Antibody-drug conjugates (ADCs) present unique pharmacokinetic (PK) challenges due to analyte heterogeneity, dynamic drug-to-antibody ratio (DAR) distributions, target-mediated disposition, and systemic release of cytotoxic payloads. Unlike monoclonal antibodies or small molecules alone, ADCs require integrated modeling approaches capable of simultaneously describing total antibody, conjugated species, DAR-specific subpopulations, and free drug exposure. Over the past decade, pharmacometric strategies have evolved from empirical population PK models toward semi-mechanistic and system-based frameworks incorporating deconjugation kinetics, intracellular processing, and target-mediated drug disposition. In this review, we examine current PK modeling paradigms for ADCs, evaluate their strengths and limitations in clinical development, and discuss how quantitative frameworks inform dose selection, exposure-response relationships, and translational prediction. As ADC platforms diversify, rigorous mechanistic PK modeling is increasingly central to optimizing therapeutic index and regulatory decision-making.

Therapeutic oligonucleotides (ONs), including antisense-oligonucleotides, small interfering RNA, aptamers, and conjugated modalities, have emerged as an important class of drugs with increasing clinical impact. Unlike small molecules, ONs undergo metabolism primarily through nuclease-mediated cleavage, generating complex profiles of shortened metabolites that often differ by a single nucleotide and may retain pharmacological or toxicological relevance. Comprehensive metabolite identification is therefore essential for understanding ONs pharmacokinetics (PK), tissue exposure, and safety. Liquid chromatography coupled to mass spectrometry (LC-MS) has become the principal analytical platform for ONs metabolite identification. Recent advances in chromatographic separation, high-resolution mass spectrometry, fragmentation strategies, and data processing tools have substantially improved the depth, confidence, and throughput of metabolite characterization. This review provides an overview of ONs biotransformation pathways and critically examines modern LC-MS strategies used for metabolite separation, detection, and structural elucidation. Emphasis is placed on high-resolution MS acquisition approaches, charge-state management, complementary fragmentation techniques, and software-assisted metabolite annotation. Emerging trends and future directions in ONs metabolite analysis are also discussed, with a focus on supporting translational PK and regulatory decision making.

This study builds on our previous findings on the role of salivary lysophosphatidic acid (LPA) species in humans to investigate their presence, together with salivary gland LPA receptor (LPAR) expression in a Porphyromonas gingivalis- infected murine (C57BL/6J) model of periodontal disease (PD). Utilizing LC-MS/MS for LPA analysis alongside confocal LPAR imaging and second harmonic (SHG) imaging for collagen visualization, we compared mouse salivary LPA levels and gland LPAR expression to previously established human and mouse data. The findings reveal that while healthy mouse saliva maintains low homeostatic LPA levels, PD triggers an ~ 10-fold increase, mirroring the elevation we observed in PD patients. Furthermore, the study confirmed the presence of LPA1, LPA3, and LPA4 within submandibular gland (SMG) tissue. Notably, LPA3 was identified as the most widely distributed subtype, while providing the first evidence of LPA4 expression in adult mouse salivary glands. The presence of multiple LPARs suggests that LPA signaling is a critical factor in salivary gland biology. The documented existence of multiple LPARs within salivary glands indicates that they must be taken into consideration in future research concerning autoimmune conditions, and in pharmacological studies involving drugs that impact salivary gland biology and secretory function.

