5-Aminolevulinic Acid (5-ALA) is the only fluorophore approved by the FDA as an intraoperative optical imaging agent for fluorescence-guided surgery in patients with glioblastoma. The dosing regimen is based on rodent tests where a maximum signal occurs around 6 hours after drug administration. Here, we construct a computational framework to simulate the transport of 5-ALA through the stomach, blood, and brain, and the subsequent conversion to the florescent agent protoporphyrin IX at the tumor site. The framework combines compartmental models with spatially-resolved partial differential equations, enabling one to address questions regarding quantity and timing of 5-ALA administration before surgery. Numerical tests in two spatial dimensions indicate that, for tumors exceeding the detectable threshold, the time to peak flouresent concentration is 2-7 hours, broadly consistent with the current surgical guidelines. Moreover, the framework enables one to examine the specific effects of tumor size and location on the required dose and timing of 5-ALA administration before glioblastoma surgery. less

Despite a record number of clinical studies investigating various anti-cancer drugs, the 5-year survival rate for multiple myeloma (MM) patients in the United States is only 55%, and nearly all patients relapse. Poor patient outcomes demonstrate that myeloma cells are \"born to survive,\" which means they can adapt and evolve following treatment. As a result, new therapeutic approaches to combat this survival mechanism and target treatment-resistant malignant cells are required. Mcl-1, an anti-apoptotic protein, is required for the development of MM and resistance to therapy. This study looks at the possibility of KS18, a Mcl-1 inhibitor derived from pyoluteorin, to treat resistant MM. We show that KS18 inhibits Mcl-1 selectively and promotes post-translational modifications, resulting in UPS-dependent Mcl-1 degradation. Our findings show that KS18-induced Mcl-1 degradation results in caspase-dependent apoptosis. Importantly, KS18 triggered apoptosis in MM patient samples and bortezomib-resistant cells, synergizing with venetoclax to boost apoptosis. Furthermore, KS18 inhibits colony formation in bortezomib-resistant cells. KS18 treated NSG mice displayed significant tumor shrinkage without significant toxicity after four weeks of therapy with a single acceptable dose each week, indicating its powerful anti-neoplastic and anti-resistance characteristics. This study strongly implies that KS18 may treat MM and provide new hope to patients who are experiencing recurrence or resistance. less

Using plants in the biological production of silver nanoparticles (AgNPs) is becoming increasingly important as a practical and environmentally benign method. In the current work, silver nanoparticles were made utilizing the significant Ocimum kilimandscharicum, and their potential to combat certain bacteria was discussed. Both aqueous and methanol plant extracts were used for reducing silver nitrate at different time intervals (30 to 150 minutes) and pH (2 to 11). The UV-visible absorption spectrum recorded for both methanol and aqueous extracts revealed successful synthesis of AgNPs. The antimicrobial activity of the AgNPs was evaluated against Escherichia coli ATCC 25922, Salmonella choleraesuius ATCC 10708, and Staphylococcus aureus ATCC 25923. 50mg/ml was the concentration of the extracts tested. The best zone of inhibition was recorded for the methanol and aqueous-mediated AgNPs, ranging from 12{+/-}1 to 16{+/-}1mm. The methanol and aqueous extract silver nanoparticles had the same Minimum Inhibitory Concentration (6.25{+/-}0.00 mg/ml), whereas the Minimum Bactericidal Concentrations were 12.5{+/-}0.00 and 25{+/-}0.00 mg/ml, respectively. The highest inhibition zone of 16{+/-}1 mm was observed against Salmonella choleraesuius with 50{+/-}0.00 mg/ml aqueous silver nanoparticles. The results show that the silver nanoparticles made with Ocimum kilimandscharicum have potent antibacterial action against those microorganisms. less

