A two-photon fluorescent probe records the intracellular pH through ‘OR’ logic operation via internal calibration

Podder, Arup,Won, Miae,Kim, Soobin,Verwilst, Peter,Maiti, Mrinmoy,Yang, Zhigang,Qu, Junle,Bhuniya, Sankarprasad,Kim, Jong Seung Elsevier 2018 Sensors and actuators. B Chemical Vol.268 No.-

<P><B>Abstract</B></P> <P>Mapping the intracellular location and concentration of hydronium ions (H<SUB>3</SUB>O<SUP>+</SUP>) and their dynamics could be a useful diagnostic tool in modern clinical science. Current needs motivated us to develop a molecular pH probe <B>1</B>, operating as a logic gate, and its analogue <B>2</B>. The pyridyl moiety in <B>1</B> played a significant role in proton capture and release, in acidic to alkaline pH environments. In contrast, <B>2</B> failed to show a similar spectroscopic behavior. <B>1</B> shows emission maximum at 450 nm that is independent on the pH, with excitation at 353 nm or 410 nm in acid and alkaline pH, respectively. <B>1</B> was employed to provide input-dependent (excitation wavelength) fluorescence images in a cellular milieu to detect pH changes in cellular organelles such as lysosomes and mitochondria. Furthermore, <B>1</B> provided information on the variation of the pH in the presence of cellular ROS. <B>1</B> was also found to enable the real-time monitoring of cell acidification due to nutrient starvation, which is closely associated with mitochondrial malfunction, fusion and mitophagy processes. We envision that in due course <B>1</B> can open up new research avenues in the diagnostic sector for validating the pH in the cellular milieu.</P> <P><B>Highlights</B></P> <P> <UL> <LI> This is the first example of a “OR” Logic operative two-photon probe for pH determination. </LI> <LI> Probe <B>1</B> enables the tracking of a wide pH range in cellular organelles without interference from cellular autofluorescence. </LI> <LI> This nontoxic probe can detect pH fluctuation due to ROS, such as H<SUB>2</SUB>O<SUB>2</SUB> and others. </LI> <LI> Probe <B>1</B> can provide dual-mode emission/cell images either by single photon or two-photon modes. Therefore, it is a unique cellular pH probe. </LI> <LI> Finally, <B>1</B> revealed pH fluctuations due to nutrient starvation, which is related to mitochondrial malfunction, fusion, and mitophagy. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>“OR” logic gated pH probe (1) detects the pH in cell organelles in the presence of ROS and under nutrient starvation conditions.</P> <P>[DISPLAY OMISSION]</P>

A rhodamine based fluorescent probe validates substrate and cellular hypoxia specific NADH expression

Podder, Arup,Koo, Seyoung,Lee, Jiyeong,Mun, Sora,Khatun, Sabina,Kang, Hee-Gyoo,Bhuniya, Sankarprasad,Kim, Jong Seung The Royal Society of Chemistry 2019 Chemical communications Vol.55 No.4

<P>Herein, we report a rhodamine-based redox probe (MQR) to visualize cytosolic NADH in the cellular milieu. Its high sensitivity and selectivity allowed it to track the alteration of the NADH level under metabolic perturbation, suggesting its potential as a useful tool to study the association between the NADH level and metabolic abnormalities with clinical significance.</P>

Naringenin Exerts Cytoprotective Effect Against Paraquat-Induced Toxicity in Human Bronchial Epithelial BEAS-2B Cells Through NRF2 Activation

( Biswajit Podder ),( Ho Yeon Song ),( Yong Sik Kim ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.5

We have previously shown that paraquat (PQ)-induced oxidative stress causes dramatic damage in various human cell lines. Naringenin (NG) is an active flavanone, which has been reported to have beneficial bioactivities, including antioxidative, anti-inflammatory, and antitumorigenic activities, with a relatively low toxicity to normal cells. In this study, we intended to assess the cytoprotective effect of NG against PQ-induced toxicity in the human bronchial epithelial BEAS-2B cell line. Co-treatment with NG in PQ-treated BEAS-2B cells can reduce PQ-induced cellular toxicity. NG can also decrease the generation of intracellular ROS caused by PQ treatment. We also observed that treatment with NG in PQ-exposed BEAS-2B cells can significantly induce the expression of antioxidant-related genes, including GPX2, GPX3, GPX5, and GPX7. NG co-treatment can also activate the NRF2 transcription factor and promote its nuclear translocation. In addition, NG co-treatment can induce the expression of NRF2-downstream target genes such as that of heme oxygenase-1 (HO-1) and NAD(P)H:quinone oxidoreductase 1 (NQO1). A small interfering RNA study revealed that the knockdown of NRF2 can abrogate NG-mediated protection of the cells from PQ-induced cellular toxicity. We propose that NG effectively alleviates PQ-induced cytotoxicity in human bronchial epithelial BEAS-2B cells through the NRF2-regulated antioxidant defense pathway, and NG might be a good therapeutic candidate molecule in oxidative stress-related diseases.

