Prolonged Administration Research Articles

Gefitinib Reverses PD-L1-Mediated Immunosuppression Induced by long-term Glutamine blockade in Bladder Cancer.

Glutamine is a major energy source for tumor cells and blocking glutamine metabolism is being investigated as a promising strategy for cancer therapy. However, the antitumor effect of glutamine blockade in bladder cancer remains unclear, necessitating further investigation. Here, we demonstrated that glutamine metabolism was involved in the malignant progression of bladder cancer. Treatment with the glutamine antagonist 6-Diazo-5-oxo-L-norleucine (DON) inhibited the growth of bladder cancer cells in vitro in several ways. In addition, we observed inhibition of tumor growth in bladder cancer-bearing mice using JHU083, a prodrug that was designed to prevent DON-induced toxicity. However, the antitumor immune effect of T cells changed from activation to inhibition as the administrated time extended. We found that both in vitro treatment with DON and in vivo prolonged administration of JHU083 led to the upregulation of PD-L1 in bladder cancer cells. Mechanistically, glutamine blockade up-regulated PD-L1 expression in bladder cancer cells by accumulating ROS, subsequently activating the EGFR/ERK/C-Jun signaling pathway. Combination treatment of JHU083 and gefitinib reversed the up-regulation of PD-L1 in bladder cancer cells induced by prolonged glutamine blockade, resulting in the alleviation of T-cell immunosuppression and a significant improvement in therapeutic outcome. These preclinical findings show promise for glutamine metabolism targeting as a viable therapeutic strategy for bladder cancer, with the potential for further enhancement through combined treatment with gefitinib.

PROTOCOLOS TERAPÊUTICOS PARA O TRATAMENTO DA ESPOROTRICOSE FELINA REFRATÁRIA E INOVAÇÕES DECORRENTES DAS PESQUISAS

Sporotrichosis in felines is much more pathogenic than in other animals or in humans, even if immunocompetent, which makes treatment a challenge in clinical medicine, as many of them are refractory to the preferred therapy such as Itraconazole or Ketoconazole. This situation also arises from the prolonged oral administration of the medication, the animal's reluctance to take the drug and the guardian's abandonment of the prescription when he notices an improvement in the lesions caused by the disease, which generates relapses and resistance to the antifungals available on the market. Itraconazole, as well as Ketoconazole, act by causing permeability of the plasma membrane and cell necrosis, however, they have adverse effects such as nausea and vomiting in the first and liver disease as relevance in the second. Terbinafine works by interrupting the synthesis of ergosterol and as a consequence the cell membrane ruptures and death, however, it also has side effects such as gastrointestinal and skin problems. Potassium iodide can be used in combination with azoles or Terbinafine, although the literature does not agree with the issue, as not all felines respond to treatment, adverse effects are due to iodism such as depression, anorexia, vomiting and diarrhea. There are also alternative treatments that can enhance oral medication, such as laser therapy, ozone therapy, cryotherapy and cryosurgery. Due to the issue of non-responsive felines, there is research being developed aiming to provide new therapeutic horizons for these animals, such as the use of intranasal infusion of 1% Clotrimazole spray associated with Itraconazole added with Potassium iodide or not, with good results, no relapses and no side effects. NikZ, the result of a substance produced by the bacterium Streptomyces tendae, called Nikkomycin Z, also appears as a new therapeutic potential for the treatment of Sporotrix spp., with a blocking effect on chitin synthase present in the cell wall of fungi, were maide research with dogs, rats and is in Phase 1 of experiments on humans. NikZ represents a new form of prescription, as it can be placed in the animals' drinking water, avoiding the desertion of those responsible for the felines, it has no observable adverse effects in the analyzes carried out and represents an alternative for cats refractory to existing medical protocols in veterinary.

Differential Effects of Intrahippocampal Administration of Ceftriaxone on Morphine Dependence and Withdrawal Syndrome in Rats.

