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Metal‐organic framework materials (MOFs) are highly porous and are the subject of growing interest among scientists globally for their utilization in drug delivery. In this work, iron‐based metal‐organic frameworks (MOFs) MIL‐100(Fe) were synthesized by ultrasonic method and studied for the loading of the chloroquine phosphate (CQP). The CQP‐release behavior of the material was investigated in water media with various pH solutions of 1.2, 2, 4.5, and phosphate‐buffered saline (PBS) at temperatures of 25 °C to 45 °C. MIL‐100(Fe) material had a high surface area of up to 1080 m2/g and could completely release the CQP at a pH of 1.2 after 25 hours. At other pH conditions, the drug exhibited an initial rapid release, followed by a gradual slowdown, ultimately achieving complete release after 96 hours. The maximal loading capacity for the CQP by the MIL‐100(Fe) was determined to be approximately 30 % at the pH solution of 2 (stomach environment). The findings from the activity assessment against the K1 malaria parasite strain (P. falciparum) indicated that the concentration at which the active ingredient, when carried by the material, exhibited inhibition was 193 nM/L. The evaluation outcomes for acute toxicity of the drug carrier material revealed an LD50 value exceeding 5000 mg/kg (equivalent to 1500 mg of CQP base). In contrast, when using the active substance alone, acute toxicity resulted in an LD50 value of 675 mg/kg. [ABSTRACT FROM AUTHOR]

Background: Plasmodium vivax is the second most common malaria parasite in Ethiopia. It has been treated with chloroquine (CQ) for the past seven decades. However, the emergence of CQ-resistant strains in the nation urged the Federal Ministry of Health of Ethiopia to review its national malaria treatment guideline in 2018. In the revised guideline, the first-line treatment for uncomplicated P. vivax infection is a combination of CQ and primaquine (PQ). Thus, the present study was designed to evaluate the in vivo efficacy of CQ and PQ combination therapy against clinical P. vivax mono-infection in one of the malaria-endemic areas of Ethiopia. Methods: An open-label prospective clinical trial was conducted in the Limmu Kossa District, Jimma zone, Southwest Ethiopia, from September 2023 to March 2024. A total of 108 patients were recruited for the study. All participants received treatment with CQ at a dosage of 25 mg/kg over three days, followed by PQ at 0.25 mg/kg for 14 consecutive days. Patients were monitored for 42 days for any signs of treatment failure and malaria clinical symptoms, as per the World Health Organization (WHO) guidelines for anti-malarial drug evaluation. Additionally, haemoglobin (Hb) levels, body temperature, any adverse events, and signs of haemolysis were assessed. Data was analysed using R-software (version 4.0.0) and a significant level was considered at p < 0.05. Results: The median age of the patients was 23 years, ranging from 2.5 to 62 years. Of the 108 patients initially recruited, 100 completed the 42-day follow-up period. The combination therapy of CQ and PQ for uncomplicated clinical P. vivax malaria demonstrated excellent therapeutic efficacy, with a 100% cure rate observed at both day 28 and day 42. Additionally, the recommended low dose of PQ (0.25 mg/kg) was well-tolerated, with no signs of. Additionally, most common malaria symptoms were disappeared early in the follow-up period. Conclusion: The combination of CQ plus PQ has exhibited excellent efficacy against uncomplicated P. vivax malaria mono-infections. To preserve this efficacy, it is critical to ensure patients adhere to the full course of PQ treatment, despite its extended duration. Therefore, health authorities should put emphasis on the boosting of the public on the importance of finishing the prescribed medication regimen. [ABSTRACT FROM AUTHOR]

An in vivo mouse model of Clonorchis sinensis (C. sinensis) infection with or without the administration of autophagy inhibitor chloroquine (CQ) stimulation was established to assess the possible involvement of autophagic response during C. sinensis infection. Abnormal liver function was observed at 4, 6, and 8 weeks post-infection, as indicated by elevated levels of ALT/GPT, AST/GOT, TBIL, and α-SMA in the infected groups. These findings indicated that C. sinensis infection activated autophagy, as shown by a decreased LC3II/I ratio and accumulated P62 expression in infected mice. Interestingly, CQ administration exhibited dual and opposing effects during the infection. In the early stage of infection, the engagement of CQ appeared to mitigate symptoms by reducing inflammation and fibrotic responses. However, in the later stage of infection, CQ might contribute to parasite survival by evading autophagic targeting, thereby exacerbating hepatic impairment and worsening liver fibrosis. Autophagy in liver was suppressed throughout the infection. These observations attested that C. sinensis infection triggered autophagy, and highlighted a complex role for CQ, with both protective and detrimental effects, in the in vivo process of C. sinensis infection. [ABSTRACT FROM AUTHOR]

In this paper, we describe our discovery of burnettiene A (1) as an antimalarial compound from the culture broth of Lecanicillium primulinum (current name: Flavocillium primulinum) FKI-6715 strain utilizing our original multidrug-sensitive yeast system. This polyene-decalin polyketide natural product was originally isolated as an antifungal active compound from Aspergillus burnettii. However, the antifungal activity of 1 has been revealed in only one fungal species, and the mechanism of action of 1 remains unknown. After the validation of mitochondrial function inhibitory of 1 , we envisioned a new antimalarial drug discovery platform based on mitochondrial function inhibitory activity. We evaluated antimalarial activity and 1 showed antimalarial activity against Plasmodium falciparum FCR3 (chloroquine sensitive) and the K1 strain (chloroquine resistant). Our study revealed the utility of our original screening system based on a multidrug-sensitive yeast and mitochondrial function inhibitory activity for the discovery of new antimalarial drug candidates. [ABSTRACT FROM AUTHOR]

Simple Summary: The restoration of HLA-class-I expression in lung cancer cells may reverse immune evasion, enhance cytotoxic T-cell activity, and improve immune checkpoint inhibitors' therapeutic efficacy. In the current study, we provide experimental evidence that autophagy blockage either at the late stage (chloroquine and bafilomycin) or early stage of the autophagic process (ULK1 inhibitors and MAP1LC3A silencing) can up-regulate the expression of HLA-class-I molecules in the A549 and H1299 NSCLC cell lines and their CD133+ stem cells. This upregulation enhances the cytotoxic activity of activated CD8+ T-cells, in particular after pre-incubation with anti-PD-L1 monoclonal antibodies. The restoration of HLA-class-I-mediated antigen presentation with autophagy blockers in lung cancer is a promising avenue that urgently requires further exploration in clinical trials. Background/Objectives: Immune checkpoint inhibitors have an established role in non-small cell lung cancer (NSCLC) therapy. The loss of HLA-class-I expression allows cancer cell evasion from immune surveillance, disease progression, and failure of immunotherapy. The restoration of HLA-class-I expression may prove to be a game-changer in current immunotherapy strategies. Autophagic activity has been recently postulated to repress HLA-class-I expression in cancer cells. Methods: NSCLC cell lines (A549 and H1299) underwent late-stage (chloroquine and bafilomycin) and early-stage autophagy blockage (ULK1 inhibitors and MAP1LC3A silencing). The HLA-class-I expression was assessed with flow cytometry, a Western blot, and RT-PCR. NSCLC tissues were examined for MAP1LC3A and HLA-class-I expression using double immunohistochemistry. CD8+ T-cell cytotoxicity was examined in cancer cells pre-incubated with chloroquine and anti-PD-L1 monoclonal antibodies (Moabs); Results: A striking increase in HLA-class-I expression following incubation with chloroquine, bafilomycin, and IFNγ was noted in A549 and H1299 cancer cells, respectively. This effect was further confirmed in CD133+ cancer stem cells. HLA-class-I, β2-microglobulin, and TAP1 mRNA levels remained stable. Prolonged exposure to chloroquine further enhanced HLA-class-I expression. Similar results were noted following exposure to a ULK1 and a PIKfyve inhibitor. Permanent silencing of the MAP1LC3A gene resulted in enhanced HLA-class-I expression. In immunohistochemistry experiments, double LC3A+/HLA-class-I expression was seldom. Pre-incubation of H1299 cancer cells with chloroquine and anti-PD-L1 MoAbs increased the mean % of apoptotic/necrotic cells from 2.5% to 18.4%; Conclusions: Autophagy blockers acting either at late or early stages of the autophagic process may restore HLA-class-I-mediated antigen presentation, eventually leading to enhanced immunotherapy efficacy. [ABSTRACT FROM AUTHOR]