ABSTRACT Background Group 3 (MYC-driven) medulloblastoma (MB) is a highly aggressive brain tumor with poor-prognosis and limited treatment options. We previously identified protein-arginine methyltransferase-5 (PRMT5) as a promising target in Group 3 MB with its control on MYC protein stability. In this follow up study, we further mechanistically investigated PRMT5 control on MYC transcription and targeted it pharmacologically for therapeutic proof-of-concept. Methods Using pharmacogenetic inhibition approaches against PRMT5 in MYC-amplified (Group 3) MB cell line and neurosphere models in vitro and in vivo , we investigated molecular mechanism(s) and anti-cancer efficacy of PRMT5 inhibition. Results Our experiments demonstrated that PRMT5 epigenetically regulates MYC transcription in MYC-amplified MB cells by binding to the proximal-promoter region of the MYC gene and contributing to the enriched symmetric-dimethylation of histone H4R3 in the same region. We further showed that PRMT5 is recruited to the MYC promoter by its interaction with BRD4, the major BET-protein responsible for MYC transcription. PRMT5 inhibition caused the suppression of MYC-induced transcriptional programs and target genes, with widespread disruption of splicing across the transcriptome, particularly affecting metabolism-related gene products. Pharmacologic inhibition of PRMT5 using a panel of selective small-molecule inhibitors demonstrates suppression of cell growth/survival in a MYC-dependent manner in MB cells. Moreover, our in vivo analyses of PRMT5 inhibition, in mice treated with one of the potent pharmacologic inhibitors, particularly a lipid-decorated form of it, demonstrated reduced cerebellar tumor growth with suppressed MYC expression and prolonged survival of mice with MYC-amplified MB xenografts. Conclusions Our findings establish a functional link between PRMT5 and MYC-mediated transcriptional regulation, suggesting a promising therapeutic approach targeting the PRMT5-MYC axis for MYC-driven MB. Key Points PRMT5 acts as an epigenetic regulator of MYC transcription, RNA splicing and associated energy metabolism in MYC-driven MB. PRMT5 inhibition selectively suppresses cell growth/survival in MYC-driven MB. PRMT5 inhibition reduces tumor burden and prolongs survival in a MYC-driven MB mouse model. Importance of the Study Group 3 medulloblastoma is a highly aggressive pediatric brain tumor marked by MYC amplification, malignant clinical behavior, and poor survival outcomes despite intensive multimodal therapy. Because MYC remains largely undruggable, there is an urgent need for effective and less toxic treatment options for affected children. This study identifies protein arginine methyltransferase 5 (PRMT5) as a key epigenetic regulator of MYC transcription and MYC-dependent oncogenic programs in Group 3 MB. We show that PRMT5 is recruited to the MYC promoter via BRD4, sustains MYC-driven transcription and RNA splicing networks associated with metabolism, and supports MB tumor growth. Importantly, pharmacologic inhibition of PRMT5 using a selective brain-penetrant inhibitor suppresses MYC expression, reduces cerebellar tumor burden, and prolongs survival in MYC-amplified MB models. These findings provide a strong translational rationale for PRMT5 inhibition as a targeted therapeutic strategy for high-risk MB, with the potential to improve outcomes while reducing treatment-related toxicity.

Antibody-drug conjugates (ADCs) are complex biotherapeutics that combine the selectivity of monoclonal antibodies with the potency of cytotoxic payloads. Their structural heterogeneity, arising from variable conjugation sites, drug-to-antibody ratios (DAR), linker stability, and post-translational modifications, presents significant challenges for characterization and quality control. Multi-Attribute Methods (MAM), based on high-resolution mass spectrometry, have emerged as powerful tools to simultaneously identify and quantify critical quality attributes at multiple levels of analysis, including intact protein, subunits, and peptides. Compared with traditional assays such as capillary electrophoresis-sodium dodecyl sulfate, hydrophobic interaction chromatography, and Ion exchange chromatography, MAM offers higher specificity, sensitivity, and the ability to detect unexpected modifications in a single workflow. This review summarizes recent advances in the application of MAM to ADC development, highlighting strategies for monitoring DAR distribution, conjugation site occupancy, and stability of linkers and payloads. We also discuss challenges in method qualification, data handling, and regulatory acceptance, along with the potential of automation and machine learning to support large-scale implementation. Finally, we explore the future outlook of MAM as an integrated platform method for ADCs characterization and quality control, with emphasis on its role in regulatory submissions and quality control release testing.

Antibody-Drug Conjugates (ADCs) represent a rapidly growing class of targeted therapeutics, combining the specificity of monoclonal antibodies with the potency of cytotoxic payloads. The structural complexity and heterogeneity of ADCs arising from variations in drug to antibody ratio (DAR), conjugation sites, and post translational modifications demand advanced analytical strategies for comprehensive characterization and quantification throughout development. Liquid chromatography coupled with mass spectrometry (LC-MS) has emerged as an indispensable platform for ADC analysis, offering high sensitivity, selectivity, and structural resolution across multiple levels. This review highlights recent advances in LC-MS workflows, including intact mass analysis, subunit/middle-down profiling, peptide mapping, and bioanalytical assays for free payloads and catabolites. We discuss emerging technologies such as multi-attribute methods (MAM), native MS, ion mobility, and hybrid ligand-binding assay (LBA)-LC-MS platforms that enhance throughput and analytical depth. Special focus is given to quantification strategies in biological matrices and regulatory expectations, including International Council for Harmonization (ICH) M10 and Food and Drug Administration (FDA) guidance on method validation. As ADC pipelines expand into new therapeutic areas, the integration of automation and AI-driven data processing is poised to transform LC-MS into a high throughput, intelligent tool for both product characterization and clinical monitoring. These innovations collectively support safer, more effective ADC development from discovery through approval.

