Good Manufacturing Practice Research Articles

Microbiological safety of dry-aged meat: a critical review of data gaps and research needs to define process hygiene and safety criteria

Dry-aged meat is gaining popularity among food business operators and private consumers. The process is carried out in aerobic conditions by hanging beef carcasses or placing subprimal or primal cuts in a dedicated cabinet for several weeks or even months while controlling the environment through the management of process parameters such as temperature, relative humidity, and airflow. In this review, we present a critical evaluation of the literature to evaluate tools to manage the process to guarantee food safety and identify critical control points, as well as good hygienic and manufacturing practices. In controlled aging conditions, only Listeria monocytogenes and Yersinia enterocolitica can multiply, while a reduction in the number of Salmonella spp. and Escherichia coli O157:H7 is generally reported. Enterobacteriaceae usually decrease on the surface of the meat during maturation; thus, for the purpose of the hygienic evaluation of the production process, a count no higher than that of unmatured meat is expected. Besides, various studies report that the total bacterial count and the spoilage microorganisms significantly increase on the surface of the meat, up to 5-6 Log CFU/g in the absence of visible spoilage. Bacteria of the Pseudomonas genus tend to progressively replace other microorganisms during maturation; thus, the total mesophilic or psychrophilic bacterial load is not a good indicator of process hygiene for matured meat. Critical parameters for the control of the process are temperature, relative humidity, and ventilation, which should be monitored during the process. For this reason, equipment designed and certified for dry aging must be used, and the manufacturer must validate the process. Food business operators must apply general good manufacturing practices (GMP) and good hygiene practices (GHP) for meat processing and some GMP and GHP specific for dry aging. Several research needs were identified, among them the evolution of the populations of L. monocytogenes and Y. enterocolitica and the microbiology of the inner parts of the dry-aged meat.

A GMP-compliant manufacturing method for Wharton’s jelly-derived mesenchymal stromal cells

BackgroundWharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) hold great therapeutic potential in regenerative medicine. Therefore, it is crucial to establish a Good Manufacturing Practice (GMP)-compliant methodology for the isolation and culture of WJ-MSCs. Through comprehensive research, encompassing laboratory-scale experiments to pilot-scale studies, we aimed to develop standardized protocols ensuring the high yield and quality of WJ-MSCs manufacturing.MethodsFirstly, optimization of parameters for the enzymatic digestion method used to isolate WJ-MSCs was conducted. These parameters included enzyme concentrations, digestion times, seeding densities, and culture media. Additionally, a comparative analysis between the explant method and the enzymatic digestion method was performed. Subsequently, the consecutive passaging of WJ-MSCs, specifically up to passage 9, was evaluated using the optimized method. Finally, manufacturing processes were developed and scaled up, starting from laboratory-scale flask-based production and progressing to pilot-scale cell factory-based production. Furthermore, a stability study was carried out to assess the storage and use of drug products (DPs).ResultsThe optimal parameters for the enzymatic digestion method were a concentration of 0.4 PZ U/mL Collagenase NB6 and a digestion time of 3 h, resulting in a higher yield of P0 WJ-MSCs. In addition, a positive correlation between the weight of umbilical cord tissue and the quantities of P0 WJ-MSCs has been observed. Evaluation of different concentrations of human platelet lysate revealed that 2% and 5% concentrations resulted in similar levels of cell expansion. Comparative analysis revealed that the enzymatic digestion method exhibited faster outgrowth of WJ-MSCs compared to the explant method during the initial passage. Passages 2 to 5 exhibited higher viability and proliferation ability throughout consecutive passaging. Moreover, scalable manufacturing processes from the laboratory scale to the pilot scale were successfully developed, ensuring the production of high-quality WJ-MSCs. Multiple freeze-thaw cycles of the DPs led to reduced cell viability and viable cell concentration. Subsequent thawing and dilution of the DPs resulted in a significant decrease in both metrics, especially when stored at 20–27 °C.ConclusionThis study offers valuable insights into optimizing the isolation and culture of WJ-MSCs. Our scalable manufacturing processes facilitate the large-scale production of high-quality WJ-MSCs. These findings contribute to the advancement of WJ-MSCs-based therapies in regenerative medicine.