Simple Summary: Recent advancements in anticancer drug treatments face significant challenges due to drug resistance, often leading to only partial or short-term responses in patients. Our previous in vitro studies indicated that autophagy is linked to drug resistance in prostate cancer (PCa). Consequently, combination therapies targeting multiple pathways could help overcome resistance and extend treatment efficacy. To validate our earlier findings, we conducted in vivo experiments. Our data demonstrated that combining Abiraterone (Abi) with an autophagy inhibitor such as Chloroquine (Chl) not only reduces autophagy but also suppresses tumors more effectively than Abi alone. Thus, the combination of Abi with autophagy inhibitors represents a promising therapeutic strategy that could potentially offer a novel approach for patients. Objective: Abi, when used in conjunction with prednisone, is an established treatment for advanced PCa. Our goal was to explore the level of autophagy induced by Abi treatment, both alone and in combination with the autophagy inhibitor Chl, in a castrated mouse xenograft model. Methods: LNCaP cells were injected into the left and right sides of the back of nude mice that had been previously castrated. Mice were divided into four groups and treated daily with intraperitoneal injections of vehicle (control), Abi (10 mg/kg), Abi (10 mg/kg) combined with Chl (10 mg/kg), or Chl (10 mg/kg), and were monitored for periods of 2 and 3 weeks. Results: A significant reduction in tumor weight was observed in mice treated with the combination therapy, as opposed to those receiving vehicle control, Abi, or Chl alone. Mice receiving Abi + Chl exhibited reduced expression of ATG5, Beclin 1, and LC3 punctuations, along with an increase in P62, as determined by immunofluorescence and WES analysis. AR expression decreased significantly in all treatment groups compared to the control. PSMA expression was highest in the vehicle and combined treatment groups after 3 weeks, with a significant reduction observed with Chl treatment. Conclusions: These findings demonstrate that Abi + Chl treatment lowers autophagy levels and suppresses tumors more effectively than Abi alone. [ABSTRACT FROM AUTHOR]

Objective: To evaluate whether hydroxychloroquine (HCQ) or chloroquine (CQ) are effective for the treatment of oral lichen planus (OLP). Materials and Methods: A literature search was conducted in four databases. Clinical studies investigating the effect of HCQ/CQ in patients with OLP were included. Results: Eleven studies were included. Four were RCTs and seven quasi‐experimental studies. The studies included 390 patients diagnosed with OLP, of which 326 and 7 received HCQ and CQ, respectively. 46 patients received topical dexamethasone, 5 placebo and 6 griseofulvin as controls. Five studies assessed pain, and all of them obtained pain reduction with the use of HCQ. Six studies reported objective clinical improvement of OLP with the use of HCQ. Five studies that used a subjective scale obtained that 24%–100% of the patients achieved a complete/almost complete improvement of OLP lesions and its symptomatology. The most frequent side effects were vision problems, gastric discomfort, rash, nauseas, headaches, skin pigmentation, and elevated kidney function. 17 patients had to withdraw from the studies. Conclusions: Current evidence is scarce to confirm HCQ as a therapeutic option for OLP. More RCTs are needed to compare its efficacy with topical corticosteroids and to evaluate whether HCQ reduces relapses of OLP. [ABSTRACT FROM AUTHOR]

Background: Rheumatoid arthritis (RA) is an autoimmune disorder requiring long-term combination therapy with diseasemodifying antirheumatic drugs (DMARDs), non-steroidal anti-inflammatory agents (NSAIDS), low-dose steroids, proton pump inhibitors, calcium, and folic acid and these drugs cause multitude of side effects. This study on the efficacy and tolerability of drugs used in RA among South Indian patients can guide in tailoring a treatment regimen more suited for South Indian population. Aims and Objectives: The main objective of this study was to evaluate the efficacy and tolerability of drugs used in the treatment of RA among South Indian patients. Materials and Methods: In this prospective observational study, 120 newly diagnosed RA patients attending rheumatology outpatient department in a tertiary care center in South India were recruited after obtaining Institutional Ethics Committee approval and informed consent. Sociodemographic details and baseline investigations were done. Each patient was followed up for a period of 6 months wherein, response to treatment was evaluated and adverse drug reactions (ADRs) reported were collected, analyzed and appropriate treatment was instituted. Data analysis was done using SPSS software. Results: One hundred patients completed the study. There was female preponderance (85%) and 38% were rheumatoid factor positive. Efficacy of combination therapy was evidenced falling erythrocyte sedimentation rate (P < 0.001), modified health assessment questionnaire,and disease activity score 28 scorings after 1st, 3rd, and 6th month of therapy (P < 0.001) and improvement in hemoglobin count (P < 0.001). The number of swollen joints was also significantly reduced (P < 0.001). Common ADRs reported were gastritis (29%), mucosal ulcers (10%), macrocytic anemia (3%), chloroquine-related ocular toxicity (2%). Conclusion: NSAIDS and steroids helped to achieve early remission and caused most of adverse effects. Methotrexate and chloroquine had good tolerability and were preferred DMARDs in our center. Thus, we conclude that the combination therapy used in our center is efficacious and well tolerated by South Indian population. [ABSTRACT FROM AUTHOR]

