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Methods in Molecular Biology (Clifton,... 2024We present an approach based on photoacoustic lifetime imaging (PALI) to map the distribution of oxygen partial pressure (pO) in tissue. This method utilizes methylene...
We present an approach based on photoacoustic lifetime imaging (PALI) to map the distribution of oxygen partial pressure (pO) in tissue. This method utilizes methylene blue, a dye widely used in clinical applications, as an oxygen-sensitive imaging agent. PALI measurement of oxygen relies upon pO-dependent excitation lifetime of the dye. The technique maps the excited triplet state of oxygen-sensitive dye, thus reflecting the spatial and temporal distributions of tissue oxygen.
Topics: Humans; Diagnostic Imaging; Spectrum Analysis; Hypoxia; Oxygen; Methylene Blue
PubMed: 38319575
DOI: 10.1007/978-1-0716-3633-6_10 -
Cell Death & Disease Dec 2023Glioblastoma (GBM) is the most frequent and lethal brain tumor, whose therapeutic outcome - only partially effective with current schemes - places this disease among the...
Glioblastoma (GBM) is the most frequent and lethal brain tumor, whose therapeutic outcome - only partially effective with current schemes - places this disease among the unmet medical needs, and effective therapeutic approaches are urgently required. In our attempts to identify repositionable drugs in glioblastoma therapy, we identified the neuroleptic drug chlorpromazine (CPZ) as a very promising compound. Here we aimed to further unveil the mode of action of this drug. We performed a supervised recognition of the signal transduction pathways potentially influenced by CPZ via Reverse-Phase Protein microArrays (RPPA) and carried out an Activity-Based Protein Profiling (ABPP) followed by Mass Spectrometry (MS) analysis to possibly identify cellular factors targeted by the drug. Indeed, the glycolytic enzyme PKM2 was identified as one of the major targets of CPZ. Furthermore, using the Seahorse platform, we analyzed the bioenergetics changes induced by the drug. Consistent with the ability of CPZ to target PKM2, we detected relevant changes in GBM energy metabolism, possibly attributable to the drug's ability to inhibit the oncogenic properties of PKM2. RPE-1 non-cancer neuroepithelial cells appeared less responsive to the drug. PKM2 silencing reduced the effects of CPZ. 3D modeling showed that CPZ interacts with PKM2 tetramer in the same region involved in binding other known activators. The effect of CPZ can be epitomized as an inhibition of the Warburg effect and thus malignancy in GBM cells, while sparing RPE-1 cells. These preclinical data enforce the rationale that allowed us to investigate the role of CPZ in GBM treatment in a recent multicenter Phase II clinical trial.
Topics: Humans; Glioblastoma; Chlorpromazine; Pyruvate Kinase; Cell Line, Tumor; Energy Metabolism
PubMed: 38092755
DOI: 10.1038/s41419-023-06353-3 -
Indian Journal of Pathology &... 2023The Fine needle aspiration cytology (FNAC) is considered as a valuable and distinguished diagnostic test in the initial assessment of the patients presenting with a mass...
BACKGROUND
The Fine needle aspiration cytology (FNAC) is considered as a valuable and distinguished diagnostic test in the initial assessment of the patients presenting with a mass in the head and neck region or when a recurrence is suspected after previous treatment.
AIMS
This study was therefore designed to elucidate the efficacy of FNAC as an alternate diagnostic tool to histopathology in head and neck swellings and evaluation of staining efficacy of PAP and MGG stain over Haematoxylin and eosin (H and E) in routine cytopathological smears.
SETTINGS AND DESIGN
The study was conducted in the Department of Oral and Maxillofacial Pathology, where FNAC samples were collected from 150 patients with head and neck swellings.
MATERIALS AND METHODS
All the slides were stained with H and E, Papanicolaou (PAP), and May Grunewald Giemsa (MGG) stains. The cytopathological diagnosis was compared with histopathological diagnosis based on H and E stained sections obtained from paraffin-embedded formalin-fixed biopsy specimen of benign and malignant neoplasms.
STATISTICAL ANALYSIS USED
The resulting data were analyzed using SPSS software version 19. Differences between the variables were analyzed using Pearson Chi-square test and Kruskal-Wallis test wherever applicable.