Tracking progress on Antimicrobial Resistance by the Quadripartite Country Self-Assessment Survey (TrACSS) in G7 countries, 2017-2023: opportunities and gaps

Antimicrobial resistance (AMR) poses serious challenges to the healthcare systems worldwide. Multiple factors and activities contribute to the development and spread of antimicrobial-resistant microorganisms. Monitoring progress in combating AMR is fundamental at both global and national levels to drive multisectoral actions, identify priorities, and coordinate strategies. Since 2017, the World Health Organization (WHO) has collected data through the Tracking AMR Country Self-Assessment Survey (TrACSS). TrACSS data are published in a publicly-available database. In 2023, 71 (59.9%) out of 177 responding countries reported the existence of a monitoring and evaluation plan for their National Action Plan (NAP) on AMR, and just 20 countries (11.3%) the allocation of funding to support NAP implementation. Countries reported challenges including limited financial and human resources, lack of technical capacity, and variable political commitment. Even across the Group of Seven (G7) countries, which represent some of the world's most advanced economies, many areas still need improvement, such as full implementation of infection prevention and control measures, adoption of WHO access/watch/reserve (AWaRe) classification of antibiotics, effective integration of laboratories in AMR surveillance in the animal health and food safety sectors, training and education, good manufacturing and hygiene practices in food processing, optimising pesticides use and environmental residues of antimicrobial drugs. Continuous and coordinated efforts are needed to strengthen multisectoral engagement to fight AMR.

Advancing 6-bromo-7-[11C]methylpurine to clinical use: improved regioselective radiosynthesis, non-clinical toxicity data and human dosimetry estimates

Background6-Bromo-7-[11C]methylpurine ([11C]BMP) is a radiotracer for positron emission tomography (PET) to measure multidrug resistance-associated protein 1 (MRP1) transport activity in different tissues. Previously reported radiosyntheses of [11C]BMP afforded a mixture of 7- and 9-[11C]methyl regioisomers. To prepare for clinical use, we here report an improved regioselective radiosynthesis of [11C]BMP, the results of a non-clinical toxicity study as well as human dosimetry estimates based on mouse PET data.Results[11C]BMP was synthesised by regioselective N7-methylation of 6-bromo-7H-purine (prepared under good manufacturing practice) with [11C]methyl triflate in presence of 2,2,6,6-tetramethylpiperidine magnesium chloride in a TRACERlab™ FX2 C synthesis module. [11C]BMP was obtained within a total synthesis time of approximately 43 min in a decay-corrected radiochemical yield of 20.5 ± 5.2%, based on starting [11C]methyl iodide, with a radiochemical purity > 99% and a molar activity at end of synthesis of 197 ± 130 GBq/μmol (n = 28). An extended single-dose toxicity study conducted in male and female Wistar rats under good laboratory practice after single intravenous (i.v.) administration of unlabelled BMP (2 mg/kg body weight) revealed no test item related adverse effects. Human dosimetry estimates, based on dynamic whole-body PET data in female C57BL/6J mice, suggested that an i.v. injected activity amount of 400 MBq of [11C]BMP will deliver an effective dose in the typical range of 11C-labelled radiotracers.Conclusions[11C]BMP can be produced in sufficient amounts and acceptable quality for clinical use. Data from the non-clinical safety evaluation showed no adverse effects and suggested that the administration of [11C]BMP will be safe and well tolerated in humans.

