1.In-vitro cancer cell cytotoxicity and alpha amylase inhibition effect of seven tropical fruit residues
Gupta Priti ; Bhatnagar Ira ; Kim Se-Kwon ; Verma Kumar Ajay ; Sharma Anubhuti
Asian Pacific Journal of Tropical Biomedicine 2014;(z2):665-671
Objective:To determine quantitative phytochemical, anticancer and antidiabetic effect of seven Indian tropical fruit residues.
Methods:In-vitro cytotoxic activity (IC50) was evaluated against cervical cancer cells (HeLa), breast cancer cells (MCF-7), hepatocellular carcinoma cells (HepG-2) and bone sarcoma cells (MG-63) and alpha amylase inhibition assay was used for antidiabetic activity.
Results: Results of phytochemical analysis revealed that all residues contained remarkable amount of alkaloid, saponin, tannin and flavonoid. Notable cancer cell growth inhibition was observed for the extract from Carissa carandas pomace and Litchi sinensis seeds with IC50 values ranged from 56.72 to 89.24 μg/mL. Alpha amylase inhibition assay was measured at six different concentrations (5, 10, 25, 50, 100 and 200 mg/mL) by using different solvent extract. Results showed that Carissa carandas possessed best activity with IC50 value as 29.66 mg/mL followed by other residues in methanol extract.
Conclusions:Study suggests that these fruit residues demonstrate promising antidiabetic and anticancer activity that substantiated its ethno medicinal use and may provide new molecules for the treatment of these diseases.
2.Non-homologous End Joining Inhibitor SCR-7 to Exacerbate Low-dose Doxorubicin Cytotoxicity in HeLa Cells.
Ajay KUMAR ; Devyani BHATKAR ; Devashree JAHAGIRDAR ; Nilesh Kumar SHARMA
Journal of Cancer Prevention 2017;22(1):47-54
Among the genotoxic drug regimens, doxorubicin (DOX) is known for its high-dose side effects in several carcinomas, including cervical cancer. This study reports on testing the combined use of a DOX genotoxic drug and SCR-7 non-homologous end joining (NHEJ) inhibitor for HeLa cells. An in vitro DNA damaging assay of DOX was performed on plasmid and genomic DNA substrate. In vitro cytotoxicity was investigated using trypan blue dye exclusion, DNA metabolizing, and propidium iodide-based flow cytometric assays. DOX (between 20–100 μM) displayed clear DNA binding and interaction, such as the shearing and smearing of plasmid and genomic DNA. DNA metabolizing assay data indicate that HeLa lysate with DOX and SCR-7 treatment exhibited better in vitro plasmid DNA stability compared with DOX treatment alone. SCR-7 augmented the effects of low-dose DOX by demonstrating enhanced cell death from 15% to 50%. The flow cytometric data also supported that the combination of SCR-7 with DOX lead to a 23% increase in propidium iodide-based HeLa staining, thus indicating enhanced death. In summary, the inhibition of NHEJ DNA repair pathway can potentiate low-dose DOX to produce appreciable cytotoxicity in HeLa cells.
Cell Death
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DNA
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DNA Damage
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DNA End-Joining Repair
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DNA Repair
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Doxorubicin*
;
Drug Therapy
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Genomic Instability
;
HeLa Cells*
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Humans
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In Vitro Techniques
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Plasmids
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Propidium
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Trypan Blue
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Uterine Cervical Neoplasms
3.Molecular heterogeneity of plpE gene in Indian isolates of Pasteurella multocida and expression of recombinant PlpE in vaccine strain of P. multocida serotype B: 2.
Ajay Pratap SINGH ; Satparkash SINGH ; Rajeev RANJAN ; Santosh Kumar GUPTA ; Vijendra Pal SINGH ; Bhaskar SHARMA
Journal of Veterinary Science 2010;11(3):227-233
Outer membrane proteins of Pasteurella (P.) multocida have been known to be protective immunogens. Pasteurella lipoprotein E (PlpE) has been reported to be an important cross reactive outer membrane protein in P. multocida. The gene encoding the PlpE of P. multocida serotypes A: 3, B: 2 and D: 1 was amplified from the genomic DNA. The amplified products were cloned and the nucleotide sequence was determined. Sequence analysis of the recombinant clones revealed a single open reading frame of 1,011 bp, 1,008 bp and 1,017 bp encoding a protein with a calculated molecular mass of 37.829 kDa, 37.389 kDa and 37.965 kDa for serotypes A: 3, B: 2 and D: 1 respectively. The comparison of the plpE sequence in different capsular types revealed a high degree (>90%) of homology. Furthermore, the plpE gene of Haemorhhagic septicaemia causing serotype (B: 2) was expressed in E. coli and recombinant PlpE was strongly immunostained by antiserum against whole cell antigen, indicating that the protein is expressed in vivo.