Intraarticular (IA) administration of ProGel-Dex was previously found to provide sustained joint pain relief with excellent safety in arthritis animal models. To explore ProGel-Dex’ working mechanisms, we conducted a comprehensive pharmacokinetics/biodistribution (PK/BD) study of IA ProGel-Dex and the free Dex released in an osteoarthritis mouse model. An initial “burst” release and distribution of ProGel-Dex was observed in all organs/tissues post IA administration. The higher-than-1.5 AUCinf_obs/AUCall ratios for both ProGel-Dex and free Dex support their long-term presence within the DMM joint beyond the experimental endpoint. The overall systemic organ/tissue exposures to ProGel-Dex and free Dex released were found to be much lower than those detected within the OA joint with IA ProGel-Dex. Together, these data support that the potent and long-sustained OA joint pain relief and the excellent safety of IA ProGel-Dex can be attributed to its prolonged retention in OA joint and the pathology-driven local activation.

Therapeutic oligonucleotides have emerged as a transformative drug class, yet their physicochemical complexity poses unique analytical challenges in bioanalysis. Liquid chromatography mass spectrometry (LC-MS) has become a powerful platform for their quantification, offering high specificity and structural insight. However, accurate measurement requires addressing challenges such as nonspecific binding, matrix effects, nuclease degradation, and ion-pairing interferences from sample preparation to LC-MS analysis. This review provides a practical roadmap for establishing robust LC-MS workflows for oligonucleotides bioanalysis, emphasizing optimized sample preparation, column and mobile phase selection, ionization control, and fragmentation tuning. Key strategies to minimize analytical artifacts, improve recovery, and ensure regulatory compliance are discussed in the context of current FDA or EMA bioanalytical validation guidelines. Collectively, this work outlines the critical considerations and systematic optimizations needed to achieve reliable, reproducible, and sensitive quantification of therapeutic oligonucleotides in complex biological matrices, supporting their successful clinical translation by informing pharmacokinetics, therapeutic potential, and safety profiles.

MYC is one of the most deregulated oncogenic transcription factors in human cancers. MYC amplification/or overexpression is most common in Group 3 medulloblastoma and is positively associated with poor prognosis. MYC is known to regulate the transcription of major components of protein synthesis (translation) machinery, leading to promoted rates of protein synthesis and tumorigenesis. MTOR signaling-driven deregulated protein synthesis is widespread in various cancers, including medulloblastoma, which can promote the stabilization of MYC. Indeed, our previous studies demonstrate that the key components of protein synthesis machinery, including mTOR signaling and MYC targets, are overexpressed and activated in MYC-amplified medulloblastoma, confirming MYC-dependent addiction of enhanced protein synthesis in medulloblastoma. Further, targeting this enhanced protein synthesis pathway with combined inhibition of MYC transcription and mTOR translation by small-molecule inhibitors, demonstrates preclinical synergistic anti-tumor potential against MYC-driven medulloblastoma in vitro and in vivo. Thus, inhibiting enhanced protein synthesis by targeting the MYC indirectly and mTOR pathways together may present a highly appropriate strategy for treating MYC-driven medulloblastoma and other MYC-addicted cancers. Evidence strongly proposes that MYC/mTOR-driven tumorigenic signaling can predominantly control the translational machinery to elicit cooperative effects on increased cell proliferation, cell cycle progression, and genome dysregulation as a mechanism of cancer initiation. Several small molecule inhibitors of targeting MYC indirectly and mTOR signaling have been developed and used clinically with immunosuppressants and chemotherapy in multiple cancers. Only a few of them have been investigated as treatments for medulloblastoma and other pediatric tumors. This review explores concurrent targeting of MYC and mTOR signaling against MYC-driven medulloblastoma. Based on existing evidence, targeting of MYC and mTOR pathways together produces functional synergy that could be the basis for effective therapies against medulloblastoma.