Quality control in SARS-CoV-2 RBD-Fc vaccine production using LC–MS to confirm strain selection and detect contaminations from other strains

Coronavirus disease of 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is an ongoing outbreak, disrupting human life worldwide. Vaccine development was prioritized to obtain a biological substance for combating the viral pathogen and lessening disease severity. In vaccine production, biological origin and relevant materials must be carefully examined for potential contaminants in conformity with good manufacturing practice. Due to fast mutation, several SARS-CoV-2 variants and sublineages have been identified. Currently, most of COVID-19 vaccines are developed based on the protein sequence of the Wuhan wild type strain. New vaccines specific for emerging SARS-CoV-2 strains are continuously needed to tackle the incessant evolution of the virus. Therefore, in vaccine development and production, a reliable method to identify the nature of subunit vaccines is required to avoid cross-contamination. In this study, liquid chromatography-mass spectrometry using quadrupole-time of flight along with tryptic digestion was developed for distinguishing protein materials derived from different SARS-CoV-2 strains. After analyzing the recombinantly produced receptor-binding domain (RBD) of the SARS-CoV-2 spike protein, nine characteristic peptides were identified with acceptable limits of detection. They can be used together to distinguish 14 SARS-CoV-2 strains, except Kappa and Epsilon. Plant-produced RBD-Fc protein derived from Omicron strains can be easily distinguished from the others with 4–5 unique peptides. Eventually, a peptide key was developed based on the nine peptides, offering a prompt and precise flowchart to facilitate SARS-CoV-2 strain identification in COVID-19 vaccine manufacturing.

Development and Implementation of an Ultraviolet-Dye-Based Qualification Procedure for Hand Washing and Disinfection to Improve Quality Assurance of Pharmacy Preparations and Compounding, Especially in Cleanrooms: A Pilot Study

Even though, nowadays, most medicines are manufactured industrially, patients may have medical needs that can only be met by a tailor-made approach. This requires the availability of pharmacy preparations made under Good Manufacturing Practice (GMP) conditions. An efficient hand hygiene practice is essential herewith, especially if sterile products that are prepared in a cleanroom are concerned. The effectiveness of hand washing and hand disinfection procedures greatly relies on adequate training. We carried out an observational cross-sectional pilot study aimed at optimizing hand hygiene training with objective and measurable quality assessments using an ultraviolet (UV) dye. Practical acceptance criteria for qualifying personnel through this method were set and evaluated. In total, 25 GMP-qualified cleanroom operators washed and disinfected their hands with UV dye hand wash lotion and UV dye hand alcohol, respectively. To obtain a proof-of-concept, the results were judged based on adherence to the WHO six-step protocol and associated acceptance criteria. Commonly missed areas were brought to light, and the influence of procedure duration was investigated. UV-dye-based assessments appeared to be more valuable in hand disinfection than in hand washing. In both procedures, the back of the hands and the thumbs were frequently missed. This underpins the need for enhanced and repeated education on hand washing and disinfection. Additionally, a dry skin gave rise to extra cleaning challenges. From this pharmacy practice pilot study with a focus on pharmaceutical product care, it may be concluded that the application of UV-dye-based assessments offers valuable insights for pharmacists to optimize hand hygiene, thereby increasing the safety of tailor-made medicines and on-site preparations.

On-Chip Neural Induction Boosts Neural Stem Cell Commitment: Toward a Pipeline for iPSC-Based Therapies.

The clinical translation of induced pluripotent stem cells (iPSCs) holds great potential for personalized therapeutics. However, one of the main obstacles is that the current workflow to generate iPSCs is expensive, time-consuming, and requires standardization. A simplified and cost-effective microfluidic approach is presented for reprogramming fibroblasts into iPSCs and their subsequent differentiation into neural stem cells (NSCs). This method exploits microphysiological technology, providing a 100-fold reduction in reagents for reprogramming and a ninefold reduction in number of input cells. The iPSCs generated from microfluidic reprogramming of fibroblasts show upregulation of pluripotency markers and downregulation of fibroblast markers, on par with those reprogrammed in standard well-conditions. The NSCs differentiated in microfluidic chips show upregulation of neuroectodermal markers (ZIC1, PAX6, SOX1), highlighting their propensity for nervous system development. Cells obtained on conventional well plates and microfluidic chips are compared for reprogramming and neural induction by bulk RNA sequencing. Pathway enrichment analysis of NSCs from chip showed neural stem cell development enrichment and boosted commitment to neural stem cell lineage in initial phases of neural induction, attributed to a confined environment in a microfluidic chip. This method provides a cost-effective pipeline to reprogram and differentiate iPSCs for therapeutics compliant with current good manufacturing practices.

