serum eye drops alleviate ocular symptoms of diseases such as sicca syndrome, or chronic graft-versus-host disease. This study was designed for good manufacturing practice validation of our standard manufacturing, storage and transport processes for both autologous and allogenic SEDs. Specifications of quality parameters are lacking and were aimed to be defined. Using sterile collected, coagulated whole blood, serum was separated by centrifugation and filled into single-use eye drop applicator vials. Quality control tests included visual inspection, sterility, leukocyte concentration, pH, vitamin A, TGF-ß and VEGF-A. Samples were collected after manufacture and after 24 h and 6 months of frozen storage (-20°C). Sterility testing was performed after opening the SED applicators at specified intervals. For transport validation, SEDs were packed in insulated transport bags and stored at 20-24°C and 30-32°C for 8 h. Vitamin A, TGF-ß and VEGF-A assays showed no difference in concentration between fresh and 24 h frozen serum. All specifications for pH (aim 7.4) and cellular contamination were met and microbiological contamination tests were negative. Shelf-life was defined as 6 months at -20°C. Once opened, the product must be used within 24 h to avoid bacterial outgrowth. Transporting frozen SEDs from the manufacturer via a local pharmacy to the patient within a maximum of 4 h was demonstrated. The GMP compliance of our production, storage and transport processes for autologous and allogenic SEDs was successfully validated. 100% serum eye drops in single-use applicators can be safely used for up to 24 h after opening.

An Alternative Medium to Support Sterility Testing using the Growth Direct® Rapid Sterility System

Recently, it has been shown that there are other media that perform equivalently or better than the compendial sterility test media, TSB and FTM1. The results presented here demonstrate that a new single medium, Rapid Sterility Medium (RSM) Broth, performs as well as or better for the growth promotion of test organisms compared to compendial sterility test media. In side-by-side tests of aerobic bacteria, yeast and molds and anaerobic organisms, RSM broth supported growth promotion comparable to TSB or FTM for a panel of microorganisms relevant to the pharmaceutical market. These results demonstrate that RSM broth can serve as a valid alternative for both TSB and FTM for sterility testing being performed by the Growth Direct® Rapid Sterility System. Keywords: Sterility test, Alternative media, Rapid Microbial Method, Growth Direct System®

Bacterial species in cord blood and their significance in the context of clinical use

Approximately 90 % of infants born before 28 full weeks(extremely-preterm-infants) receive erythrocyte transfusions in early life. Umbilical cord blood(UCB) has been investigated as an alternative source for erythrocyte transfusions to preterm neonates. This retrospective study aimed to compile/evaluate spectrum of bacteria groups/species intermittently detected in processed UCB at National-Swedish-Cord blood bank, (NS-CBB) during the years 2008–2020.Consecutive data from the years 2008–2020 were investigated. UCB from healthy newborns born after 37 full weeks of gestation was collected following clamping of cord (1 min) through cannulation of umbilical vein(vaginal-and C-section-deliveries). In total, 5194 cord blood units (UCBUs) that met NS-CBB-guidelines for total nucleated-cell-content(TNC) were manufactured from 8875 collections. Of 5194 UCBUs,77,6 % were from vaginal-and 22,4 % from C-section deliveries.Samples(10 mL) were collected from surplus eryhtrocyte fraction post-processing(n = 5194), transferred into BACT/ALERT® aerobic/anaerobic culture flasks and monitored 10 days using BACT/ALERT®−3D-Microbial-Detection-Systems. Positive samples were subcultured and typed for bacterial groups and/or species.Out of 5194 processed sampled UCB units,186 (3,6 %) were discarded due to positive sterility tests, 92 % were detected in samples from vaginal-deliveries and 8 % from C-section-deliveries. In all,16 different groups of bacteria and 27 species were identified. Common bacterial/groups and species were anaerobe gram-negative rods(n = 28),coagulase-negative-staphylococci(n = 21),gram-positive rods(n = 21),anaerobe-gram-positive cocci(n = 20) and viridans-streptococci(n = 13). Extracted from these results,in positive samples(n = 13) from C-section deliveries, bacteria were found:viridans-streptococci(n = 7),Aerococcus-urinae(n = 1), Staphylococcus lugdunensis(n = 1),other coagulase-negative staphylococci(n = 1) or a mix of aerobic/anaerobic bacteria(n = 3).Our results are in alignment with previously published contamination rates in processed UCBUs. Still, results point towards importance of strict microbial monitoring when manufacturing UCBUs to achieve patient-safe- products for stem-cell transplantation/transfusion.

Pilot Study for Isolation of Stromal Vascular Fraction with Collagenase Using an Automated Processing System.