RESULTS
The FNAC as a diagnostic tool has sensitivity of 84.8%, 72.72%, and 78.78%, specificity of 62.5%, 75%, and 75%, and accuracy of 80.48%, 73.14%, and 78.04% in H and E, MGG, and PAP stain, respectively. PAP stain was the most efficient stain when all qualitative parameters are taken into consideration with maximum sensitivity and specificity for achieving definitive cytodiagnosis.
CONCLUSIONS
The FNAC is an inexpensive and minimally invasive technique to diagnose different types of head and neck swellings and complement histopathological diagnosis.
Topics: Humans; Coloring Agents; Pathology, Oral; Staining and Labeling; Neck; Cytological Techniques; Azure Stains; Hematoxylin
PubMed: 37530331
DOI: 10.4103/ijpm.ijpm_1254_21 -
Redox Biology Jul 2024Inflammation and subsequent mitochondrial dysfunction and cell death worsen outcomes after revascularization in ischemic stroke. Receptor-interacting protein kinase 1...
Modulation of NLRP3 inflammasome-related-inflammation via RIPK1/RIPK3-DRP1 or HIF-1α signaling by phenothiazine in hypothermic and normothermic neuroprotection after acute ischemic stroke.
BACKGROUND
Inflammation and subsequent mitochondrial dysfunction and cell death worsen outcomes after revascularization in ischemic stroke. Receptor-interacting protein kinase 1 (RIPK1) activated dynamin-related protein 1 (DRP1) in a NLRPyrin domain containing 3 (NLRP3) inflammasome-dependent fashion and Hypoxia-Inducible Factor (HIF)-1α play key roles in the process. This study determined how phenothiazine drugs (chlorpromazine and promethazine (C + P)) with the hypothermic and normothermic modality impacts the RIPK1/RIPK3-DRP1 and HIF-1α pathways in providing neuroprotection.
METHODS
A total of 150 adult male Sprague-Dawley rats were subjected to 2 h middle cerebral artery occlusion (MCAO) followed by 24 h reperfusion. 8 mg/kg of C + P was administered at onset of reperfusion. Infarct volumes, mRNA and protein expressions of HIF-1α, RIPK1, RIPK3, DRP-1, NLRP3-inflammation and cytochrome c-apoptosis were assessed. Apoptotic cell death, infiltration of neutrophils and macrophages, and mitochondrial function were evaluated. Interaction between RIPK1/RIPK3 and HIF-1α/NLRP3 were determined. In SH-SY5Y cells subjected to oxygen/glucose deprivation (OGD), the normothermic effect of C + P on inflammation and apoptosis were examined.
RESULTS
C + P significantly reduced infarct volumes, mitochondrial dysfunction (ATP and ROS concentration, citrate synthase and ATPase activity), inflammation and apoptosis with and without induced hypothermia. Overexpression of RIPK1, RIPK3, DRP-1, NLRP3-inflammasome and cytochrome c-apoptosis were all significantly reduced by C + P at 33 °C and the RIPK1 inhibitor (Nec1s), suggesting hypothermic effect of C + P via RIPK1/RIPK3-DRP1pathway. When body temperature was maintained at 37 °C, C + P and HIF-1α inhibitor (YC-1) reduced HIF-1α expression, leading to reduction in mitochondrial dysfunction, NLRP3 inflammasome and cytochrome c-apoptosis, as well as the interaction of HIF-1α and NLRP3. These were also evidenced in vitro, indicating a normothermic effect of C + P via HIF-1α.
CONCLUSION
Hypothermic and normothermic neuroprotection of C + P involve different pathways. The normothermic effect was mediated by HIF-1α, while hypothermic effect was via RIPK1/RIPK3-DRP1 signaling. This provides a theoretical basis for future precise exploration of hypothermic and normothermic neuroprotection.
