Scant Extracellular NAD Cleaving Activity of Human Neutrophils is Down-Regulated by fMLP via FPRL1

Hasan, Md. Ashraful,Sultan, Md. Tipu,Ahn, Won-Gyun,Kim, Yeon-Ja,Jang, Ji-Hye,Hong, Chang-Won,Song, Dong-Keun The Korean Society of Pharmacology 2014 The Korean Journal of Physiology & Pharmacology Vol.18 No.6

Extracellular nicotinamide adenine dinucleotide (NAD) cleaving activity of a particular cell type determines the rate of the degradation of extracellular NAD with formation of metabolites in the vicinity of the plasma membrane, which has important physiological consequences. It is yet to be elucidated whether intact human neutrophils have any extracellular NAD cleaving activity. In this study, with a simple fluorometric assay utilizing $1,N^6$-ethenoadenine dinucleotide (etheno-NAD) as the substrate, we have shown that intact peripheral human neutrophils have scant extracellular etheno-NAD cleaving activity, which is much less than that of mouse bone marrow neutrophils, mouse peripheral neutrophils, human monocytes and lymphocytes. With high performance liquid chromatography (HPLC), we have identified that ADP-ribose (ADPR) is the major extracellular metabolite of NAD degradation by intact human neutrophils. The scant extracellular etheno-NAD cleaving activity is decreased further by N-formyl-methionine-leucine-phenylalanine (fMLP), a chemoattractant for neutrophils. The fMLP-mediated decrease in the extracellular etheno-NAD cleaving activity is reversed by WRW4, a potent FPRL1 antagonist. These findings show that a much less extracellular etheno-NAD cleaving activity of intact human neutrophils compared to other immune cell types is down-regulated by fMLP via a low affinity fMLP receptor FPRL1.

N-acetyl-L-cysteine and cysteine increase intracellular calcium concentration in human neutrophils

Hasan, Md. Ashraful,Ahn, Won-Gyun,Song, Dong-Keun The Korean Society of Pharmacology 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.5

N-acetyl-L-cysteine (NAC) and cysteine have been implicated in a number of human neutrophils' functional responses. However, though $Ca^{2+}$ signaling is one of the key signalings contributing to the functional responses of human neutrophils, effects of NAC and cysteine on intracellular calcium concentration ($[Ca^{2+}]_i$) in human neutrophils have not been investigated yet. Thus, this study was carried out with an objective to investigate the effects of NAC and cysteine on $[Ca^{2+}]_i$ in human neutrophils. We observed that NAC ($1{\mu}M{\sim}1mM$) and cysteine ($10{\mu}M{\sim}1mM$) increased $[Ca^{2+}]_i$ in human neutrophils in a concentration-dependent manner. In NAC pre-supplmented buffer, an additive effect on N-formyl-methionine-leucine-phenylalanine (fMLP)-induced increase in $[Ca^{2+}]_i$ in human neutrophils was observed. In $Ca^{2+}$-free buffer, NAC- and cysteine-induced $[Ca^{2+}]_i$ increase in human neutrophils completely disappeared, suggesting that NAC- and cysteine-mediated increase in $[Ca^{2+}]_i$ in human neutrophils occur through $Ca^{2+}$ influx. NAC- and cysteine-induced $[Ca^{2+}]_i$ increase was effectively inhibited by calcium channel inhibitors SKF96365 ($10{\mu}m$) and ruthenium red ($20{\mu}m$). In $Na^+$-free HEPES, both NAC and cysteine induced a marked increase in $[Ca^{2+}]_i$ in human neutrophils, arguing against the possibility that $Na^+$-dependent intracellular uptake of NAC and cysteine is necessary for their $[Ca^{2+}]_i$ increasing activity. Our results show that NAC and cysteine induce $[Ca^{2+}]_i$ increase through $Ca^{2+}$ influx in human neutrophils via SKF96365- and ruthenium red-dependent way.

N-acetyl-L-cysteine and cysteine increase intracellular calcium concentration in human neutrophils

Md. Ashraful Hasan,안원균,송동근 대한약리학회 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.5