Pelargonium graveolens L., frequently known as rose-scented geranium (family: Geraniaceae; 2n = 88), is an herbaceous medicinal and aromatic plant cultivated for its highly valued and significant volatile essential oil. It widely excels in the food, cosmetics, aromatherapy, and medicinal industries. The present study focuses on the perennial cultivation of geranium for taking multi-harvest in a year by the novel shed-saving agro technology (SSAT) technology for generating a higher yield of essential oils. A 5 year field experiment was carried out at the research farm of CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow, India. During experimentation, two geranium varieties, V1 ( CIM-Pawan ) and V2 (Bourbon ), were grown under a semi-protected polythene shed and harvested 14 times at various intervals. The study found that the maximum fresh herbage yield (14.04 t ha-1) was recorded in variety V1 at IIIrd harvest. The highest essential oil yield (23.76 kg ha-1) was recorded in variety V1 at the Ist harvest. After carefuly examining the trial results, it is suggested that geranium cultivation can be continued as a perennial crop for multiple harvests under a semi-protective shed to obtain higher yields with better financial returns. In contrast, only 1 or 2 harvests are possible under open (natural) conditions of northern Indian plains. Keywords: Pelargonium graveolens, Production technology, Perennial crop, North Indian plains, Low-cost novel technology, Shed-saving agro-technology.

In the worldwide healthcare system, medicinal herbs have become an important source of therapeutic assistance for both people and animals, not only in the case of illness but also as a possible resource for preserving good health.In India, almost 70% of people would rather use herbal cosmetics for medical purposes.The current situation indicates that the use of herbal cosmetics has increased in the personal care system and that they are greatly needed in day-to-day living.Herbal cosmetics are regularly used in human life and the cosmetics industry includes a wide range of items, including creams, lotions, fragrances, and skin-cleansing treatments.Cosmetic preparations often contain a lot of natural components.Medicinal herbs have gained a substantial place in the global health system for both humans and animals, not only in the treatment of sickness but also as a potential resource for preserving good health.In the Future cosmetics and skin care product formulas are expected to be more advanced than ever.In East Asia, the use of traditional medicine has been demonstrated to improve the survival rate of patients with stomach cancer when combined with current medications.Due to their greater availability and fewer side effects, herbal remedies are preferred over artificial ones.The goal of this research is to create a scientific framework for assessing the potential toxicity of traditional herbal medicines (THMs) using publicly available regulatory documentation.The Indonesia a is one of the biggest supplier's countries for herbal raw materials in cosmetic products.Many plants are available naturally as well as different uses that can be made as basic ingredients for cosmetic preparations.For generations, people have utilized herbs in cosmetics and medications.It is generally known that they have the ability to beautify, enhance the look of the skin, and treat a variety of skin conditions.UV radiation protection and side effect avoidance are always necessary since UV radiation can result in sunburns, wrinkles, lowered immunity to infections, accelerated aging, and cancer.Over the last several decades, there has been a growing demand for herbal medicines due to the increased public awareness of living a healthy lifestyle by going "back to nature."CLIN acanthus nutans, also known as Sabah snake grass or "belalai gajah" locally in Malaysia, is one of the therapeutic plants that is becoming more and more popular.It is a member of the Acantharean family.

Background Medulloblastoma (MB) patients with MYC oncogene amplification or overexpression exhibit extremely poor prognoses and therapy resistance. However, MYC itself has been one of the most challenging targets for cancer treatment. Here, we identify a novel marinopyrrole natural derivative, MP1, that shows desirable anti-MYC and anti-cancer activities in MB. Methods In this study, using MYC-amplified (Group 3) and non-MYC amplified MB cell lines in vitro and in vivo, we evaluated anti-cancer efficacies and molecular mechanism(s) of MP1. Results MP1 significantly suppressed MB cell growth and sphere counts and induced G2 cell cycle arrest and apoptosis in a MYC-dependent manner. Mechanistically, MP1 strongly downregulated the expression of MYC protein. Our results with RNA-seq revealed that MP1 significantly modulated global gene expression and inhibited MYC-associated transcriptional targets including translation/mTOR targets. In addition, MP1 inhibited MYC-target metabolism, leading to declined energy levels. The combination of MP1 with an FDA-approved mTOR inhibitor temsirolimus synergistically inhibited MB cell growth/survival by downregulating the expression of MYC and mTOR signaling components. Our results further showed that as single agents, both MP1 and temsirolimus, were able to significantly inhibit tumor growth and MYC expression in subcutaneously or orthotopically MYC-amplified MB bearing mice. In combination, there were further anti-MB effects on the tumor growth and MYC expression in mice. Conclusion These preclinical findings highlight the promise of marinopyrrole MP1 as a novel MYC inhibition approach for MYC-amplified MB.