Snake envenoming is a major, but neglected, tropical disease. Among venomous snakes, those inducing neurotoxicity such as kraits (Bungarus genus) cause a potentially lethal peripheral neuroparalysis with respiratory deficit in a large number of people each year. In order to prevent the development of a deadly respiratory paralysis, hospitalization with pulmonary ventilation and use of antivenoms are the primary therapies currently employed. However, hospitals are frequently out of reach for envenomated patients and there is a general consensus that additional, non-expensive treatments, deliverable even long after the snake bite, are needed. Traumatic or toxic degenerations of peripheral motor neurons cause a neuroparalysis that activates a pro-regenerative intercellular signaling program taking place at the neuromuscular junction (NMJ). We recently reported that the intercellular signaling axis melatonin-melatonin receptor 1 (MT1) plays a major role in the recovery of function of the NMJs after degeneration of motor axon terminals caused by massive Ca2+ influx. Here we show that the small chemical MT1 agonists: Ramelteon and Agomelatine, already licensed for the treatment of insomnia and depression, respectively, are strong promoters of the neuroregeneration after paralysis induced by krait venoms in mice, which is also Ca2+ mediated. The venom from a Bungarus species representative of the large class of neurotoxic snakes (including taipans, coral snakes, some Alpine vipers in addition to other kraits) was chosen. The functional recovery of the NMJ was demonstrated using electrophysiological, imaging and lung ventilation detection methods. According to the present results, we propose that Ramelteon and Agomelatine should be tested in human patients bitten by neurotoxic snakes acting presynaptically to promote their recovery of health. Noticeably, these drugs are commercially available, safe, non-expensive, have a long bench life and can be administered long after a snakebite even in places far away from health facilities. less

Introduction: Endometriosis is a chronic inflammatory disease that affects ~10% of women. A significant fraction of patients experience limited or no efficacy with current therapies. Tissue adjacent to endometriosis lesions often exhibits increased neurite and vascular density, suggesting that disease pathology involves neurotrophic activity and angiogenesis. Objectives: We aim to evaluate the potential for key tyrosine-kinase-receptor-coupled neurotrophic molecules to contribute to endometriosis-associated pain in mice. Methods: The levels of VEGFR1 regulators (VEGFA, VEGFB, PLGF, and sVEGFR1) were quantified by ELISA in peritoneal fluid from endometriosis patients undergoing surgery and used to calculate VEGFR1 occupancy. We used genetic depletion, neutralizing antibody, and pharmacological approaches to specifically block ligand (NGF or BDNF) or neurotrophic receptor (VEGFR1, TRKs) in a murine model of endometriosis-associated pain. Endometriosis-associated pain was determined using the von Frey filaments method, quantification of spontaneous abdominal pain-related behavior, and thermal discomfort. Diseases parameters were evaluated by lesion size and prevalence. Results: We found that entrectinib (pan-Trk inhibitor) or anti-NGF treatments reduced evoked pain, spontaneous pain, and thermal discomfort. In contrast, even though receptor occupancy revealing that VEGFR1 agonist levels are sufficient to support pain, blocking VEGFR1 signaling via antibody or tamoxifen-induced knockout did not reduce pain or lesion size in mice. Targeting BDNF-TrkB with an anti-BDNF antibody also proved ineffective. Conclusions: This suggests NGF-TrkA signaling, but not BDNF-TrkB or VEGF-VEGFR1, mediates endometriosis-associated pain. Moreover, entrectinib blocks endometriosis-associated pain and reduces lesion sizes. Our results also indicated that entrectinib-like molecules are promising candidates for endometriosis treatment. less

Desensitisation of the mu-opioid receptor (MOR) is proposed to underlie the initiation of opioid analgesic tolerance and previous work has shown that agonist-induced phosphorylation of the MOR C-tail contributes to this desensitisation. Moreover, we and others have shown that phosphorylation is important for {beta}-arrestin recruitment to the receptor, and that ligands of different efficacies induce distinct patterns, or barcodes, of receptor phosphorylation. Within the MOR C-tail, the 370TREHPSTANT379 motif harbours Ser/Thr residues important for these regulatory functions. 375Ser acts as a primary phosphorylation site of a ligand-dependent, hierarchical, and sequential process, whereby flanking 370Thr, 376Thr and 379Thr residues can get subsequently phosphorylated. Here we used HEK293 GRK KO cells, in combination with phosphosite specific antibodies and site-directed mutagenesis of the MOR, to evaluate the contribution of the different GRK subfamilies to ligand-induced phosphorylation barcodes and {beta}-arrestin2 recruitment. We show that both GRK subfamilies (GRK2/3 and GRK5/6) promote phosphorylation of Thr370 and Ser375. However, only GRK2/3 induce phosphorylation of Thr376 and Thr379, which is required to promote robust {beta}-arrestin recruitment to the receptor. Moreover, while DAMGO and fentanyl can engage all kinases to promote Thr370 and Ser375 phosphorylation, under endogenous GRK expression conditions, morphine-induced phosphorylation of these residues is specifically mediated by GRK5/6. These data provide insight into the mechanisms of MOR regulation and suggest that the cellular complement of the different GRK subfamilies plays an important role in determining the tissue responses of distinct opioid agonists. less

