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  1. Article ; Online: Free spermidine evokes superoxide radicals that manifest toxicity

    Vineet Kumar / Rajesh Kumar Mishra / Debarghya Ghose / Arunima Kalita / Pulkit Dhiman / Anand Prakash / Nirja Thakur / Gopa Mitra / Vinod D Chaudhari / Amit Arora / Dipak Dutta

    eLife, Vol

    2022  Volume 11

    Abstract: Spermidine and other polyamines alleviate oxidative stress, yet excess spermidine seems toxic to Escherichia coli unless it is neutralized by SpeG, an enzyme for the spermidine N-acetyl transferase function. Thus, wild-type E. coli can tolerate applied ... ...

    Abstract Spermidine and other polyamines alleviate oxidative stress, yet excess spermidine seems toxic to Escherichia coli unless it is neutralized by SpeG, an enzyme for the spermidine N-acetyl transferase function. Thus, wild-type E. coli can tolerate applied exogenous spermidine stress, but ΔspeG strain of E. coli fails to do that. Here, using different reactive oxygen species (ROS) probes and performing electron paramagnetic resonance spectroscopy, we provide evidence that although spermidine mitigates oxidative stress by lowering overall ROS levels, excess of it simultaneously triggers the production of superoxide radicals, thereby causing toxicity in the ΔspeG strain. Furthermore, performing microarray experiment and other biochemical assays, we show that the spermidine-induced superoxide anions affected redox balance and iron homeostasis. Finally, we demonstrate that while RNA-bound spermidine inhibits iron oxidation, free spermidine interacts and oxidizes the iron to evoke superoxide radicals directly. Therefore, we propose that the spermidine-induced superoxide generation is one of the major causes of spermidine toxicity in E. coli.
    Keywords Escherichia coli ; SpeG ; spermidine toxicity ; superoxide radical ; iron metabolism ; oxidative stress ; Medicine ; R ; Science ; Q ; Biology (General) ; QH301-705.5
    Subject code 500
    Language English
    Publishing date 2022-04-01T00:00:00Z
    Publisher eLife Sciences Publications Ltd
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article: Mass Spectrometry-Based Diagnosis of Hemoglobinopathies: A Potential Tool for the Screening of Genetic Disorder

    Das, Rajdeep / Amit Kumar Mandal / Boby Mathew / Cecil Ross / Debnath Pal / Gopa Mitra / Vijay Bhat

    Biochemical genetics. 2016 Dec., v. 54, no. 6

    2016  

    Abstract: Hemoglobinopathies are caused by point mutation in globin gene that results in structural variant of hemoglobin. While 7 % of world populations are carrier of hemoglobinopathies, the prevalence of the disease varies between 3 to 17 % across different ... ...

    Abstract Hemoglobinopathies are caused by point mutation in globin gene that results in structural variant of hemoglobin. While 7 % of world populations are carrier of hemoglobinopathies, the prevalence of the disease varies between 3 to 17 % across different population groups in India. In a diagnostic laboratory, alkaline gel electrophoresis and cation exchange-based HPLC (CE-HPLC) are most widely used techniques for characterization of hemoglobin variants. In the above methods, the differential surface charge of hemoglobin molecule in variants is exploited for their characterization. Sometime, co-migration of variants in gel electrophoresis and co-elution or elution with unknown retention time in automated CE-HPLC might lead to ambiguity in the analysis of hemoglobinopathies. Under such circumstances, it is necessary to use other analytical methods that provide unambiguous results. Mass spectrometry-based proteomics approach and DNA sequence analysis are examples of such alternative methods. In the present study, liquid chromatography coupled to mass spectrometry has been used for three commonly observed variants in India, e.g., HbE, HbQ India and HbD Punjab that appeared with inappropriate results in the conventional analysis. A customized hemoglobin variant database has been used in the mass spectrometry-based analysis of those three variants. Mass spectrometry-based proteomics approach was used to analyze above variant sample accurately.
    Keywords databases ; DNA ; gel electrophoresis ; genes ; genetic disorders ; hemoglobin ; high performance liquid chromatography ; mass spectrometry ; nucleotide sequences ; point mutation ; proteomics ; screening ; sequence analysis ; India
    Language English
    Dates of publication 2016-12
    Size p. 816-825.
    Publishing place Springer US
    Document type Article
    ZDB-ID 2168-4
    ISSN 1573-4927 ; 0006-2928
    ISSN (online) 1573-4927
    ISSN 0006-2928
    DOI 10.1007/s10528-016-9758-5
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  3. Article: Effects of different weaning strategies on survival, growth and digestive system development in butter catfish Ompok bimaculatus (Bloch) larvae

