Premix technologies for drug delivery: manufacturing, applications, and opportunities in regulatory filing.

Active pharmaceutical ingredients (APIs) and excipients can be carefully combined in premix-based materials before being added to dosage forms, providing a flexible platform for the improvement of drug bioavailability, stability, and patient compliance. This is a promising and transformative approach in novel and generic product development, offering both the potential to overcome challenges in the delivery of complex APIs and viable solutions for bypassing patent hurdles in generic product filing. We discuss the different types of premixes; manufacturing technologies such as spray drying, hot melt extrusion, wet granulation, co-crystal, co-milling, co-precipitation; regulatory filing opportunities; and major bottlenecks in the use of premix materials in different aspects of pharmaceutical product development.

Surface engineered nanodiamonds: mechanistic intervention in biomedical applications for diagnosis and treatment of cancer.

In terms of biomedical tools, nanodiamonds (ND) are a more recent innovation. Their size typically ranges between 4 to 100 nm. ND are produced via a variety of methods and are known for their physical toughness, durability, and chemical stability. Studies have revealed that surface modifications and functionalization have a significant influence on the optical and electrical properties of the nanomaterial. Consequently, surface functional groups of NDs have applications in a variety of domains, including drug administration, gene delivery, immunotherapy for cancer treatment, and bio-imaging to diagnose cancer. Additionally, their biocompatibility is a critical requisite for theirin vivoandin vitrointerventions. This review delves into these aspects and focuses on the recent advances in surface modification strategies of NDs for various biomedical applications surrounding cancer diagnosis and treatment. Furthermore, the prognosis of its clinical translation has also been discussed.

3D bioprinting of stromal cells-laden artificial cornea based on visible light-crosslinkable bioinks forming multilength networks.

3D bioprinting has the potential for the rapid and precise engineering of hydrogel constructs that can mimic the structural and optical complexity of a healthy cornea. However, the use of existing light-activated bioinks for corneal printing is limited by their poor cytocompatibility, use of cytotoxic photoinitiators (PIs), low photo-crosslinking efficiency, and opaque/colored surface of the printed material. Herein, we report a fast-curable, non-cytotoxic, optically transparent bioprinting system using a new water-soluble benzoyl phosphinate-based PI and photocrosslinkable methacrylated hyaluronic acid (HAMA). Compared with commercially available PIs, the newly developed PI, lithium benzoyl (phenyl) phosphinate (BP), demonstrated increased photoinitiation efficiency under visible light and low cytotoxicity. Using a catalytic amount of BP, the HA-based bioinks quickly formed 3D hydrogel constructs under low-energy visible-light irradiation (405 nm, <1 J cm-2). The mechanical properties and printability of photocurable bioinks were further improved by blending low (10 kDa) and high (100 kDa) molecular weight (MW) HAMA by forming multilength networks. For potential applications as corneal scaffolds, stromal cell-laden dome-shaped constructs were fabricated using MW-blended HAMA/BP bioink and a digital light processing printer. The HA-based photocurable bioinks exhibited good cytocompatibility (80%-95%), fast curing kinetics (<5 s), and excellent optical transparency (>90% in the visible range), potentially making them suitable for corneal tissue engineering.

Disease spectrum and its molecular characterisation in the lentil production system of lower-Indo Gangetic plains.

