Manufacture Of Medicines Research Articles

Identifying and visualising multimorbidity and comorbidity patterns in patients in the English National Health Service: a population-based study

Globally, there is a paucity of multimorbidity and comorbidity data, especially for minority ethnic groups and younger people. We estimated the frequency of common disease combinations and identified non-random disease associations for all ages in a multiethnic population. In this population-based study, we examined multimorbidity and comorbidity patterns stratified by ethnicity or race, sex, and age for 308 health conditions using electronic health records from individuals included on the Clinical Practice Research Datalink linked with the Hospital Episode Statistics admitted patient care dataset in England. We included individuals who were older than 1 year and who had been registered for at least 1 year in a participating general practice during the study period (between April 1, 2010, and March 31, 2015). We identified the most common combinations of conditions and comorbidities for index conditions. We defined comorbidity as the accumulation of additional conditions to an index condition over an individual's lifetime. We used network analysis to identify conditions that co-occurred more often than expected by chance. We developed online interactive tools to explore multimorbidity and comorbidity patterns overall and by subgroup based on ethnicity, sex, and age. We collected data for 3 872 451 eligible patients, of whom 1 955 700 (50·5%) were women and girls, 1 916 751 (49·5%) were men and boys, 2 666 234 (68·9%) were White, 155 435 (4·0%) were south Asian, and 98 815 (2·6%) were Black. We found that a higher proportion of boys aged 1-9 years (132 506 [47·8%] of 277 158) had two or more diagnosed conditions than did girls in the same age group (106 982 [40·3%] of 265 179), but more women and girls were diagnosed with multimorbidity than were boys aged 10 years and older and men (1 361 232 [80·5%] of 1 690 521 vs 1 161 308 [70·8%] of 1 639 593). White individuals (2 097 536 [78·7%] of 2 666 234) were more likely to be diagnosed with two or more conditions than were Black (59 339 [60·1%] of 98 815) or south Asian individuals (93 617 [60·2%] of 155 435). Depression commonly co-occurred with anxiety, migraine, obesity, atopic conditions, deafness, soft-tissue disorders, and gastrointestinal disorders across all subgroups. Heart failure often co-occurred with hypertension, atrial fibrillation, osteoarthritis, stable angina, myocardial infarction, chronic kidney disease, type 2 diabetes, and chronic obstructive pulmonary disease. Spinal fractures were most strongly non-randomly associated with malignancy in Black individuals, but with osteoporosis in White individuals. Hypertension was most strongly associated with kidney disorders in those aged 20-29 years, but with dyslipidaemia, obesity, and type 2 diabetes in individuals aged 40 years and older. Breast cancer was associated with different comorbidities in individuals from different ethnic groups. Asthma was associated with different comorbidities between males and females. Bipolar disorder was associated with different comorbidities in younger age groups compared with older age groups. Our findings and interactive online tools are a resource for: patients and their clinicians, to prevent and detect comorbid conditions; research funders and policy makers, to redesign service provision, training priorities, and guideline development; and biomedical researchers and manufacturers of medicines, to provide leads for research into common or sequential pathways of disease and inform the design of clinical trials. UK Research and Innovation, Medical Research Council, National Institute for Health and Care Research, Department of Health and Social Care, Wellcome Trust, British Heart Foundation, and The Alan Turing Institute.

A survey of Japanese people's attitude to and experience of Kampo medicines

AbstractAimKampo medicine is traditional Japanese herbal medicine derived from ancient Chinese medicine. In the past, the Japan Kampo Medicines Manufacturers Association conducted surveys and reported on the awareness and experience of Kampo medicines among those participating in Kampo workshops as well as the local community. Here, we investigated the Japanese public's awareness and experience of using Kampo medicines through an online questionnaire. In addition, we analyzed how Kampo medicines are perceived and presented a favorable prospect for Kampo medicines.MethodsIn 2020, we conducted an online survey of registered Japanese members (aged over 25 years) of a research company.ResultsA total of 2060 respondents answered the questionnaire. Of these, 71% had taken Kampo medicines. Among this group, 63.5% obtained Kampo medicines through a doctor's prescription, 46.3% purchased Kampo medicines through pharmacies, drugstores, over the Internet, or by mail order, and only 11.3% had purchased Kampo medicines at Kampo specialty pharmacies. Although many people have taken Kampo medicines, they do not always have an accurate understanding of Kampo medicines as medicines. Especially in the case of Kampo medicines for prescription, people take them passively and do not have a strong awareness of Kampo medicines. In contrast, over‐the‐counter Kampo medicines are widely recognized as drugs.ConclusionMedical personnel, manufacturers, and pharmacists need to provide accurate information about Kampo medicines.