Introduction Extrahepatic metabolism/conjugation is characteristic for carboxylic acid drugs. Other challenges for this group of compounds include deconjugation of their metabolites and low and varying unbound fraction in plasma (fu). Thus, it is comparably difficult to estimate their in vivo intrinsic hepatic metabolic clearance (CLint) and hepatic CL (CLH) and to predict their in vivo CLint, CLH and CL. One objective was to investigate the laboratory variability of fu and CLint for carboxylic acids. Another objective was to compare human hepatocytes, measured log D and the software ANDROMEDA with regards to prediction of human in vivo CLint of carboxylic acids. Materials and Methods Measured unbound hepatocyte CLint, non-renal CL (surrogate for CLH), non-renal CLint (surrogate for hepatic metabolic CLint), log D and fu data were taken from studies in the literature. ANDROMEDA (by Prosilico; version 1.0) was used for in silico predictions of CLint for carboxylic acids not used in the training set of the CLint-model. Results and Discussion 13- and 51-fold mean and maximum ratios were observed for highest and lowest reported hepatocyte predicted in vivo CLint, respectively. Corresponding estimates for in vitro fu were 18- and 50-fold, respectively. The data set with the apparently highest number of carboxylic acids contains 39 carboxylic acids with in vitro CLint and log D (both measured at the same laboratory), in vivo CLint and in vitro fu. 18 carboxylic acids were excluded as their in vitro CLint was below the limit of quantification. The correlation coefficient (R2) for log hepatocyte predicted in vivo CLint vs log in vivo CLint was 0.34. The corresponding R2 for log D vs log in vivo CLint was 0.40 (0.47 for 64 carboxylic acids). The Q2 (forward-looking R2) between in silico (ANDROMEDA) predicted and measured log in vivo CLint for 12 carboxylic acids was 0.86. The corresponding R2 for hepatocytes and log D were 0.67 and 0.66, respectively. ANDROMEDA produced a lower maximum prediction error compared to hepatocytes and also predicted the in vivo CLint for all carboxylic acids out of reach for the hepatocyte assay. Conclusion Large interlaboratory variability was demonstrated for fu and hepatocyte assays. Log D, and especially ANDROMEDA, outperformed the hepatocyte assay for the prediction of CLint of carboxylic acids in vivo in man. less

Papillomaviruses infect cutaneous tissue in various species including bovines and from benign warts to malignant squamous cell carcinoma causing severe economic losses to the farmers. The mechanisms by which bovine papillomaviruses interact with host tissue are unclear. Hence in this study using classical network analysis tools, we evaluated interactions of Bovine papilloma (BPV) variants, with markers and receptors implicated in squamous cell carcinoma. Additionally, the thuja phytoconstituents were also evaluated for its potential to target the BPV and squamous cell carcinoma network interactions to understand the mechanism of its clinical benefits. Various protein composition of 14 different virus variants of BPV were assessed against 24 markers of squamous cell carcinoma. Among these interactions EGFR consistently exhibited high-affinity interactions with the E1 protein in all isoforms of BPV. Type 4 BPV displayed the maximum number of binding sites (14) with a binding pocket score ranging from 15.47 to 141.34 and a probability score of 0.75 to 0.99. The comparison of the binding pockets identified that BPV types 2 and 13 had the highest number of common amino acid sequences. Further the alpha helix structure of specific common amino acid sequences, contribute to a more robust and widespread affinity interaction with both E1 of various BPV types and EGFR. Analysis of thuja phytochemicals suggested superior efficacy of Beyerene and Terpinene-4-ol towards all ten BPV targets and bEGFR. In conclusion, our comprehensive study leading to identification of E1 protein of BPV as a major interacting network with bEGFR, their key binding sites, and efficacy of thuja phytoconstituents offer valuable insight into further experimental validation and development of novel therapeutic strategies against BPV-associated diseases. less