Lentil is a food legume grown in the Indo-Gangetic plains including lower Gangetic Bengal (LGB). Lentil productivity in this zone is severely impeded because of the prevalence of several biotic cues. Plausible reports regarding the status of disease scenario and the associated risk factors are missing. Therefore, judicious crop management strategies are lacking. An intensive survey of 267 farmers' fields was conducted over 3 years in major lentil-growing districts of LGB to evaluate the disease incidence and prevalence. Additional insights were generated, apprehending isolation and characterisation of associated pathogens through spore morphology and molecular markers as well as elucidating the role of biophysical factors in influencing disease development. Climate change has shifted the disease dimension of lentil and precipitated new disease complexes of great risk, which was reflected through geospatial mapping results in the present study. The prevalence of three major diseases, namely collar rot (Sclerotium rolfsii), lentil blight complex (LBC) incited by both Alternaria and Stemphylium, and lentil rust (Uromyces viciae-fabae), was ascertained through cultural and molecular studies and contextualised through pathogenicity appraisal. This study is the first to investigate the complex mixed infection of Alternaria alternata and Stemphylium botryosum, successfully isolating S. botyrosum in India, and confirming the pathogens through sequencing by using internal transcribed spacer (ITS) primers and Stemphylium-specific Glycerol-3-phosphate dehydrogenase 1 (gpd1) and gpd2 primers. Unlike late planting, early planting promoted collar rot infestation. LBC and rust incidence were magnified in late planting. Soil texture resulted in the spatial distribution of collar rot disease. The surveyed data also highlighted the potential role of resistant cultivars and cropping pattern intervention to ensure associational resistance towards addressing the disease bottleneck in lentil.

Role of Angiogenesis and Its Biomarkers in Development of Targeted Tumor Therapies.

Angiogenesis plays a significant role in the human body, from wound healing to tumor progression. "Angiogenic switch" indicates a time-restricted event where the imbalance between pro- and antiangiogenic factors results in the transition from prevascular hyperplasia to outgrowing vascularized tumor, which eventually leads to the malignant cancer progression. In the last decade, molecular players, i.e., angiogenic biomarkers and underlying molecular pathways involved in tumorigenesis, have been intensely investigated. Disrupting the initiation and halting the progression of angiogenesis by targeting these biomarkers and molecular pathways has been considered as a potential treatment approach for tumor angiogenesis. This review discusses the currently known biomarkers and available antiangiogenic therapies in cancer, i.e., monoclonal antibodies, aptamers, small molecular inhibitors, miRNAs, siRNAs, angiostatin, endostatin, and melatonin analogues, either approved by the U.S. Food and Drug Administration or currently under clinical and preclinical investigations.

Assessing the advantages of 3D bioprinting and 3D spheroids in deciphering the osteoarthritis healing mechanism using human chondrocytes and polarized macrophages.

The molecular niche of an osteoarthritic microenvironment comprises the native chondrocytes, the circulatory immune cells, and their respective inflammatory mediators. Although M2 macrophages infiltrate the joint tissue during osteoarthritis (OA) to initiate cartilage repair, the mechanistic crosstalk that dwells underneath is still unknown. Our study established a co-culture system of human OA chondrocytes and M2 macrophages in 3D spheroids and 3D bioprinted silk-gelatin constructs. It is already well established that Silk fibroin-gelatin bioink supports chondrogenic differentiation due to upregulation of the Wnt/ß-catenin pathway. Additionally, the presence of anti-inflammatory M2 macrophages significantly upregulated the expression of chondrogenic biomarkers (COL-II, ACAN) with an attenuated expression of the chondrocyte hypertrophy (COL-X), chondrocyte dedifferentiation (COL-I) and matrix catabolism (MMP-1 and MMP-13) genes even in the absence of the interleukins. Furthermore, the 3D bioprinted co-culture model displayed an upper hand in stimulating cartilage regeneration and OA inhibition than the spheroid model, underlining the role of silk fibroin-gelatin in encouraging chondrogenesis. Additionally, the 3D bioprinted silk-gelatin constructs further supported the maintenance of stable anti-inflammatory phenotype of M2 macrophage. Thus, the direct interaction between the primary OAC and M2 macrophages in the 3D context, along with the release of the soluble anti-inflammatory factors by the M2 cells, significantly contributed to a better understanding of the molecular mechanisms responsible for immune cell-mediated OA healing.

Exploring the interaction between extracellular matrix components in a 3D organoid disease model to replicate the pathophysiology of breast cancer.