Medicinal Plants Used by Traditional Practitioners for the Treatment of Diabetes, Obesity and Arterial Hypertension in the Dja and Lobo Department of Cameroon

Background/Purpose: Diabetes is a metabolic disease characterised by chronic hyperglycaemia, with disturbance of carbohydrate, lipid and protein metabolism, resulting from a defect in insulin secretion and/or action. The International Diabetes Federation (IDF) estimated in 2017 that 425 million people were living with diabetes worldwide. It is estimated that by 2045, 629 million adults will have diabetes. In Cameroon, a prevalence of 5.59% was recorded in 2018, regardless of gender [1]. In the Department of Dja and Lobo, an ethnopharmacological study was set up to identify and characterize the medicinal flower used in the management of metabolic syndrome. Material and methods: Surveys were conducted among 135 people. Data on medicinal recipes used in the management of diabetes, hypertension and obesity were collected according to a standardized framework. The plant samples mentioned were collected and identified at the National Herbarium of Cameroon and characterized. Phytochemical screening was performed on each species. Results: A total of 135 natives from six different villages were interviewed about plants used in the management of metabolic syndrome in the Department of Dja and Lobo. The study identified 85 species in 49 families. The fabaceae, asteraceae and flacourthiaceae families each had at least five species cited by the stakeholders who participated in our investigation. The medicinal plants inventoried totaled 7 biological types. In total 41 species were involved in the treatment of diabetes. The most used preparation methods were decoction and trituration. Phytochemical screening revealed the presence of flavonoids, Phenols, Polyphenols, Tannins, Saponins in almost all the extracts collected. Anthocyanins were extremely abundant in almost all extracts. Conclusion: The results obtained constitute a very valuable source of information for the region studied. The valorization of these plants and the determination of the health profile of the local populations of the Dja and Lobo department requires the establishment of modern processing units specialised in the manufacture of medicines.

Controlling the Polymorphism of Indomethacin with Poloxamer 407 in a Gas Antisolvent Crystallization Process.

The polymorphic control of active pharmaceutical ingredients (APIs) is a major challenge in the manufacture of medicines. Crystallization methods that use supercritical carbon dioxide as an antisolvent can create unique solid forms of APIs, with a particular tendency to generate metastable polymorphic forms. In this work, the effects of processing conditions within a gas antisolvent (GAS) crystallization method, such as pressure, stirring rate, and temperature, as well as the type of solvent used and the presence of an additive, on the polymorphism of indomethacin were studied. Consistent formation of the X-ray powder diffraction-pure α polymorphic form of indomethacin by GAS was only achieved when a polymer, poloxamer 407, was used as an additive. Using the GAS method in combination with poloxamer 407 as a molecular additive enabled full control over the polymorphic form of indomethacin, regardless of the processing conditions employed, such as pressure, temperature, stirring rate, and type of solvent. A detailed molecular modeling study provided insight into the role of poloxamer 407 in the polymorphic outcome of indomethacin and concluded that it favored the formation of the α polymorph.

Autonomous DataFactory: High-throughput screening for large-scale data collection to inform medicine manufacture

Using small-scale crystallisation to inform downstream processes, we can reduce time and material costs in medicine manufacturing. This work introduces a preliminary workflow for information-rich data collection of crystallisation parameters including solubility, induction time, growth rate, secondary nucleation rate, particle shape and size. Large-scale data collection was achieved for 6 active pharmaceutical ingredients (APIs) in 31 solvents in less than 9 months with the results for aspirin presented here. Highlights include the identification of 24 potential alternative crystallisation solvents for manufacturing aspirin, all of which yield the biorelevant polymorph. Automation of this workflow will enable the use of robotics to further reduce time and material usage when conducting crystallisation experiments for future APIs.