Ursolic Acid Activates Intracellular Killing Effect of Macrophages During Mycobacterium tuberculosis Infection

( Biswajit Podder ),( Woong Sik Jang ),( Kung Woo Nam ),( Byung Eui Lee ),( Ho Yeon Song ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.5

Tuberculosis is one of the most threatening infectious diseases to public health all over the world, for which Mycobacterium tuberculosis (MTB) is the etiological agent of pathogenesis. Ursolic acid (UA) has immunomodulatory function and exhibits antimycobacterial activity. However, the intracellular killing effect of UA has yet to be elucidated. The aim of this study was to evaluate the intracellular killing effect of UA during mycobacterial infection. The intracellular killing activity of UA was evaluated in the macrophage cell line THP-1 by the MGIT 960 system as well as by CFU count. The production of reactive oxygen species (ROS) and the level of nitric oxide (NO) were measured using DCF-DA and Griess reagent, respectively. Phagocytosis was observed by a fluorescence-based staining method, and the colony forming units were enumerated on 7H11 agar medium following infection. In addition, MRP8 mRNA expression was measured by qRT-PCR. UA significantly decreased the number of intracellular Mycobacterium through generation of ROS and NO. In addition, it profoundly activated the phagocytosis process of THP-1 cells during MTB-infection. Furthermore, our data demonstrated that UA activated the phagocytosis process in human monocyte cells through MRP8 induction. These data suggest that UA firmly contributes to the intracellular killing effect of macrophages during mycobacterial infection.

Selective monitoring of vascular cell senescence via β-Galactosidase detection with a fluorescent chemosensor

Kim, Eun-Joong,Podder, Arup,Maiti, Mrinmoy,Lee, Jong Min,Chung, Bong Geun,Bhuniya, Sankarprasad Elsevier 2018 Sensors and actuators. B Chemical Vol.274 No.-

<P><B>Abstract</B></P> <P>A new senescence-responsive fluorescent probe (<B>SRP</B>) was developed for the detection of β-galactosidase activity in senescent cells. UV-absorption of t probe <B>SRP</B> at 495 nm was increased in the presence of β-galactosidase. Its fluorescence at λ<SUB>em</SUB> 545 nm increased ∼27-fold upon incubation with β-galactosidase (0.1 U/mL). The <B>SRP</B> probe is non-toxic and highly chemoselective for β-galactosidase. The high cell viability and chemoselcivity of probe <B>SRP</B> offer it to be a suitable marker for assessing cell senescence in live cells. Using this probe, H<SUB>2</SUB>O<SUB>2</SUB>-induced cellular senescence of human umbilical vein cells (HUVECs) could be distinguished from normal cells based on the extent of fluorescent labeling of the cells. Moreover, it was preferentially localized in acidic lysosomes. Overall, <B>SRP</B> is a unique chemosensor that can provide preclinical information on cell senescence in vascular endothelial cells.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The probe <B>SRP</B> is nontoxic and biocompatible. </LI> <LI> The probe <B>SRP</B> is highly selective toward β-galactosidase. </LI> <LI> The probe <B>SRP</B> selectively is localized in lysosomes. </LI> <LI> The probe <B>SRP</B> enables to detect vascular cell senescence. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Anti-Mycobacterial Activity of Tamoxifen Against Drug-Resistant and Intra-Macrophage Mycobacterium tuberculosis

Jang, Woong Sik,Kim, Sukyung,Podder, Biswajit,Jyoti, Md. Anirban,Nam, Kung-Woo,Lee, Byung-Eui,Song, Ho-Yeon The Korean Society for Microbiology and Biotechnol 2015 Journal of microbiology and biotechnology Vol.25 No.6

Recently, it has become a struggle to treat tuberculosis with the current commercial antituberculosis drugs because of the increasing emergence of multidrug-resistant (MDR) tuberculosis and extensively drug-resistant (XDR) tuberculosis. We evaluated here the antimycobacterial activity of tamoxifen, known as a synthetic anti-estrogen, against eight drugsensitive or resistant strains of Mycobacterium tuberculosis (TB), and the active intracellular killing of tamoxifen on TB in macrophages. The results showed that tamoxifen had antituberculosis activity against drug-sensitive strains (MIC, 3.125-6.25 µg/ml) as well as drugresistant strains (MIC, 6.25 to 12.5 µg/ml). In addition, tamoxifen profoundly decreased the number of intracellular TB in macrophages in a dose-dependent manner.