Glutamate is a key factor in opiate addiction. Glial glutamate transporter-1 (GLT-1) plays a prominent role in glutamate homeostasis. Therefore, different regimens of ceftriaxone as a GLT-1 activator were prescribed to determine whether modulating GLT-1 prevents morphine dependence or withdrawal syndrome. Rats received 10 mg/kg morphine subcutaneously for ten consecutive days. Intrahippocampal ceftriaxone (0.5 μL of 0.5 mM solution) was injected 30 min before morphine administration to assess its effect on dependence process. In the next experiment, after the animals became dependent, ceftriaxone was injected before or after the last morphine administration, and its effect on withdrawal symptoms was evaluated. The reversibility of developed dependence was evaluated in the conditions when morphine and ceftriaxone were administered simultaneously. Two hours after the last morphine injection, naloxone hydrochloride (1.5 mg/kg) was administered, and morphine withdrawal syndrome was recorded for 25 min. Ceftriaxone administration before each morphine injection caused a decrease in the occurrence of withdrawal symptoms. Single dose of ceftriaxone after or before the last dose of morphine did not change the withdrawal symptoms significantly. Ceftriaxone injection for 5 days after becoming dependent could decrease the occurrence of some withdrawal symptoms. Modulation of glutamate with ceftriaxone during morphine injection may be able to prevent dependence. However, a single dose of ceftriaxone after becoming dependent could not decrease withdrawal syndrome. More prolonged administration of ceftriaxone could alleviate the induced dependence.

The Design Features, Quality by Design Approach, Characterization, Therapeutic Applications, and Clinical Considerations of Transdermal Drug Delivery Systems—A Comprehensive Review

Transdermal drug delivery systems (TDDSs) are designed to administer a consistent and effective dose of an active pharmaceutical ingredient (API) through the patient’s skin. These pharmaceutical preparations are self-contained, discrete dosage forms designed to be placed topically on intact skin to release the active component at a controlled rate by penetrating the skin barriers. The API provides the continuous and prolonged administration of a substance at a consistent rate. TDDSs, or transdermal drug delivery systems, have gained significant attention as a non-invasive method of administering APIs to vulnerable patient populations, such as pediatric and geriatric patients. This approach is considered easy to administer and helps overcome the bioavailability issues associated with conventional drug delivery, which can be hindered by poor absorption and metabolism. A TDDS has various advantages compared to conventional methods of drug administration. It is less intrusive, more patient-friendly, and can circumvent first pass metabolism, as well as the corrosive acidic environment of the stomach, that happens when drugs are taken orally. Various approaches have been developed to enhance the transdermal permeability of different medicinal compounds. Recent improvements in TDDSs have enabled the accurate administration of APIs to their target sites by enhancing their penetration through the stratum corneum (SC), hence boosting the bioavailability of drugs throughout the body. Popular physical penetration augmentation methods covered in this review article include thermophoresis, iontophoresis, magnetophoresis, sonophoresis, needle-free injections, and microneedles. This review seeks to provide a concise overview of several methods employed in the production of TDDSs, as well as their evaluation, therapeutic uses, clinical considerations, and the current advancements intended to enhance the transdermal administration of drugs. These advancements have resulted in the development of intelligent, biodegradable, and highly efficient TDDSs.

Effect of hydrocortisone on the course of leprosic infection in an experiment

Leprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae), accompanied by a violation of various parts of the immune system. In this regard, the development of an experimental model of leprosy may be based on the creation of an artificial defect of the macrophage link, in particular, using glucocorticoids. Aim: To develop and characterize a model of generalized leprosy infection in mice against the background of prolonged administration of hydrocortisone.Material and methods. To conduct the study, the following groups were formed: the control group – 50 mice infected by the Shepard method; the experimental group – 50 mice similarly infected against the background of hydrocortisone administration. Hydrocortisone was administered intramuscularly in a single daily dose of 25 mg/kg on the first day 2 hours before infection with M. leprae suspension, then in the form of four courses of 2 weeks at two-week intervals.Results. The administration of hydrocortisone led to an acceleration of the reproduction of mycobacteria in the paw pad compared with the control (without hydrocortisone). The generalization of leprosy infection is confirmed by the presence of lepromatous structures in the form of granulomas from macrophages with a high content of M. leprae in liver, spleen, lungs and kidneys.Conclusion. This model allows us to study the properties of the pathogen and the mechanisms of pathogenesis of the disease, and to screen pharmacological drugs with potential anti-leprosy activity in the future.