Background: During the coronavirus disease (COVID)-19 pandemic several drugs were used to manage the patients mainly those with a severe phenotype. Potential drugs were used off-label and major concerns arose from their applicability to managing the health crisis highlighting the importance of clinical trials. In this context, we described the mechanisms of the three repurposed drugs [Ivermectin-antiparasitic drug, Chloroquine/Hydroxychloroquine-antimalarial drugs, and Azithromycin-antimicrobial drug]; and, based on this description, the study evaluated the clinical efficacy of those drugs published in clinical trials. The use of these drugs reflects the period of uncertainty that marked the beginning of the COVID-19 pandemic, which made them a possible treatment for COVID-19. Methods: In our review, we evaluated phase III randomized controlled clinical trials (RCTs) that analyzed the efficacy of these drugs published from the COVID-19 pandemic onset to 2023. We included eight RCTs published for Ivermectin, 11 RCTs for Chloroquine/Hydroxychloroquine, and three RCTs for Azithromycin. The research question (PICOT) accounted for P—hospitalized patients with confirmed or suspected COVID-19; I—use of oral or intravenous Ivermectin OR Chloroquine/Hydroxychloroquine OR Azithromycin; C—placebo or no placebo (standard of care); O—mortality OR hospitalization OR viral clearance OR need for mechanical ventilation OR clinical improvement; and T—phase III RCTs. Results: While studying these drugs' respective mechanisms of action, the reasons for which they were thought to be useful became apparent and are as follows: Ivermectin binds to insulin-like growth factor and prevents nuclear transportation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), therefore preventing cell entrance, induces apoptosis, and osmotic cell death and disrupts viral replication. Chloroquine/Hydroxychloroquine blocks the movement of SARS-CoV-2 from early endosomes to lysosomes inside the cell, also, this drug blocks the binding between SARS-CoV-2 and Angiotensin-Converting Enzyme (ACE)-2 inhibiting the interaction between the virus spike proteins and the cell membrane and this drug can also inhibit SARS-CoV-2 viral replication causing, ultimately, the reduction in viral infection as well as the potential to progression for a higher severity phenotype culminating with a higher chance of death. Azithromycin exerts a down-regulating effect on the inflammatory cascade, attenuating the excessive production of cytokines and inducing phagocytic activity, and acts interfering with the viral replication cycle. Ivermectin, when compared to standard care or placebo, did not reduce the disease severity, need for mechanical ventilation, need for intensive care unit, or in-hospital mortality. Only one study demonstrated that Ivermectin may improve viral clearance compared to placebo. Individuals who received Chloroquine/Hydroxychloroquine did not present a lower incidence of death, improved clinical status, or higher chance of respiratory deterioration compared to those who received usual care or placebo. Also, some studies demonstrated that Chloroquine/Hydroxychloroquine resulted in worse outcomes and side-effects included severe ones. Adding Azithromycin to a standard of care did not result in clinical improvement in hospitalized COVID-19 participants. In brief, COVID-19 was one of the deadliest pandemics in modern human history. Due to the potential health catastrophe caused by SARS-CoV-2, a global effort was made to evaluate treatments for COVID-19 to attenuate its impact on the human species. Unfortunately, several countries prematurely justified the emergency use of drugs that showed only in vitro effects against SARS-CoV-2, with a dearth of evidence supporting efficacy in humans. In this context, we reviewed the mechanisms of several drugs proposed to treat COVID-19, including Ivermectin, Chloroquine/Hydroxychloroquine, and Azithromycin, as well as the phase III clinical trials that evaluated the efficacy of these drugs for treating patients with this respiratory disease. Conclusions: As the main finding, although Ivermectin, Chloroquine/Hydroxychloroquine, and Azithromycin might have mechanistic effects against SARS-CoV-2 infection, most phase III clinical trials observed no treatment benefit in patients with COVID-19, underscoring the need for robust phase III clinical trials. [ABSTRACT FROM AUTHOR]

Chemodynamic therapy represents a novel tumor therapeutic modality via triggering catalytic reactions in tumors to yield highly toxic reactive oxygen species (ROS). Nevertheless, low efficiency catalytic ability, potential systemic toxicity and inefficient tumor targeting, have hindered the efficacy of chemodynamic therapy. Herein, a rationally designed catalytic nanoplatform, composed of folate acid conjugated liposomes loaded with copper peroxide (CP) and chloroquine (CQ; a clinical drug) (denoted as CC@LPF), could power maximal tumor cytotoxicity, mechanistically via maneuvering endogenous and exogenous copper for a highly efficient catalytic reaction. Despite a massive autophagosome accumulation elicited by CP-powered autophagic initiation and CQ-induced autolysosomal blockage, the robust ROS, but not aberrant autophagy, underlies the synergistic tumor inhibition. Otherwise, this combined mode also elicits an early onset, above all, long-term high-level existence of immunogenic cell death markers, associated with ROS and aberrant autophagy -triggered endoplasmic reticulum stress. Besides, CC@LPF, with tumor targeting capability and selective tumor cytotoxicity, could elicit intratumor dendritic cells (mainly attributed to CQ) and tumor infiltrating CD8+ T cells, upon combining with PD-L1 therapeutic antibody, further induce significant anti-tumor effect. Collectively, the rationally designed nanoplatform, CC@LPF, could enhance tumor chemoimmunotherapy via deploying endogenous plus exogenous copper and remodeling tumor microenvironment. [ABSTRACT FROM AUTHOR]

Introduction: Many drugs are irritating to the gastric mucosa and induce gastric erosion and when these side effects are coupled with additional parameters, such as bacterial infection, stress, and gastric pH, these together induce gastric ulcer. The present study aimed to evaluate the gastric erosion effects of chloroquine using mice models. Methods: A total of 20 mice were used for this study, divided into two groups of 10 each; the control group was administered only standard food and water, and the chloroquine group was given standard food with water with additional chloroquine solution of 1.2 mg/kg daily orally for a month. The stomachs were dissected and sliced for histological staining and analysis. Results: Chloroquine has remarkably induced tissue degeneration and villi sloughing alongside white blood cell infiltration with patchy areas of stomach erosion compared to the normal architecture of the stomach tissue of the control group. Conclusion: Chloroquine-induced gastric erosion with potential involvement of the many regions of the stomach reaching deep tissue layers. [ABSTRACT FROM AUTHOR]

Currently, a diagnosis with KRAS mutant pancreatic ductal adenocarcinoma (PDAC) means a death warrant, so finding efficient therapeutic options is a pressing issue. Here, we presented that pharmacologic ascorbate, chloroquine and resveratrol co‐treatment exerted a synergistic cytotoxic effect on PDAC cell lines. The observed synergistic cytotoxicity was a general feature in all investigated cancer cell lines independent of the KRAS mutational status and seems to be independent of the autophagy inhibitory effect of chloroquine. Furthermore, it seems that apoptosis and necroptosis are also not likely to play any role in the cytotoxicity of chloroquine. Both pharmacologic ascorbate and resveratrol caused double‐strand DNA breaks accompanied by cell cycle arrest. It seems resveratrol‐induced cytotoxicity is independent of reactive oxygen species (ROS) generation and accompanied by a significant elevation of caspase‐3/7 activity, while pharmacologic ascorbate‐induced cytotoxicity shows strong ROS dependence but proved to be caspase‐independent. Our results are particularly important since ascorbate and resveratrol are natural compounds without significant harmful effects on normal cells, and chloroquine is a known antimalarial drug that can easily be repurposed. [ABSTRACT FROM AUTHOR]