Production of [211At]NaAt solution under GMP compliance for investigator-initiated clinical trial.

The alpha emitter astatine-211 (211At) is garnering attention as a novel targeted alpha therapy for patients with refractory thyroid cancer resistant to conventional therapy using beta emitter radioiodine (131I). Herein, we aimed to establish a robust method for the manufacturing and quality control of [211At]NaAt solution for intravenous administration under the good manufacturing practice guidelines for investigational products to conduct an investigator-initiated clinical trial. 211At was separated and purified via dry distillation using irradiated Bi plates containing 211At obtained by the nuclear reaction of 209Bi(4He, 2n)211At. After purification, the 211At trapped in the cold trap was collected in a reaction vessel using 15 mL recovery solution (1% ascorbic acid and 2.3% sodium hydrogen carbonate). After stirring the 211At solution for 1h inside a closed system, the reaction solution was passed through a sterile 0.22μm filter placed in a Grade A controlled area and collected in a product vial to prepare the [211At]NaAt solution. According to the 3-lot tests, decay collected radioactivity and radiochemical yield of [211At]NaAt were 78.8 ± 6.0 MBq and 40 ± 3%, respectively. The radiochemical purity of [211At]At- obtained via ion-pair chromatography at the end of synthesis (EOS) was 97 ± 1%, and remained > 96% 6h after EOS; it was detected at a retention time (RT) 3.2-3.3min + RT of I-. LC-MS analysis indicated that this principal peak corresponded with an astatide ion (m/z = 210.988046). In gamma-ray spectrometry, the 211At-related peaks were identified (X-ray: 76.9, 79.3, 89.3, 89.8, and 92.3keV; γ-ray: 569.7 and 687.0keV), whereas the peak at 245.31keV derived from 210At was not detected during the 22h continuous measurement. The target material, Bi, was below the 9 ng/mL detection limit in all lots of the finished product. The pH of the [211At]NaAt solution was 7.9-8.6; the concentration of ascorbic acid was 9-10mg/mL. Other quality control tests, including endotoxin and sterility tests, confirmed that the [211At]NaAt solution met all quality standards. We successfully established a stable method of [211At]NaAt solution that can be administered to humans intravenously as an investigational product.

RNA-Based vaccine manufacturing: infrastructure, regulations, and global implications

RNA-based vaccines have emerged as a powerful tool in the fight against infectious diseases, including the recent COVID-19 pandemic. However, their successful production requires properly designed facilities for RNA synthesis, purification, and formulation. This review article explores the facility requirements, regulatory aspects, and global implications of specialized RNA vaccine production. It highlights the importance of Good Manufacturing Practice (GMP), equipment and technology in the production process, providing a critical analysis of collaborations, investments, and the establishment of RNA plants around the world. Finally, it addresses the challenges faced by low- and middle-income countries and the role of the "tripod" production model involving collaboration between university, industry, and government. Overall, this document clarifies the most relevant aspects of RNA-based vaccine manufacturing, emphasizing the need for quality, safety, and scalability to meet global health demands.

Perspectives on systematic capacity building in pharmaceutical regulation for regulators of medical products.