Topics: Animals; Receptor-Interacting Protein Serine-Threonine Kinases; NLR Family, Pyrin Domain-Containing 3 Protein; Rats; Ischemic Stroke; Hypoxia-Inducible Factor 1, alpha Subunit; Male; Signal Transduction; Inflammasomes; Dynamins; Rats, Sprague-Dawley; Phenothiazines; Inflammation; Neuroprotection; Humans; Disease Models, Animal; Hypothermia, Induced
PubMed: 38692093
DOI: 10.1016/j.redox.2024.103169 -
International Immunopharmacology Jul 2023Methylene blue (MB) has anti-inflammatory properties, however, its underlying molecular mechanism remains elusive. This study aimed to investigate whether and how MB...
Methylene blue (MB) has anti-inflammatory properties, however, its underlying molecular mechanism remains elusive. This study aimed to investigate whether and how MB could attenuate lipopolysaccharide (LPS)-induced microglial activation, neuroinflammation, and neurobehavioral deficits. We measured the expression of pro-inflammatory factors and performed three neurobehavioral tests to assess the effect of MB on neuroinflammation and neurocognitive dysfunction in LPS-treated adult C57BL/6N male mice or LPS-stimulated microglia cells. In vitro and in vivo experiments were further performed to investigate the molecular mechanism underlying MB inhibition of neuroinflammation using various experimental methods, including western blot, RT-qPCR, immunofluorescence, seahorse measurement, positron emission tomography (PET) scan, and flow cytometry analyses. Our results demonstrated that microglial activation and M1 polarization were induced by LPS exposure, resulting in an inflammatory response and neuronal apoptosis. Furthermore, LPS induced metabolic reprogramming in microglial cells. However, MB treatment substantially inhibited LPS-induced elevated levels of pro-inflammatory factors and reversed metabolic activation in vivo, which eventually led to the resolution of neuroinflammation and neurobehavioral improvement. Mechanistically, MB specifically inhibited the LPS-induced overexpression of PHD3 in vitro and in vivo. The pharmacological and genetic manipulations unveiled that the Siah2/Morg1/PHD3 signaling pathway may mediate MB protection against LPS-induced neuroinflammation and neurotoxicity. Therefore MB inhibited PHD3-dependent neuroinflammation may via Siah2/Morg1/PHD3 pathway, and that PHD3 expressed in microglia may be a drug target for the treatment of neuroinflammation-related brain disorders.
Topics: Mice; Animals; Male; Inflammation; Microglia; Lipopolysaccharides; Neuroinflammatory Diseases; Methylene Blue; Mice, Inbred C57BL; Mice, Inbred Strains; Ubiquitin-Protein Ligases
PubMed: 37210913
DOI: 10.1016/j.intimp.2023.110349 -
International Journal of Molecular... Nov 2023The use of a combination of nanoparticles as antimicrobial agents can be one strategy to overcome the tendency of microbes to become resistant to antibiotic action....
The use of a combination of nanoparticles as antimicrobial agents can be one strategy to overcome the tendency of microbes to become resistant to antibiotic action. Also, the optimization of nano-photocatalysts to efficiently remove persistent pollutants from wastewater is a hot topic. In this study, two composites ZnO/Au (1% wt.) and ZnO/Ag (1% wt.) were synthesized by simple aqueous solution methods. The structure and morphology of the r nanocomposites were analyzed by structural and optical characterization methods. The formation of AuNPs and AgNPs in these experiments was also discussed. The antimicrobial properties of ZnO, ZnO/Au, and ZnO/Ag nanomaterials were investigated against Gram-negative bacteria () and Gram-positive bacteria (). The results showed an increase of 80% in the antimicrobial activity of ZnO/Au against compared with 30% in the case of ZnO/Ag. Similarly, in the case of the strain tests, ZnO/Au increased the antimicrobial activity by 55% and ZnO/Ag by 33%. The photocatalytic tests indicated an improvement in the photocatalytic degradation of methylene blue (MB) under UV irradiation using ZnO/Au and ZnO/Ag nanocomposites compared to bare ZnO. The photocatalytic degradation efficiency of ZnO after 60 min of UV irradiation was ∼83%, while the addition of AuNPs enhanced the degradation rate to ∼95% (ZP2), and AgNP presence enhanced the efficiency to ∼98%. The introduction of noble metallic nanoparticles into the ZnO matrix proved to be an effective strategy to increase their antimicrobial activity against and , and their photocatalytic activity was evaluated through the degradation of MB dye. Comparing the enhancing effects of Au and Ag, it was found that ZnO/Au was a better antimicrobial agent while ZnO/Ag was a more effective photocatalyst under UV irradiation.