Metformin is the most common first-line oral therapeutic agent used in the treatment of type-2 diabetes, one of the most prevalent chronic diseases in North America. Post excretion, the compound enters wastewater treatment plants where it is partially bio-transformed by bacteria into guanylurea. Both metformin and guanylurea enter freshwater environments in wastewater effluent where they are available for uptake by aquatic biota. However, our understanding of the effects of metformin and guanylurea on aquatic life is limited. We tested the hypothesis that metformin and guanylurea can influence the development of zebrafish (Danio rerio), by assessing morphometrics, cardiac development, energetic state, and behaviour of early larvae. Embryos were exposed to environmentally relevant (0.4, 4, 40 g{middle dot}L-1) and supra-environmental (400 and 4000 g{middle dot}L-1) concentrations of metformin and guanylurea from the 4-cell stage (3 hours post fertilization; hpf), until first feed (120 hpf). Exposures to 40 g{middle dot}L-1 and higher of both metformin and guanylurea increased mortality. Metformin delayed hatching at the highest concentration tested (4000 g{middle dot}L-1). The incidence of spinal curvature increased with exposure to both chemicals at supra-environmental levels (400 and 4000 g{middle dot}L-1 for metformin; 400 g{middle dot}L-1 for guanylurea). Metformin and guanylurea exposure imposed slight bradycardia in early development, but did not alter oxygen consumption, ATP levels, carbohydrate levels, general swimming, light-dark movement, startle response, or thigmotaxis, irrespective of exposure concentration. The results suggest similar and low sensitivities of larval fish to both metformin and guanylurea. Apart from a small increase in mortality, these compounds impart a modest impact to the early-life stages of zebrafish that are largely limited to supra-environmental concentrations. less

Objective: Drug-induced liver injury (DILI) is a major failure mode in pharmaceutical development. This study aims to address the limitations of existing preclinical models by introducing a high-throughput, microfluidic liver-on-a-chip system, termed as \"Curio Barrier Liver Chips,\" seeded with human liver organoids to enable metabolic and phenotypic morphologic characterization. Methods: Curio Barrier liver chips, fabricated in an 8x2 well configuration, were utilized to establish 3D liver organoid cultures. Human-induced pluripotent stem cells (iPSCs) were differentiated into hepatic-like organoids (HLOs), and their viability, liver-specific functions, and pharmacological responses were assessed over 28 days. Results: The Curiochips successfully maintained liver physiology and function, showing strong albumin secretion and cytochrome P450 (CYP) activities through 28 days. Unlike traditional models requiring millimolar drug concentrations to detect hepatotoxicity, this platform showed increased sensitivity for APAP and FIAU at micromolar concentrations. In situ differentiation of foregut spheroids to liver organoids was also achieved, further simplifying the establishment of liver chips. Furthermore, the chips demonstrated viability, function and DILI responsiveness for 28 days making this an improved model for studying idiosyncratic DILI with prolonged drug exposure and high-throughput capabilities compared to other available systems or primary human hepatocytes. Conclusions: The Curiochips offer an advanced, miniaturized in vitro model for early-stage drug development and a sensitive, responsive and cost-effective means to detect direct hepatotoxicity. The high-throughput capability, coupled with robust functionality and pharmacological responses make it a promising tool for improving the prediction and understanding of DILI mechanisms in general and those that required prolonged drug exposure. The model also opens new avenues for research in other chronic liver diseases. Keywords: Drug-induced liver injury, hepatotoxicity, microfluidic liver chips, microphysiologic liver model, high-throughput screening, iPSC-derived hepatic-like organoids, chronic DILI model, Curiochips. less