Malaria caused by Plasmodium protozoa, transmitted by the Anopheles mosquitoes' is one ofthe most important diseases. Current antimalarial drugs target vital parasite progressions ormetabolic pathways essential for parasitic development, thus aiding the host in overcomingthe infection by inhibiting protozoa growth. However, at the same time, it also produces adisruptive consequence on the host cells. These cause severe adverse effects on the host andlead to drug resistance. The urgent need for novel, non‐toxic anti‐protozoal compounds isevident due to the resistance developed against drugs like chloroquine andhydroxychloroquine. Metal complexes of various elements like iron, gold, ruthenium, and othershave shown their anti‐malarial potential. We have reviewed the research ongoing globally on the developments of new molecules of chloroquine coupled with different transition elements and describe the structure–activity relationship of chloroquine, which also provides insights into the chemistry of these compounds. [ABSTRACT FROM AUTHOR]

Pyruvate dehydrogenase kinase (PDK), which phosphorylates the pyruvate dehydrogenase complex, regulates glucose metabolism in skeletal muscle. PDK1, an isozyme whose expression is controlled by hypoxia-inducible factor-1α (HIF-1α), is thought to play a role in muscle adaptation to hypoxia. While transcriptional upregulation of PDK1 by HIF-1α is well characterised, mechanisms controlling proteolysis of PDK1 in skeletal muscle have not been thoroughly investigated. Proteasome inhibitor MG132 paradoxically reduced the abundance of PDK1 in human cancer cells and rat L6 myotubes, suggesting that MG132 might direct PDK1 towards autophagic degradation. The objectives of our current study were to determine (1) whether MG132 suppresses PDK1 levels in primary human myotubes, (2) whether chloroquine, an inhibitor of autophagy, prevents MG132-induced suppression of PDK1 in L6 myotubes, and (3) whether PYR-41, an inhibitor of ubiquitination, suppresses PDK1 in L6 myotubes. Using qPCR and/or immunoblotting, we found that despite markedly upregulating HIF-1α protein, MG132 did not alter the PDK1 expression in cultured primary human myotubes, while it suppressed both PDK1 mRNA and protein in L6 myotubes. The PDK1 levels in L6 myotubes were suppressed also during co-treatment with chloroquine and MG132. PYR-41 markedly increased the abundance of HIF-1α in primary human and L6 myotubes, while reducing the abundance of PDK1. In L6 myotubes treated with PYR-41, chloroquine increased the abundance of the epidermal growth factor receptor, but did not prevent the suppression of PDK1. Collectively, our results suggest that cultured myotubes degrade PDK1 via a pathway that cannot be inhibited by MG132, PYR-41, and/or chloroquine. [ABSTRACT FROM AUTHOR]

Abstract Chloroquine and hydroxychloroquine, while effective in rheumatology, pose risks of retinal toxicity, necessitating regular screening to prevent visual disability. The gold standard for screening includes retinal imaging and automated perimetry, with multifocal electroretinography (mfERG) being a recognized but less accessible method. This study explores the efficacy of Artificial Intelligence (AI) algorithms for detecting retinal damage in patients undergoing (hydroxy-)chloroquine therapy. We analyze the mfERG data, comparing the performance of AI models that utilize raw mfERG time-series signals against models using conventional waveform parameters. Our classification models aimed to identify maculopathy, and regression models were developed to predict perimetric sensitivity. The findings reveal that while regression models were more adept at predicting non-disease-related variation, AI-based models, particularly those utilizing full mfERG traces, demonstrated superior predictive power for disease-related changes compared to linear models. This indicates a significant potential to improve diagnostic capabilities, although the unbalanced nature of the dataset may limit some applications.

Abstract Background Plasmodium vivax is the second most common malaria parasite in Ethiopia. It has been treated with chloroquine (CQ) for the past seven decades. However, the emergence of CQ-resistant strains in the nation urged the Federal Ministry of Health of Ethiopia to review its national malaria treatment guideline in 2018. In the revised guideline, the first-line treatment for uncomplicated P. vivax infection is a combination of CQ and primaquine (PQ). Thus, the present study was designed to evaluate the in vivo efficacy of CQ and PQ combination therapy against clinical P. vivax mono-infection in one of the malaria-endemic areas of Ethiopia. Methods An open-label prospective clinical trial was conducted in the Limmu Kossa District, Jimma zone, Southwest Ethiopia, from September 2023 to March 2024. A total of 108 patients were recruited for the study. All participants received treatment with CQ at a dosage of 25 mg/kg over three days, followed by PQ at 0.25 mg/kg for 14 consecutive days. Patients were monitored for 42 days for any signs of treatment failure and malaria clinical symptoms, as per the World Health Organization (WHO) guidelines for anti-malarial drug evaluation. Additionally, haemoglobin (Hb) levels, body temperature, any adverse events, and signs of haemolysis were assessed. Data was analysed using R-software (version 4.0.0) and a significant level was considered at p

INTRODUCTION: Potentially inappropriate medications (PIMs) for older adults cause more adverse effects than benefits. The 2019 American Geriatrics Society Beers Criteria (2019BC) considered five clinical situations as PIM use in older adults. Can drug analysis, according to these situations, assist in the act of making prescriptions for older people? Seeking a practical example for this question, we assessed drugs currently questioned as to their safe use among older people. OBJECTIVE: To check if chloroquine and hydroxychloroquine fit the PIM criteria for older adults and whether this analysis is clinically applicable. METHODS: We systematized the objective based on the five clinical situations defined as PIM use in older adults by the 2019BC. RESULTS: Chloroquine and hydroxychloroquine fulfill, respectively, four and five of these clinical situations. This evaluation allowed the likely definition of these drugs as PIMs for older adults in a simple way, based on a brief analysis of the available literature. CONCLUSION: Chloroquine and hydroxychloroquine may be considered PIMs for older adults. We expect that this analysis can be replicated with other drugs and reduce iatrogenesis in older people.

Abstract Gambogic acid is a natural bioactive compound from the brownish resin of the Garcinia hanburyi trees, which has been shown to induce cell death of cancer cells with a synergistic effect together with other compounds. In the present study, we aim to investigate whether the combination of gambogic and chloroquine, one of the inhibitor of autophagy, could increase cell death in human NSCLC cells. As a result, gambogic acid and chloroquine synergistically suppress the growth of NSCLC cells (CI

Abstract Chemodynamic therapy represents a novel tumor therapeutic modality via triggering catalytic reactions in tumors to yield highly toxic reactive oxygen species (ROS). Nevertheless, low efficiency catalytic ability, potential systemic toxicity and inefficient tumor targeting, have hindered the efficacy of chemodynamic therapy. Herein, a rationally designed catalytic nanoplatform, composed of folate acid conjugated liposomes loaded with copper peroxide (CP) and chloroquine (CQ; a clinical drug) (denoted as CC@LPF), could power maximal tumor cytotoxicity, mechanistically via maneuvering endogenous and exogenous copper for a highly efficient catalytic reaction. Despite a massive autophagosome accumulation elicited by CP-powered autophagic initiation and CQ-induced autolysosomal blockage, the robust ROS, but not aberrant autophagy, underlies the synergistic tumor inhibition. Otherwise, this combined mode also elicits an early onset, above all, long-term high-level existence of immunogenic cell death markers, associated with ROS and aberrant autophagy -triggered endoplasmic reticulum stress. Besides, CC@LPF, with tumor targeting capability and selective tumor cytotoxicity, could elicit intratumor dendritic cells (mainly attributed to CQ) and tumor infiltrating CD8+ T cells, upon combining with PD-L1 therapeutic antibody, further induce significant anti-tumor effect. Collectively, the rationally designed nanoplatform, CC@LPF, could enhance tumor chemoimmunotherapy via deploying endogenous plus exogenous copper and remodeling tumor microenvironment. Graphical abstract