Having a robust, integrated regulatory system is important for ensuring the availability of safe and efficacious medical products of good quality and for protecting public health. However, less than 30% of countries globally have reached the required regulatory maturity level three, with low- and middle-income countries facing challenges in attracting and retaining qualified staff. World Health Organization (WHO) advocates for systematic workforce development, including competency-based education, to address these gaps. We provide perspectives on a systematic approach to capacity building of medicine regulators based on the experience and lessons learnt in developing and piloting the WHO global competency framework for medicine regulators through three scenarios. A systematic approach to capacity building, such as the human performance technology model, can be used to implement the WHO competency framework as part of organizational performance improvement while ensuring that initiatives are well-defined, targeted, and aligned with organizational goals. The competency framework can be used in different contexts, such as improving organization performance for individual regulatory authorities, strengthening regional collaborations, harmonization and reliance on medical products assessment and joint good manufacturing practices inspections of pharmaceutical manufacturers, and developing learning programs for medicine regulators. A competency-based learning approach for regulatory professionals ensures the transfer of learning to the workplace by integrating real-world practices in learning activities and assessments. Further work is required to develop and validate the assessment instruments, apply the competency framework in other contexts, expanding the learning programmes while continuously providing feedback for further refinement of the competency framework and implementation support tools.

Theoretical Approaches to Process Validation in Pharmaceutical Manufacturing Process

Under the more general statutory CGMP provisions of section 501(a)(2)(B) of the Federal Food, Drug and Cosmetic Act, validation of manufacturing processes is deemed a legal part of current good manufacturing practice for active pharmaceutical ingredients (APIs). Validation of manufacturing processes is required by the Current Good Manufacturing Practice (CGMP) regulations for finished pharmaceuticals (21 CFR 211.100 and 211.110). A medication should be created that is suitable for its intended purpose, according to the fundamental tenet of quality assurance. Procedure Validation is the collection and estimation of data that proves a process can reliably produce high-quality goods, starting with process design and continuing through commercial production. Three stages of validation are suggested by the 2011 USFDA process validation guideline - Process design, Process qualification, Continued Process qualification. An introduction and broad review of process validation in the pharmaceutical manufacturing process, specifically in the tablet manufacturing process, are provided in this article. Process validation ensures that a process will consistently create a product that meets its predefined quality features and qualities. It is a crucial step in the design, prototyping, and manufacturing processes. Since quality is always a necessary precondition for any product, pharmaceuticals must be produced to the greatest standards of quality. Furthermore, end-product testing does not ensure the product's quality on its own; rather, quality assurance methods need to be applied to incorporate the product's quality throughout the whole process, rather than only testing it at the conclusion.

Impact of Cosmetics and Cleansers in Atopic Dermatitis—How to Advise Patients

Abstract Purpose of Review Atopic dermatitis (AD) or eczema is a frequent chronic inflammatory skin disease. Taking care of the eczematous skin is important to reduce the inflammation and return it to a healthier looking nourished skin. Beyond recommending topical anti-inflammatory drugs and the use of emollients, recommendations on the correct use of other everyday skin products, as cleansers or make-up products, are lacking. Recent Findings The dry, itchy, and inflamed eczematous skin causes anxiety, poor self-image, low self-esteem, decreased social skills, and an overall decrease in a patients’ quality of life. The use of make-up has been shown to ameliorate these impacts. Knowledge on the components of cosmetic products can help suit the products to an eczematous skin. Existing data identifies agents more likely to cause allergic contact dermatitis and shows how to identify cosmetics that follow the principles of “hurdle technology,” how rubbing during cleansing is a significant and previously unrecognized exacerbation factor, and how cleansers interact with eczematous skin. An adequate choice of all these products allows a patient to enjoy its benefits, while preserving a healthier skin. Summary Guiding healthcare professionals on the composition of the cosmetics used, avoiding products with high allergenic properties, identifying products that follow the principles of “hurdle technology,” and educating patients on the appropriate use of make-up and cleansing products have a positive impact on the care of atopic dermatitis and should be part of a holistic approach to a patient. Key messages - Adverse reactions to cosmetics can come from allergic sensitization or irritant stimuli (like the rubbing associated with washing the skin). - Optimizing the composition of the cosmetics used is essential. Ideally, the patients should distinguish the multiple components of a cosmetic, identifying potential risks. - Avoid products with high allergenic properties, like fragrances or preservatives. Select products which follow the principles of “hurdle technology”: sterile cosmetic technology, good manufacturing practices, appropriate packaging, emulsion form, water activity, and pH control. - Appropriate use of make-up is important for better care and quality of life of AD patients. Guidance on their use and appropriate cleansing products and techniques are essential to prevent exacerbations.