In vitro models are necessary to study the pathophysiology of the disease and the development of effective, tailored treatment methods owing to the complexity and heterogeneity of breast cancer and the large population affected by it. The cellular connections and tumor microenvironments observed in vivo are often not recapitulated in conventional two-dimensional (2D) cell cultures. Therefore, developing 3D in vitro models that mimic the complex architecture and physiological circumstances of breast tumors is crucial for advancing our understanding of the illness. A 3D scaffold-free in vitro disease model mimics breast cancer pathophysiology by allowing cells to self-assemble/pattern into 3D structures, in contrast with other 3D models that rely on artificial scaffolds. It is possible that this model, whether applied to breast tumors using patient-derived primary cells (fibroblasts, endothelial cells, and cancer cells), can accurately replicate the observed heterogeneity. The complicated interactions between different cell types are modelled by integrating critical components of the tumor microenvironment, such as the extracellular matrix, vascular endothelial cells, and tumor growth factors. Tissue interactions, immune cell infiltration, and the effects of the milieu on drug resistance can be studied using this scaffold-free 3D model. The scaffold-free 3D in vitro disease model for mimicking tumor pathophysiology in breast cancer is a useful tool for studying the molecular basis of the disease, identifying new therapeutic targets, and evaluating treatment modalities. It provides a more physiologically appropriate high-throughput platform for screening large compound library in a 96-384 well format. We critically discussed the rapid development of personalized treatment strategies and accelerated drug screening platforms to close the gap between traditional 2D cell culture and in vivo investigations.

The accuracy of ultrasound scan in diagnosing retained products of conception: a systematic review and meta-analysis.

OBJECTIVE: This study aimed to analyze and summarize the evidence on the accuracy of different ultrasound methods in the diagnosis of retained products of conception. DATA SOURCES: We searched Ovid SP, the Cumulative Register to Nursing & Allied Health Literature, EBSCO, and grey literature including Core, Trip, Networked Digital Library of Theses and Dissertations Global ETD search, BMJ Best Practice, PubMed, GreyLit report website (http://www.greylit.org/), Cochrane Central Register of Controlled Trials, and Google scholar (https://scholar.google.com/). STUDY ELIGIBILITY CRITERIA: We included prospective and retrospective cross-sectional or Cohort studies that evaluated both ultrasound findings (before management of retained products of conception) and histopathologic results of retained products of conception at all gestational ages. METHODS: We used Covidence for data extraction from the studies and quality assessment. The meta-analysis was performed using RevMan 5.4 (forest plot), MetaDTA version 2.01, and Meta-DiSc 2.0 online software. RESULTS: In total, 11 studies were eligible for data extraction and meta-analysis. The total number of study participants from these 11 studies were 1567. Of these, 9 studies were included to test the accuracy of an echogenic mass, 4 studies analyzed the accuracy of endometrial thickness, and 5 studies analyzed the accuracy of color Doppler flow to predict retained products of conception. We found that echogenic mass had the highest sensitivity, specificity, and diagnostic odds ratio for predicting retained products of conception. The sensitivity, specificity, and diagnostic odds ratio were 0.915 (95% confidence interval, 0.844-0.955), 0.843 (95% confidence interval, 0.615-0.947), and 57.787 (95% confidence interval, 15.171-220.112), respectively. The diagnostic threshold for endometrial thickness was set at 10 mm with a sensitivity, specificity, and diagnostic odds ratio of 0.667 (95% confidence interval, 0.072-0.981), 0.866 (95% confidence interval, 0.375-0.986), and 12.927 (95% confidence interval, 0.23-726.582). The sensitivity, specificity, and diagnostic odds ratio of color Doppler flow were 0.850 (95% confidence interval, 0.756-0.913), 0.406 (95% confidence interval, 0.198-0.655), and 3.893 (95% confidence interval, 1.005-15.081). CONCLUSION: Our review concluded that an echogenic mass is the most sensitive and specific predictor of retained products of conception after any pregnancy event. The most important limitation of our review is that the design of the studies included led to significant statistical heterogeneity.

Lactate acidosis and simultaneous recruitment of TGF-ß leads to alter plasticity of hypoxic cancer cells in tumor microenvironment.