Management of a rare case of extra hepatic portal vein obstruction with temporomandibular joint ankylosis and review of literature

Kailash Chand Kurdia,Ambuj Aggarwal,Cherring Tandup,Divya Dahiya,Subrata Podder,Arunanshu Behera 한국간담췌외과학회 2021 Annals of hepato-biliary-pancreatic surgery Vol.25 No.2

Extrahepatic portal venous obstruction (EHPVO) and temporomandibular joint (TMJ) ankylosisis are significant problems in Asian countries. Both EHPVO and bilateral TMJ ankylosis may have rare association due to protein C and S deficiency which may cause hypercoagulability as well as reduced fibrinolytic activity. Ankylosis arising in early childhood is associated with facial asymmetry, feeding difficulty and speech development alterations. It is also associated with great challenges of endoscopic management in extra hepatic portal vein obstruction (EHPVO) with variceal bleed as well as air way management during surgical management and post-operative recovery. Recently a case series had shown association of TMJ ankylosis with EHPVO due to protein C deficiency which might be an etiological factor for both EHPVO as well as TMJ ankylosis. This case report documents a case of 14 year young girl who had TMJ ankylosis due to ear infection and EHPVO with esophageal varices had multiple episodes of upper GI bleed with mild deficiency of protein C and S, successfully managed with proximal splenorenal shunt to prevent further episodes of upper GI bleed, as endoscopic management is not feasible due to TMJ ankylosis.

A network-biology approach for identification of key genes and pathways involved in malignant peritoneal mesothelioma

Mahfuz, A.M.U.B.,Zubair-Bin-Mahfuj, A.M.,Podder, Dibya Joti Korea Genome Organization 2021 Genomics & informatics Vol.19 No.2

Even in the current age of advanced medicine, the prognosis of malignant peritoneal mesothelioma (MPM) remains abysmal. Molecular mechanisms responsible for the initiation and progression of MPM are still largely not understood. Adopting an integrated bioinformatics approach, this study aims to identify the key genes and pathways responsible for MPM. Genes that are differentially expressed in MPM in comparison with the peritoneum of healthy controls have been identified by analyzing a microarray gene expression dataset. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses of these differentially expressed genes (DEG) were conducted to gain a better insight. A protein-protein interaction (PPI) network of the proteins encoded by the DEGs was constructed using STRING and hub genes were detected analyzing this network. Next, the transcription factors and miRNAs that have possible regulatory roles on the hub genes were detected. Finally, survival analyses based on the hub genes were conducted using the GEPIA2 web server. Six hundred six genes were found to be differentially expressed in MPM; 133 are upregulated and 473 are downregulated. Analyzing the STRING generated PPI network, six dense modules and 12 hub genes were identified. Fifteen transcription factors and 10 miRNAs were identified to have the most extensive regulatory functions on the DEGs. Through bioinformatics analyses, this work provides an insight into the potential genes and pathways involved in MPM.

A bioorthogonal `turn-on' fluorescent probe for tracking mitochondrial nitroxyl formation

Sunwoo, K.,Bobba, K.,Lim, J. Y.,Park, T.,Podder, A.,Heo, J.,Lee, S.,Bhuniya, S.,Kim, J. unknown 2017 Chemical communications Vol. No.

<P>A bioreductant-resistant 'turn-on' chemodosimetric fluorescent probe Mito-1 has been developed for the detection of mitochondrial HNO in live cells. Mito-1 enables the detection of HNO as low as similar to 18 nM. It has the capability to detect both exogenous and endogenous mitochondrial HNO formations in cellular milieus by providing fluorescence images. Its two-photon imaging ability fosters its use as a non-invasive imaging tool for the detection of mitochondrial nitroxyl.</P>

Azo-based small molecular hypoxia responsive theranostic for tumor-specific imaging and therapy

Zhou, Ying,Maiti, Mrinmoy,Sharma, Amit,Won, Miae,Yu, Le,Miao, Lan Xi,Shin, Jinwoo,Podder, Arup,Bobba, Kondapa Naidu,Han, Jiyou,Bhuniya, Sankarprasad,Kim, Jong Seung Elsevier 2018 Journal of controlled release Vol.288 No.-

<P><B>Abstract</B></P> <P>We report herein, an azo-derivative (<B>AzP1</B>) of FDA approved antineoplastic drug SN-38 (irinotecan analogue) as a theranostic agent with a potential for both tumor hypoxia-specific activation and therapy. The theranostic <B>AzP1</B> was found to be stable within a biologically relevant pH scale and was chemically inert towards other competitive biological analytes. However, upon treatment with rat-liver microsomes, <B>AzP1</B> showed a self-calibrated fluorescence enhancement at λ<SUB>em</SUB> = 560 nm. The cytotoxicity profile of <B>AzP1</B> was tested in various cancer lines. Under hypoxic conditions, prodrug <B>AzP1</B> exhibited activation to release the parent drug (SN-38) and enhanced cytotoxicity in cancer cells with concomitant fluorescence enhancement at 560 nm, which served to monitor both the drug activation and tracing purposes. The therapeutic potential of <B>AzP1</B> for both tumor-specific activation and suppression of tumor weights was validated in xenograft mouse model. Collectively, the synthetic ease and hypoxia-sensitive activation along with promising therapeutic properties highlight the potential of theranostic <B>AzPI</B> in future cancer treatment programs.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>