Subchronic Toxicity Assessment of Zingiber roseum Rhizome in Mice Model: Safety Evaluation at Various Doses

Aims:: The aim of this study is to determine the potential adverse effects associated with the prolonged administration of Zingiber roseum rhizome extract. Background:: This study aimed to evaluate the sub-chronic toxicity of Z. roseum, commonly known as rosy ginger, using a mouse model. Z. roseum has been traditionally used for its medicinal properties; however, there is limited information regarding its potential toxic effects. Objective:: The objective of this study is to assess the safety profile of ZRR extract at various doses and conduct a detailed analysis of hematological, biochemical, and histological parameters regarding sub-chronic toxicity. Methods:: Mice were orally administered ZRR methanolic extract at doses of 300, 600, and 1200 mg/kg for 14 days as per the guidelines of ‘The Brazilian Agency of National Health Surveillance.’ Subchronic toxicity was conducted by monitoring multiple indicators, including changes in body weight, food and water consumption, blood profile (HB, RBC, WBC, and PLT), and biochemical markers (ALT, AST, ALP, TP, ALB, TC, TG, HDL, LDL, Creatinine, and Urea) and histopathological examination of the liver. Results:: Throughout the study, the mice showed normal behavior and appeared healthy. The administration of Z. roseum at all tested doses did not significantly affect body weight, food, and water intake, blood, biochemical markers, or liver. Z. roseum at these doses was safe, with no fatalities or harm. Conclusion:: Lastly, the sub-chronic administration of Z. roseum at doses of 300, 600, and 1200 mg/kg in a mice model did not elicit any toxic effects, indicating its potential safety as a therapeutic agent.

Perioperative Antibiotic Prophylaxis Duration in Patients Undergoing Cystectomy With Urinary Diversion

Perioperative antibiotic prophylaxis (PAP) reduces the incidence of surgical site infections (SSIs). Guidelines recommend 24 hours or less of PAP for cystectomy with urinary diversion; however, evidence specifying optimal PAP duration for this surgery is lacking, and prolonged administration of PAP is common practice. To establish noninferiority of 24-hour PAP vs extended-duration PAP in preventing SSIs within 90 days after cystectomy with urinary diversion. This single-center, noninferiority randomized clinical trial was performed from April 18, 2018, to August 18, 2022, in patients aged older than 18 years undergoing elective open cystectomy with urinary diversion. Exclusion criteria were contraindications to administered drugs and inability to follow study procedures. PAP administered for 24 hours (24-hour PAP group) vs PAP until all catheters and stents were removed (extended PAP group). The primary end point was the rate of SSI, and the secondary end points included all-cause mortality, both within 90 days after surgery. Noninferiority of the 24-hour PAP treatment was assessed by comparing the 90% CI (corresponding to a significance level of α = .05) with the predefined noninferiority margin of 10%. A total of 95 patients were randomly assigned to the 24-hour PAP group (median [IQR] age, 69.3 [63.1-76.8] years; 66 males [69.5%]) and 98 to the extended PAP group (median [IQR] age, 69.5 [60.8-75.5] years; 68 males [69.4%]). Patients in the 24-hour PAP group received PAP for a median of 1 day (IQR, 1-1 day), and patients in the extended PAP group received PAP for a median of 8 days (IQR, 7-10 days). No significant differences in SSIs occurring within 90 days were found (24-hour PAP group, 8 patients [8.4%]; extended PAP group, 12 patients [12.2%]; P = .53). The risk difference for 90-day cumulative SSI incidence was -3.8% (90% CI, -11.1% to 3.4%), establishing noninferiority of 24-hour PAP vs extended PAP to prevent SSI. Mortality was not significantly different between groups. The findings of this randomized clinical trial demonstrate noninferiority of 24-hour PAP vs extended-duration PAP in preventing SSIs within 90 days after cystectomy with urinary diversion and may contribute to antibiotic stewardship efforts in urology. ClinicalTrials.gov Identifier: NCT03305627.