Current state and prospects for the development of the domestic pharmaceutical industry

The main stages of the development of the Russian medical and pharmaceutical industry in 2010 are presented in the context of the dominance of large foreign companies producing medicines and medical devices in the pharmaceutical market. This reflects the political and economic restrictive measures illegally taken against Russia by the European Union, the United States of America, and some other unfriendly countries and has influenced the dynamics of the development of the medical and pharmaceutical industry in our country. To harmonize approaches to the production and quality control of medicines, this paper presents the stages of large-scale reforms on the transition of the Russian pharmaceutical industry to standards of good manufacturing practice and the implementation of state programs aimed at modernizing the industrial segment of the pharmaceutical industry. As part of the improvement of the standardization system of medicines, the study also presents the characteristics of the XIII, XIV, and XV editions of the State Pharmacopoeia of the Russian Federation published during the study period, in the development of which employees of the Military Medical Academy named after S.M. Kirov participated. The role and purpose of the creation of the developed Pharmacopoeia of the Eurasian Economic Union, approved by the Board of the Eurasian Economic Commission, is also reflected in this paper. This article describes the phased implementation of federal target programs, state programs, and other strategic plans and measures of nonfinancial support from the state, which have significantly expanded competencies in the field of development and production of medicines and medical devices and increased their output during the period analyzed. The development of medicines and increasing their production volumes with the help of pharmaceutical clusters, which acted as the “locomotives” of the industry, and various projects in establishing the production of innovative medical products with the help of the Industrial Development Fund, which increased the investment attractiveness of these sectors of the national economic complex, played an important role. The results revealed the essence of the changes concerning the revival of the activities of industrial pharmacies, considering a personalized approach to the treatment of patients. The study also presents an assessment of the Pharmaceutical industry Development Strategy for the period up to 2030.

A retrospective regulatory analysis of FDA recalls carried out by pharmaceutical companies from 2012 to 2023

To introduce products in the US market, pharmaceutical companies must first obtain FDA clearance. Manufacturers might recall a product if it poses a risk of damage or violates FDA regulations. This study investigates the types, causes and consequences of recalls, as well as FDA participation and suitable recall strategies. We relied on the FDA website to gather recall data sets from 2012 to 2023, collecting information on the date of issuance, company and type of violation. The most frequent causes for recalls were sterility issues and inadequate compliance with current good manufacturing practices (cGMP). An examination of sterility recalls revealed two primary causes: a lack of assurance in sterility (accounting for 48% of recalls) and instances of non-sterility (making up 45% of recalls). A thorough examination of cGMP recalls revealed five primary types of violations: process control issues, inadequate storage practices, manufacturing problems, the presence of nitroso-amine impurities and concerns regarding stability. The findings demonstrate that sterility and cGMP compliance are FDA priorities. Pharmaceutical companies must, therefore, enhance quality compliance and create effective quality management systems that oversee the manufacturing process, quality control, personnel training and documentation to avoid these recalls. Companies should establish an internal compliance checklist and be prepared for the rectification process.