Geometry in Pharmaceutical Systems

The aim of the paper is to brief out geometry in pharmaceutical systems, i.e., influence of tablet surface area/volume (SA/Vol) on drug release from controlled-release matrix tablets containing hydroxyl propyl methyl cellulose (HPMC) and describes a situation where the erosion rates of the tablet are different in the radial and axial directions, 3D printing technologies available for the manufacture of modern medicines and so on.

Efforts at building capacity for manufacturing and testing the quality of medicines in Sub-Saharan Africa: Historical evidence from the BIRS programme

Background: Some countries in sub-Saharan Africa combat the proliferation of substandard medicines using anti-counterfeiting devices. An alternative measure involves advanced pharmaceutical education. The Biotechnology Innovation and Regulatory Science (BIRS) educate students in pharmaceutical Good Manufacturing Practices (GMP) and quality control (QC). Objective: This study assessed the African BIRS programme’s efforts to initiate and sustain the manufacturing of quality medicines. Method: Qualitative case studies were used to assess the impact of the BIRS professional training intervention. A convenient purposeful sample of alumni students working in pharmaceutical manufacturing and quality control (QC) participated. Quantitative parametric methods were used to test the hypothesis that participants would significantly increase over the years if the programme met its objectives in the region. Results: Alumni from ten countries implemented and sustained projects in Good Manufacturing Practice (GMP) and QC. Participants from 2016 (17 males, 8 females) and 2018 (14 males, 8 females) increased in year 2020 (31 males, 22 females), p-values < 0.05, α = 0.05. Conclusion: Advanced education has improved efforts in maintaining quality healthcare in the region.

Integrating pressure sensor control into semi-solid extrusion 3D printing to optimize medicine manufacturing

Semi-solid extrusion (SSE) is a three-dimensional printing (3DP) process that involves the extrusion of a gel or paste-like material via a syringe-based printhead to create the desired object. In pharmaceuticals, SSE 3DP has already been used to manufacture formulations for human clinical studies. To further support its clinical adoption, the use of a pressure sensor may provide information on the printability of the feedstock material in situ and under the exact printing conditions for quality control purposes. This study aimed to integrate a pressure sensor in an SSE pharmaceutical 3D printer for both material characterization and as a process analytical technology (PAT) to monitor the printing process. In this study, three materials of different consistency were tested (soft vaseline, gel-like mass and paste-like mass) under 12 different conditions, by changing flow rate, temperature, or nozzle diameter. The use of a pressure sensor allowed, for the first time, the characterization of rheological properties of the inks, which exhibited temperature-dependent, plastic and viscoelastic behaviours. Controlling critical material attributes and 3D printing process parameters may allow a quality by design (QbD) approach to facilitate a high-fidelity 3D printing process critical for the future of personalized medicine.

Quadratic and linear radiation impact on 3D convective hybrid nanofluid flow in a suspension of different temperature of waters: Transpiration and Fourier Fluxes