Edible ultrasmall polyphenolic nanozymes for oral treatment of alcohol-induced acute gastritis

Excessive alcohol consumption and prolonged administration of non-steroidal anti-inflammatory drugs (NSAIDs) lead to gastritis which affects millions globally, yet safe and effective treatment options remain limited. While a range of nanozymes have been employed in the oral treatment of inflammatory bowel disease (IBD), the research directed toward gastritis therapy remains unexplored. The challenges of rapid gastric emptying, harsh gastric acid erosion, and the non-edible ingredient constitution hinder the practical use of nanozymes in this context. Herein, three types of sub-10 nm ultrasmall iron-polyphenol nanozymes (UIPNs) were synthesized by using fully edible natural active polyphenols, iron ions and PVP. UIPNs rapidly penetrated the gastric mucosa, resided in the stomach for over 12 h, and remained stable in the harsh acidic environment. They exhibited excellent enzyme-like activities of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD). Compared to omeprazole, all three types of UIPNs showed superior activity, with Fis UIPNs demonstrating the best performance in preventing and protecting against alcohol-induced gastric mucosal injury in mice. The fully food-sourced components, constructed into ultrasmall nanozymes via non-covalent interactions, ensure long-term safety and hold promise for playing significant roles in various gastrointestinal diseases.

Acute Hematogenous Osteomyelitis in Pediatric Patients.

This article focuses on the advancements made in diagnostic techniques and drug interventions of acute hematogenous osteomyelitis. A diagnosis necessitates a combination of factors, including inflammatory markers and imaging findings, as well as the collection of specimens for culture when feasible. Subsequently, treatment should be based on epidemiology, mechanisms of resistance, and susceptibility findings. A brief course of intravenous (IV) antibiotics, followed by oral antibiotics, may be employed for uncomplicated infections if there is improvement in the clinical condition and a decline in C-reactive protein levels. However, for complex infections caused by methicillin-resistant Staphylococcus aureus, prolonged administration of IV antibiotics is recommended, along with surgical intervention if necessary. [Pediatr Ann. 2024;53(10):e392-e395.].

Pharmacokinetic comparison of subcutaneously administered CT-P13 (biosimilar of infliximab) via autoinjector and pre-filled syringe in healthy participants.

CT-P13, a biosimilar of infliximab, is used to treat inflammatory diseases that arise from immune system complications, resulting in excessive and persistent inflammation. The subcutaneous (SC) formulation of CT-P13 overcomes the drawback of prolonged administration associated with the intravenous (IV) infliximab biosimilar, necessitating autoinjector (AI) administration. This randomized, open-label, two-arm, parallel-group, single-dose clinical pharmacology study aimed to evaluate the pharmacokinetics (PK) and safety of CT-P13 SC administration via AI compared with the existing pre-filled syringe (PFS) method. A total of 147 healthy participants were randomized into two groups, of which 139 completed the study. Blood samples were collected from before CT-P13 SC administration to 2016 h after the start of the administration. Serum concentrations were analyzed using the Meso Scale Discovery electrochemiluminescence method. Geometric mean ratios (90% confidence interval) of the AUCinf (areas under the concentration-time curve from zero to infinity) and Cmax (The maximum serum concentration) for CT-P13 SC AI versus CT-P13 SC PFS groups, were 94.15% (85.02%-104.26%), 92.48% (84.66%-101.01%), respectively. CT-P13 SC AI and CT-P13 SC PFS achieved comparable PK because the 90% CI was within the predefined equivalence margin. At the end of the study, immunogenicity results revealed that 70 (97.22%) and 73 (98.65%) participants tested positive for anti-drug antibody (ADA) in the CT-P13 SC AI and CT-P13 SC PFS groups, respectively. They were tested positive for neutralizing antibodies. No other significant safety differences were observed between the treatment groups. In conclusion, both administrations demonstrated PK equivalence and were both safe and well-tolerated.

Clinical characteristics and risk factors associated with diverse manifestations of COVID-19 in patients aged 85 years and older.