LD14b is an amyloid-beta (A beta) 17 beta-hydroxysteroid dehydrogenase type 10 (A beta-17 beta-HSD10) protein-protein interaction modulator that shows promising in vitro and ex vivo activity to rescue A beta-induced mitochondrial dysfunction, A beta-induced toxicity, and A beta-mediated inhibition of estradiol synthesis. The current study investigated in vitro human S9 fractions metabolic stability, apparent permeability, human and mouse plasma protein binding, in vivo pharmacokinetics, and tissue distribution in Balb/cJ mice. A fast (8-min), sensitive, reliable, and reproducible LC-MS/MS method was developed and validated over the dynamic range of 1-1000 ng/mL for the quantification of LD14b in different biological matrices (plasma, liver, kidney, brain, lungs, heart). LD14b was metabolically stable in human liver S9 fractions with 70% remaining after 90 minutes of incubation, showed intermediate apparent permeability of 3.55 x 10(-06) cm/s and 6.16 x 10(-06) cm/s for apical-to-basolateral (A-to-B) and basolateral-to-apical (B-to-A), respectively across the Caco-2 monolayer, and was medium/highly bound to human plasma proteins (84.1%), mouse plasma proteins (85.7%), and mouse brain homogenate (95.4%). LD14b showed an in vivo predicted % absorption of 52% in Balb/cJ mice and was well-distributed to the peripheral tissues (liver, kidney, lungs, and heart) including the brain.

Abstract Medulloblastoma (MB) is the most common type of childhood brain cancer worldwide. Although current treatment with surgery and extensive chemoradiation has led to increased survival rates, many MB patients still die from the disease. Moreover, surviving patients suffer severe long-term side effects as a consequence of treatment. It is therefore crucial to develop more effective and less toxic therapies. The most aggressive subtype of MB tumors often exhibits amplification or overexpression of the MYC oncogene. Patients with MYC-amplified MB exhibit a high frequency of cerebrospinal tumor dissemination, often experience treatment resistance and have extremely poor prognoses. While the MYC oncogene is established as the oncogenic driver in Group 3 MB, it has remained undruggable. Thus, targeting regulatory components of MYC and the signaling pathways regulated by it is of great potential therapeutic value. Studies have revealed that MB has very few germline mutations in cancer predisposition genes, suggesting that dysregulated epigenetic pathways might be critical in MB pathogenesis. Particularly, dysregulation of epigenetic modifiers, including histone methyltransferases and histone demethylases, is very common in Group 3 MB, compared to other MB subgroups. Therefore, it is important to identify such epigenetic modifiers that may have controls on MYC and its tumorigenic activities and explore these as the epigenetic drug-candidate targets in Group 3 (MYC-driven) MB. In this regard, we found that protein arginine demethylase Jumonji C domain-containing protein 6 (JMJD6), an emerging key epigenetic enzyme in cancers, is a novel regulator of MYC expression in MYC-driven MB. We observed high levels of JMJD6 that not only mirror MYC expression in the most aggressive MB but also correlate with poor outcomes in these patients. Knockdown of JMJD6 decreased MYC expression, cellular proliferation/survival and stemness in MYC-amplified MB cells. Mechanistically, our results revealed that JMJD6 forms complexes with proteins involved in maintaining and regulating the promoter-pause at super-enhancers, suggesting that JMJD6 can regulate MYC at the transcription level. Moreover, our in vivo analyses of JMJD6 inhibition, either with inducible gene knockdown or a pharmacologic small molecule inhibitor, demonstrated anti-MB potential with suppressed MYC expression. Based on this background and preliminary observations, we hypothesize that JMJD6 plays crucial roles in the most aggressive MB by regulating MYC expression and hence MYC-driven tumorigenesis. Accordingly, we hypothesize that targeting the JMJD6-MYC axis by JMJD6 inhibition can serve as a powerful therapeutic strategy for MYC-driven MB. Citation Format: Matthew Kling, Devendra Kumar, Sutapa Ray, Shantaram Joshi, Don Coulter, Nagendra K. Chaturvedi. JMJD6 as a novel tumorigenic factor and therapeutic target in group 3 (MYC-driven) medulloblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3346.