Recently, the word Hybrid is mostly used due to its high impact on various systems like medicinal manufacturing, automotive experimental designs, electrical designs of where heat transport is vital and naval equipment designs etc. The underlying intension of this study is to oversee flow and heat transmission on the three- dimensional boundary layer flow owing to the stretching along x−direction and y−direction of an incompressible ternary hybrid nanofluid through a rectangular closed domain is considered. Nanoparticles magnetite (Fe3O4) in spherical shape, copper (Cu) in cylindrical shape and silver (Ag) in platelet shape were considered along with water as base fluid. The proposed system is numerically solved with shooting procedure (R-K 4th order). Outcomes obtained for velocity along x-direction f′(ζ), velocity along y-direction g(ζ), and temperature distribution θ(ζ) are elucidated via the construction of graph. Comparative studies of obtained results with previously published results are discoursed with different compositions (Composition-I, Composition-II, and Composition-III). The Compsition-1 of mixture as magnetite (Fe3O4) in spherical shape, copper (Cu) in cylindrical shape and silver (Ag) in platelet shape with base water at 00c, Composition-II mixture as magnetite (Fe3O4) in spherical shape, copper (Cu) in cylindrical shape and silver (Ag) in platelet shape with base water at 100c and Composition-III mixture as magnetite (Fe3O4) in spherical shape, copper (Cu) in cylindrical shape and silver (Ag) in platelet shape with base water at 500c.The main outcome of this study is found to be - The important note is the friction factor is higher in normal water compared to water at 100c, water at500c.the heat transfer rate is more in quadratic thermal radiation case compared to linear radiation case. The significant note is intermediate temperature water at 100c is more equated to other two cases water at 500c and normal water.

Исследование генотоксичности и окислительного стресса лекарственных растений

Biologically active substances (BAS) contained in medicinal plants are used in the manufacture of medicines, and, accordingly, have certain properties, such as, for example, antioxidant, anti-inflammatory effect, bacteriostatic, bactericidal, the ability to increase the body’s resistance to genotoxicants, etc. Infusions of medicinal plants Oregano (Origanum vulgare) and Greater celandine (Chelidonium majus), selected in their places of growth: in the mountains of Nozhai-Yurtovsky (1090 m above sea level) and Shatoisky (1200 m above sea level) areas of the Chechen Republic, traditionally used for medicinal purposes, were studied for the induction of oxidative stress on strains of luminescent bacteria E. coli. Artificially created Escherichia coli MG1655 biosensor strains containing specially designed plasmids of the pBR322 variant carrying the Photorhabdus luminescens bacterium luxCDABE operon placed under an inducible promoter activated only when certain chemical agents appear in the medium were used in the work. E. coli strains with hybrid plasmids: pSoxS-lux or pKatG-lux were used for detection of substances causing oxidative stress in cells, and with hybrid plasmids: pColD-lux or pRecA-lux for detection of genotoxic substances. It was found that the studied medicinal infusions can have a synergistic effect in combination with the genotoxic drug dioxidine and the oxidant hydrogen peroxide. At the same time, they can act both as an antioxidant and as an antigenotoxicant, depending on the concentration. The concentrations of infusions of Origanum vulgare and Chelidonium majus at the concentration of Greater Celandine – 6 g (CN2) and 3 g (CN3) caused oxidative stress. All other concentrations of Origanum vulgaris and Celandine greater on all bioluminescent strains of E. coli had a slight bactericidal effect. Further studies of biologically active substances of medicinal plants will make it possible to propose them as a protective barrier to the genotoxic, mutagenic or toxic effects of various environmental pollutants

Technical-legal aspects of the normative consolidation of the signs of the objective side of the crime provided for in Article 238.1 of the Criminal Code of the Russian Federation

The article analyzes the peculiarities of the regulation of criminal liability for the circulation of falsified, substandard and unregistered medicines, medical devices and the turnover of falsified biologically active additives (Article 238.1 of the Criminal Code of the Russian Federation). First of all, the author highlights violations of the general rules of legislative technique, which provide for compliance with the requirements of saving regulatory material, the use of similar terms in the construction of homogeneous compositions of crimes, etc. The author notes that it is necessary to normalize the level of punishability of single-order elements of crimes enshrined in Articles 235.1 and 238.1 of the Criminal Code of the Russian Federation by simultaneously indicating the large size of the acts committed in these criminal law prohibitions. The article notes the need for additional criminological analysis of other possible illegal actions with falsified, substandard or unregistered medicines, medical products and biologically active additives, such as - purchase, storage, transportation. The author also offers various options for interpreting the signs of the objective side, such as production and marketing, examines the features and controversial aspects of the qualification of an unfinished crime, explores the issues of excluding criminal liability in the manufacture of medicines and medical devices in the process of conducting clinical trials and research, and carrying out export operations.