The goal of this study is to assess the clinical attributes exhibited by patients aged 85 years and older who present different manifestations of COVID-19, and to examine the factors influencing the classification of the disease severity. This retrospective study was conducted at a single center, encompassing an analysis of clinical data obtained from patients with COVID-19 admitted to a general geriatric hospital in Hangzhou, Zhejiang, China, during the period from December 20, 2022, to February 1, 2023. The study focused on 91 eligible patients whose disease severity was compared based on the imaging findings. A total of 91 patients aged 85 years and older, with a median age of 92, including 46 males, 10 exhibiting mild symptoms, 48 moderate cases, and 33 severe cases met the inclusion criteria. Notably, disease severity displayed a significant correlation with age (p < 0.011). All patients presented with complicated chronic underlying conditions and a history of prolonged medication use. Rheumatic immune diseases (p = 0.040) and endocrine medications, primarily hypoglycemic agents (p = 0.034), exhibited statistical significance. Additionally, markers such as lactate dehydrogenase (LDH) (p = 0.030), interleukin 6 (IL-6) (p = 0.014), and D-dimer (p = 0.007) revealed significant associations with disease severity. Chest computed tomography scans predominantly revealed inflammatory features (n = 81, 89.0%). Notably, patients classified as having mild or moderate conditions exhibited eventual improvement, while 13 patients (39.4%) among the severe cases succumbed to the disease. The classification of disease among patients aged 85 years or older old is correlated with advanced age, concurrent rheumatic immune diseases, and prolonged administration of endocrine medications. Furthermore, patients with elevated levels of LDH, IL-6, and D-dimer demonstrated a higher propensity for developing severe diseases.

Extracts and Terpenoids from Stevia Species as Potential Anthelmintics for Neglected Tropical Diseases Caused by Cestode Parasites.

Cestodes are etiological agents of neglected diseases such as echinococcosis and cysticercosis, which are major public health problems. Antiparasitic treatment relies on a small number of approved drugs, which are often only partially effective, poorly tolerated and require prolonged administration. Thus, the discovery of novel potential treatments is critical. The Stevia genus (Asteraceae) includes species that are recognized as a source of bioactive compounds, with many species associated with medicinal uses. In this study, the cestocidal activity of four South American Stevia species that previously showed antiprotozoal activity was analyzed using a motility assay on the laboratory cestode model, Mesocestoides vogae. The four Stevia extracts showed cestocidal activity, with S. alpina var. alpina as the most active. The sesquiterpene lactones estafietin and eupatoriopicrin were purified from S. alpina var. alpina and S. maimarensis, respectively, and tested on M. vogae. Estafietin showed cestocidal activity, inhibiting parasite viability in a dose-dependent manner, even from the first day of incubation. Consistent with the motility effects, the extract of S. alpina var. alpina and estafietin induced marked alterations in the morphology of the parasite. The results of this report show that Stevia species represent a source of new molecules with potential for the treatment of neglected tropical diseases caused by cestodes.

The Effect of Subchronic Methamphetamine Administration on the NLRP3 Inflammasome, Memory Functions, and Hippocampal Morphology

Background: While it is established that addictive doses of methamphetamine correlate with inflammation-mediated neurotoxic pathways, the extent of toxicity resulting from subchronic administration at lower doses remains uncertain. Objectives: This study aimed to investigate the subtle effects of daily subchronic methamphetamine (MA) administration on neuroinflammatory processes, cognitive dimensions, gene expressions, and hippocampal morphology. Methods: The experimental study employed a longitudinal design with three groups (G1B, G2B, G3B) receiving methamphetamine (5 mg/kg, intraperitoneally, once daily) for 1, 2, or 3 weeks, respectively. Corresponding control groups (G1A, G2A, G3A) received 0.2 mL of normal saline. Spatial learning, novel object recognition, and passive avoidance tests were conducted to assess spatial, recognition, and fear avoidance memories. Hippocampal morphology was evaluated using Nissl staining, and the expressions of NLRP3, ASC, and caspase-1 genes were measured as markers of neuroinflammation. Results: Statistical analyses, including one-way and two-way ANOVA, showed that subchronic low-dose administration of MA led to significant activation of the inflammasome (NLRP3, ASC, and caspase-1), which may have resulted in pyramidal cell death in the hippocampus. The hippocampal structure in the CA1 region was completely disrupted. Spatial memory and passive avoidance learning were impaired in the MA groups, while recognition memory remained unaffected. Conclusions: The findings suggest that prolonged administration of 5 mg/kg of methamphetamine may be associated with significant inflammasome activation, pyramidal cell death, and mild cognitive decline. Contrary to previous evidence, even lower doses of methamphetamine taken over an extended period could be neurotoxic.