Abstract Background Gastric cancer (GC) is a prevalent malignant tumor, and the RNA-binding protein polypyrimidine tract-binding protein 1 (PTBP1) has been identified as a crucial factor in various tumor types. Moreover, abnormal autophagy levels have been shown to significantly impact tumorigenesis and progression. Despite this, the precise regulatory mechanism of PTBP1 in autophagy regulation in GC remains poorly understood. Methods To assess the expression of PTBP1 in GC, we employed a comprehensive approach utilizing western blot, real-time quantitative polymerase chain reaction (RT–qPCR), and bioinformatics analysis. To further identify the downstream target genes that bind to PTBP1 in GC cells, we utilized RNA immunoprecipitation coupled with sequencing (si-PTBP1 RNA-seq). To evaluate the impact of PTBP1 on gastric carcinogenesis, we conducted CCK-8 assays, colony formation assays, and GC xenograft mouse model assays. Additionally, we utilized a transmission electron microscope, immunofluorescence, flow cytometry, western blot, RT–qPCR, and GC xenograft mouse model experiments to elucidate the specific mechanism underlying PTBP1’s regulation of autophagy in GC. Results Our findings indicated that PTBP1 was significantly overexpressed in GC tissues compared with adjacent normal tissues. Silencing PTBP1 resulted in abnormal accumulation of autophagosomes, thereby inhibiting GC cell viability both in vitro and in vivo. Mechanistically, interference with PTBP1 promoted the stability of thioredoxin-interacting protein (TXNIP) mRNA, leading to increased TXNIP-mediated oxidative stress. Consequently, this impaired lysosomal function, ultimately resulting in blockage of autophagic flux. Furthermore, our results suggested that interference with PTBP1 enhanced the antitumor effects of chloroquine, both in vitro and in vivo. Conclusion PTBP1 knockdown impairs GC progression by directly binding to TXNIP mRNA and promoting its expression. Based on these results, PTBP1 emerges as a promising therapeutic target for GC. Graphical Abstract

The authors have advised there are errors in the Y-axis of Figures 1, 2 and 4 on pages 1097, 1099 and 1101, respectively. The Y-axis label “Body weight (kg)” in figure parts 1B, 2C and 4c should read “Body weight (g)”. The Y-axis label “Th2 cell counts (×103)” in figure parts 1G, 2H and 4H should read “Proportion of Th2 cells (%)”. The correct figures are as follows. Figure 1 Continued. Figure 1 CQ relieved MC903-induced type 2 inflammatory response in AD mice. (A) The severity of ear skin lesions; (B) dermatitis severity score; (C) left ear thickness; (D) H&E staining; (E) TB staining; (F) ELISA measurements of serum TSLP, IL-4, IL-13, IFN-γ, and IgE levels; (G) flow cytometry to determine the content of peripheral blood Th2 cells (CD3+CD4+CD193+). N = 6. Data were represented as mean ± standard deviation, one-way ANOVA was for data comparisons in panels A/D-G, with Tukey’s test serving for post hoc testing. ns represented P > 0.05, *P < 0.05, **P < 0.01, and ***P < 0.001. Two-way ANOVA was adopted in panels B-C, and Tukey’s multiple comparisons test was implemented for post hoc test. *Represented comparisons with the Control group, *P < 0.05, ***P < 0.001, #Represented comparisons with the AD group, #P < 0.05, ##P < 0.01. Figure 2 Continued. Figure 2 CQ abated type 2 inflammatory response in AD mice by inactivating TLR3. (A) Western blot detection of TLR3 protein expression; (B) the severity of ear skin lesions; (C) dermatitis severity score; (D) left ear thickness; (E) H&E staining; (F) TB staining; (G) ELISA detections of serum TSLP, IL-4, IL-13, IFN-γ, and IgE levels; (H) flow cytometry to assess the content of Th2 cells (CD3+CD4+CD193+). N = 6. Data were represented as mean ± standard deviation. Data in panel A were tested by one-way ANOVA, followed by post hoc testing using Tukey’s test. Two-way ANOVA was used for panels (C and D), and Šídák’s multiple comparisons test was used for post hoc test. Data in panels B/E-H were examined by independent sample t-test. ns represented P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001. Figure 4 Continued. Figure 4 Activation of NLRP3 partially averted the alleviating effect of CQ on type 2 inflammatory response in AD mice. (A) Western blot to test the expression levels of NLRP3, ASC and cleaved caspase-1 proteins; (B) the severity of ear skin lesions; (C) dermatitis severity score; (D) left ear thickness; (E) H&E staining; (F) TB staining; (G) ELISA detection of serum TSLP, IL-4, IL-13, IgE, IL-1β and IL-18 levels; (H) flow cytometry to assess the content of Th2 cells (CD3+CD4+CD193+). N = 6. Data were represented as mean ± standard deviation, and independent sample t-test was employed for comparisons in panels A-B/E-H. Two-way ANOVA was used in panels C-D, and Šídák’s multiple comparisons test was conducted for post hoc test. *P < 0.05, **P < 0.01. These corrections do not significantly impact the overall findings and conclusions of the paper. We would like to assure readers that the corrected values and labels do not alter the interpretations or validity of the research. The authors sincerely apologize for any confusion these errors may have caused.

Abstract Background Plasmodium vivax malaria is still an important public health problem in Ethiopia. Unlike Plasmodium falciparum, P. vivax has a dormant liver stage (hypnozoite) that can be a risk of recurrent vivax malaria unless treated by radical cure with primaquine. Drug resistance to chloroquine is threatening malaria control and elimination efforts. This study assessed the therapeutic efficacy and safety of chloroquine plus 14 days of primaquine on P. vivax infection based on parasitological, clinical, and haematological parameters. Methods A single-arm in vivo prospective therapeutic efficacy study was conducted to assess the clinical and parasitological response to the first-line treatment of P. vivax in Ethiopia, chloroquine plus 14 days low dose of (0.25 mg/kg/day) primaquine between December 2022 and March 2023 at Hamusit Health Centre using the standard World Health Organization (WHO) protocol. A total of 100 study participants with P. vivax mono-infection who were over 6 months old were enrolled and monitored for adequate clinical and parasitological responses for 42 days. The WHO double-entry Excel sheet and SPSS v.25 software were used for Kaplan–Meier survival analysis, and a paired t-test was used for analysis of haemoglobin improvements between follow up days. Results A total of 100 patients were enrolled among those, 96% cases were rural residents, 93% had previous malaria exposure, and predominant age group was 5–15 years (61%). 92.6% (95% CI 85.1–96.4%) of enrolled patients were adequate clinical and parasitological response, and 7.4% (95% CI 3.6–14.9%) recurrences were observed among treated patients. The fever and parasite clearance rate on day 3 were 98% and 94%, respectively. The baseline haemoglobin levels improved significantly compared to those days 14 and 42 (p

