Curcumin Conjugates Induce Apoptosis Via a Mitochondrion Dependent Pathway in MCF-7 and MDA-MB-231 Cell Lines

Singh, Durg Vijay,Agarwal, Shikha,Singh, Preeti,Godbole, Madan Madhav,Misra, Krishna Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.10

In order to enhance the bioavailability of curcumin its conjugates with piperic acid and glycine were synthesized by esterifying the 4 and 4' phenolic hydroxyls, the sites of metabolic conjugation. Antiproliferative and apoptotic efficacy of synthesized conjugates was investigated in MCF-7 and MDA-MB-231 cell lines. $IC_{50}$ values of di-O-glycinoyl (CDG) and di-O-piperoyl (CDP) esters of curcumin were found to be comparable with that of curcumin. Both conjugates induced chromatin condensation fragmentation and apoptotic body formation. CDP exposure to MCF-7 cells induced apoptosis initiating loss of mitochondrial membrane potential (${\Delta}{\Psi}m$) followed by inhibition of translocation of transcription factor NF-${\kappa}B$ and release of Cytochrome-C. Reactive oxygen species (ROS) production was evaluated by fluorescent activated cell sorter. Change in ratio of Bcl2/Bclxl was observed, suggesting permeablization of mitochondrial membrane leading to the release of AIF, Smac and other apoptogenic molecules. DNA fragmentation as a hallmark for apoptosis was monitored by TUNEL as well as agrose gel electrophoresis. Thus, it was proven that conjugation does not affect the therapeutic potential of parent molecule in vitro, while these could work in vivo as prodrugs with enhanced pharmacokinetic profile. Pharmacokinetics of these molecules under in vivo conditions is a further scope of this study.

Tumor associated mast cells: biological roles and therapeutic applications

Shikha Saxena,Anil Singh,Priyanka Singh 대한해부학회 2020 Anatomy & Cell Biology Vol.53 No.3

Mast cells (MCs) are immune cells of the myeloid lineage and are present in connective tissues throughout the body. The activation and degranulation of MCs significantly modulates many aspects of physiological and pathological conditions in various settings. Recent data have expanded the concept that inflammation is a critical component for tumor progression. Interestingly, three of the most aggressive human cancers, malignant melanoma, breast carcinoma and colorectal adenocarcinoma, are commonly associated with a marked host response comprising of various inflammatory cells, but especially MCs around the tumor periphery. A systematic review of the literature was performed based on the English titles listed in the PubMed, EBSCO, Cochrane, Science Direct, ISI web Science, and SciELO databases using the keywords. Abstracts and full-text articles were assessed. This review summarizes the current understanding of the role of MCs in tumor progression.

Do Human Papilloma Viruses Play Any Role in Oral Squamous Cell Carcinoma in North Indians?

Singh, Vineeta,Husain, Nuzhat,Akhtar, Naseem,Kumar, Vijay,Tewari, Shikha,Mishra, Sridhar,Misra, Sanjeev,Khan, M.Y. Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.16

Background: Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy among males in India. While tobacco and alcohol are main aetiological factors, human papilloma virus (HPV) presence has surprisingly increased in head and neck Squamous Cell Carcinoma (HNSCC) in the past two decade but its frequency in OSCCS is still uncertain. We aim to explore the frequency of HPV and its major genotypes in North Indian patients and their association with clinicopathological and histopathological features and p16 expression pattern. Materials and Methods: The study group comprised 250 histologically proven cases of OSCC. HPV was detected by real time PCR in tumor biopsy specimens and confirmed by conventional PCR with PGMY09/PGMY11 primers. Genotyping for high-risk types 16/18 was conducted by type specific PCR. p16 expression was assessed by immunohistochemsitry. Results: HPV presence was confirmed in 23/250 (9.2%) OSCC cases, of which 30.4% had HPV 16 infection, 17.4%were positive for HPV 18 and 26.1% had co-infections. HPV presence was significantly associated with male gender (p=0.02) and habit of pan masala chewing (p=0.01). HPV positive cases also had a history of tobacco consumption in 91.3% cases. p16 over expression was observed in 39.1% of HPV positive cases but this was not significantly different from negative cases (p=0.54). Conclusions: The frequency of HPV in OSCC is low in North-India and majority of cases are associated with a tobacco habit. It appears that tobacco shows a confounding effect in HPV positive cases and use of p16 protein as a reliable marker to assess the potential etiological role of HPV in OSCC in our population is not suggested.