Lactate acidosis is often observed in the tumor microenvironment (TME) of solid tumors. This is because glucose breaks down quickly via glycolysis, causing lactate acidity. Lactate is harmful to healthy cells, but is a major oncometabolite for solid cancer cells that do not receive sufficient oxygen. As an oncometabolite, it helps tumor cells perform different functions, which helps solid hypoxic tumor cells spread to other parts of the body. Studies have shown that the acidic TME contains VEGF, Matrix metalloproteinases (MMPs), cathepsins, and transforming growth factor-ß (TGF-ß), all of which help spread in direct and indirect ways. Although each cytokine is important in its own manner in the TME, TGF-ß has received much attention for its role in metastatic transformation. Several studies have shown that lactate acidosis can cause TGF-ß expression in solid hypoxic cancers. TGF-ß has also been reported to increase the production of fatty acids, making cells more resistant to treatment. TGF-ß has also been shown to control the expression of VEGF and MMPs, which helps solid hypoxic tumors become more aggressive by helping them spread and create new blood vessels through an unknown process. The role of TGF-ß under physiological conditions has been described previously. In this study, we examined the role of TGF-ß, which is induced by lactate acidosis, in the spread of solid hypoxic cancer cells. We also found that TGF-ß and lactate work together to boost fatty acid production, which helps angiogenesis and invasiveness.

Multi compartmental 3D breast cancer disease model-recapitulating tumor complexity in in-vitro.

Breast cancer is the most common ailment among women. In 2020, it had the highest incidence of any type of cancer. Many Phase II and III anti-cancer drugs fail due to efficacy, durability, and side effects. Thus, accelerated drug screening models must be accurate. In-vivo models have been used for a long time, but delays, inconsistent results, and a greater sense of responsibility among scientists toward wildlife have led to the search for in-vitro alternatives. Stromal components support breast cancer growth and survival. Multi-compartment Transwell models may be handy instruments. Co-culturing breast cancer cells with endothelium and fibroblasts improves modelling. The extracellular matrix (ECM) supports native 3D hydrogels in natural and polymeric forms. 3D Transwell cultured tumor spheroids mimicked in-vivo pathological conditions. Tumor invasion, migration, Trans-endothelial migration, angiogenesis, and spread are studied using comprehensive models. Transwell models can create a cancer niche and conduct high-throughput drug screening, promising future applications. Our comprehensive shows how 3D in-vitro multi compartmental models may be useful in producing breast cancer stroma in Transwell culture.

Development of a biomimetic arch-like 3D bioprinted construct for cartilage regeneration using gelatin methacryloyl and silk fibroin-gelatin bioinks.

In recent years, engineering biomimetic cellular microenvironments have been a top priority for regenerative medicine. Collagen II, which is arranged in arches, forms the predominant fiber network in articular cartilage. Due to the shortage of suitable microfabrication techniques capable of producing 3D fibrous structures,in vitroreplication of the arch-like cartilaginous tissue constitutes one of the major challenges. Hence, in the present study, we report a 3D bioprinting approach for fabricating arch-like constructs using two types of bioinks, gelatin methacryloyl (GelMa) and silk fibroin-gelatin (SF-G). The bioprinted SF-G constructs displayed increased proliferation of the encapsulated human bone marrow-derived mesenchymal stem cells compared to the GelMA constructs. Biochemical assays, gene, and protein expression exhibited the superior role of SF-G in forming the fibrous collagen network and chondrogenesis. Protein-protein interaction study using Metascape evaluated the function of the proteins involved. Further GeneMANIA and STRING analysis using Col 2A1, SOX 9, ACAN, and the genes upregulated on day 21 in RT-PCR, i.e.ß-catenin, TGFßR1, Col 1A1 in SF-G and PRG4, Col 10A1, MMP 13 in GelMA validated ourin vitroresults. These findings emphasized the role of SF-G in regulating the Wnt/ß-catenin and TGF-ßsignaling pathways. Hence, the 3D bioprinted arch-like constructs possess a substantial potential for cartilage regeneration.

Mechanistic crosstalk of extracellular calcium-mediated regulation of maturation and plasticity in human monocytes.