There are many potential therapeutic applications for autologous adipose-derived stromal cells. These cells are found in a heterogeneous population isolated from adipose tissue called the stromal vascular fraction (SVF). Closed automated systems are available to release cells from the adherent stroma. Here, we test one system to evaluate the heterogeneous output for yield, purity, cellular characterization, and stemness criteria. The SVF was isolated from three donors using the Automated Cell Station (ACS) from BSL Co., Ltd., Busan, Republic of Korea. The SVF cellular output was characterized for cell yield and viability, immunophenotyping analysis, pluripotent differentiation potential, adhesion to plastic, and colony-forming units. Additionally, the SVF was tested for endotoxin and collagenase residuals. The SVF yield from the ACS system was an average volume of 7.9 ± 0.5 mL containing an average of 19 × 106 nucleated cells with 85 ± 12% viability. Flow cytometry identified a variety of cells, including ASCs (23%), macrophages (24%), endothelial cells (5%), pericytes (4%), and transitional cells (0.5%). The final concentrated product contained cells capable of differentiating into adipogenic, chondrogenic, and osteogenic phenotypes. Furthermore, tests for SVF sterility and purity showed no evidence of endotoxin or collagenase residuals. The ACS system can efficiently process cells from adipose tissue within the timeframe of a single surgical procedure. The cellular characterization indicated that this system can yield a sterile and concentrated SVF output, providing a valuable source of ASCs within the heterogeneous cell population.

An Alternative Microbiological Validation for an Online Water Bioburden Analyzer.

The Mettler-Toledo 7000RMS analyzer is a Bio-Fluorescent Particle Counter (BFPC) used to monitor real-time bioburden results from Purified Water (PW). Validation of the analyzer using 13 microorganisms and a low-intensity, fluorescent, polystyrene bead. During the execution of the validation, a laboratory water system that met Purified Water (PW) quality standards was connected to the 7000RMS, and a syringe pump was used to introduce various concentrations of microorganisms and fluorescent polystyrene beads to the analyzer. Samples were collected and tested via the traditional Membrane Filtration (MF) method and the Colony Forming Unit (CFU) plate count results were compared to the Auto-Fluorescent Unit (AFU) of the 7000RMS analyzer. The validation study was designed to follow the guidance in United States Pharmacopeia (USP) Chapter <1223 > (1), European Pharmacopeia (EP) Chapter 5.1.6 (2), PDA Technical Report 33 (3). Concepts and strategies were adapted from EP Chapter 2.6.12 Microbiological Examination of Non-sterile Products: Microbial Enumeration Tests, EP 10.2 (4), European Pharmacopeia Chapter 2.6.1 Sterility, EP 10.2 (5), USP Chapter <61> Microbiological Examination of Non-sterile Products: Microbial Enumeration Tests (6), USP Chapter <71> Sterility Tests (7), Japanese Pharmacopoeia (JP) General Information Chapter G8 Water: Quality Control of Water for Pharmaceutical Use (8). All pre-determined validation acceptance criteria for Accuracy, Specificity, Precision, Limit of Detection (LOD), Limit of Quantitation (LOQ), Linearity, and Range were met. Further, the 7000RMS demonstrated Performance Equivalence to the MF method per USP <1223> but characteristically lacked correlation to the CFU. This validation approach highlights the superior capabilities of the 7000RMS when compared against the traditional compendial MF testing method for PW.

Analysis of reconstitution process, stability, stability of intravenous antibiotics in hospitalised patients in a private hospital

Introduction: reconstitution of sterile preparations is a series of changes in the drug from its original condition to a new product by dissolving or adding other ingredients carried out aseptically by the pharmacy. Aim: determine the suitability of the reconstitution technique, the suitability of stability (solvent type and solvent volume), and the suitability of the sterility of the reconstitution product. Methods: data collection obtained by observation is carried out with the help of a checklist that is adapted to hospital standard operating procedures (SOPs), Inject-able Drug Guide, guidelines for mixing inject-able drugs, and preparations. 48 processes of reconstitution of intravenous antibiotics were carried out in hospitals that could be observed as samples. Results: conformity to the preparation stage were 99%, the mixing stage was 73%, the labelling was 100%, and the labelling was 90%. In evaluating the stability of the solvent type category, the suitability of 100%. In the volume category of the solvent used, 90% were suitable, and 10% were unsuitable for the antibiotic’s cefotaxime 1 gram, vancomycin 1 gram, and sulbactam + ampicillin Na 1 gram. From the sterility test results, the reconstitution of ceftriaxone intravenous antibiotic preparations showed that the bacterial preparations were free of microorganisms (sterile). Conclusion: the personnel performing the reconstitution are nurses trained in intravenous antibiotic recovery. One of the factors that caused the reconstitution process not to be under the hospital's KSB was the lack of training and supervision from the hospital management. SPO recovery is not installed in an intravenous drug mixing room.