Abstract A070: Time-Dependent Oxidation and Aggregation of Vimentin upon Binding by β-lap Induced Immunogenic Cell Death

Abstract β-lapachone (β-lap) has been demonstrated as an NAD(P)H:quinone oxidoreductase1 (NQO1) bioactivatable compound. It initiated a futile redox cycle catalyzed by NQO1, resulting in reactive oxygen species (ROS) generation, with consumption of the reductive NAD(P)H. After a 4-h pulse of sufficiently lethal dose of β-lap, high levels of ROS diffused into nuclei to induce tumor-selective DNA damage and programmed necrosis in cancers overexpressing NQO1. Recently, our experiments showed that prolonged low dose β-lap administration exerted significant and NQO1 independent antitumor efficacy in mouse models with no overt signs of toxicity. Thus, we synthesized a photoactive β-lap probe to identify the molecular target of β-lap. Vimentin, a major constituent of the intermediate filament family of proteins, was identified as the direct binding target of β-lap. Vimentin was highly upregulated in multiple cancers including PDAC, and was a binding partner of β-lap with a dissociation constant at micromolar levels. Ligand binding assays in Panc1 cells employing β-lap as a specific competitor showed that vimentin was bound by β-lap-probe in a β-lap-competitive manner, and further confirmed that the in vivobinding target of β-lap was vimentin. Moreover, it was manifested that endogenous vimentin mRNA levels directly correlated with sensitivity to prolonged exposure of β-lap in a panel of pancreatic cancer cell lines, and knockout of vimentin in Panc1 cells markedly decreased lethality after prolonged exposure of β-lap. Mechanistically, we revealed that β-lap bound to vimentin and induced disulfide bonds that participated in disulfide cross-linking between a pair of vimentin dimers in a concentration and time dependent manner, forming vimentin oligomers of approximately 150 kDa. Prolonged exposure of β-lap promoted the disruption of filament architecture and vimentin degradation, ultimately resulting in cell death. In addition, the cell death by targeting vimentin exhibited a more inflammatory condition and enhanced DC activation. Together, our findings unveiled an unknown molecular target for β-lap and provided valuable insights into its mechanism of action. Targeting vimentin also presented a new strategy that inhibited the tumor growth and synergized with immunotherapy to disrupt immune evasion and reduce immune suppression against pancreatic cancers. Citation Format: Feng Qian, Lei Dou, Kaixin Wang, Yi Xia, Zhidong Chen, Yumeng Zu, Shuang Xi. Time-Dependent Oxidation and Aggregation of Vimentin upon Binding by β-lap Induced Immunogenic Cell Death [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A070.

Acute and chronic effects of methanolic extract of Teucrium polium on blood parameters and histopathology of liver and kidney in Female Rats

Teucrium polium L. is commonly used as a medicinal plant in Algeria to fight various human diseases. In this study, extracting aerial parts of T. polium with methanol gave a dry matter yield of 8.24 %. The effects of methanolic extract of T. polium "TPME" were examined per os on female rats Albino Wistar for six weeks. Biochemical and hematological serum parameters and morphology and histopathology of organs of treated rats were studied. Acute toxicity showed low toxicity with a lethal dose of 50% LD50 &gt; 2400 mg / Kg of body weight. These data can be used to classify these plants as slightly toxic. However, sub-acute treatment for six weeks of rats with 75, 150, and 300 mg of methanolic extract / Kg of body weight significantly increased the most studied hematological parameters. Biochemical analysis revealed a significant increase of renal parameters (urea, creatinine, uric acid, Na, and K), accompanied by an increase in the relative weight of kidney, lipidic (cholesterol), and hepatic glutamate oxaloacetate transaminase (GOT) values in all treated rats. Histopathological examination confirmed the biochemical tests by the observation of perilobular necrosis areas, bile duct and inflammatory infiltration of the liver, and the presence of marked intracytoplasmic vacuoles in the kidney with the dose 300 mg of TPME Kg of animal body weight. Use of Teucrium polium L. may cause hepatotoxicity and nephrotoxicity after prolonged herb administration. Keywords: Teucrium polium L, TPME, LD50, biochemical parameters, toxicity

Abstract A051: Innate immune cell dynamics and pancreatic tumor progression in alcoholic pancreatitis

