Pharmaceutical Industry Research Articles

Evaluating the biomedical potential of phytomediated silver doped zinc oxide nanoparticles derived from n-butanol fraction of Adiantum venustum D. Don

Cancer and bacterial antibiotic resistance are the first two pressing issues confronting the pharmaceutical industry today. With this in mind, the current goal of this research is to create eco-friendly, green synthesized nanostructures that have the potential to at least partially eliminate these issues. The research aims to synthesize ZnO (zinc oxide) and Ag-doped ZnO (silver doped zinc oxide) nanoparticles with two different concentrations (AgxZn1-xO) and variations in their biological potential. To the best of our knowledge, the n-butanol fraction of A. venustum is being used as a first attempt to synthesize Ag-doped ZnO nanoparticles that too with varying concentrations. The XRD (X-ray diffraction) technique endorses the materialization of wurtzite structure for zinc oxide (ZnO–B) and hexagonal wurtzite configuration for the silver-doped complexes (AgZnO–B1 and AgZnO–B2). The Scherrer formula was utilized to examine the average crystallite sizes of ZnO–B (29.21 nm), AgZnO–B1 (22.03 nm), and AgZnO–B2 (27.76 nm). Using transmission electron microscopy micrographs, the grain sizes of AgZnO–B1 (57.72 ± 1.84 nm), AgZnO–B2 (75.31 ± 2.03 nm), and ZnO–B (45.84 ± 0.57 nm) were measured. The XPS (X-ray Photoelectron Spectroscopy) revealed the Zn2+ and Ag + oxidation state of green synthesized nanoparticles. Additionally, the antioxidant, anti-inflammatory, antimicrobial and anticancer activities were assessed in relation to the n-butanol fraction of A. venustum and the synthesized nanomaterials. Of all the synthesized nanoparticles, AgZnO–B1 has proven to be a potent biomedical agent in all biological activities. AgZnO–B1 was found to have potential as an anti-inflammatory (IC50- EAA 33.63 ± 0.05 μg/mL; BSA 28.02 ± 0.80 μg/mL) and anticancer (IC50 85.27 μg/mL) agent in the study. Therefore, it should be underlined that nanoparticles have enormous biological efficacy and used in a range of therapeutic contexts.

How do corporate patenting strategies adapt to public health crises?

ABSTRACT By exploiting the health emergencies in China, we investigate the strategic reactions concerning patenting strategies to public health crises. Results show that firms maintained innovation input levels during the Severe Acute Respiratory Syndrome (SARS) period. Moreover, firms relied more on past technological paths and became technologically more specialized, as responses to SARS. In addition, we find that the main effects are weaker among pharmaceutical companies and that there is a positive treatment effect on invention patent applications and grants. Our findings remain valid after accounting for potential endogeneity problems with a variety of robustness tests.

Effect of cephalexin on chlorophyll and carotenoid content of Chlorella pyrenoidosa-2378 and its biodegradation in BG-11 medium

Cephalexin (CFX), a β-lactam antibiotic which is identified for the treatment of different disease infections, has been reported as a micropollutant in wastewater released from sewage, hospital, or pharmaceutical industries. Chlorella pyrenoidosa-2378, a green microalgal strain, is recognized for its degradation ability against wastewater pollutants and the potential of the biomass it produces. In this present study, the cultivation process of Chlorella pyrenoidosa-2378 strain with CFX concentration of 50 mg/L, 100 mg/L, 150 mg/L, and 200 mg/L added to its culture medium labeled as test I, test II, test III, and test IV, respectively, is the process being observed for evaluation of its degradation ability and as sustainable approach for antibiotic degradation. chlorophyll a in test IV, chlorophyll b in test III, and carotenoid content in test I were observed to be higher in amount than control by 0.775 mg/L, .069 mg/L, and 0.356 mg/L, respectively. Using the HPLC method, the total removal efficiency was observed to be 79.09%, 86.26%, 89.62%, and 88.03% against 50, 100, 150, and 200 mg/L concentrations of CFX, respectively. The observations that C. pyrenoidosa-2378 remained stable while being used as an alternative bioremediation method, provide an alibi for its novelty and potentiality to be used commercially for biomass production and at industrial scale for degradation.Graphical

Strategies for producing probiotic biomass and postbiotics from Akkermansia muciniphila in submerged cultivations incorporating prebiotic sources.