Reproductive performance of genetically engineered mice housed in different housing systems

Shikha Yadav,Inderjeet Yadav,Kunal Pratap,Pradeep Kumar Tiwari,Vijay Pal Singh 한국실험동물학회 2017 Laboratory Animal Research Vol.33 No.2

The genetically engineered mice require special husbandry care and are mainly housed in Individually Ventilated Cage (IVC) systems and Static Micro Isolator Cages (SMIC) to minimize the risk for spreading undesirable microorganisms. However, the static micro isolation cage housing like SMIC are being replaced with IVC systems in many facilities due to a number of benefits like a higher density housing in limited space, better protection from biohazards and allergens and decreased work load due to decreased frequency of cage changing required in this system. The purpose of this study was to examine the reproductive performance of genetically engineered mice housed in individually ventilated cages (IVC) and Static Micro Isolator Cages (SMIC). When the B6C3-Tg (APPswe, PSEN1dE9) 85Dbo/Mmjax transgenic mice were housed in these two housing systems, the number of litters per dam, number of pups born per dam and number of pups weaned per dam were found to be slightly higher in the IVC as compared to the SMIC but the difference was not significant (P<0.05). In case of Growth Associated Protein 43 (GAP-43) knockout mice, the number of litters born per dam and the number of pups born per dam were marginally higher in the IVC as compared to those housed in SMIC but the difference was not significant (P<0.05). Only the number of pups weaned per dam were found to be significantly higher as compared to those housed in the SMIC system at P<0.05.

In-vitro Studies on Copper Nanoparticles and Nano-hydroxyapatite Infused Biopolymeric Composite Scaffolds for Bone Bioengineering Applications

Shikha Kumari,Abha Mishra,Divakar Singh,Chenzhong Li,Pradeep Srivastava 한국생물공학회 2023 Biotechnology and Bioprocess Engineering Vol.28 No.1

This research study deals with the development of copper nanoparticles (CN) and nano-hydroxyapatite (nHAP) infused chitosan (C) and gelatin (G) based nanocomposite scaffolds for bone tissue engineering applications. Human-origin osteoblast cells (MG-63) were seeded over the scaffolds to investigate the novel biomimetic extracellular matrix system. The scanning electron microscopy (SEM) showed an average pore size between 100-146 μm for all the C-G-nHAP-CN based scaffolds. The in-vitro degradation study showed 74-83% degradation for CN-based scaffolds. For 0.03% CN based scaffold degradation rate (84%) was very close to the control scaffold. Swelling ratio was highest for the chitosan scaffold and it was in the range between 5.25-5.93 mg/mL for other scaffolds. Compressive moduli were highest for 0.03% CN scaffold (3.32 MPa) which was relatively very high in comparison to C-G-nHAP scaffold with 2.4 MPa strength in a wet state. Stress-strain graphs also show the maximum displacement by 0.03% CN scaffold. The functional and structural analysis for the scaffolds showed the presence of nHAP in the scaffold and CN peaks within the composite structure. Differential scanning colorimetry testing showed reduced crystallinity in CN-based scaffolds with a melting temperature of 320ºC. Their 2D cell behaviour in the Electrical Cell Impedance System (ECIS) study showed maximum cell spreading and growth in 0.02% CN-based scaffold. The cell-seeded composite was tested for 3-(4,5-dimethylthiazolyl-2)-2,5- diphenyltetrazolium bromide (MTT), 4,6-diamidino-2- phenylindole (DAPI), and acridine orange and propidium iodide (AOPI) assay for testing its cytocompatibility for MG-63 cell line. Cell proliferation and cell spreading was observed by SEM in all the CN-based scaffolds. Alkaline phosphatase (ALP) activity was highest in 0.03% CN scaffold with 2.0 optical density (OD) value. Alizarin Red Stain (ARS) staining was performed to support this study. It can be statistically depicted that nHAP and 0.03% CNbased scaffold could be potential biomaterial for minor to severe bone-related tissue regeneration applications.