Abstract Introduction: Persistent inflammation due to alcoholic chronic pancreatitis (ACP) is associated with pancreatic ductal adenocarcinoma (PDAC). Myeloid cell infiltration within the pancreas, particularly with oncogenic mutant KrasG12D/+ (*Kras), is crucial for driving PDAC carcinogenesis. Our previous work demonstrated that hyperactivation of cyclic AMP response element binding protein (CREB) accelerates ADM/PanIN progression with heightened fibroinflammation. However, the association between CREB in precursor neoplastic lesions and innate myeloid cells remains unclear. Methods: We established an ACP mouse model using an alcohol diet along with prolonged caerulein administration in Ptf1aCreERTM/+; LSL-KrasG12D/+ (KC), and CREB deleted [Crebfl/fl in KC (KCC-/-)] mice. Single-cell RNA sequencing (scRNA-seq) of the mouse pancreas, multiparametric immunophenotyping of tumor-infiltrating myeloid cells and chromatin immunoprecipitation (ChIP-seq) with CREB pulldown were performed. Cytokine arrays and qPCR studies followed co-culture assays using mouse neutrophils (J774M) and macrophages (RAW 264.7) with epithelial cell lines harboring *Kras with in vitro ACP induction were conducted. Results: ACP induction in KC mice led to increased infiltration of neutrophils and alternatively activated macrophages with the pancreas. Conversely, Creb-deleted mice exhibited significantly reduced myeloid cell infiltration despite ACP induction. ChIP-Seq and qPCR identified CREB-regulated chemokines and immunomodulators as mediators of myeloid cell trafficking. Co-culture of mouse neutrophils and macrophages with KC-ACP conditioned media significantly increased IL-1 receptor antagonist (Il-1ra) expression and secretion. Single-cell transcriptomic analysis revealed heightened Il1rn mRNA levels in neutrophils and macrophages in KC-ACP when compared with KC pancreatic tumors. Creb-deleted PDAC models showed strong downregulation of Il1rn in macrophages, confirming CREB’s role in modulating myeloid cell interactions via epithelial cell signaling. Conclusions: Our findings highlight that CREB activation in epithelial cells under ACP conditions facilitates crosstalk with myeloid cells, promoting IL-1ra upregulation in neutrophils and macrophages. This CREB-driven regulation of IL-1ra in myeloid cells contributes to inflammation-associated tumorigenesis. Targeting this axis may offer a novel therapeutic approach to curb pancreatic cancer progression. Citation Format: Siddharth Mehra, Sudhakar Jinka, Varun Krishnamoorthy, Supriya Srinivasan, Anna Bianchi, Andrew Adams, Haleh Amirian, Manan Patel, Jashodeep Datta, Nipun Merchant, Nagaraj Nagathihalli. Innate immune cell dynamics and pancreatic tumor progression in alcoholic pancreatitis [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A051.

Design, Synthesis, molecular dynamic analysis, and In-Vivo anti-diabetic evaluation of novel hydrazine carboximidamide derivatives

Diabetes is one of the most rapidly growing health problems globally, and researchers working hard to develop novel efficient therapies against it. In this regard, hydrazine carboximidamide has a great deal of attention owing to its diverse biological activities. Herein, a new series of N, N-disubstituted hydrazine carboximidamide derivatives was designed, synthesized, and assessed for their antidiabetic activity. Docking studies were performed to initially evaluate the binding of the designed compounds to the target enzyme. It was found that all of the pivotal ligand-enzyme interactions, including the interactions with Glu205/Glu206 aminoacids were observed in docking studies. The molecular dynamic analysis showed that the antidiabetic activity of the newly developed compounds may be implemented through the inhibition of dipeptidyl peptidase 4 (DPP-4), an important target that involves enhancing insulin release and suppressing glucagon discharge from the pancreas. The target molecules were synthesized practically and all the newly-developed chemical structures were confirmed via spectrophotometric methods, including IR, LC-MS, 1H NMR, and 13C NMR techniques. Finally, the in-vivo studies were carried out to confirm the results obtained from the docking and molecular dynamics studies. Compounds 6b and 6d demonstrated improved glucose tolerance through oral glucose tolerance tests in NMRI mice (at a dose of 10 mg/kg). Furthermore, prolonged administration of compounds 6b and 6d to diabetic Wistar rats for 14 days led to a noteworthy reduction in blood levels of glucose, akin to the effects observed with sitagliptin, a standard diabetes medication.