Animals
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Bacterial Outer Membrane Proteins/*genetics/immunology/metabolism
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Base Sequence
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Blotting, Western
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Cattle
;
Cattle Diseases/*microbiology
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Cloning, Molecular
;
Electrophoresis, Polyacrylamide Gel
;
Escherichia coli
;
*Genetic Variation
;
Hemorrhagic Septicemia/microbiology/*veterinary
;
India
;
Lipoproteins/*genetics/immunology/metabolism
;
Molecular Sequence Data
;
Open Reading Frames/genetics
;
Pasteurella multocida/*genetics/immunology
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Sequence Analysis, DNA
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Sequence Homology
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Serotyping
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Species Specificity
4.COVID-19 Rapid Antigen Test: Role in Screening Prior to Gastrointestinal Endoscopy
Ashok DALAL ; Ujjwal SONIKA ; Manish KUMAR ; Roshan GEORGE ; Ajay KUMAR ; Siddharth SRIVASTAVA ; Sanjeev SACHDEVA ; Barjesh Chander SHARMA
Clinical Endoscopy 2021;54(4):522-525
Background/Aims:
The severe acute respiratory syndrome coronavirus 2 pandemic has affected the gastrointestinal (GI) endoscopy units globally owing to the risk of transmission. We present our data on the use of rapid antigen test (RAT) as a screening tool prior to endoscopy to prevent the transmission of coronavirus disease (COVID-19).
Methods:
This study was a retrospective analysis of patients who underwent any GI endoscopic procedure from July 2020 to October 2020 at a tertiary referral center in New Delhi, India. All patients underwent screening for COVID-19 using RAT, and endoscopy was performed only when the RAT was negative. The data are presented as numbers and percentages.
Results:
A total of 3,002 endoscopic procedures were performed during the study period. Only one endoscopic procedure was performed in a COVID-19 positive patient. A total of 53 healthcare workers were involved in conducting these procedures. Only 2 healthcare workers (3.8%) were diagnosed COVID-19 positive, presumably due to community-acquired infection, during this period.
Conclusions
The COVID-19 RAT is easily usable as a simple screening tool prior to GI endoscopy during the COVID-19 pandemic.
5.COVID-19 Rapid Antigen Test: Role in Screening Prior to Gastrointestinal Endoscopy
Ashok DALAL ; Ujjwal SONIKA ; Manish KUMAR ; Roshan GEORGE ; Ajay KUMAR ; Siddharth SRIVASTAVA ; Sanjeev SACHDEVA ; Barjesh Chander SHARMA
Clinical Endoscopy 2021;54(4):522-525
Background/Aims:
The severe acute respiratory syndrome coronavirus 2 pandemic has affected the gastrointestinal (GI) endoscopy units globally owing to the risk of transmission. We present our data on the use of rapid antigen test (RAT) as a screening tool prior to endoscopy to prevent the transmission of coronavirus disease (COVID-19).
Methods:
This study was a retrospective analysis of patients who underwent any GI endoscopic procedure from July 2020 to October 2020 at a tertiary referral center in New Delhi, India. All patients underwent screening for COVID-19 using RAT, and endoscopy was performed only when the RAT was negative. The data are presented as numbers and percentages.
Results:
A total of 3,002 endoscopic procedures were performed during the study period. Only one endoscopic procedure was performed in a COVID-19 positive patient. A total of 53 healthcare workers were involved in conducting these procedures. Only 2 healthcare workers (3.8%) were diagnosed COVID-19 positive, presumably due to community-acquired infection, during this period.
Conclusions
The COVID-19 RAT is easily usable as a simple screening tool prior to GI endoscopy during the COVID-19 pandemic.
6. Ebola virus disease: Past, present and future
Harish RAJAK ; Deepak Kumar JAIN ; Avineesh SINGH ; Ajay Kumar SHARMA ; Anshuman DIXIT
Asian Pacific Journal of Tropical Biomedicine 2015;5(5):337-343
Ebola virus disease is one of the most deadly ailments known to mankind due to its high mortality rate (up to 90%) accompanying with the disease. Ebola haemorrhagic fever (EHF) is an infectious disease of animal that can be transmitted to both human and non-human primates. The first epidemic of EHF occurred in 1976 in the Democratic Republic of the Congo. The incubation period of ebola is less than 21 days. Ebola virus infections are depicted by immune suppression and a systemic inflammatory response that leads to damage of the vascular, coagulation and immune systems, causing multi-organ failure and shock. Five genetically distinct members of the Filoviridae family responsible for EHF are as follows: Zaire ebolavirus, Sudan ebolavirus, Côte d'Ivoire ebolavirus, Bundibugyo ebolavirus and Reston ebolavirus. The ongoing 2014 West Africa ebola epidemic has been considered as the most serious panic in the medical field with respect to both the number of human cases and death toll. The natural host for ebola virus is unknown, thus it is not possible to carry out programs to regulate or abolish virus from transmission to people. The ebola virus infection provides little chance to develop acquired immunity causing rapid progression of the disease. It is pertinent to mention that at present, there is no antiviral therapy or vaccine that is helpful against ebola virus infection in humans. The impediment of EHF necessitates much better understanding of the epidemiology of the disease, particularly the role of wildlife, as well as bats, in the spread of ebola virus to humans.