Yanqing Liu,1,&ast; Yuqing Meng,1,&ast; Junzhe Zhang,1 Liwei Gu,1 Shengnan Shen,1 Yongping Zhu,1 Jigang Wang1,2 1State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-Di Herbs, Artemisinin Research Center, and Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, People’s Republic of China; 2Department of Pharmacological Sciences, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 117600, Singapore&ast;These authors contributed equally to this workCorrespondence: Jigang Wang; Yongping Zhu, State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Artemisinin Research Center, and Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, People’s Republic of China, Email jgwang@icmm.ac.cn; ypzhu@icmm.ac.cnAbstract: Chloroquine is a common antimalarial drug and is listed in the World Health Organization Standard List of Essential Medicines because of its safety, low cost and ease of use. Besides its antimalarial property, chloroquine also was used in anti-inflammatory and antivirus, especially in antitumor therapy. A mount of data showed that chloroquine mainly relied on autophagy inhibition to exert its antitumor effects. However, recently, more and more researches have revealed that chloroquine acts through other mechanisms that are autophagy-independent. Nevertheless, the current reviews lacked a comprehensive summary of the antitumor mechanism and combined pharmacotherapy of chloroquine. So here we focused on the antitumor properties of chloroquine, summarized the pharmacological mechanisms of antitumor progression of chloroquine dependent or independent of autophagy inhibition. Moreover, we also discussed the side effects and possible application developments of chloroquine. This review provided a more systematic and cutting-edge knowledge involved in the anti-tumor mechanisms and combined pharmacotherapy of chloroquine in hope of carrying out more in-depth exploration of chloroquine and obtaining more clinical applications.Keywords: Chloroquine, pharmacology, applications, combined pharmacotherapy

Objective To evaluate the effects of chloroquine alone and in combination with traditional antifungal agents on the Candida albicans biofilms and its drug resistance. Methods This study used standard strains of Candida albicans, and drug-resistant strains of Candida albicans. The inhibitory effects of chloroquine alone and in combination with antifungal drugs on biofilms of Candida albicans were detected by XTT reduction method and chessboard dilution method respectively. The morphological characteristics of biofilms were observed under scanning electron microscopy. Results Chloroquine at the concentration of 50 μmol/L or above showed a direct inhibitory effect and increased with concentration. Chloroquine combined with amphotericin B had a synergistic inhibitory effect. Results of the time-killing curve showed that the growth trends of biofilms treated with chloroquine alone and combined with amphotericin B varied in different time periods during the experimental culture. Morphological observation also revealed that chloroquine alone and in combination with amphotericin B could reduce the ability of Candida albicans to form hyphae and biofilms. Conclusion Chloroquine has an inhibitory effect on Candida albicans biofilms and can reduce its drug resistance. Furthermore, chloroquine shows a synergistic anti-fungal effect when combined with amphotericin B.

Pharmaceutically active compounds are the most widely produced and consumed consumer products that pose a substantial threat to the environment and living organisms owing to their pharmacokinetics, side effects, and contraindications. In this study, the degradation of chloroquine (CQ), a popular antimalarial and recently proposed COVID-19 drug, was investigated under electron beam (EB) irradiation of aqueous solutions. Both the hydroxyl radical and hydrated electron generated in the radiolysis of water contribute to the degradation of CQ in aqueous solution. The overall removal efficiency for 125 mg·L-1 of the CQ solution under EB treatment is reported to be >80% at neutral pH at a maximum irradiation dose of 7 kGy. Removal efficiency is further favored by acidic and slightly alkaline conditions where reactions with hydroxyl radicals and hydrated electrons are favored, respectively. Additionally, increments in the applied dose resulted in the increased removal efficiency for the same concentration of CQ. Conversely, the removal efficiency decreased with increasing concentration of CQ at the same irradiation dose. The initial solution pH, applied irradiation dose, and initial pollutant concentration play an important role in the EB-induced degradation of CQ by influencing the available oxidizing and reducing species. The chemical oxygen demand (COD) and total organic carbon (TOC) were not significantly decreased during the treatment process and indicated the formation of organic byproducts, which were not further degraded under the current experimental conditions.

Abstract Background Plasmodium vivax malaria is a leading cause of morbidity in Ethiopia. The first-line treatment for P. vivax is chloroquine (CQ) and primaquine (PQ), but there have been local reports of CQ resistance. A clinical study was conducted to determine the efficacy of CQ for the treatment of P. vivax malaria in southern Ethiopia. Methods In 2021, patients with P. vivax mono-infection and uncomplicated malaria were enrolled and treated with 25 mg/kg CQ for 3 consecutive days. Patients were followed for 28 days according to WHO guidelines. The data were analysed using per-protocol (PP) and Kaplan‒Meier (K‒M) analyses to estimate the risk of recurrent P. vivax parasitaemia on day 28. Results A total of 88 patients were enrolled, 78 (88.6%) of whom completed the 28 days of follow-up. Overall, 76 (97.4%) patients had adequate clinical and parasitological responses, and two patients had late parasitological failures. The initial therapeutic response was rapid, with 100% clearance of asexual parasitaemia within 48 h. Conclusion Despite previous reports of declining chloroquine efficacy against P. vivax, CQ retains high therapeutic efficacy in southern Ethiopia, supporting the current national treatment guidelines. Ongoing clinical monitoring of CQ efficacy supported by advanced molecular methods is warranted to inform national surveillance and ensure optimal treatment guidelines.

AbstractPurposeMethodsResultsConclusionAutophagy dysregulation triggers extracellular matrix remodeling via changes in cellular collagen levels and protease secretion. However, the effect of autophagy on scleral extracellular matrix remodeling in the context of myopia is not fully understood. In this study, we measured the level of autophagy in sclera of form deprivation myopic guinea pigs; we also sought a correlation between the level of autophagy in human scleral fibroblasts and the extent of COL1A1 synthesis.We measured the level of COL1A1 expression and the levels of autophagic protein markers in scleral tissues in vivo using a form deprivation myopic guinea pig model. Rapamycin and chloroquine were respectively used to activate and inhibit autophagy in cultured human scleral fibroblasts. COL1A1 gene and protein expression levels were analyzed via quantitative real-time polymerase chain reaction, Western blotting, and immunofluorescence. Levels of autophagy-related proteins were assessed via Western blotting.The sclera of form deprivation myopic guinea pig eyes exhibited decreased expression of COL1A1 and increased expression level of autophagy. After chloroquine exposure, human scleral fibroblasts exhibited decreased autophagy and increased COL1A1 expression.Inhibition of scleral fibroblast autophagy increased COL1A1 expression at the gene and protein levels, thus explaining the effect of autophagy on collagen synthesis by scleral fibroblasts. [ABSTRACT FROM AUTHOR]