This research propounds an innovative technology focused on sustainability to increase the biomass yield of Akkermansia muciniphila, the next-generation probiotic, using prebiotic sources to replace or reduce animal mucin levels. A series of experimental design approaches were developed aiming to optimize the growth of Akkermansiamuciniphila by incorporating extracts of green leafy vegetables and edible mushroom into the cultivation media. Experiments using kale extract (KE), Brassica oleracea L., associated with lyophilized mushroom extract (LME) of Pleurotus ostreatus were the most promising, highlighting the assays with 0.376% KE and 0.423% LME or 1.05% KE and 0.5% LME, in which 3.5 × 1010 CFU (Colony Forming Units) mL- 1 was achieved - higher than in experiments in optimized synthetic media. Such results enhance the potential of using KE and LME not only as mucin substitutes, but also as a source to increase Akkermansia muciniphila biomass yields and release short-chain fatty acids. The work is relevant to the food and pharmaceutical industries in the preparation of the probiotic ingredient.

Determining Fermentation Conditions to Enhance Antioxidant Properties and Nutritional Value of Basil Seeds Using Lactobacillus plantarum

Fermented seeds and their bioactive compounds have captivated global interest due to their functional properties. Basil seeds are used worldwide in the food, cosmetic, and pharmaceutical industries, but their functional and nutritional properties after fermentation are not known. The aim of this study was to optimize the effect of fermentation on the improvement in the functional properties of basil seeds by Lactobacillus plantarum. Basil seed samples were categorized into seven water volumes (named A–G) and fermented for 24, 48, 72, and 96 h with L. Plantarum. The results show that the pH and total microbial content (TMC) significantly changed after 24 h of fermentation (p < 0.05). Fermentation significantly increased the antioxidant properties and niacin content of basil seeds compared with non-fermented control samples (p < 0.05). Fermented samples G-24, G-48, and G-72 (samples with a ratio of 1 g (basil):10 mL (water) fermented for 24,48 and 72 h), exhibited the highest DPPH and ABTS• scavenging activity. The total polyphenol content (TPC) was most prominent in the samples G-72, G-24, and G-48, respectively. Sample G-48 showed the highest antioxidant activity. Notably, sample G-24 showed a significant increase in niacin content (64 µg/mL). These results underscore that varying moisture levels and fermentation durations have a significant impact on the nutritional/functional value of basil seeds. Overall, fermenting basil seeds with L. plantarum increased their functional properties with greater antioxidant and TPC activities as well as increased nutritional value.

Production of Cellulolytic Enzymes by Paecilomyces TP08 Using Grape Stalk and Wheat Bran as Substrates in Solid State Culture

Objective: The objective of this study is to investigate the use of grape stems and wheat bran as substrates in solid state fermentation, for the production of cellulolytic enzymes using the fungus Paecilomyces TP08 sp. Theoretical Framework: In grape production, driven by increased consumption and exports, by-products represent 30% of the total volume of grapes processed for wine, requiring efficient management to ensure sustainability. Method: The methodology adopted for this research involves solid-state cultivation, with a substrate made up of 50% grape stems and 50% wheat bran, with samples taken every 24 hours. Each sample taken went through a filtration process, followed by enzyme activity analysis using the DNS method. Results and Discussion: The results obtained showed that the cultivation with the association of wheat bran and grape stalks presented good total cellulase activity during the process, value of 0.846 U/g in 144h, followed by the activity of 0.663U/g observed in 24h. Research Implications: The practical and theoretical implications of this research are discussed, providing insights into how the results can be applied or influence practices in the field of industrial biotechnology. These implications could cover the food, textile and pharmaceutical industries. Originality/Value: This study contributes to the literature by associating grape stalks with wheat bran for the production of fungal enzymes. The relevance and value of this research are evidenced by the use of an innovative substrate (grape stalks) associated with a conventional substrate (wheat bran) for induction of the microorganism little used in the literature for enzyme production.

