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MTHFR C677T GENE MUTATION ASSOCIATED WITH SEVERE RISK FOR MENTAL RETARDATION IN CHILDREN
AJIT KUMAR SAXENA
International Journal of Genetics , 2011,
Abstract: MTHFR gene polymorphism regulate folate metabolism, required for normal development of central nervoussystem and any error in metabolism either due to hereditary or sporadic gene mutation in the family lead to the developmentof mental retardation in heterozygous condition. In the present study the five families having severe congenital mentalretardation was evaluated for C677T genotype variations i.e. CC, CT & TT in probands, mother & father using PCR-RFLPanalysis. Highest frequency (60%) was observed in heterozygous condition in the mother of proband. Statistical analysisshowing significant difference (p=0.024) were observed in the father of the proband. Interestingly, the insertion of 68bp ofCβS gene mutation was also observed in father of one family suggesting paternal or maternal factors influencing for severemental retardation in children of consanguineous families as an independent risk factor. However, the high degree of geneticheterogeneity is quite striking because of severe mutation in two families. The pedigree analysis showing that the severityof disease transmission probably due to the penetrance of gene and their mode of inheritance is autosomal recessive innature.
Evaluation of Cassia occidentalis for in vitro cytotoxicity against human cancer cell lines and antibacterial activity
Bhagat Madhulika,Saxena Ajit
Indian Journal of Pharmacology , 2010,
Abstract: Objective : To evaluate the in vitro cytotoxicity and antibacterial properties of Cassia occidentalis (whole plant) via alcoholic, hydro-alcoholic, and aqueous extracts against eight human cancer cell lines from six different tissues and four bacterial strains. Material and Methods : In vitro cytotoxicity against the human cancer cells, cultured for 48h in presence of different concentrations C. occidentalis extracts and percentage of cell viability, was evaluated using the sulforhodamine-B (SRB) assay. The antibacterial activity was performed using the standard protocol against bacterial strains. Results : It was observed that aqueous extract of C. occidentalis (whole plant) had more potential than hydro-alcoholic and alcoholic extracts against HCT-15, SW-620, PC-3, MCF-7, SiHa, and OVCAR-5 human cancer cell lines at 100, 30, and 10 μg/ml in a dose-dependent manner. The hydro-alcoholic extract showed potential against Bacillus subtillis. Conclusion : The plant can be explored for the possible development of lead molecules for drug discovery.
Biological evaluation of 8-alkyl xanthines as potential cytotoxic agents  [PDF]
Renuka Suravajhala, Nitasha Suri, Madhulika Bhagat, Ajit Kumar Saxena
Advances in Biological Chemistry (ABC) , 2013, DOI: 10.4236/abc.2013.33035
Abstract:

A series of 8-substituted alkyl xanthines were evaluated in vitro to test the cytotoxocity in cells. For this experiment, we utilized different mammalian cancer cell lines primarily representing prostrate and lung. One of the compounds synthesized, viz. 8-tertbutyl caffeine showed potent anticancer activity at low concentrations against DU145 when compared to adriamycin. Further experiments were carried out to check the cell cycle arrest in the DU145 cells treated with adriamycin, caffeine and 8-tert butyl caffeine. We observed that there was an arrest in G1 phase of cell cycle at 24 hours while at 48 hours of incubation, the cells were constantly distributed (59.71% -70.79%). We conclude that the effect of 8-tertbutyl caffeine is relatively comparable to caffeine whereas in adriamycin treated cells, we observed the cells underwent G2 arrest. We evaluate the studies on these effects by showing potent analogues which could be used as promising anticancer agents.

Isolation, purification and characterization of an N-acetyl-D-lactosamine binding mitogenic and anti-proliferative lectin from tubers of a cobra lily Arisaema utile Schott  [PDF]
Vikram Dhuna, Kshitija Dhuna, Jatinder Singh, Ajit Kumar Saxena, Satyam Kumar Agrawal, Sukhdev Singh Kamboj
Advances in Bioscience and Biotechnology (ABB) , 2010, DOI: 10.4236/abb.2010.12012
Abstract: Lectins are the carbohydrate-binding proteins of non-immune origin which have been the subject of intense investigation over the last few decades owing to the variety of interesting biological properties. Most of the lectins which have been purified and characterized from plants have been obtained from dicotyledons. In the present study a lectin was purified from tubers of a monocot plant Arisaema utile (AUL) Schott by affinity chromatography on asialofetuin-linked amino activated silica beads. AUL gave a single band in SDS-PAGE at pH 8.3 corresponding to subunit Mr 13.5 kDa. The native molecular mass of AUL was 54 kDa suggesting a homotetrameric structure. AUL gave multiple bands in isoelectric focusing and in native PAGE at pH 8.3. AUL was inhibited by N-acetyl-D-lactosamine (Lac NAc), a disaccharide and asialofetuin, a complex desialylated serum glycoprotein. When treated with denaturing agents, the lectin was stable in the presence of urea (3 M), thiourea (4 M) and guanidine HCl (4 M). AUL was a glycoprotein with a carbohydrate content of 1.2%. Complete loss of activity was observed upon modification of tryptophan residues of the lectin. The activity was reduced to 25% after modification of tyrosine. Chemical modification of arginine, histidine, serine and cysteine residues of AUL did not affect its activity. Using Far UV CD spectra the estimated secondary structure was 37% α-helix, 25% β-sheet and 38% random contributions. The lectin showed potent mitogenic response towards human lymphocytes. In vitro anti-proliferative assay using 11 human cancer cell lines resulted in 50% inhibition of six cell lines viz. SW-620, HCT-15, SK-N-SH, IMR-32, Colo-205 and HT-29 at 38, 42, 43, 49, 50 and 89 µg/ml, respectively.