N-acetyl-L-cysteine (NAC) and cysteine have been implicated in a number of human neutrophils’ functional responses. However, though Ca2+ signaling is one of the key signalings contributing to the functional responses of human neutrophils, effects of NAC and cysteine on intracellular calcium concentration ([Ca2+]і) in human neutrophils have not been investigated yet. Thus, this study was carried out with an objective to investigate the effects of NAC and cysteine on [Ca2+]і in human neutrophils. We observed that NAC (1 mM ~ 1 mM) and cysteine (10 mM ~ 1 mM) increased [Ca2+]і in human neutrophils in a concentration-dependent manner. In NAC pre-supplmented buffer, an additive effect on N-formyl-methionine-leucinephenylalanine (fMLP)-induced increase in [Ca2+]i in human neutrophils was observed. In Ca2+-free buffer, NAC- and cysteine-induced [Ca2+]i increase in human neutrophils completely disappeared, suggesting that NAC- and cysteine-mediated increase in [Ca2+]i in human neutrophils occur through Ca2+ influx. NAC- and cysteine-induced [Ca2+]і increase was effectively inhibited by calcium channel inhibitors SKF96365 (10 mM) and ruthenium red (20 mM). In Na+-free HEPES, both NAC and cysteine induced a marked increase in [Ca2+]i in human neutrophils, arguing against the possibility that Na+-dependent intracellular uptake of NAC and cysteine is necessary for their [Ca2+]і increasing activity. Our results show that NAC and cysteine induce [Ca2+]і increase through Ca2+ influx in human neutrophils via SKF96365- and ruthenium reddependent way.

Magnetite Loaded Cross-linked Polystyrene Composite Particles Prepared by Modified Suspension Polymerization and Their Potential Use as Adsorbent for Arsenic(III)

Hasan Ahmad,Mostafa Kaiyum Sharafat,Mohammad Ashraful Alam,Mohammad Mahbubor Rahman,Klaus Tauer,Hideto Minami,Mosammat Sharmin Sultana,Basudev Kumar Das,Rukhsana Shabnam 한국고분자학회 2017 Macromolecular Research Vol.25 No.7

Magnetic particles have attracted much interest because they can be easily guided under the influence of magnetic field and are considered suitable in biomedical as well as in separation technology. In this work, magnetite (Fe3O4) loaded cross-linked polystyrene (PS) composite particles are prepared by a novel suspension polymerization of styrene and divinylbenzene (DVB) in presence of nanosized iron oxide (Fe3O4) particles dispersed in n-octane. The structure and surface properties are systematically characterized using FTIR, scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD) and thermogravimetric analyses (TGA). The morphology and surface structure reveal that Fe3O4 nanoparticles are mostly dispersed within the P(S-DVB) copolymer layer. Finally the applicability of Fe3O4/P(S-DVB) composite polymer particles as adsorbent for toxic element like trivalent arsenic (As) has been investigated. Fe3O4/P(S-DVB) composite polymer particles exhibited good sorption efficiency compared to reference P(S-DVB) copolymer particles.

Antinociceptive and gastro-protective effect of the ethanolic extract of the flowering top of Anthocephalus Cadamba Roxb

Subhan, Nusrat,Hasan, Raquibul,Hossain, Mokarram,Akter, Raushanara,Majumder, Muntasir Mamun,Rahman, Mostafizur,Ahmed, Kamaluddin,Ghani, Abdul,Alam, Ashraful Kyung Hee Oriental Medicine Research Center 2009 Oriental pharmacy and experimental medicine Vol.9 No.4

The effect of alcoholic extract of Anthocephalus (A.) Cadamba Roxb. was evaluated in experimental models of pain and ulcer. Hot tail flick test, hot plate test and acetic acid induced writhing test were employed for evaluating the peripheral as well as central analgesic mechanism exerted by the extracts. Gastroprotective activity was examined by HCl and ethanol induced gastric damage test. Test group received crude extract 500 mg/kg showed maximum time needed for the response against thermal stimuli (6.26 ${\pm}$ 0.439 s) which is comparable to diclofenac sodium (6.56 ${\pm}$ 0.381 s) in hot tail flick method. These experimental results also followed the experimental results of hot plate test where crude extract 500 mg/kg showed maximum time needed for the response against thermal stimuli (4.74 ${\pm}$ 0.234 s) which is comparable to diclofenac sodium (5.58 ${\pm}$ 0.585 s). The crude extract at 500 and 250 mg/kg showed significant reduction in acetic acid induced writhing in mice with a maximum effect of 68.026% reduction at 500 mg/kg dose which is comparable to standard diclofenac sodium (79.93%). In gastroprotective study the extract of A. Cadamba (250 and 500 mg/kg) significantly inhibited ulceration induced by both HCl and ethanol dose dependently. Results of the study suggest that the extract possesses both analgesic and gastroprotective activity on mice.