Phase Transfer Catalysis: New Technology to Boost Organic Farming

Introduction of Phase Transfer Catalysis (PTC) technology, which was developed in the 1970s has revolutionised the dyes and pharmaceutical industries in many ways. PTC is a process that facilitates the organic compounds to move from one phase to another phase, without any change in the chemical qualities. Application of this technology was explored in the 1980s for agriculture, by using different plant extracts as PT catalysts. The objective was to facilitate increased absorption of nutrients by plants, by stimulating plant growth and easy availability of nutrients in the soil. Introduction of nano-carbon materials with nutrients and herbal growth stimulants could further support the crop growth by facilitating increased uptake of nutrients, apart from enabling the plants to develop resilience to harsh environmental conditions, resulting in higher crop yields. Some organic bio-stimulants developed recently by Snehasrusthi Group in India have been very successful in improving the growth, yield and quality of several seasonal and perennial crops in India. These bio-stimulants have excellent potential to boost organic agricultural production and income of farmers in the future.

Drug Shortages and Patient Harms

What Is the Issue? Drug shortages are a global issue with complex dynamics. Shortages can occur because of disruption at any point along the drug supply chain. Several strategies are used in Canada to prevent or alleviate the effects of drug shortages, including mandatory reporting by drug manufacturers. An understanding of the amount and types of real or potential harms caused to patients can inform policy decisions around drug shortage management and prevention. What Did We Do? We searched for literature providing evidence on patient outcomes associated with supply chain disruptions of pharmaceuticals and vaccines. An information specialist conducted a search of peer-reviewed literature sources published between January 1, 2003, and September 13, 2023. Documents were excluded if the objective was to investigate the potential effects of a drug shortage in the absence of an actual drug shortage or if the outcomes were not direct patient harms. What Did We Find? One scoping review and 33 nonrandomized studies were identified that evaluated patient outcomes associated with supply chain disruptions of pharmaceuticals and vaccines. We identified a wide variety of drug classes experiencing shortages. The most frequently reported shortages were anesthetics, oncology drugs, vaccines, drugs for the treatment of COVID-19, antimicrobials, and small-volume parenteral solutions. Most of the included primary studies concluded that the replacement drug or protocol was a safe or acceptable alternative to the shortage drug. The subset of primary studies that concluded that the replacement drug or protocol was not a safe or acceptable alternative to the shortage drug reported worse outcomes in health system use (including length of hospital stay), adverse events, disease progression, and mortality. What Does This Mean? Drug shortages have the potential to cause harm to patients and some drug shortages may have a greater impact on patients than others. The ability to predict which drugs could cause the greatest harm during a supply disruption would be a great benefit for future planning. The diversity of drugs experiencing shortages and their associated harms emphasizes that decision-makers may need to take a case-by-case approach when developing policies meant to lessen the impact of drug shortages.

Using Artificial Intelligence to Advance the Research and Development of Orphan Drugs

While artificial intelligence has successful and innovative applications in common medicine, could its application facilitate research on rare diseases? This study explores the application of artificial intelligence (AI) in orphan drug research, focusing on how AI can address three major barriers: high financial risk, development complexity, and low trialability. This paper begins with an overview of orphan drug development and AI applications, defining key concepts and providing a background on the regulatory framework of and AI’s role in medical research. Next, it examines how AI can lower financial risks by streamlining drug discovery and development processes, analyzing complex data, and predicting outcomes to improve our understanding of rare diseases. This study then explores how AI can enhance clinical trials through simulations and virtual trials, compensating for the limited patient populations available for rare disease research. Finally, it discusses the broader implications of integrating AI in orphan drug development, emphasizing the potential for AI to accelerate drug discovery and improve treatment success rates, and highlights the need for ongoing innovation and regulatory support to maximize the benefits of AI-driven research in healthcare. Based on those results, we discuss the implications for traditional and AI-powered business in the drug industry.

Characterization of Lomofungin Gene Cluster Enables the Biosynthesis of Related Phenazine Derivatives.