Big data analytics and artificial intelligence technologies based collaborative platform empowering absorptive capacity in health care supply chain: An empirical study

The healthcare supply chain involves the manufacturing and delivery of medicines at the right time, at the right place, and in the correct quantity. In the world of uncertainties, especially deadly pandemics, the digitalization of the healthcare supply chain has emerged as one of the urgent phenomena to implement, for which organizations are focussing on the omnichannel healthcare approach. This paper explores (a) the antecedents of big data analytics and artificial intelligence (BDA-AI) technology-based collaborative platform for empowering absorptive capacity in omnichannel health care processes; (b) the effect of BDA-AI collaborative platform powered absorptive capacity in omnichannel health care processes and organization performance. The data is collected using a structured questionnaire from healthcare supply chain executives working in South Africa. The findings indicate that the involvement of managerial factors will improve the capacity of health care organizations to develop a BDA-AI technology-driven collaborative platform to assimilate, transfer and exploit critical information from large data sets. It will capacitate healthcare supply chains to deliver innovative performance to healthcare businesses. This work is the first of its kind to examine big data-based knowledge gained in the context of the omnichannel supply chain.

Microscopic analysis of Trichosanthes dioica

One of the most nutrient-dense cucurbit vegetables is the pointed gourd (Trichosanthes dioica Roxb.), which enjoys a prized spot in the Indian market during the warm and wet months. It is a perennial crop that is quite popular because it is available for eight months out of the year (February–September). It has certain medical effects due to its high protein and vitamin A content, and there are numerous studies about its involvement in decreasing blood sugar levels. In this paper, an effort was made to examine the microscopic characteristics of Trichosanthes dioica. Leaf exhibited uniceriate glandular epidermis with a single cellular head. Spongy parenchymatous cells, collenchymatous cells, pericycle, phloem, and xylem were all present after the epidermis. The Xylem vessels were spiral and reticulate. The collenchymatous cells' inside. The existence of uniseriate multicellular trichomes was shown by powder microscopy. crystal sheath oil globules, anomocytic stomata, liginified spiral arteries, and epidermis cells. The powders were subjected to various reagent treatments during the powder analysis, and various colours were visible to the naked eye and under UV light. There was quantitative microscopy. Vein termination and vein islet averages were 13.6 and 12, respectively. The upper and lower epidermis of the leaf had average stomatal counts of 12.5 and 16.33, respectively. The upper and lower epidermis of the leaf had average stomatal indices of 22.87 and 24.62, respectively. Data from these studies were helpful for evaluating the raw materials used in the manufacturing of herbal medicines and for anatomical research.

The Future of Pharmaceuticals: AI Drug Discovery & COVID-19 Treatments

AI Drug Discovery (AIDD) is the implementation of artificial intelligence and computational modeling in the process of finding the ideal medicinal drug for a given ailment; its primary benefit is exponentially speeding up the drug discovery process to about 1 to 2 years, whereas the traditional steps of lead optimization, clinical trials, regulatory approval, and medicinal manufacturing would span approximately 10 years. The COVID-19 pandemic could have been far worse and for a far longer duration if AIDD was not utilized, demonstrating great technological advancement in the field of medicine from the Spanish Flu pandemic in 1918. The benefits of AIDD in areas outside of COVID-19 can be seen in how Wisecube AI developed treatments for Alzheimer’s disease, Schrodinger AI was able to suppress tumors through protein analysis, and the drug discovery database of the scientific community is expanded through AIDD algorithms produced by companies like Standigm and Cytoreason. The idea of repurposing, the use of preexisting drugs to be slightly altered to cover completely different ailments, is also enhanced. In terms of COVID-19 treatments, however, AIDD is used to attack the virus through all forms of vaccinology, including protein-based, RNA-based, and mRNA-based vaccines. The Pfizer and Moderna vaccines are recommended the most by the CDC because of their effective innovation with AIDD in mRNA vaccinology. Simply, AIDD accelerates the R&D timeline of finding drug treatments for the prominent diseases that concern global intervention, and the world nears medicinal mastery with the utilization of computers.