Innate immune cells play a pivotal role in controlling tissue repair and rejection after biomaterial implantation. Calcium supplementation regulates cellular responses and alter the pathophysiology of various diseases. A series of macrophage activations through differential plasticity has been observed after cell-to-material interactions. We investigated the role of calcium supplementation in controlling macrophage phenotypes in pro-inflammatory and pre-reparative states. Oxidative defence and mitochondria involvement in cellular plasticity and the sequential M0 to M1 and M1 to M2 transitions were observed after calcium supplementation. This study describes the molecular mechanism of reactive oxygen species and drives the interconnected cellular plasticity of macrophages in the presence of calcium. Gene expression, and immunostaining, revealed a relationship between MHC class II maturation and cellular plasticity. This study elucidated the role of controlled calcium supplementation under various conditions. These findings underscore the molecular mechanism of calcium-mediated immune induction and its favourable use in different calcium-containing biomaterials., essential for tissue regeneration.

A lung targeted miR-29 mimic as a therapy for pulmonary fibrosis.

BACKGROUND: MicroRNAs are non-coding RNAs that negatively regulate gene networks. Previously, we reported that systemically delivered miR-29 mimic MRG-201 reduced fibrosis in animal models, supporting the consideration of miR-29-based therapies for idiopathic pulmonary fibrosis (IPF). METHODS: We generated MRG-229, a next-generation miR-29 mimic based on MRG-201 with improved chemical stability due to additional sugar modifications and conjugation with the internalization moiety BiPPB (PDGFbetaR-specific bicyclic peptide)1. We investigated the anti-fibrotic efficacy of MRG-229 on TGF-ß1 treated human lung fibroblasts (NHLFs), human precision cut lung slices (hPCLS), and in vivo bleomycin studies; toxicology was assessed in two animal models, rats, and non-human primates. Finally, we examined miR-29b levels in a cohort of 46 and 213 patients with IPF diagnosis recruited from Yale and Nottingham Universities (Profile Cohort), respectively. FINDINGS: The peptide-conjugated MRG-229 mimic decreased expression of pro-fibrotic genes and reduced collagen production in each model. In bleomycin-treated mice, the peptide-conjugated MRG-229 mimic downregulated profibrotic gene programs at doses more than ten-fold lower than the original compound. In rats and non-human primates, the peptide-conjugated MRG-229 mimic was well tolerated at clinically relevant doses with no adverse findings observed. In human peripheral blood from IPF patients decreased miR-29 concentrations were associated with increased mortality in two cohorts potentially identified as a target population for treatment. INTERPRETATION: Collectively, our results provide support for the development of the peptide-conjugated MRG-229 mimic as a potential therapy in humans with IPF. FUNDING: This work was supported by NIH NHLBI grants UH3HL123886, R01HL127349, R01HL141852, U01HL145567.

Modeling crack propagation in heterogeneous materials: Griffith's law, intrinsic crack resistance, and avalanches.

Various kinds of heterogeneity in solids, including atomistic discreteness, affect the fracture strength as well as the failure dynamics remarkably. Here we study the effects of an initial crack in a discrete model for fracture in heterogeneous materials, known as the fiber bundle model. We find three distinct regimes for fracture dynamics depending on the initial crack size. If the initial crack is smaller than a certain value, it does not affect the rupture dynamics and the critical stress, while for a larger initial crack, the growth of the crack leads to breakdown of the entire system, and the critical stress depends on the crack size in a power-law manner with a nontrivial exponent. The exponent, as well as the limiting crack size, depend on the strength of heterogeneity and the range of stress relaxation in the system.

Regulation of Transactivation at C-TAD Domain of HIF-1α by Factor-Inhibiting HIF-1α (FIH-1): A Potential Target for Therapeutic Intervention in Cancer.