Phenazine-based small molecules are nitrogen-containing heterocyclic compounds with diverse bioactivities and electron transfer properties that exhibit promising applications in pharmaceutical and electrochemical industries. However, the biosynthetic mechanism of highly substituted natural phenazines remains poorly understood. In this study, we report the direct cloning and heterologous expression of the lomofungin biosynthetic gene cluster (BGC) from Streptomyces lomondensis S015. Reconstruction and overexpression of the BGCs in Streptomyces coelicolor M1152 resulted in eight phenazine derivatives including two novel hybrid phenazine metabolites, and the biosynthetic pathway of lomofungin was proposed. Furthermore, gene deletion suggested that NAD(P)H-dependent oxidoreductase gene lomo14 is a nonessential gene in the biosynthesis of lomofungin. Cytotoxicity evaluation of the isolated phenazines and lomofungin was performed. Specifically, lomofungin shows substantial inhibition against two human cancer cells, HCT116 and 5637. These results provide insights into the biosynthetic mechanism of lomofungin, which will be useful for the directed biosynthesis of natural phenazine derivatives.

Environmental analysis of biotechnologies for biofuels, bioplastics, and bioproducts: a greenhouse gas (GHG) emissions review

Biotechnology and biomanufacturing development has the potential to strengthen the bioeconomy as new opportunities in many areas such as the energy, chemical, agriculture, pharmaceutical, and food industries can be unlocked. Biotechnology and biomanufacturing refer to the technologies that use microorganisms, molecular biology, metabolic engineering, and chemical processing to transform biobased resources (e.g., biomass) into new products. These microorganisms are genetically engineered in such a way that the production of new products happens more efficiently. Creating new products through biotechnology and biomanufacturing will promote shifting from a fossil-based economy to a bioeconomy. However, these new technologies will need to be evaluated from the accessibility, affordability, and sustainability point of view. In this paper, a review of recent studies evaluating the carbon footprint of biotechnologies to produce fuel, bioplastics, and bioproducts is presented. The assumptions, biogenic and coproduct credit subtraction, and co-product treatment methods in the life-cycle assessment (LCA) showed an important impact on the results of the different studies. Besides, integrated biorefineries presented an alternative to improve the environmental impact of bioproducts compared to single-product refineries.

A Comprehensive Review of the Ethnobotanical Uses, Pharmacological, and Toxicological Profiles of Piper capense L.f. (Piperaceae)

Commonly known as wild pepper, Piper capense (P. capense) is a culinary herb mainly used as a secret in preparation of “Nkui” and “Nah poh” in Bayangam, West Cameroon. However, it also has many interesting pharmacological properties, which is why the people of sub-Saharan Africa so highly prize it for the treatment of multiple human pathologies. This study aimed to highlight the traditional uses, phytochemical composition, biological activities, and toxicological profile of the P. capense plant, to draw the attention of pharmaceutical companies to its enormous potential for the development of future phyto- or pharmaceutical products. Documentary research was meticulously carried out in the Web of Sciences, Scopus, Pubmed/Medline, and Google Scholar databases according to PRISMA 2020 guidelines. The results show that extracts and compounds isolated from Piper capense have interesting anticancer, antibacterial, antimalarial, hypoglycemic, anti-epileptic, and antidepressant activities. Methanolic extracts and essential oils from P. capense exhibit no harmful effects when directly applied to normal human hepatocytes, umbilical cord cells, intestinal cells, and keratinocyte cell lines. Additionally, methanolic extracts administered acutely or subchronically at low doses (≤250 mg/kg body weight) in Wistar rats also demonstrate no adverse effects. In conclusion, given its interesting activities, P. capense is a viable option for developing new antimalarial, anticancer, antibacterial, hypoglycemic, anti-epileptic, and antidepressant drugs. However, many avenues still need to be explored before translation into drugs.

The AMPK/cAMP Metabolic Signaling Axis as a Possible Therapeutic Target for Diabetes.

Diabetes is a complex disease, despite the availability of numerous treatments, its progression and complications can only be mitigated and managed to a certain extent. After the onset, diabetes cannot be reversed. Its global expansion makes it challenging for governments to control the considerable costs of treating people with diabetes. Many studies have been carried out by widely recognized pharmaceutical companies that are considering the development of new drugs for diabetic treatments. Diets, sedentary habits, and lifestyles that are currently prevalent have an enormous influence on the global spread of diabetes. The tools available to clinicians for therapy do not solve the problem. It is known that a patient, when diagnosed, would already have had diabetes for more than three years. Studies on diabetes signaling consider the effects of hyperglycemia but also highlight the roles of insulin receptor activation and resistance. Understanding the intricate signaling network and its interactions with hyperglycemiainduced pathways is crucial. In this context, the cyclic AMP/AMPK axis emerges as a promising therapeutic target for diabetes. However, there is a noticeable lack of literature exploring the metabolic network induced by hyperglycemia and its interconnected pathways. Therefore, investigating the cyclic cAMP/AMPK axis could provide valuable insights, given its complex connections with various metabolic pathways. This mini-review aims to delve into the metabolic signaling of the AMPK/cAMP axis in the context of diabetes, highlighting its metabolic interactions and potential implications.