7.Elevated N1-Acetylspermidine Levels in Doxorubicintreated MCF-7 Cancer Cells: Histone Deacetylase 10 Inhibition with an N1-Acetylspermidine Mimetic
Ajay Kumar RAJ ; Kiran Bharat LOKHANDE ; Kratika KHUNTETA ; Sachin Chakradhar SARODE ; Nilesh Kumar SHARMA
Journal of Cancer Prevention 2024;29(2):32-44
Cancer drug resistance is associated with metabolic adaptation. Cancer cells have been shown to implicate acetylated polyamines in adaptations during cell death. However, exploring the mimetic of acetylated polyamines as a potential anticancer drug is lacking.We performed intracellular metabolite profiling of human breast cancer MCF-7 cells treated with doxorubicin (DOX), a well known anticancer drug. A novel and in-house vertical tube gel electrophoresis assisted procedure followed by LC-HRMS analysis was employed to detect acetylated polyamines such as N1-acetylspermidine. We designed a mimetic N1-acetylspermidine (MINAS) which is a known substrate of histone deacetylase 10 (HDAC10). Molecular docking and molecular dynamics (MDs) simulations were used to evaluate the inhibitory potential of MINAS against HDAC10. The inhibitory potential and the ADMET profile of MINAS were compared to a known HDAC10 inhibitor Tubastatin A. N1-acetylspermidine, an acetylated form of polyamine, was detected intracellularly in MCF-7 cells treated with DOX over DMSO-treated MCF-7 cells. We designed and curated MINAS (PubChem CID 162679241). Molecular docking and MD simulations suggested the strong and comparable inhibitory potential of MINAS (–8.2 kcal/ mol) to Tubastatin A (–8.4 kcal/mol). MINAS and Tubastatin A share similar binding sites on HDAC10, including Ser138, Ser140, Tyr183, and Cys184. Additionally, MINAS has a better ADMET profile compared to Tubastatin A, with a high MRTD value and lower toxicity. In conclusion, the data show that N1-acetylspermidine levels rise during DOX-induced breast cancer cell death. Additionally, MINAS, an N1-acetylspermidine mimetic compound, could be investigated as a potential anticancer drug when combined with chemotherapy like DOX.
8.Anatomical study of the suprascapular notch: quantitative analysis and clinical considerations for suprascapular nerve entrapment.
Ajay KUMAR ; Anu SHARMA ; Poonam SINGH
Singapore medical journal 2014;55(1):41-44
INTRODUCTIONDetailed anatomical knowledge of the suprascapular notch (SSN) is important for the management of entrapment neuropathy and interventional procedures. The objective of the present study was to collect data on the morphological features and anatomical variations of the SSN in an Indian population.
METHODSWe studied 268 human scapulae of unknown sex (126 right-sided, 142 left-sided) taken from the Department of Anatomy, Dayanand Medical College and Hospital, India. SSNs were classified as either type I, II, III, IV or V, based on the shape of the inferior border of the incisura, and comparison of the SSN's vertical and transverse diameters. The shape of the SSN (i.e V- or U-shaped), if present, was also recorded.
RESULTSType II SSN was the most common (50.00%), followed by type I, type IV and type III (32.46%, 9.70% and 7.84%, respectively). For right-sided type II SSNs, the transverse and vertical diameters were 9.1 ± 3.2 mm and 5.2 ± 1.9 mm, respectively, while those for left-sided type ll SSNs were 9.2 ± 2.4 mm and 5.1 ± 1.8 mm, respectively. Generally, the transverse diameter of type II SSN was found to be greater than that of type III SSN. The incidence of U-shaped SSN was 51.49%, while that of V-shaped SSN was 2.99%.
CONCLUSIONThis study of the morphometrical characteristics and anatomical variations of SSN provides an anatomical database of SSN in the Indian context. This database will be of use in surgical procedures, as the information can be used to ensure adequate access to and complete decompression of the suprascapular nerve.
Humans ; India ; Nerve Compression Syndromes ; diagnosis ; Scapula ; anatomy & histology ; surgery ; Shoulder Joint ; injuries