Topics: Zinc Oxide; Staphylococcus aureus; Gold; Metal Nanoparticles; Anti-Bacterial Agents; Nanocomposites; Methylene Blue
PubMed: 38069261
DOI: 10.3390/ijms242316939 -
PeerJ 2023Amultigenerational study on was carried out by exposing three subsequent generations to pharmaceuticals chlorpromazine (CPZ) and diclofenac (DCF), and two lanthanide...
Amultigenerational study on was carried out by exposing three subsequent generations to pharmaceuticals chlorpromazine (CPZ) and diclofenac (DCF), and two lanthanide chlorides, gadolinium as GdCl and europium as EuCl. As the treatments, environmentally relevant concentrations were chosen (0.001, 0.01 and 0.1 mg/L for CPZ; 0.1, 1 and 10 mg/L for DCF; 0.425, 4.25 and 42.5 µg/L for Gd and 0.41, 4.1 and 41 µg/L for Eu). Survival, population growth and reproduction success were evaluated at 21 and 30 days of exposure, and the whole observation period lasted 40 days. The least sensitive to all selected substances was the first daphnid generation (F1). Within 21-day exposure, no significant effects of the psychotropic drug CPZ on survival were observed in generations F1-F3. The anti-inflammatory drug DCF did not affect survival in the F1 generation; however, it significantly reduced survival in the F3 generation at 1-10 mg/L. Both lanthanides did not affect survival in the F1 and F2 generations of but considerably decreased survival in the F3 at 4-42 µg/L. Both pharmaceuticals stimulated the reproduction of in the F1 generation, while inhibition occurred at the highest tested concentrations in generations F2 and F3. The inhibitory effect on the reproductive success of lanthanides in the F2 generation resembled that for CPZ but not for DCF. The dynamics of adverse effects during the 21-30-day period revealed that despite increased mortality in the controls (up to 30%), concentrations used in the study minified, in most instances, the survival and aggravated population growth and reproduction success of . Our data suggest that as a test organism can be used for 21 days in multigenerational investigations, especially when testing close to environmental concentrations. In this respect, the standard chronic toxicity assay seems limited since prolonged observations and several generations of daphnids are required to obtain reliable information for the risk assessment of potentially aggressive chemicals.
Topics: Animals; Chlorpromazine; Cladocera; Diclofenac; Lanthanoid Series Elements
PubMed: 38025671
DOI: 10.7717/peerj.16472 -
Reproductive Sciences (Thousand Oaks,... Feb 2024Endometriosis is an immune chronic inflammatory disease, and there are currently no more effective drugs for treating endometriosis due to its unknown etiology....
Endometriosis is an immune chronic inflammatory disease, and there are currently no more effective drugs for treating endometriosis due to its unknown etiology. Salbutamol is a β2-adrenergic receptor (β2AR) agonist commonly used to treat asthma by selectively activating β2 receptors on airway smooth muscle and leukocytes, exerting bronchial dilation and synergistic anti-inflammatory effects. In recent years, β2AR agonists have been used in endometriosis studies, and we speculate that salbutamol may have a therapeutic effect on endometriosis. The purpose of this research was to explore the therapeutic effect of salbutamol on endometriosis mice. The mouse endometriosis model was established and treated with different doses of salbutamol. Endometrial lesions were harvested for pathological diagnosis, immunohistochemistry (IHC), Masson staining, and toluidine blue analysis. We found that the number and size of endometriotic lesions were all significantly decreased after 3 weeks of treatment with different doses of salbutamol on endometriosis model mice (P < 0.05). After Salbutamol treatment, the amount of mast cells (toluidine blue) and macrophages (F4/80) in the lesions as well as the expressions of interleukin (IL)-1β, tumor necrosis factor (TNF)-ɑ, platelet-derived growth factor subunit B (PDGFB), CD31, transforming growth factor (TGF)-β, Masson staining, BCL2, TUBB3, substance P (SP), and nerve growth factor (NGF) were significantly reduced (P < 0.05). These results suggested that salbutamol could effectively treat endometriosis in mice by reducing immune inflammatory cells and factors, angiogenesis, and fibrosis, increasing apoptosis of endometriotic lesions, and decreasing neurogenesis.