AI Powered Bioinformatics - Expediting Diagnostic Testing: A Survey

Research has demonstrated the positive impact of artificial intelligence and Bioinformatics in the field of clinical diagnosis. The integration of AI methodologies into bioinformatics has opened new avenues for breakthroughs in genomics, proteomics, and personalized medicine. The document emphasizes the role of AI in early disease detection, improving patient outcomes, and enhancing healthcare systems by avoiding the need for expensive and time-consuming operations as illnesses worsen. The methodology section provides insights into the approach utilized, including the review of 30 articles from highly regarded journals about AI and bioinformatics that expedite diagnostic testing in the medical field. using survey to gather information and divide it into sub-sections focusing on diagnostic cancer diseases, COVID-19, and genetic and chronic diseases. The survey gathered 52 responses, and the results revealed significant agreements with the findings in the papers, particularly in the importance of developing novel biosensors and diagnostic tools for rapid and accessible detection of SARS-CoV-2, and the potential of AI in laboratory settings, pharmaceutical industry, and disease diagnosis. Overall, the document provides a comprehensive overview of the transformative role of AI in bioinformatics, emphasizing its potential to revolutionize disease diagnosis, treatment, and public health decision-making, while also addressing the challenges and opportunities associated with the integration of AI technologies in the healthcare industry. The rigorous methodology and alignment of survey results with the research findings validate the significance of AI-powered bioinformatics in expediting diagnostic testing and improving patient safety in healthcare.

Regulatory Compliance and Risk Management in Pharmaceuticals and Healthcare

Purpose: This study aims to examine the evolving regulatory landscape and its impact on compliance and risk management within the pharmaceutical and healthcare industries. It seeks to identify effective strategies for maintaining data integrity and traceability in accelerated pharmaceutical development (APD), and to understand how these strategies improve patient care and healthcare system responsiveness. Methodology: The research involves a comprehensive review of current regulations, emerging trends, and industry practices. It utilizes an analysis of regulatory updates, risk management frameworks, and case studies to illustrate successful compliance strategies and the challenges faced by organizations. Findings: The study highlights key regulatory requirements, identifies common risks and mitigation strategies, and provides insights into effective practices for ensuring data integrity and traceability. It demonstrates that robust compliance and risk management practices contribute to enhanced patient outcomes and improved healthcare system efficiency. Unique Contribution to Theory, Policy, and Practice: This research offers a detailed examination of the intersection between regulatory compliance and risk management, providing a valuable framework for understanding and addressing the complexities of the pharmaceutical and healthcare sectors. It contributes to theory by integrating recent regulatory changes with practical risk management approaches, informs policy by highlighting critical areas for regulatory focus, and offers actionable recommendations for industry practitioners to optimize compliance and data management processes.

Plasticization of a stiff pharmaceutical solid for better tabletability via cocrystallization: Shape synthons as supramolecular protecting groups

Extremely high particle stiffness and very low hardness is a serious concern in various mechanical processes in pharmaceutical manufacturing. Here we report an exceptionally high Young’s modulus (E) of ∼ 18 GPa in a drug, isoniazid (INH). This is one of the highest experimentally determined values among all reported pharmaceutical molecular crystals, which we attribute to the presence of a strong three-dimensional (3D) hydrogen bonding network (HBN). Further, we successfully reduced the 3D HBN in INH to 2D in its cocrystal using a co-former, 3,4-dimethylbenzoic acid (DMBA), where its two hydrophobic groups act like protecting groups at supramolecular level and prevent the extension of HBN. This reduced the E in the 1:1 cocrystal, INH-DMBA, by many folds and markedly improved its powder tabletability. To the best of our knowledge, this is the first reliable molecular level approach to alter the stiffness of pharmaceutical crystals and tabletability improvement in a predictable manner, hence, is important in the context of crystal engineering.