Awareness of cervical cancer screening among nursing staff in a tertiary institution of rural India

Ekta Singh,Shikha Seth,Vidya Rani,Dhiraj Kumar Srivastava 대한부인종양학회 2012 Journal of Gynecologic Oncology Vol.23 No.3

Objective: To determine the baseline information about the knowledge of cervical cancer and explore attitude and practice of Pap smear screening among staff nurses. Methods: A pretested structured self administered questionnaire based survey was done on 205 staff nurses working in Rural Institute of Medical Sciences & Research, Saifai, Etawah, containing mostly recognition and some recall type questions about demographics, knowledge about cervical cancer, its risk factors, screening techniques, attitudes towards cervical cancer screening and its practices. Results: In this study, 74% knew that Pap smear is used for detection of cervical cancer, but only 59% knew that it can detect both cancerous as well as precancerous lesions of the cervix. Only 18% of the respondents knew about human papillomavirus vaccine. A 47% of respondents had never taken a Pap smear; 63% never referred patients for the screening. Most nurses (79%) thought that a speculum examination and Pap smear are procedures to be performed by the doctors. Among all the respondents, only 11% had ever undergone a Pap smear on themselves. Conclusion: Despite knowledge of the gravity of cervical cancer and prevention by screening, attitudes and practices towards screening were negative. Objective: To determine the baseline information about the knowledge of cervical cancer and explore attitude and practice of Pap smear screening among staff nurses. Methods: A pretested structured self administered questionnaire based survey was done on 205 staff nurses working in Rural Institute of Medical Sciences & Research, Saifai, Etawah, containing mostly recognition and some recall type questions about demographics, knowledge about cervical cancer, its risk factors, screening techniques, attitudes towards cervical cancer screening and its practices. Results: In this study, 74% knew that Pap smear is used for detection of cervical cancer, but only 59% knew that it can detect both cancerous as well as precancerous lesions of the cervix. Only 18% of the respondents knew about human papillomavirus vaccine. A 47% of respondents had never taken a Pap smear; 63% never referred patients for the screening. Most nurses (79%) thought that a speculum examination and Pap smear are procedures to be performed by the doctors. Among all the respondents, only 11% had ever undergone a Pap smear on themselves. Conclusion: Despite knowledge of the gravity of cervical cancer and prevention by screening, attitudes and practices towards screening were negative.

P-VEP as Predictor of Occlusion Therapy

Satendra Singh,Shikha Gautam 대한안과학회 2009 Korean Journal of Ophthalmology Vol.23 No.2

Isolation of a new strain of Aspergillus and molecular structure elucidation of unknown metabolite produced from castor oil

Sumit Sharma,Shikha Singh,Saurabh Jyoti Sarma 한국공업화학회 2023 Journal of Industrial and Engineering Chemistry Vol.120 No.-

A new Aspergillus strain BU20S was isolated and identified by sequence analysis of ribosomal internaltranscribed spacer (ITS) regions. Unknown metabolite produced by the strain was identified by elucidationof molecular structure using a combination of 13C and 1H NMR, MS, FTIR, XRD, and HPLC analysis. Themetabolite was found to be kojic acid, which has the potential to treat methicillin-resistantStaphylococcus aureus (MRSA) infection, among other industrial applications. It was observed that the isolatedstrain can produce kojic acid from castor oil and product titer can be increased by supplementingthe medium with rice straw-derived depolymerized lignin. The substrate concentrations were optimizedby response surface methodology using Design-Expert software. As high as 6.69 ± 0.24 g/L of kojic acidwas obtained using castor oil (20 g/L) and depolymerized rice straw lignin (3 g/L). This is the first reporton kojic acid production from castor oil.