In vitro cytotoxicity of extracts and fractions of Calotropis procera (Ait.) roots against human cancer cell lines
Bhagat Madhulika,Arora Jatinder,Saxena Ajit
International Journal of Green Pharmacy , 2010,
Abstract: This study was designed to determine the antiproliferative activity of three extracts (alcoholic, hydro-aqueous and aqueous) and their fractions from the root part of Calotropis procera using human oral (KB) and central nervous system (SNB-78) cancer cell lines as a model system. KB and SNB-78 cells were cultured in the presence of extracts and fractions at various concentrations (10, 30 and100 μg/ml) for 48 h, and the percentage of cell viability was evaluated by the sulforhodamine-B (SRB) assay. Our result indicates that out of the three extracts of C. procera (root), alcoholic extract had shown greater potential for growth inhibition followed by hydro-aqueous extract at three different concentration of 10 μg/ml, 30 μg/ml and 100 μg/ml in a dose-dependent manner, whereas aqueous extract was found to be least active against both oral and CNS human cancer lines. On evaluation of the fractions prepared from alcoholic and hydro-aqueous extracts, it was observed that chloroform fraction from alcoholic extract was antiproliferative for oral (KB) cancer cell line and n-butanol fraction from alcoholic extract was antiproliferative for CNS cancer cell line than remaining fractions at three different concentration of 10 μg/ml, 30 μg/ml, 100 μg/ml in a dose-dependent manner. Thus, our result indicates that the root part of C. procera possess in vitro cytotoxicity against oral and CNS human cancer cell lines. Further investigations are required to obtain the clinically important lead molecules for the drug development.
OPTIMIZATION OF CHANNEL ALLOCATION ALGORITHM FOR HOT SPOT CELLS IN WIRELESS NETWORKS
Kriti Saraswat,Ajit Kumar Shrivastava,Amit Saxena
International Journal of Computers & Technology , 2012,
Abstract: Dense deployment of cellular networks is leading to scarcity of communication bandwidth or what we call as channel. If compared to its wired counterparts, wireless cellular network have limited number of channels available, which gives rise to problem of efficient channel allocation. Here, in this piece of work, the main objective is to put an effort to improve existing channel allocation scheme. In earlier existing hybrid allocation scheme, the base station notifies about the hot-spots to the Mobile Switching Centre (MSC) and if MSC has available channels in its central pool then it satisfies the request. Now, the novelty of this work starts where central pool gets exhausted and request of channels from base station (BS) still arrives and is served by returning the unused channels by different cells back to MSC on its request. The simulation of this approach is expounded and evaluated over OMNeT++ in a scenario with fixed channel allocation and hybrid approach by varying the proportion of dynamic channels to total number of channels available and the effectiveness is evaluated in terms of Call blocked and Call dropped versus System load.
Primary primitive neuroectodermal tumor: An unusual cause of right ventricular intracavitary obstruction in a child
Thachil Ajit,Saxena Anita,Choudhary Ujjwal,Ray Ruma
Annals of Pediatric Cardiology , 2008,
Abstract: A six-year-old boy presented with a brief history suggestive of right heart failure. Investigations revealed a mass filling almost the entire right ventricle. Palliative resection of the mass was done. The operative specimen revealed a primary primitive neuroectodermal tumor of the heart, the first of its kind reported in the pediatric age group .
Anti-Tumoral and Toxicological Studies with 2-[4-{3-(2-Chloroethyl)-3-Nitrosoureido}butyl]-Naphthalimide, a Novel Nitrosourea Molecule
Asama Mukherjee,Sushanta Dutta,Gousia Chashoo,Ajit Kumar Saxena
Journal of Cancer Molecules , 2010,
Abstract: AIM: In the present study, a novel nitrosourea compound was prepared. Its anti-tumor efficacy was evaluated on human tumor cell lines in vitro and murine tumors in vivo. Drug-induced toxicities in normal and tumor-bearing mice at its optimum dose and in human peripheral blood mononuclear cells were investigated.METHODS: In vitro cytotoxicity was determined by MTT or SRB (sulphorhodamine B) colorimetric assay. In vivo activity was assessed by measuring the increase in the median survival time of drug-treated (T) over untreated control (C) mice. Drug-induced toxicities in respect of hematological parameters, femoral bone marrow and splenic cellularities as well as biochemical parameters and histopathology of liver and kidney were assessed in vivo in normal and sarcoma-180 bearing mice sequentially on days 9, 14 and 19 following drug treatment at 50 mg/kg dose at the schedule QD1-7. Inhibition of DNA/RNA synthesis was measured by the incorporation of 3H-thymidine or 3H-uridine uptake by Ehrlich ascites carcinoma (EAC) cells in vitro.RESULTS: Results indicated a significant cytotoxicity of 2-[4-{3-(2-chloroethyl)-3-nitrosoureido}butyl]-naphthalimide in histiocytic lymphoma U-937 cells in vitro and an excellent tumor regression effect in the mice with sarcoma-180 (T/C > 250) and EAC (T/C = 217) tumors in vivo. No cardiotoxicity, hepatotoxicity or nephrotoxicity was reflected in biochemical parameters at the optimum dose. An initial hyposplenic cellularity and the femoral bone marrow suppression effect observed on day 9 reached normalcy by day 19. No adverse effect was observed on peripheral blood mononuclear cells. It inhibited the synthesis of DNA/RNA in EAC cells comparable to standards at the same concentration of 8 microM.CONCLUSION: Results warranted a promising therapeutic potential of 2-[4-{3-(2-chloroethyl)-3-nitrosoureido}butyl]-naphthalimide for further investigation.