Entrapment of glucose oxidase from Aspergillus niger ISL-09 in poly(acrylamide-co-acrylic acid) hydrogels for improved stability and catalytic efficiency towards industrial applications

The present study highlights the true potential of Aspergillus niger ISL-09 to produce glucose oxidase (GOx) and focuses on improving the catalytic properties of GOx by entrapping it in poly acrylamide-co-acrylic acid (AAm-co-AAc) hydrogels, fabricated by free-radical polymerization method. For this purpose, the optimum conditions for GOx activity were found to be: 15 g soybean meal (SBM) substrate, 20 mL moisture content, 96 h incubation period, and 1.5 mL of fungal spore suspension. The purified extracellular GOx was observed to have 32.4% yield, with 0.02 U/mg specific activity and approximately 80 kDa subunit molecular weight. The GOx was most active at pH 5.5 and 40 °C. The immobilization of GOx in poly (AAm-co-AAc) hydrogels led towards improved stability and catalytic efficiency, resulting in a 21.7% increase in activity compared to free enzyme. The study also examined the potential of GOx in the pharmaceutical and textile industries as Ca-gluconate producer and bleaching agent, respectively. The study concludes that A. niger ISL-09 is a promising source for GOx production under optimal conditions. Furthermore, the immobilization of GOx in poly (AAm-co-AAc) hydrogels can significantly improve its catalytic properties, making it suitable for different industrial applications. However, further scaling up is required for the better implementation in industry.

The Impact Of Job Characteristics On Employee Turnover Intention Through Psychological Contract In The Pharmaceutical Industry Of Pakistan

Introduction: The purpose of this research work is to examine the effect of job characteristics on intended turnover in the pharmaceutical industry of Pakistan with the ‘Psychological contract’ as a mediator amidst specialized knowledge that is pertinent to the sector’s growth. The characteristics included in this paper are autonomy, task identity, and skill variety, whereby studies were done to determine the effects of these characteristics on employee turnover intentions. Thus, the psychological contract acts as a pivotal link to breach, thereby increasing dissatisfaction and emergence of turnover. Objective: The study aims at establishing correlation between job content and turnover intention, Leader Member Exchange (LMX) as the moderator and psychological contract as mediator. Therefore, this research aims at filling this knowledge gap in relation to strategies of employee retention within the pharmaceutical industry of Pakistan and the way through which workforce stability could be boosted. Methods: The quantitative method was adopted, and based on the PLS-SEM technique to assess the hypothesis regarding the interaction[1] of job characteristics, psychological contract, LMX and turnover intention. The data were collected form 328 full time employees of pharmaceutical industries established in Pakistan. Recording of overall fit indices also supported these results. Results: The results show that among the job characteristics, autonomy was statistically significant and positive with β = 0. 058 and p < 0. 05 with corresponding tests showing significance for the-job skill variety (β = 0. 184, p < 0. 01) and feedback (β = 0. 166 p < 0. 01) to positively influence the psychological contract, thus translating to reduced turnover intention. LMX mitigates this relationship with high-quality LMX being particularly beneficial in enhancing the effect of the psychological contract on retention; (β = 0. 329, t < 0. 001). Conclusion: Consequently, the study shown that job characteristics should be encouraged and leader member relations should be promoted in order to minimize turnover intention in the context of Pakistan’s pharmaceutical industry. These findings are relevant to the organizational leaders who wish to enhance the retention rates of their employees and increase productivity through the effective job design and leadership practices. This is an important step in the validation of the model and has implications for the discussion on workforce stability within growing high demand sectors for application across the industries.