A Portable Fluid Administration System for Prolonged Intravenous Fluid Administration in Subadult and Adult White Rhinoceroses (Ceratotherium simum).

While translocations of white rhinoceroses have become an important conservation tool, dehydration during long-distance transports has been identified as a welfare concern. Intravenous (iv) fluid administration might therefore be useful to mitigate dehydration; however, special requirements need to be met to make iv fluid administration suitable for large, wild rhinoceroses during transport. Requirements include a portable and robust system that is capable of delivering high flow rates, is easy to set up, and remains patent and operating for long periods of time while allowing the animals to freely stand or lay down in the transport crates. Due to the lack of suitable fluid administration systems, we developed a custom-made system consisting of 8 L drip bags, a three-part, 4.4-m-long, large bore and partially coiled administration set, and a robust, battery-operated infusion pump, which allowed us to successfully administer iv fluids at a maintenance rate of 1-2 mL/kg/h to eight rhinoceroses for 24 h during a mock transport. While iv fluid administration in transported rhinoceroses is time intensive and the large amount of drip bags required during lengthy transports might pose a limitation, the developed system may be useful for the long-distance transport of small groups of rhinoceroses. Furthermore, this system would be of value for injured or sick rhinoceroses, which require parenteral fluid therapy when commercially available infusion pumps cannot provide the large fluid volumes needed.

Prolonged administration of oral phenylbutazone and firocoxib in horses has no impact on selected cytokine and growth factor concentrations in platelet-rich plasma and autologous protein solution.

To determine the effects of prolonged administration of the oral NSAIDs phenylbutazone and firocoxib on concentrations of cytokines and growth factors in platelet-rich plasma (PRP) and autologous protein solution (APS). 6 adult University owned horses. Horses were randomized to receive phenylbutazone (1 g, orally, q 12 h) or firocoxib (57 mg, orally, q 24 h) for 6 days. Blood was obtained and processed for APS (Pro-Stride) and PRP (Restigen) before the administration of NSAIDs and at 7 days (1 day following cessation of NSAIDs). Horses underwent a two-week washout period, during which blood was obtained at 14 days and 21 days. The protocol was repeated with a crossover design. PRP and APS were analyzed for concentrations of platelets, leukocytes, and several cytokines (IL-1β, IL-10, IL-6, IL-8, and tumor necrosis factor-α) and growth factors (PDGF, FGF-2, and TGF-β1) using immunoassays. Plasma was evaluated for drug concentrations. No significant differences existed in concentrations of growth factors and cytokines before or after prolonged administration of NSAIDs. There were significant differences in concentrations of leukocytes and platelets in PRP compared to APS, with higher concentrations of leukocytes at the day 7 time point (T) in APS (phenylbutazone) and in concentrations of platelets in APS at T0 (firocoxib) and in APS at T7 (phenylbutazone). Veterinarians can recommend the administration of these oral NSAIDs prior to obtaining blood for PRP and APS provided a single-day washout period is instituted.

Caffeic acid derivative MPMCA suppresses osteoclastogenesis and facilitates osteoclast apoptosis: implications for the treatment of bone loss disorders.

Osteoclast activity plays a crucial role in the pathological mechanisms of osteoporosis and bone remodeling. The treatment of these disorders involves the use of pharmacological medicines that work by inhibiting the activity of osteoclasts. Nevertheless, the prevalent and infrequent negative consequences of current antiresorptive and bone anabolic treatments pose significant drawbacks, hence restricting their prolonged administration in patients, particularly those who are elderly and/or suffer from many medical conditions. We are currently in the process of creating a new molecule called N-(4-methoxyphen) methyl caffeamide (MPMCA), which is a derivative of caffeic acid. This compound has shown potential in preventing the production of osteoclasts and causing existing osteoclasts to undergo cell apoptosis. Our investigation discovered that MPMCA hinders osteoclast function via suppressing the MAPK pathways. The expectation is that the findings of this study will stimulate the advancement of a novel approach to treating anti-resorption.