In vitro anti-oxidant activity of the leaves of Dillenia indica

Moni Rani Saha,Md Ashraful Alam,SM Raquibul Hasan,Raushanara Akter,Md Mokarram Hossain,Ehsanul Hoque Mazumder,Md Sohel Rana 경희대학교 융합한의과학연구소 2009 Oriental Pharmacy and Experimental Medicine Vol.9 No.4

The methanol extract of Dillenia indica was tested for antioxidant activity as determined by free radical scavenging of DPPH radical scavenging assay, reducing power, total antioxidant capacity measured by phosphomolybdenum method, total phenolic content and total flavonoids content determination assays. The extract showed significant activities in all antioxidant assays compared to the standard antioxidant in a dose dependent manner. In DPPH radical scavenging assay the IC50 value of the extract was found to be 100.53 μg/ml while ascorbic acid has the IC50 value 58.92 μg/ml. Dillenia indica extract showed strong reducing power and total antioxidant capacity. Moreover, methanol extracts also possess high amount of phenolics and flavovonoids and expressed as gallic acid and rutin equivalent respectively. The remarkable activities exhibited in reactive oxygen species scavenging may attributed to the high amount of hydrophilic phenolics present in Dillenia indica. The methanol extract of Dillenia indica was tested for antioxidant activity as determined by free radical scavenging of DPPH radical scavenging assay, reducing power, total antioxidant capacity measured by phosphomolybdenum method, total phenolic content and total flavonoids content determination assays. The extract showed significant activities in all antioxidant assays compared to the standard antioxidant in a dose dependent manner. In DPPH radical scavenging assay the IC50 value of the extract was found to be 100.53 μg/ml while ascorbic acid has the IC50 value 58.92 μg/ml. Dillenia indica extract showed strong reducing power and total antioxidant capacity. Moreover, methanol extracts also possess high amount of phenolics and flavovonoids and expressed as gallic acid and rutin equivalent respectively. The remarkable activities exhibited in reactive oxygen species scavenging may attributed to the high amount of hydrophilic phenolics present in Dillenia indica.

In vitro anti-oxidant activity of the leaves of Dillenia indica

Saha, Moni Rani,Alam, Ashraful,Hasan, Raquibul,Akter, Raushanara,Hossain, Mokarram,Mazumder, Ehsanul Hoque,Rana, Sohel Kyung Hee Oriental Medicine Research Center 2009 Oriental pharmacy and experimental medicine Vol.9 No.4

The methanol extract of Dillenia indica was tested for antioxidant activity as determined by free radical scavenging of DPPH radical scavenging assay, reducing power, total antioxidant capacity measured by phosphomolybdenum method, total phenolic content and total flavonoids content determination assays. The extract showed significant activities in all antioxidant assays compared to the standard antioxidant in a dose dependent manner. In DPPH radical scavenging assay the $IC_{50}$ value of the extract was found to be 100.53${\mu}g/ml$ while ascorbic acid has the $IC_{50}$ value 58.92${\mu}g/ml$. Dillenia indica extract showed strong reducing power and total antioxidant capacity. Moreover, methanol extracts also possess high amount of phenolics and flavovonoids and expressed as gallic acid and rutin equivalent respectively. The remarkable activities exhibited in reactive oxygen species scavenging may attributed to the high amount of hydrophilic phenolics present in Dillenia indica.

Ellagic acid rich Momordica charantia fruit pulp supplementation prevented oxidative stress, fibrosis and inflammation in liver of alloxan induced diabetic rats

Syed Yeasin Arafat,Ashraful Alam,Mohammad Nayeem,Sharfiat Jahan,Zarifa Karim,Hasan Mahmud Reza,Hemayet Hossain,Mohammad Shohel 경희대학교 융합한의과학연구소 2016 Oriental Pharmacy and Experimental Medicine Vol.31 No.4

The present study was undertaken to investigate the hepatoprotective effect of Momordica charantia in alloxan induced diabetic rats. The experimental rats were randomly divided into four groups. Oral glucose tolerance test (OGTT) and biochemical analysis for oxidative stress parameters and liver function marker enzymes were analyzed for each group of rats. Moreover, histological staining was also performed on liver sections. Momordica charantia supplementation prevented the rise of blood glucose level and improved oral glucose tolerance test in alloxan induced diabetic rats. Liver marker enzymes such as aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP) activities were increased in alloxan induced rats which were normalized in Momordica charantia supplemented group. In addition, elevated level of oxidative stress markers were also observed in alloxan induced diabetic rats. Momordica charantia supplementation significantly restored the superoxide dismutase (SOD), catalase (CAT) enzymes activities and increased reduced glutathione (GSH) concentration in diabetic rats. Histological assessments confirm the mononuclear cell infiltration and fibrosis in liver of alloxan induced diabetic rats which were ameliorated by Momordica charantia supplementation. The present investigation suggests the hepatoprotective nature of Momordica charantia in diabetic rats probably by attenuating oxidative stress and improving the antioxidant competence in hepatic tissues.