Validating human induced pluripotent stem cell-specific quality control tests for the release of an intermediate drug product in a Good Manufacturing Practice quality system

One of the challenges in Good Manufacturing Practice (GMP)-compliant human induced pluripotent stem cell (hiPSC) production is the validation of quality control (QC) tests specific for hiPSCs, which are required for GMP batch release. This study presents a comprehensive description of the validation process for hiPSC-specific GMP-compliant QC assays; more specifically, the validation of assays to assess the potential presence of residual episomal vectors (REVs), the expression of markers of the undifferentiated state and the directed differentiation potential of hiPSCs. Critical aspects and specific acceptance criteria were formulated in a validation plan prior to assay validation. Assay specificity, sensitivity and reproducibility were tested, and the equipment used for each assay was subjected to performance qualification. A minimum input of 20 000 cells (120 ng of genomic DNA) was defined for accurate determination of the presence of REVs. Furthermore, since vector loss in hiPSC lines is a passage-dependent process, we advocate screening for REVs between passages eight and 10, as testing at earlier passages might lead to unnecessary rejection of hiPSC lines. The cutoff value for assessment of markers of the undifferentiated state was set to the expression of at least three individual markers on at least 75% of the cells. When multi-color flow cytometry panels are used, a fluorescence minus one control is advised to ensure the control for fluorescent spread. For the assay to assess the directed differentiation potential, the detection limit was set to two of three positive lineage-specific markers for each of the three individual germ layers. All of our assays proved to be reproducible and specific. Our data demonstrate that our implemented analytical procedures are suitable as QC assays for the batch release of GMP-compliant hiPSCs.

Production of donor-derived cytotoxic T lymphocytes with potent anti-leukemia activity for adoptive immunotherapy in high-risk pediatric patients given haploidentical hematopoietic stem cell transplantation

Background aimsSomatic cell therapy based on the infusion of donor-derived cytotoxic T lymphocytes (CTL) able to recognize patients’ leukemia blasts (LB) is a promising approach to control leukemia relapse after allogeneic HSCT. The success of this approach strongly depends on the ex vivo generation of high-quality donor-derived anti-leukemia CTL in compliance with Good Manufacturing Practices (GMP). We previously described a procedure for generating large numbers of donor-derived anti-leukemia CTL through stimulation of CD8-enriched lymphocytes with dendritic cells (DCs) pulsed with apoptotic LB in the presence of interleukin (IL)-12, IL-7 and IL-15. Here we report that the use of IFN-DC and the addition of IFNα2b during the priming phase significantly improve the generation of an efficient anti-leukemia T cells response in vitro. MethodsUsing this approach, 20 high-risk pediatric patients given haploidentical HSCT for high-risk acute leukemia were enrolled and 51 batches of advanced therapy medical products (ATMP), anti-leukemia CTL, were produced. ResultsQuality controls demonstrated that all batches were sterile, free of mycoplasma and conformed to acceptable endotoxin levels. Genotype analysis confirmed the molecular identity of the ATMP based on the starting biological material used for their production. The majority of ATMP were CD3+/CD8+ cells, with a memory/terminal activated phenotype, including T-central memory populations. ATMP were viable after thawing, and most ATMP batches displayed efficient capacity to lyse patients’ LB and to secrete interferon-γ and tumor necrosis factor-α. ConclusionsThese results demonstrated that our protocol is highly reproducible and allows the generation of large numbers of immunologically safe and functional anti-leukemia CTL with a high level of standardization.

Inactivation efficacy of four commercial post-process treatments against Listeria monocytogenes and impact on the commercial quality of leafy greens

The problem of product contamination with human pathogenic microorganisms is still unsolved. Although prevention through the implementation of good agricultural practices and good manufacturing practices is the gold standard, product decontamination strategies can be used to complement preventive measures. Different bioprotective treatments have been suggested to reduce the risk of listeriosis in fresh produce, but more information is needed before industrial implementation. This study focused on the efficacy of four commercial treatments against Listeria monocytogenes growth and their effect on the organoleptic quality of baby spinach, cut romaine, cut iceberg, and shredded cabbage. The treatments included SafePro® (CHR Hansen, DK) and Holdbac® (DowDuPont, US), based on the use of lactic acid bacteria (LAB) cultures as well as two different bacteriophage solutions, ListShield (Intralytix, US) and PhageGuard Listex™ (PhageGuard, US). The selected treatments were tested under simulated commercial conditions. The application was performed after processing and before packaging using a fine mist spray. In all the trials, Listex™ showed the best results in reducing L. monocytogenes growth during shelf life in the four leafy greens chosen. Microbial reductions were close to 2 log units. L. monocytogenes reductions were observed 1 day after Listex™ application and the efficacy was maintained during storage (15 days at 7°C). Organoleptic quality, including flavor, texture, browning/yellowing, decay, and visual appearance of the four leafy greens was not significantly affected by the application of Listex™ when compared to the untreated products. Post-process treatment with Listex™ based on bacteriophages effectively reduced L. monocytogenes and can be recommended as a control tool to reduce the risks for the presence of L. monocytogenes in leafy greens.