Silkworm Gut Microflora as a Source: Biotechnological Applications

The term "microflora" has gained popularity as researchers explore the symbiotic relationships between microorganisms and their hosts, from our skin to our gut. Insects, including the silkworm, have mutualistic ties with their gut microflora, making them a valuable resource for studying essential molecules. The silkworm, with its rapid generation time, abundant genetic resources, well-defined genetic background, and numerous homologous genes, is an ideal model organism for various life science studies. Silkworm rearing is simple, inexpensive, and does not require ethical clearance, allowing the gut microflora of Bombyx mori to be harnessed for biotechnological applications. Silkworm gut microbes can synthesize and produce extracellular enzymes, such as cellulases, proteases, and lipases, which are crucial for industries ranging from biofuel production to waste management and textile processing. Additionally, these microbes can produce vitamins, such as vitamin B12, and metabolites like short-chain fatty acids (SCFAs), which have potential applications in the food and pharmaceutical industries. Antimicrobial substances produced by the gut microflora, including bacteriocins and lactic acid, are of particular therapeutic importance, offering natural alternatives to traditional antibiotics and contributing to the development of novel antimicrobial therapies. Moreover, the antioxidants generated by these symbionts, such as glutathione and superoxide dismutase (SOD), have significant implications for health supplements and cosmetics, owing to their ability to combat oxidative stress and support immune function. Beyond their metabolic capacities, silkworm symbionts can be used to enhance health, target and manage agricultural pests, and control vectors of human diseases in an environmentally friendly manner. Recent research has also highlighted the potential of mutualistic symbiotic bacteria for plastic disposal, with enzymes like PETases showing promise in degrading synthetic polymers. Despite extensive knowledge on the biology and physiology of the silkworm Bombyx mori, few studies have focused on its gut microflora. Modern genetic techniques, such as the genetic transformation of silkworms with genes of interest, may help overcome these research gaps. Increasing research efforts on silkworm gut microflora will not only reveal new symbiotic relationships but also identify new sources of biotechnologically important molecules and enzymes for therapeutic and industrial applications.

An Automated pre-Dilution Setup for Von Willebrand Factor Activity Assays.

Accurate quantification of von Willebrand factor ristocetin cofactor activity (VWF:RCo) is critical for the diagnosis and classification of von Willebrand disease, the most common hereditary and acquired bleeding disorder in humans. Moreover, it is important to accurately assess the function of von Willebrand factor (VWF) concentrates within the pharmaceutical industry to provide consistent and high-quality biopharmaceuticals. Although the performance of VWF:RCo assay has been improved by using coagulation analyzers, which are specialized devices for blood and blood plasma samples, scientists still report a high degree of intra- and inter-assay variation in clinical laboratories. Moreover, high, manual sample dilutions are required for VWF:RCo determination of VWF concentrates within the pharmaceutical industry, which are a major source for assay imprecision. For the first time, we present a precise and accurate method to determine VWF:RCo, where all critical pipetting and mixing steps are automated. A pre-dilution setup was established on CyBio FeliX (Analytik-Jena) liquid handling system, and an adapted VWF:RCo method on BCS-XP analyzer (Siemens) is used. The automated pre-dilution method was executed on three different, most frequently used coagulation analyzers and compared to manual pre-dilutions performed by an experienced operator. Comparative sample testing revealed a similar assay precision (coefficient of variation = 5.9% automated, 3.1% manual pre-dilution) and no significant differences between the automated approach and manual dilutions of an expert in this method. While no outliers were generated with the automated procedure, the manual pre-dilution resulted in an error rate of 8.3%. Overall, this operator-independent protocol enables standardization and offers an efficient way of fully automating VWF activity assays, while maintaining the precision and accuracy of an expert analyst. Key features • Automated pre-dilution setup for von Willebrand factor concentrates of various natures. • Combination of a liquid handling system (CyBio FeliX) with a coagulation analyzer (BCS-XP). • Simplifies method transfer to other laboratories. • Basic training for CyBio FeliX and BCS-XP is required. Graphical overview VWF:RCo assay principle and measurement setup. Platelets (yellow ellipsoids) with negative surface charge (- - -) are treated with formaldehyde, which partly denatures the cell surface and thus stabilizes platelets for use as assay reagents. Stabilized platelets (dark-yellow-framed yellow ellipsoids) are then brought in contact with ristocetin A (chemical structure shown; black dots), which binds to the platelet surface and facilitates binding of VWF (green circles). The graphs show an example of quantitative determination of platelet agglutination by measurement of light transmission, where increasing amounts of VWF increase light transmission over time. The photo in the left-bottom corner shows the CyBio FeliX setup for VWF sample dilution and the photo in the right-bottom corner displays the BCS-XP system, which is used for VWF:RCo measurements.