Topics: Humans; Female; Mice; Animals; Endometriosis; Albuterol; Tolonium Chloride; Substance P
PubMed: 37814201
DOI: 10.1007/s43032-023-01371-0 -
Indian Journal of Cancer 2023Diagnostic adjuncts such as toluidine blue have been investigated as screening tools that improve visual examination of potentially malignant disorders (PMD) and oral...
BACKGROUND
Diagnostic adjuncts such as toluidine blue have been investigated as screening tools that improve visual examination of potentially malignant disorders (PMD) and oral cancer. Acetic acid has been reported to be of value in the early detection of cervical cancers. This study assessed the utility of 5% acetic acid as a diagnostic adjunct in oral PMD and compared the accuracy of acetic acid with toluidine blue in the detection of dysplastic PMD and high-risk lesions.
MATERIALS AND METHODS
This cross-sectional study was conducted at a dental hospital in a rural setting. Thirty-one patients with oral PMD formed the study group. Five percent acetic acid was applied to the lesions, followed by toluidine blue application and biopsy. Sensitivity, specificity, and positive and negative predictive values were computed considering true positives as stain uptake in dysplastic and high-risk PMD.
RESULTS
The sensitivity, specificity, and positive and negative predictive values of acetic acid for identifying dysplastic or malignant lesions were 100%, 13.3%, 51.2%, and 100%, respectively, and that for toluidine blue were 75%, 100%, 100%, and 78.9%, respectively. The corresponding values for identifying high-risk PMD (lesions with moderate and severe dysplasia) using acetic acid were 100%, 9.1%, 25.9%, and 100%, respectively, and for toluidine blue were 85.7%, 81.8%, 60%, and 94.7%, respectively.
CONCLUSION
The utility of acetic acid in detecting dysplasia and high-risk PMD is severely limited due to its poor specificity. Compared with acetic acid, toluidine blue is a superior screening tool.
Topics: Humans; Tolonium Chloride; Acetic Acid; Cross-Sectional Studies; Precancerous Conditions; Mouth Diseases
PubMed: 36861696
DOI: 10.4103/ijc.IJC_42_21 -
Advanced Science (Weinheim,... Apr 2024Tumors often overexpress glucose-regulated proteins, and agents that interfere with the production or activity of these proteins may represent novel cancer treatments....
Tumors often overexpress glucose-regulated proteins, and agents that interfere with the production or activity of these proteins may represent novel cancer treatments. The chlorpromazine derivative JX57 exhibits promising effects against endometrial cancer with minimal extrapyramidal side effects; however, its mechanisms of action are currently unknown. Here, glucose-regulated protein 75 kD (GRP75) is identified as a direct target of JX57 using activity-based protein profiling and loss-of-function experiments. The findings show that GRP75 is necessary for the biological activity of JX57, as JX57 exhibits moderate anticancer properties in GRP75-deficient cancer cells, both in vitro and in vivo. High GRP75 expression is correlated with poor differentiation and poor survival in patients with endometrial cancer, whereas the knockdown of GRP75 can significantly suppress tumor growth. Mechanistically, the direct binding of JX57 to GRP75 impairs the structure of the mitochondria-associated endoplasmic reticulum membrane and disrupts the endoplasmic reticulum-mitochondrial calcium homeostasis, resulting in a mitochondrial energy crisis and AMP-activated protein kinase activation. Taken together, these findings highlight GRP75 as a potential prognostic biomarker and direct therapeutic target in endometrial cancer and suggest that the chlorpromazine derivative JX57 can potentially be a new therapeutic option for endometrial cancer.
Topics: Female; Humans; AMP-Activated Protein Kinases; Chlorpromazine; Endometrial Neoplasms; HSP70 Heat-Shock Proteins; Membrane Proteins; Mitochondria
PubMed: 38342610
DOI: 10.1002/advs.202304203