MYC amplification or overexpression is most common in Group 3 medulloblastomas and is positively associated with poor clinical outcomes. Recently, protein arginine methyltransferase 5 (PRMT5) overexpression has been shown to be associated with tumorigenic MYC functions in cancers, particularly in brain cancers such as glioblastoma and medulloblastoma. PRMT5 regulates oncogenes, including MYC, that are often deregulated in medulloblastomas. However, the role of PRMT5-mediated post-translational modification in the stabilization of these oncoproteins remains poorly understood. The potential impact of PRMT5 inhibition on MYC makes it an attractive target in various cancers. PRMT5 inhibitors are a promising class of anti-cancer drugs demonstrating preclinical and preliminary clinical efficacies. Here, we review the publicly available preclinical and clinical studies on PRMT5 targeting using small molecule inhibitors and discuss the prospects of using them in medulloblastoma therapy.

Diabetes, the most common disease in the world, must be managed. Modern cutting-edge technology is able to help with diabetic self-management by taking secure measurements and monitoring health data. This includes the blockchain, Internet of Things, augmented reality, and virtual reality (medical sensors and wearables). The platform architecture implemented in this chapter, which is based on the Internet of Things and blockchain, makes it easy for patient to effectively control their diabetes on their own while helping doctors in more effective diabetes management. Internet of Things and blockchain are integrated in the architecture to collect patient data and rapidly and safely exchange it with the medical team while maintaining patient privacy.

ID 25340 Poster Board 79 Our laboratory has pioneered studying the biology of lysophosphatidic acid (LPA) in oral homeostasis and periodontal disease. LPA is the simplest lipid mediator and plays key homeostatic and inflammatory roles. Saliva is essential for maintaining oral health, and salivary glands are thus pivotal, as they participate in regulating immunity and inflammation, especially in oral disease prevention, oral infection control (Papagerakis et al., 2014), and autoimmunity. Salivary gland secretion and function is also affected by many pharmacological agents. We (Bathena et al. 2011) and others have reported the presence of multiple LPA species in human healthy normal and in periodontal disease saliva. Therefore, we hypothesized that the same LPA species would also be present in mouse saliva, and that mouse salivary glands would also express multiple LPARs. Methods: the simple, sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method we developed in 2011 was used to quantify LPA species (LPA 18:0, LPA 16:0, and LPA 18:1) in C57BL/6 mouse saliva. To further our work, we previously also developed whole-mount in situ LPA receptor (LPAR) subtype localization for various oral soft and hard tissues (Cerutis et al. 2016). Polyclonal antibodies against LPA1, LPA3, and LPA4 were used in indirect immunofluorescence for labeling normal mouse salivary glands using this whole-mount in situ technique. Results: As we reported for humans, LPA species are also present in low, homeostatic concentrations in mouse saliva. As periodontal disease develops, paralleling what we reported in Bathena et al. (2011) for human saliva, some of the LPA species increase approximately 10-fold as well. Confocal microscopy confirmed LPA1, LPA3, and substantial LPA4 labeling; similar but different patterns of distribution were seen for each GPCR. In conclusion, this confirms LPA1 and LPA3 expression (like in NOD mice, Park et al., (2017) but is the first report of the presence of LPA4 in mouse salivary gland tissue. The presence of multiple confirmed LPARs in mouse salivary glands suggests that these receptors need to be factored into not only studies of autoimmune conditions but also for pharmacological studies of drugs affecting salivary gland biology and secretion. Support: NIDCR/NIGMS 1R15DE028687-01(D.R.C).