    Pradhan, Pravata Kumar / Enric Gisbert / Gopa Mitra / Joykrushna Jena / Neeraj Sood

    Aquaculture. 2014 Mar. 20, v. 424-425

    2014  

    Abstract: The effects of different feeding regimes on survival, growth and morphogenesis of the digestive system in butter catfish, Ompok bimaculatus (Bloch) larvae were investigated in the present study. Eight different feeding regimes varying on the type of food ...

    Abstract The effects of different feeding regimes on survival, growth and morphogenesis of the digestive system in butter catfish, Ompok bimaculatus (Bloch) larvae were investigated in the present study. Eight different feeding regimes varying on the type of food (Artemia nauplii, zooplankton or microdiet) and the time after hatching at which those different food items were offered to larvae were evaluated in order to find the most convenient weaning strategy for butter catfish larvae. The results indicated that the larvae weaned after 7days post hatch (dph) showed similar survival rates with those fed just live prey (Artemia nauplii or zooplankton), whereas early weaning (before 7dph) resulted in poor survival. In terms of growth performance, larvae fed solely on Artemia nauplii, showed the best results followed by those fed with zooplankton and those fed on a combination of Artemia nauplii and zooplankton. In contrast, the larvae fed under the other feeding regimes including the microdiet showed similar final growth values. With regard to the effect of different weaning strategies on the morphogenesis of the digestive system, the results indicated that early weaning delayed the development of the stomach, intestine and pancreas. However, at the end of the trial at 17dph, no differences in the level of development were observed among treatments regardless of the weaning protocol tested, which indicated the high plasticity of butter catfish to different nutritional conditions once their digestive system was completely developed and larvae were adapted to the microdiet. Hence, it was concluded that it is feasible to rear butter catfish larvae with zooplankton without dependence upon Artemia nauplii, and also that larvae may be weaned onto microdiets after a short period of co-feeding when weaning takes place after 7dph. These findings would be very useful to improve the actual larval rearing techniques for this promising catfish species from the Indian sub-continent.
    Keywords Artemia ; catfish ; early weaning ; growth performance ; hatching ; intestines ; morphogenesis ; nauplii ; Ompok ; pancreas ; rearing ; stomach ; survival rate ; zooplankton
    Language English
    Dates of publication 2014-0320
    Size p. 120-130.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 185380-6
    ISSN 0044-8486 ; 0044-8516
    ISSN 0044-8486 ; 0044-8516
    DOI 10.1016/j.aquaculture.2013.12.041
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    In stock of ZB MED Bonn / Germany

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  4. Article: Mass spectrometry based characterization of Hb Beckman variant in a falsely elevated HbA1c sample

    Das, Rajdeep / Amit Kumar Mandal / Boby Mathew / Cecil Ross / Debnath Pal / Gopa Mitra / Monita Muralidharan / Vijay Bhat

    Analytical biochemistry. 2015 Nov. 15, v. 489

    2015  

    Abstract: Glycated hemoglobin (HbA1c) is a ‘gold standard’ biomarker for assessing the glycemic index of an individual. HbA1c is formed due to nonenzymatic glycosylation at N-terminal valine residue of the β-globin chain. Cation exchange based high performance ... ...