Stakeholder’s Perspective of Clinical Decision Support System  [PDF]
Ajit Kumar
Open Journal of Business and Management (OJBM) , 2016, DOI: 10.4236/ojbm.2016.41005
Abstract: Clinical Decision Support System (CDSS) has potential opportunities to improve overall safety, quality and cost-effectiveness of healthcare. The CDSS has existed for more than four decades, but its adoption rate by medical communities is not encouraging even in the countries that have been a pioneer in developing them. At many sites, it was problematic, stalled in the planning stages or never even attempted. To date, CDSS is considered as a partially successful system. Several current challenges have not been adequately addressed during the development of CDSS. As per latest research, the lists of challenges are: improve the human-computer interface, disseminate best practices in CDSS design, development, and implementation, summarize patient-level information, prioritize and filter recommendations to the user, create an architecture for sharing executable CDSS modules and services, combine recommendations for patients with co-morbidities, prioritize CDSS content development and implementation, create Internet-accessible clinical decision support repositories, use free text information to drive clinical decision support, and mine huge clinical databases to create new CDSS. The preceding list has been considered as challenges due to unmet expectations of CDSS’s stakeholders, such as CDSS product development and maintenance team (product owner, project managers, system architect, system designers, system developers, system administrators, and system maintenance team), sales and marketing personnel, end-users. We found that most of the CDSS literature talked about the challenges and their details, but they do not throw enough light from CDSS stakeholders’ perspective while building and upon using CDSS. This paper describes CDSS from various stakeholders’ perspective, highlighting the challenges faced by them in owning, building, and using them.
6-Nitro-2-(3-hydroxypropyl)-1H-benz[de]isoquinoline-1,3-dione, a potent antitumor agent, induces cell cycle arrest and apoptosis
Asama Mukherjee, Sushanta Dutta, Muthiah Shanmugavel, Dilip M Mondhe, Parduman R Sharma, Shashank K Singh, Ajit K Saxena, Utpal Sanyal
Journal of Experimental & Clinical Cancer Research , 2010, DOI: 10.1186/1756-9966-29-175
Abstract: Compounds 1a-j were initially screened in MOLT-4, HL-60 and U-937 human tumor cell lines and results were compared with established clinical drugs. Cytotoxicities of compounds 1d and 1i were further evaluated in a battery of human tumor cell lines and in normal human peripheral blood mononuclear cells. Cell cycle analysis of compound 1i treated MOLT-4 cells was studied by flow cytometry. Its apoptosis inducing effect was carried out in MOLT-4 and HL-60 cells by flow cytometry using annexin V-FITC/PI double staining method. The activities of caspase-3 and caspase-6 in MOLT-4 cells following incubation with compound 1i were measured at different time intervals. Morphology of the MOLT-4 cells after treatment with 1i was examined under light microscope and transmission electron microscope. 3H-Thymidine and 3H-uridine incorporation in S-180 cells in vitro following treatment with 8 μM concentration of compounds 1d and 1i were studied.6-Nitro-2-(3-hydroxypropyl)-1H-benz[de]isoquinoline-1,3-dione (compound 1i), has exhibited maximum activity as it induced significant cytotoxicity in 8 out of 13 cell lines employed. Interestingly it did not show any cytotoxicity against human PBMC (IC50 value 273 μM). Cell cycle analysis of compound 1i treated MOLT-4 cells demonstrated rise in sub-G1 fraction and concomitant accumulation of cells in S and G2/M phases, indicating up-regulation of apoptosis along with mitotic arrest and/or delay in exit of daughter cells from mitotic cycle respectively. Its apoptosis inducing effect was confirmed in flow cytometric study in MOLT-4 and the action was mediated by activation of both caspase 3 and 6. Light and transmission electron microscopic studies corroborated its apoptosis inducing efficacy at a concentration of 10 μM in MOLT-4 cells. Its apoptosis induction was also observed in HL-60 cells to an extent much greater than well known apoptosis inducing agents as camptothecin and cis-platin at 10 μM concentration each. It significantly inhibite
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