Background: Gastric cancer (GC) is a prevalent malignant tumor, and the RNA-binding protein polypyrimidine tract-binding protein 1 (PTBP1) has been identified as a crucial factor in various tumor types. Moreover, abnormal autophagy levels have been shown to significantly impact tumorigenesis and progression. Despite this, the precise regulatory mechanism of PTBP1 in autophagy regulation in GC remains poorly understood. Methods: To assess the expression of PTBP1 in GC, we employed a comprehensive approach utilizing western blot, real-time quantitative polymerase chain reaction (RT–qPCR), and bioinformatics analysis. To further identify the downstream target genes that bind to PTBP1 in GC cells, we utilized RNA immunoprecipitation coupled with sequencing (si-PTBP1 RNA-seq). To evaluate the impact of PTBP1 on gastric carcinogenesis, we conducted CCK-8 assays, colony formation assays, and GC xenograft mouse model assays. Additionally, we utilized a transmission electron microscope, immunofluorescence, flow cytometry, western blot, RT–qPCR, and GC xenograft mouse model experiments to elucidate the specific mechanism underlying PTBP1's regulation of autophagy in GC. Results: Our findings indicated that PTBP1 was significantly overexpressed in GC tissues compared with adjacent normal tissues. Silencing PTBP1 resulted in abnormal accumulation of autophagosomes, thereby inhibiting GC cell viability both in vitro and in vivo. Mechanistically, interference with PTBP1 promoted the stability of thioredoxin-interacting protein (TXNIP) mRNA, leading to increased TXNIP-mediated oxidative stress. Consequently, this impaired lysosomal function, ultimately resulting in blockage of autophagic flux. Furthermore, our results suggested that interference with PTBP1 enhanced the antitumor effects of chloroquine, both in vitro and in vivo. Conclusion: PTBP1 knockdown impairs GC progression by directly binding to TXNIP mRNA and promoting its expression. Based on these results, PTBP1 emerges as a promising therapeutic target for GC. [ABSTRACT FROM AUTHOR]

This research introduces a novel electrochemical sensor designed for the detection of chloroquine. The sensor was developed via a simple method to synthesize TiO2 nanoparticles on reduced graphene oxide. The modified electrodes created in this manner exhibited high electrocatalytic activity and distinct chemical reactivity. Various techniques were utilized to perform morphological characterization of the nanocomposites. These techniques revealed alterations revealed changes in functional groups and the attachment of titanium to the reduced graphene oxide present on the electrode surface, thereby elucidating the reasons for the enhanced electrochemical performance. The sensor had a broad measurement range for chloroquine, capable of detecting concentrations as low as 10−8 M. It is applicable for diverse sample analyses, including water, pharmaceuticals, human urine, and serum, with satisfaction ranging between 97 and 99%. The development strategy. An electrochemical sensor was developed using titanium oxide nanoparticles attached to reduced graphene oxide TiO2-NPs @RGO/GCE for real-time detection of chloroquine in real contaminated samples such as human urine, human serum, and tap water with low detection limit. [ABSTRACT FROM AUTHOR]

The HSP70 co-chaperone BAG3 targets unfolded proteins to degradation via chaperone assisted selective autophagy (CASA), thereby playing pivotal roles in the proteostasis of adult cardiomyocytes (CMs). However, the complex functions of BAG3 for regulating autophagy in cardiac disease are not completely understood. Here, we demonstrate that conditional inactivation of Bag3 in murine CMs leads to age-dependent dysregulation of autophagy, associated with progressive cardiomyopathy. Surprisingly, Bag3 -deficient CMs show increased canonical and non-canonical autophagic flux in the juvenile period when first signs of cardiac dysfunction appear, but reduced autophagy during later stages of the disease. Juvenile Bag3 -deficient CMs are characterized by decreased levels of soluble proteins involved in synchronous contraction of the heart, including the gap junction protein Connexin 43 (CX43). Reiterative administration of chloroquine (CQ), an inhibitor of canonical and non-canonical autophagy, but not inactivation of Atg5 , restores normal concentrations of soluble cardiac proteins in juvenile Bag3 -deficient CMs without an increase of detergent-insoluble proteins, leading to complete recovery of early-stage cardiac dysfunction in Bag3 -deficient mice. We conclude that loss of Bag3 in CMs leads to age-dependent differences in autophagy and cardiac dysfunction. Increased non-canonical autophagic flux in the juvenile period removes soluble proteins involved in cardiac contraction, leading to early-stage cardiomyopathy, which is prevented by reiterative CQ treatment. [Display omitted] • Inactivation of Bag3 in CMs initiates hypertrophic cardiomyopathy in juvenile mice • Juvenile but not late stage Bag3 -deficient CMs show increased autophagic flux • Inactivation of Atg5 does not improve cardiac function in juvenile Bag3 -cKO mice • Treatment with chloroquine normalizes cardiac function in juvenile Bag3 -cKO mice • Human BAG3P209L heart samples show similar changes as in Bag3 -cKO mice [ABSTRACT FROM AUTHOR]

Objectives To evaluate the effect of antimalarial drugs in response to therapy, incidence of LN flares, and progression of kidney disease in a large LN cohort. Methods We retrospectively studied 424 biopsy-proven LN patients followed for >3 years. We obtained demographic, clinical, laboratory, histopathological and treatment variables. Antimalarial use was approached as (i) users vs no users, (ii) according to prevalent vs incident use regarding the LN flare and (iii) according to the type of antimalarial. All outcomes were evaluated by time-to-event analyses. Adjusted hazard ratios were obtained by Cox regression. Results The cohort included 424 patients, median age of 29 years (IQR 23–37), 96% female, with a median eGFR of 81 ml/min/1.73 m2 (IQR 48–118) and proteinuria of 3.4 g/g (IQR 1.9–5.5). Antimalarial use was associated with higher complete response (aHR 1.57, 1.08–2.27), lower incidence of kidney flares (aHR 0.63, 0.43–0.92) and lower progression to kidney failure (aHR 0.37, 0.23–0.53). The effect of antimalarials on these outcomes was modified by the presentation eGFR, histological class and/or concomitant initial immunosuppressor. These protective effects were observed in patients with prevalent or incident use regarding the LN flare and patients using hydroxychloroquine. The incidence of toxic retinopathy was 1.7%, 5.7% and 8.8% by 3, 5 and 7 years of continued antimalarial use, respectively. Conclusion The use of antimalarial drugs is associated with increased response to therapy, lower incidence of kidney flares, and lower progression to kidney failure in LN patients. Conversely, this population is at high risk of toxic maculopathy, and yearly ophthalmologic examination is recommended. [ABSTRACT FROM AUTHOR]