Protective effects of Acanthopanax divaricatus extract in mouse models of Alzheimer’s disease

Ji-Jing Yan,Won-Gyun Ahn,Jun-Sub Jung,Hee-Sung Kim,Ashraful Hasan,Dong-Keun Song 한국영양학회 2014 Nutrition Research and Practice Vol.8 No.4

BACKGROUND: Acanthopanax divaricatus var. albeofructus (ADA) extract has been reported to have anti-oxidant, immunomodulatory, and anti-mutagenic activity. MATERIALS/METHODS: We investigated the effects of ADA extract on two mouse models of Alzheimer’s disease (AD); intracerebroventricular injection of β-amyloid peptide (Aβ) and amyloid precursor protein/presenilin 1 (APP/PS1)-transgenic mice. RESULTS: Intra-gastric administration of ADA stem extract (0.25 g/kg, every 12 hrs started from one day prior to injection of Aβ1-42 until evaluation) effectively blocked Aβ1-42-induced impairment in passive avoidance performance, and Aβ1-42-induced increase in immunoreactivities of glial fibrillary acidic protein and interleukin (IL)-1α in the hippocampus. In addition, it alleviated the Aβ1-42-induced decrease in acetylcholine and increase in malondialdehyde levels in the cortex. In APP/PS1-transgenic mice, chronic oral administration of ADA stem extract (0.1 or 0.5 g/kg/day for six months from the age of six to 12 months) resulted in significantly enhanced performance of the novel-object recognition task, and reduced amyloid deposition and IL-1β in the brain. CONCLUSIONS: The results of this study suggest that ADA stem extract may be useful for prevention and treatment of AD.

Antinociceptive and gastro-protective effect of the ethanolic extract of the flowering top of Anthocephalus Cadamba Roxb

Nusrat Subhan,Md Mokarram Hossain,Raushanara Akter,Muntasir Mamun Majumder,Md Mostafizur Rahman,Kamaluddin Ahmed,Abdul Ghani,Md Ashraful Alam,SM Raquibul Hasan 경희대학교 융합한의과학연구소 2009 Oriental Pharmacy and Experimental Medicine Vol.9 No.4

The effect of alcoholic extract of Anthocephalus (A.) Cadamba Roxb. was evaluated in experimental models of pain and ulcer. Hot tail flick test, hot plate test and acetic acid induced writhing test were employed for evaluating the peripheral as well as central analgesic mechanism exerted by the extracts. Gastroprotective activity was examined by HCl and ethanol induced gastric damage test. Test group received crude extract 500 mg/kg showed maximum time needed for the response against thermal stimuli (6.26 ± 0.439 s) which is comparable to diclofenac sodium (6.56 ± 0.381 s) in hot tail flick method. These experimental results also followed the experimental results of hot plate test where crude extract 500 mg/kg showed maximum time needed for the response against thermal stimuli (4.74 ± 0.234 s) which is comparable to diclofenac sodium (5.58 ± 0.585 s). The crude extract at 500 and 250 mg/kg showed significant reduction in acetic acid induced writhing in mice with a maximum effect of 68.026% reduction at 500 mg/kg dose which is comparable to standard diclofenac sodium (79.93%). In gastroprotective study the extract of A. Cadamba (250 and 500 mg/kg) significantly inhibited ulceration induced by both HCl and ethanol dose dependently. Results of the study suggest that the extract possesses both analgesic and gastroprotective activity on mice. The effect of alcoholic extract of Anthocephalus (A.) Cadamba Roxb. was evaluated in experimental models of pain and ulcer. Hot tail flick test, hot plate test and acetic acid induced writhing test were employed for evaluating the peripheral as well as central analgesic mechanism exerted by the extracts. Gastroprotective activity was examined by HCl and ethanol induced gastric damage test. Test group received crude extract 500 mg/kg showed maximum time needed for the response against thermal stimuli (6.26 ± 0.439 s) which is comparable to diclofenac sodium (6.56 ± 0.381 s) in hot tail flick method. These experimental results also followed the experimental results of hot plate test where crude extract 500 mg/kg showed maximum time needed for the response against thermal stimuli (4.74 ± 0.234 s) which is comparable to diclofenac sodium (5.58 ± 0.585 s). The crude extract at 500 and 250 mg/kg showed significant reduction in acetic acid induced writhing in mice with a maximum effect of 68.026% reduction at 500 mg/kg dose which is comparable to standard diclofenac sodium (79.93%). In gastroprotective study the extract of A. Cadamba (250 and 500 mg/kg) significantly inhibited ulceration induced by both HCl and ethanol dose dependently. Results of the study suggest that the extract possesses both analgesic and gastroprotective activity on mice.