RNA interference (RNAi) molecules have tremendous potential for cancer therapy but are limited by insufficient potency after intravenous (IV) administration. We previously found that polymer complexes (polyplexes) formed between 3′-cholesterol-modified siRNA (Chol-siRNA) or DsiRNA (Chol-DsiRNA) and the cationic diblock copolymer PLL[30]-PEG[5K] greatly increase RNAi potency against stably expressed LUC mRNA in primary syngeneic murine breast tumors after daily IV dosing. Chol-DsiRNA polyplexes, however, maintain LUC mRNA suppression for ~48 h longer after the final dose than Chol-siRNA polyplexes, which suggests that they are the better candidate formulation. Here, we directly compared the activities of Chol-siRNA polyplexes and Chol-DsiRNA polyplexes in primary murine 4T1 breast tumors against STAT3, a therapeutically relevant target gene that is overexpressed in many solid tumors, including breast cancer. We found that Chol-siSTAT3 polyplexes suppressed STAT3 mRNA in 4T1 tumors with similar potency (half-maximal ED50 0.3 mg/kg) and kinetics (over 96 h) as Chol-DsiSTAT3 polyplexes, but with slightly lower activity against total Stat3 protein (29% vs. 42% suppression) and tumor growth (11.5% vs. 8.6% rate-based T/C ratio) after repeated IV administration of equimolar, tumor-saturating doses every other day. Thus, both Chol-siRNA polyplexes and Chol-DsiRNA polyplexes may be suitable clinical candidates for the RNAi therapy of breast cancer and other solid tumors.

COVID-19 is an acute and progressive respiratory illness, which is highly contagious. In 2020, COVID-19 has become a major health issue; its prevalence has been increasing at an alarming rate across the world. Less availability of COVID-19 drugs, high treatment cost, and side effects affect the quality of life of a person infected with COVID-19 in countries with poorly developed health systems. Through infection, patients can die due to acute respiratory distress syndrome (ARDS) initiated by systemic inflammatory reactions due to the undue emancipation of chemokines and pro-inflammatory cytokines by the immune effector cells. The aim of this review is to summarize and evaluate the evidence of traditional medicine, which can facilitate the treatment options according to the clinical manifestations of COVID-19 patients and has proven effectiveness in prevention and control of disease. The systemic search for medicinal plants for the therapeutics of COVID-19 was performed considering the articles published through the different scientific databases. The results suggested that some important medicinal plants reported for antiviral and anti-allergic/anti-inflammatory activities are Withania somnifera "Ashwagandha", Asparagus racemosus "Shatavari", Ocimum sanctum "Basil", Foeniculum vulgare "Fennel", Allium Sativum "Garlic", Tinospora cordifolia "Giloy", Glycyrrhiza glabra "licorice", Organum vulgare "Oregano", Rosmarinus Officinalis "Rosemary", Salvia "Sage", Zinger officinale "Ginger", Torreya nucifera"Japenese torreya", Isatis indigotica "Ban-Lan-Gen" Echinacea, Panax ginseng, Houttuynia cordata, Cannabinoid (CBD). The traditional medicines against COVID-19, currently under clinical trials (NCT04494204, NCT04387643, NCT04395976, NCT04621903, NCT04621903, NCT04544605) and clinical application of traditional Indian and Chinese medicine for the treatment of COVID-19 are also found. This review highlights the major goal of herbal remedies and their significant role to cure antiviral diseases like COVID-19. It is suggested that promising polyherbal formulations and traditional plants must be investigated on the priority basis to solve current crisis.

There are several challenges associated with LC-MS/MS bioanalytical method development and validation. Low and variable recovery of some analytes, especially the more hydrophobic ones, is often challenging. Analytes can be lost to various extents throughout the process of sample collection, storage, before, during, and/or after sample preparation and analysis. The calculation of overall extraction recovery can detect problems of low recovery during sample preparation but does not identify the source(s) of analyte losses. Low overall analyte recovery is the net result of losses that can happen for multiple reasons at all steps of sample preparation and analysis. Therefore, identifying the source(s) of analyte loss during sample preparation can help guide the optimization the bioanalysis conditions to minimize these losses. In this article we propose a practical protocol to systematically identify and quantify the sources of low analyte recovery. This allows the proper choice of strategies to optimize the relevant bioanalytical conditions to minimize analyte losses and improve overall recovery.