Reimagining drug manufacturing paradigm in today’s pharmacy landscape

The coronavirus disease 2019 pandemic has escalated the ongoing problem of critical medication shortages, which has serious implications for the health of our patients. Currently, active pharmaceutical ingredients (APIs) are synthesized in large-scale batch operations and shipped to drug product manufacturers, where they are produced on a large scale at centralized facilities. In the centralized drug manufacturing process, the formulation components, operations, and packaging are structured to favor long-term storage and shipment of resultant medicines to the point of care, making this process vulnerable to supply chain disruptions. We propose a rethinking of the drug manufacturing paradigm with an upgraded pharmaceutical compounding-based manufacturing paradigm. This paradigm will be based on integration of continuous manufacturing of APIs and manufacturing of medicines at the point of care with application of machine learning, artificial intelligence, and 3-dimensional printing. This paradigm will support implementation of precision medicine and customization according to patients’ needs. The new model of drug manufacturing will be less dependent on the supply chain while ensuring availability of medicines in a cost-effective manner.

The Environmental Conservation, Legal and Ethical Issues concerning Herbal Products in Nigeria

It has been widely reported that there have been rising cases of communicable and non-communicable disease, given the poor level of unhealthy lifestyle and disease outbreak from poor scientific laboratory management. It must be noted that Nigeria has also had its fair share of the rising cases of most infectious diseases. However, in quest of most persons affected by various communicable diseases in Nigeria, it has led finding a better solace and cures to these communicable diseases, by resulting to taking phytomedicine or herbal product. It suffices to state that the growing demand for herbal medicine in Nigeria in the cure or treatment of communicable diseases results from its natural, medicinal, and therapeutic effects. However, the increasing demand for the herbal product has resulted in indiscriminate plant harvest and various individuals not being skilled in the art of herbal medicine production to indulge in manufacturing a poor, harmful and low-quality herbal product. It must be noted that although the international community set out ethical guidelines concerning the the conservation of plant material and manufacturing of herbal medicine. It is in this regard that this study tends to examine the international legal framework concerning environmental conservation of plant material and, local legal and ethical framework concerning the production of herbal products in Nigeria.

Development of combi-pills using the coupling of semi-solid syringe extrusion 3D printing with fused deposition modelling

Three-dimensional (3D) printing allows for the design and printing of more complex designs than traditional manufacturing processes. For the manufacture of personalised medicines, such an advantage could enable the production of personalised drug products on demand. In this study, two types of extrusion-based 3D printing techniques, semi-solid syringe extrusion 3D printing and fused deposition modelling, were used to fabricate a combi-layer construct (combi-pill). Two model drugs, tranexamic acid (water soluble, rapid release) and indomethacin (poorly water-soluble, extended release), were printed with different geometries and materials compositions. Fourier transform infrared spectroscopy results showed that there were no interactions detected between drug-drug and drug-polymers. The printed combi-pills demonstrated excellent abrasion resisting properties in friability tests. The use of different functional excipients demonstrated significant impact on in vitro drug release of the model drugs incorporated in two 3D printed layers. Tranexamic acid and indomethacin were successfully 3D printed as a combi-pill with immediate-release and sustained-release profiles, respectively, to target quick anti-bleeding and prolonged anti-inflammation functions. For the first time, this paper systematically demonstrates the feasibility of coupling syringe-based extrusion 3D printing and fused deposition modelling as an innovative platform for various drug therapy productions, facilitating a new era of personalised combi-pills development.