    Abstract Glycated hemoglobin (HbA1c) is a ‘gold standard’ biomarker for assessing the glycemic index of an individual. HbA1c is formed due to nonenzymatic glycosylation at N-terminal valine residue of the β-globin chain. Cation exchange based high performance liquid chromatography (CE–HPLC) is mostly used to quantify HbA1c in blood sample. A few genetic variants of hemoglobin and post-translationally modified variants of hemoglobin interfere with CE–HPLC-based quantification, resulting in its false positive estimation. Using mass spectrometry, we analyzed a blood sample with abnormally high HbA1c (52.1%) in the CE–HPLC method. The observed HbA1c did not corroborate the blood glucose level of the patient. A mass spectrometry based bottom up proteomics approach, intact globin chain mass analysis, and chemical modification of the proteolytic peptides identified the presence of Hb Beckman, a genetic variant of hemoglobin, in the experimental sample. A similar surface area to charge ratio between HbA1c and Hb Beckman might have resulted in the coelution of the variant with HbA1c in CE–HPLC. Therefore, in the screening of diabetes mellitus through the estimation of HbA1c, it is important to look for genetic variants of hemoglobin in samples that show abnormally high glycemic index, and HbA1c must be estimated using an alternative method.
    Keywords biomarkers ; blood glucose ; blood sampling ; cation exchange ; diabetes mellitus ; genetic variation ; glycemic index ; glycohemoglobin ; glycosylation ; hemoglobin ; high performance liquid chromatography ; mass spectrometry ; patients ; peptides ; proteolysis ; proteomics ; screening ; surface area ; valine
    Language English
    Dates of publication 2015-1115
    Size p. 53-58.
    Publishing place Elsevier Inc.
    Document type Article
    ZDB-ID 1110-1
    ISSN 1096-0309 ; 0003-2697
    ISSN (online) 1096-0309
    ISSN 0003-2697
    DOI 10.1016/j.ab.2015.07.010
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  5. Article: Ontogeny of the digestive enzymes in butter catfish Ompok bimaculatus (Bloch) larvae

    Pradhan, Pravata Kumar / Enric Gisbert / Gopa Mitra / Joykrushna Jena / Neeraj Sood

    Aquaculture. 2013 Jan. 24, v. 372-375

    2013  

    Abstract: The digestive physiology of butter catfish was studied by assessing the activity of different pancreatic (trypsin, chymotrypsin, α-amylase and lipase), gastric (pepsin) and intestinal (alkaline phosphatase) enzymes from hatching until the juvenile stage ( ...

    Abstract The digestive physiology of butter catfish was studied by assessing the activity of different pancreatic (trypsin, chymotrypsin, α-amylase and lipase), gastric (pepsin) and intestinal (alkaline phosphatase) enzymes from hatching until the juvenile stage (30dph). Larvae were reared at 27°C and fed with Artemia nauplii from 2days post hatching (dph) until 10dph, from 7–10dph with Artemia nauplii and zooplankton (Cyclopoida) and from 10dph onwards only with zooplankton. The assessment of the activity of digestive enzymes showed that enzymes involved in the digestion of proteins, lipids and carbohydrates were present in butter catfish larvae before mouth opening and increased after the onset of exogenous feeding, coinciding with the histological organization of the exocrine pancreas. The specific activity of most of the pancreatic enzymes increased until 15dph and decreased thereafter coinciding with the increase of pepsin. A progressive shift in activity from alkaline (trypsin and chymotrypsin) to acid (pepsin) proteases indicated a change in the digestive physiology of the specimen, as alkaline proteases were no longer the main digestive enzymes involved in protein digestion after the onset of acidic digestion between 15 and 21dph. The maturation of the intestine and the achievement of a juvenile-like mode of digestion were demonstrated by changes in enzyme activities from the exocrine pancreas and stomach that coincided with alterations in enzyme production occurring in the intestine (e.g. alkaline phosphatase). Considering the ontogenetic development of the digestive enzymes from the pancreas, stomach and intestine, butter catfish larvae might be weaned between 15 and 21dph, as larvae have achieved the complete maturation of their digestive capacities. These results contradict previous recommendations, which were based solely on the histological organization of the gastric gland and histochemical properties of mucous cells from the stomach, to wean butter catfish larvae at earlier ages. These findings on the functional development of the digestive system in butter catfish would be useful to improve the actual larval rearing techniques for this promising catfish species from the Indian sub-continent.
    Keywords alkaline phosphatase ; Artemia ; carbohydrates ; catfish ; chymotrypsin ; digestion ; enzyme activity ; gastric mucosa ; hatching ; intestines ; lipids ; mouth ; nauplii ; ontogeny ; pancreas ; pepsin ; proteins ; rearing ; triacylglycerol lipase ; trypsin
    Language English
    Dates of publication 2013-0124
    Size p. 62-69.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 185380-6
    ISSN 0044-8486 ; 0044-8516
    ISSN 0044-8486 ; 0044-8516
    DOI 10.1016/j.aquaculture.2012.10.024
    Database NAL-Catalogue (AGRICOLA)

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    In stock of ZB MED Bonn / Germany

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