Chloroquine (CQ) has been a good treatment for antimalarial mainstay for several decades; additionally discovered, it has a significant therapeutic impact on some instances of fungal inhibition. This study focused on the effect of novel CQ compounds on the activity of two different species of fungi, Aspergillus niger and Aspergillus falves. The activity of each CQ derivative was monitored using Nuclear magnetic resonance spectroscopy (NMR), minimal inhibitory concentration and physical parameters such as optical microscopy, UV/visible absorbance and optical band gap. NMR indicated the conjugation between the substrate and amino acid. The optical microscopic images indicated homogeneously distributed and uniform density distribution of CQ-derivative particles on the glass substrates. The samples' presented absorption peaks at 203, 207 and 220 nm wavelengths, suggesting the important electronic transition with reducing the indirect bandgap from 4.1 eV to 3.95 eV. These compounds have the best antifungal growth inhibitory properties and excellent features, indicating cosmetics use. [ABSTRACT FROM AUTHOR]

Plasmodium vivax causes the largest malaria burden in Brazil, and chloroquine resistance poses a challenge to eliminating malaria by 2035. Illegal mining in the Roraima Yanomami Indigenous territory can lead to the introduction of resistant parasites. This study aimed to investigate mutations in the pvcrt-o and pvmdr-1 genes to determine their potential as predictors of P. vivax chloroquine-resistant phenotypes. Samples were collected in two health centers of Boa Vista. A questionnaire was completed, and blood was drawn from each patient. Then, DNA extraction, PCR, amplicon purification, and DNA sequencing were performed. After alignment with the Sal-1, the amplified fragment was analyzed. Patients infected with the mutant parasites were queried in the Surveillance Information System. Among the patients, 98% (157/164) of participants were from illegal mining areas. The pvcrt-o was sequenced in 151 samples, and the K10 insertion was identified in 13% of them. The pvmdr1 was sequenced in 80 samples, and the MYF haplotype (958M) was detected in 92% of them and the TYF was detected in 8%, while the MYL was absent. No cases of recrudescence, hospitalization, or death were found. Mutations in the pvcrt-o and pvmdr-1 genes have no potential to predict chloroquine resistance in P. vivax. [ABSTRACT FROM AUTHOR]

Simple Summary: Weaning stress is a critical factor contributing to diseases and even mortality in piglets during the weaning process. Research indicates that inhibition of intestinal autophagy to a certain extent can alleviate the decline in growth performance associated with weaning stress by enhancing the management of intestinal oxidative stress and inflammation. Interestingly, gut microbes serve an irreplaceable role in the regulation of autophagy. Therefore, we speculate that intestinal microbes may play a direct or indirect role in the process of intestinal autophagy inhibition, alleviating weaning stress in piglets. In this study, we utilized autophagy inhibitors (chloroquine, CQ) or activators (rapamycin, RAPA) to modulate the intestinal autophagy level in weaned piglets. We then investigated how changes in autophagy impacted the composition of intestinal microbes and their metabolites in these piglets, allowing us to explore the effects of autophagy from a microbial perspective. We found that the level of autophagy affected the composition of intestinal microbes and metabolites of weaned piglets, which may be one of the key factors in how autophagy alleviates weaning stress in piglets. Our findings also provide some insights to may guide the future application of autophagy inhibitors in piglets' diets. Our previous study demonstrated that moderate inhibition of intestinal autophagy was beneficial to alleviate early weaning stress in piglets, but the detailed mechanism behind this was unclear. Microbiota-mediated enterocyte autophagy helps maintain intestinal homeostasis. This study investigated the effects of inhibition or activation of autophagy in intestinal microbial community compositions and metabolite profiles in piglets. Eighteen 24-day-old weaned piglets were divided into three groups (each treatment of six piglets) and treated daily with rapamycin (RAPA), chloroquine (CQ) or a control volume of normal saline (CON group). Before the formal trial, the piglets were allowed to acclimatize for 3 days, and then the trial period was 14 days. Collected samples from the ileum and colon underwent 16S rRNA gene sequencing and metabolite analysis. Significant differences in microbial composition were observed in both the ileum and colon of the RAPA and CQ groups compared to the CON group (p < 0.05). In addition, the relative levels of abundance of Peptostreptococcus, Fusobacterium, Dialister, Selenomonas and Oceanobacillus in the ileum and Porphyromonas, Bacteroides, unidentified_Lachnospiraceae, Akkermansia, Sharpea, Peptococcus, Pseudoalteromonas, Peptoclostridium and unidentified_Acidobacteria in the colon were improved in piglets fed the RAPA diet, whereas the relative levels of abundance of Turicibacter, Rickettsiella and Sarcina in the ileum and Roseburia and Kroppenstedtia in the colon were enhanced in the CQ group (p < 0.05). Meanwhile, metabolomic analysis showed that there were significant differences in metabolites among all groups (p < 0.05), and KEGG enrichment analysis revealed that differential metabolites were mainly enriched in the ABC transporters and biosynthesis of amino acids pathways. Furthermore, these metabolites were closely related to differential microorganisms (p < 0.05). Overall, autophagy inhibition regulates the composition of intestinal microorganisms and their metabolites, and these differential metabolites are significantly correlated with differential intestinal microorganisms, which may in turn affect the production performance of weaned piglets. [ABSTRACT FROM AUTHOR]

G‐protein‐coupled receptors (GPCRs) belonging to the type 2 taste receptors (TAS2Rs) family are predominantly present in taste cells to allow the perception of bitter‐tasting compounds. TAS2Rs have also been shown to be expressed in human airway smooth muscle (ASM), and TAS2R agonists relax ASM cells and bronchodilate airways despite elevating intracellular calcium. This calcium "paradox" (calcium mediates contraction by pro‐contractile Gq‐coupled GPCRs) and the mechanisms by which TAS2R agonists relax ASM remain poorly understood. To gain insight into pro‐relaxant mechanisms effected by TAS2Rs, we employed an unbiased phosphoproteomic approach involving dual‐mass spectrometry to determine differences in the phosphorylation of contractile‐related proteins in ASM following the stimulation of cells with TAS2R agonists, histamine (an agonist of the Gq‐coupled H1 histamine receptor) or isoproterenol (an agonist of the Gs‐coupled β2‐adrenoceptor) alone or in combination. Our study identified differential phosphorylation of proteins regulating contraction, including A‐kinase anchoring protein (AKAP)2, AKAP12, and RhoA guanine nucleotide exchange factor (ARHGEF)12. Subsequent signaling analyses revealed RhoA and the T853 residue on myosin light chain phosphatase (MYPT)1 as points of mechanistic divergence between TAS2R and Gs‐coupled GPCR pathways. Unlike Gs‐coupled receptor signaling, which inhibits histamine‐induced myosin light chain (MLC)20 phosphorylation via protein kinase A (PKA)‐dependent inhibition of intracellular calcium mobilization, HSP20 and ERK1/2 activity, TAS2Rs are shown to inhibit histamine‐induced pMLC20 via inhibition of RhoA activity and MYPT1 phosphorylation at the T853 residue. These findings provide insight into the TAS2R signaling in ASM by defining a distinct signaling mechanism modulating inhibition of pMLC20 to relax contracted ASM. [ABSTRACT FROM AUTHOR]