COUNTERFEIT AND FALSIFIED MEDICINES: TODAY’S CHALLENGES AND SMART SOLUTIONS

In any democratic country, special attention is paid to health protection, as a result of which, through organisational, financial, economic, and legal institutions, the state ensures and maintains the health protection of the population at the sufficient level. The purpose of the study is to examine the existing problems in the field of providing the population of Ukraine with high-quality medical drugs. The research methodology is a set of cognitive tools and methods, the use of which allowed for obtaining the most objective information about the existing problem processes and phenomena in the field of providing the population of Ukraine with high-quality medicines. Considering the critical state of the health care sector of the population of Ukraine, the study analyses legal regulations that focus on problematic issues in the health care sector of Ukraine. Attention is drawn to the need to provide the population of Ukraine with affordable and high-quality medicines that must meet the best European and international standards. Statistical data on the facts of the detection of counterfeit medicines are presented, and actors on this issue are analysed. It is noted that counterfeit and falsified medicines cause irreparable harm to the health of people all over the world, such illegal actions threaten the economy and security of the state, cause economic and financial damage to manufacturers of medical medicines, undermine their image, and harm the owners of intellectual property rights. It is noted that public relations in the field under study concern legal mechanisms related to unfair competition, which negatively affects the image of the state, and the activities of both Ukrainian and foreign business entities. It is noted that since intellectual property in its nature and content is closely linked to the growth of the economy, there is a need for more effective steps by the state to prevent violations of intellectual property rights. The results of the study, and parts of it, can be useful for both lawyers and government agencies that struggle with unfair competition in the field of medicines.

In-line spectroscopy combined with multivariate analysis methods for endpoint determination in column chromatographic adsorption processes for herbal medicine

Objective: In a chromatographic cycle, the adsorption process is a critical unit operation that has a significant impact on downstream processes and, ultimately, the quality of the final products. The development of a rapid method to determine the endpoints of adsorption processes in a large-scale manufacturing is of substantial importance for herbal medicine (HM) manufacturers. Methods: In this study, the adsorption of saponins on a macroporous resin column chromatograph, a critical unit operation in Panax notoginseng (Burkill) F.H.Chen injection manufacturing, was considered as an example. The evaluation results of in-line ultraviolet and visible spectra combined with various multivariate analysis methods, including the moving block standard deviation (MBSD), difference between the moving block average and the target spectrum (DMBA-TS), soft-independent modeling of class analogy (SIMCA), and partial least-squares discriminant analysis (PLS-DA), were compared. Results: MBSD was unsuitable for adsorption processes. The relative standard errors of prediction between the predicted and experimental endpoints were 13.2%, 4.67%, and 5.71% using DMBA-TS, SIMCA, and PLS-DA, respectively. Conclusions: Among the considered analysis methods, SIMCA and PLS-DA were more effective for endpoint determination. The results of this study provide a more comprehensive overview of the effectiveness of various multivariate analysis methods to facilitate the selection of the most suitable method. This study was also conducive to address the issues of the in-line detection of adsorption endpoints to guide practical HM manufacturing. Graphical abstract: http://links.lww.com/AHM/A23

Novel CRISPR/Cas9-mediated knockout of LIG4 increases efficiency of site-specific integration in Chinese hamster ovary cell line.

To investigate the impact of deficiency of LIG4 gene on site-specific integration in CHO cells. CHO cells are considered the most valuable mammalian cells in the manufacture of biological medicines, and genetic engineering of CHO cells can improve product yield and stability. The traditional method of inserting foreign genes by random integration (RI) requires multiple rounds of screening and selection, which may lead to location effects and gene silencing, making it difficult to obtain stable, high-yielding cell lines. Although site-specific integration (SSI) techniques may overcome the challenges with RI, its feasibility is limited by the very low efficiency of the technique. Recently, SSI efficiency has been enhanced in other mammalian cell types by inhibiting DNA ligase IV (Lig4) activity, which is indispensable in DNA double-strand break repair by NHEJ. However, this approach has not been evaluated in CHO cells. In this study, the LIG4 gene was knocked out of CHO cells using CRISPR/Cas9-mediated genome editing. Efficiency of gene targeting in LIG4-/--CHO cell lines was estimated by a green fluorescence protein promoterless reporter system. Notably, the RI efficiency, most likely mediated by NHEJ in CHO, was inhibited by LIG4 knockout, whereas SSI efficiency strongly increased 9.2-fold under the precise control of the promoter in the ROSA26 site in LIG4-/--CHO cells. Moreover, deletion of LIG4 had no obvious side effects on CHO cell proliferation. Deficiency of LIG4 represents a feasible strategy to improve SSI efficiency and suggests it can be applied to develop and engineer CHO cell lines in the future.