Hypoxia-inducible factor-1alpha (HIF-1α) is a major transcription factor that adapts to low oxygen homeostasis and regulates the expression of several hypoxic genes, which aid in cancer survival and development. It has recently piqued the interest of translational researchers in the disciplines of cancer sciences. Hypoxia triggers an ample adaptive mechanism mediated via the HIF-1α transcriptional domain. Anaerobic glycolysis, angiogenesis, metastasis, and mitophagy are adaptive mechanisms that support tumor survival by promoting oxygen supply and regulating oxygen demand in hypoxic tumor cells. Throughout this pathway, the factor-inhibiting HIF-1α is a negative regulator of HIF-1α leading to its hydroxylation at the C-TAD domain of HIF-1α under normoxia. Thus, hydroxylated HIF-1α is unable to proceed with the transcriptional events due to interference in binding of C-TAD and CBP/p300. From this review, we can hypothesize that remodeling of FIH-1 activity is a unique mechanism that decreases the transcriptional activity of HIF-1α and, as a result, all of its hypoxic consequences. Hence, this review manuscript details the depth of knowledge of FIH-1 on hypoxia-associated cellular and molecular events, a potential strategy for targeting hypoxia-induced malignancies.

3D printed hydroxyapatite promotes congruent bone ingrowth in rat load bearing defects.

3D porous hydroxyapatite (HAP) scaffolds produced by conventional foaming processes have limited control over the scaffold's pore size, geometry, and pore interconnectivity. In addition, random internal pore architecture often results in limited clinical success. Imitating the intricate 3D architecture and the functional dynamics of skeletal deformations is a difficult task, highlighting the necessity for a custom-made, on-demand tissue replacement, for which 3D printing is a potential solution. To combat these problems, here we report the ability of 3D printed HAP scaffolds forin vivobone regeneration in a rat tibial defect model. Rapid prototyping using the direct-write technique to fabricate 25 mm2HAP scaffolds were employed for precise control over geometry (both external and internal) and scaffold chemistry. Bone ingrowth was determined using histomorphometry and a novel micro-computed tomography (micro-CT) image analysis. Substantial bone ingrowth was observed in implants that filled the defect site. Further validating this quantitatively by micro-CT, the Bone mineral density (BMD) of the implant at the defect site was 1024 mgHA ccm-1, which was approximately 61.5% more than the BMD found with the sham control at the defect site. In addition, no evident immunoinflammatory response was observed in the hematoxylin and eosin micrographs. Interestingly, the present study showed a positive correlation with the outcomes obtained in our previousin vitrostudy. Overall, the results suggest that 3D printed HAP scaffolds developed in this study offer a suitable matrix for rendering patient-specific and defect-specific bone formation and warrant further testing for clinical application.

Opinion dynamics: public and private.

We study here the dynamics of opinion formation in a society where we take into account the internally held beliefs and externally expressed opinions of the individuals, which are not necessarily the same at all times. While these two components can influence one another, their difference, both in dynamics and in the steady state, poses interesting scenarios in terms of the transition to consensus in the society and characterizations of such consensus. Here we study this public and private opinion dynamics and the critical behaviour of the consensus forming transitions, using a kinetic exchange model. This article is part of the theme issue 'Kinetic exchange models of societies and economies'.

Macrophage Polarization Profiling on Native and Regenerated Silk Biomaterials.

We investigated the plasticity and polarization of THP-1 cells on native and regenerated silk-based biomaterials to address the basic paradigm of immune response. Here, we report redox kinetics, adhesion morphology, and nitric oxide release patterns to identify specific subtypes of macrophages at different time points. Water-annealed silk film and native fibrous braids from Bombyx mori silkworms showed higher anti-inflammatory cytokine profiles or M2 subtypes (as evidenced by the enhanced expression of interleukin-10, interleukin-13, and interleukin-4). Ethanol-treated Bombyx mori silk films and Antheraea mylitta braids exhibited higher levels of pro-inflammatory cytokines or the M1 subtype (as evidenced by enhanced expression of interleukin-1, interleukin-6, interleukin-8, interferon-γ, TNF-α, and GM-CSF) in contact with healthy THP monocytes for 14 days; such a long study is unprecedented. Cytokine microarray analysis revealed the transition (M0-M1, M1-M2), plasticity, and stable phenotype of THP-1 cells in a variable stage in contact with different physicochemical properties of silk-based biomaterials. The detailed immunogenicity in the context of the physicochemical properties of native and regenerative silk-based biomaterials will enable us to accurately predict the possibility of a pro-/anti-inflammatory response. It will helps to predict the in vivo reprogramming and avoid fibrosis formation to enhance their clinical translational potential.