Impact of iron deficiency anemia on chronic periodontitis and superoxide dismutase activity: a cross-sectional study

Souvik Chakraborty,Shikha Tewari,Rajinder Kumar Sharma,Satish Chander Narula,Pratap Singh Ghalaut,Veena Ghalaut 대한치주과학회 2014 Journal of Periodontal & Implant Science Vol.44 No.2

Purpose: Both chronic periodontitis (CP) and iron deficiency anemia (IDA) induce oxidativestress in the body and cause an imbalance between reactive oxygen species and antioxidants,such as superoxide dismutase (SOD). This study explored the SOD enzyme activity ofsaliva and serum in CP patients with and without IDA and analyzed the impact of IDA on CP. Methods: A total of 82 patients were divided into four groups: control group (CG, 22), periodontallyhealthy IDA patients (IDA-PH, 20), CP patients (CP, 20), and IDA patients with CP(IDA-CP, 20). After clinical measurements and samplings, serum and salivary SOD levels weredetermined using an SOD assay kit. Results: IDA-CP patients exhibited a higher gingival index, bleeding on probing, probingpocket depth, and percentage (%) of sites with a clinical attachment loss (CAL) of ≥6 mm(P<0.008) than CP patients. The mean salivary and serum SOD levels were significantly lowerin the IDA-PH, CP, and IDA-CP patients than in the CG group (P<0.008). A significantpositive correlation between salivary and serum SOD activity was observed in IDA (P<0.05). Furthermore, serum and salivary SOD levels were significantly and negatively correlated withall periodontal parameters including the percentage of sites with CAL of 4–5 and ≥6 mm(P<0.05) except the significant correlation between salivary SOD activity and mean CAL andthe percentage of sites with CAL of 4–5 mm (P>0.05) in these patients. Conclusions: Within the limits of this study, it may be suggested that IDA patients withchronic periodontitis have more periodontal breakdowns than patients with chronic periodontitis. Serum and salivary SOD activity levels were lower in the IDA-PH, CP and IDA-CPgroups than in the CG. Iron deficiency anemia influenced the serum SOD activity but did notseem to affect the salivary SOD activity in these patients.

Effectiveness of Inorganic Nitrogen on Kojic Acid Production from Fungi Aspergillus sp. BU20S

Sharma Sumit,Singh Shikha,Sarma Saurabh Jyoti 한국화학공학회 2024 Korean Journal of Chemical Engineering Vol.41 No.4

Kojic acid is a valuable compound that contributes to various therapeutic and cosmetic applications. Organic nitrogen partially contributes to fermentation and mostly prefers cell growth as well as increases production costs. This study aims to fi nd the eff ect of low-cost inorganic nitrogen in the form of ammonium chloride on kojic acid production from fungal isolate Aspergillus sp. BU20S. A 3.63-times increase in the kojic acid (4.43 ± 0.47 g/L) was found when only the ammonium chloride was supplemented in glucose (10 g/L) medium than other minimal salts. The product formation was 2.52 ± 0.56 g/L in ammonium chloride as compared to 2.02 ± 0.06 g/L yeast extract after 10 days. The carbon/nitrogen ratio (C/N) was found optimal as 15 (molecular C/N: 23.58) which gives a high titer of 5.17 ± 0.84 g/L from 10 g/L of glucose. At this optimal molecular nitrogen value, the nitrogen supplement cost can be reduced by 93–99% compared to yeast extract. The antimicrobial potential of kojic acid purifi ed from the fermented broth was also studied against methicillin-resistant Staphylococcus aureus (MRSA). The purifi ed kojic acid showed a ~ 20 mm zone of inhibition at a 2.5 mg dose loaded over 7.4 × 10 9 CFU/ mL of MRSA. This study concludes that only ammonium chloride is a suffi cient inorganic nitrogen source to produce kojic acid and is useful in reducing production costs. The purifi ed kojic acid is also an eff ective antimicrobial agent.