Calcium chloride declotted human platelet lysate (hPL) promotes the expansion of mesenchymal stromal cells and allows manufacturing of immunomodulatory active EV products

Mesenchymal stromal cells (MSCs) exert immunomodulatory effects, primarily through released extracellular vesicles (EVs). For the clinical grade manufacturing of MSC-EV products culture conditions need to support MSC expansion and allow the manufacturing of potent MSC-EV products. Traditionally, MSCs are expanded in fetal bovine serum supplemented media. However, according to good manufacturing practice (GMP) guidelines the use of animal sera should be avoided. To this end, human platelet lysate (hPL) has been qualified as animal serum replacement. Although hPL outcompetes animal sera in promoting MSC expansion, hPL typically contains components of the coagulation system that need to be inhibited or removed to avoid coagulation reactions in the cell culture. Commonly, heparin is utilized as an anticoagulant; however, higher concentrations of heparin can negatively impact MSC viability, and conventional concentrations alone do not sufficiently prevent clot formation in prepared media. To circumvent unwanted coagulation processes, this study compared various clotting prevention strategies, including different anticoagulants and a calcium chloride (CaCl2)-mediated declotting method, which in combination with heparin addition was found effective. We evaluated the influence of the differently treated hPLs on the proliferation and phenotype of primary bone marrow-derived MSCs and identified the CaCl2-mediated declotting method as the most effective option. To determine whether CaCl2 declotted hPL allows the manufacturing of immunomodulatory MSC-EV products, EVs were prepared from conditioned media of MSCs expanded with either conventional or CaCl2 declotted hPL. In addition to metric analyses, the immunomodulatory potential of resulting MSC-EV products was assessed in a recently established multi-donor mixed lymphocyte reaction assay. Our findings conclusively show that CaCl2-declotted hPLs support the production of immunomodulatory-active MSC-EV products.

Pemanfaatan Cengkeh dan Daun Salam pada berbagai Produk Kosmetik

Cloves are one of the richest sources of phenolic compounds such as eugenol, eugenol acetate and gallic acid and have great potential for pharmaceutical, cosmetic, food and agricultural applications. The aim of this community service activity is to provide knowledge regarding the benefits of cloves, which are applied in the form of cosmetic preparations, as well as providing training to partners so that their use is optimal and practical, and it is hoped that this can become a business opportunity.        The solution proposed in this service activity is (1) providing knowledge about clove plants, (2) procuring materials in the form of medicinal plants for making preparations. (3) procurement of containers for the packaging process (4) conducting Good Manufacturing Practices (GMP) training to partners so that partners can make them independently.         In service activities, the method used is an approach to the collaboration partner "SMK Mandala Bhakti Surakarta" providing counseling about clove plants and training as a transfer of science and technology (IPTEK) to make cosmetic preparations as well as evaluating the results of activities by giving partners the opportunity to practice making preparation.

[Production Risk Analysis and Key Points for Field Inspection of Medical Electron Linear Accelerator].

The medical electron linear accelerator（LINAC） has the characteristics of complex system structure, many core components and high precision control requirements, which puts forward higher requirements for product quality control and regulation. This study puts forward the main points of field inspection through the analysis of the technical characteristics and production risk of LINAC, combined with the requirements of the good manufacturing practice of medical devices. It has certain reference significance for quality management personnel and field inspectors.