Sequence variants of human mitochondrial DNA (mt DNA) have been implicated in a variety of disord... more Sequence variants of human mitochondrial DNA (mt DNA) have been implicated in a variety of disorders and conditions. Massive parallel sequencing is becoming increasingly popular due to its efficiency and cost-effectiveness. In relation to acquiring significant sequence information like levels of heteroplasmy in mt DNA, it offers a marked improvement compared to previous methods used. Here we describe a variant calling pipeline for human mitochondrial DNA using Next Generation Sequencing (NGS) data obtained by enriching the sample only for mitochondria prior to sequencing.
Gedunin is one of the major compounds found in the neem tree(Azadirachta indica). In the present ... more Gedunin is one of the major compounds found in the neem tree(Azadirachta indica). In the present study, antiproliferative potential of gedunin was evaluated in human embryonal carcinoma cells (NTERA-2, a cancer stem cell model) and peripheral blood mononuclear cells (PBMCs), using Sulforhodamine (SRB) and WST-1 assays, respectively. The effects of gedunin on expression of heat shock protein 90 (HSP90), its cochaperone Cdc37, and HSP client proteins (AKT, ErbB2, and HSF1) were evaluated by real-time PCR. Effects of gedunin on apoptosis were evaluated by (a) apoptosis associated morphological changes, (b) caspase 3/7 expression, (c) DNA fragmentation, (d) TUNEL assay, and (e) real-time PCR of apoptosis related genes (Bax,p53,andsurvivin). Gedunin showed a promising antiproliferative effect in NTERA-2 cells with IC50values of 14.59, 8.49, and 6.55 μg/mL at 24, 48, and 72 h after incubations, respectively, while exerting a minimal effect on PBMCs. Expression of HSP90, its client protein...
Garcinol (GA), a polyisoprenylated benzophenone derivative, is one of the major phytochemicals fo... more Garcinol (GA), a polyisoprenylated benzophenone derivative, is one of the major phytochemicals found in a number of fruits of Garcinia. This study aimed to develop a new nanodelivery system comprising GA, hyaluronic acid (HA), and poly(lactic-co-glycolic acid) (PLGA) to actively target breast cancer stem cells (bCSCs) grown under hypoxic conditions. HA has been reported to show higher affinity for the cluster-determinant 44 receptor (CD44), while PLGA is an FDA-approved biodegradable polymer used in clinical applications. Nanoparticles (NPs) were prepared by emulsion solvent diffusion technique using DMAB as the emulsifier. Following preparation, particle size, surface charge, and polydispersity index of NPs were characterized. Antiproliferative effects of NPs were assessed by the WST-1 cell proliferation assay. Apoptotic effects of NPs were evaluated by the caspase-3/7 assay. Effects of prepared NPs on the expression of genes related to hypoxia-inducing factors (HIF-1α and HIF-2α) ...
Additional file 1: Table S1. Optimised concentration, incubation time of the primary antibodies a... more Additional file 1: Table S1. Optimised concentration, incubation time of the primary antibodies and antigen retrieval buffer
Common genetic variations of imprinted genes that are maternally expressed could elucidate the ma... more Common genetic variations of imprinted genes that are maternally expressed could elucidate the maternal-specific inheritance of low birth weight. Human H19 is an imprinted growth regulatory gene located at chromosome 11p15.5 exclusively expressed from the maternal allele coding for a regulatory RNA implicated in the regulation of insulin-like growth factor II. We have previously described H19 1737A/G genotype frequencies in 24 healthy mother-baby pairs. PCR based RFLP method was used for genotyping. Here we report data for 167 mother-baby pairs including the previous subjects. Distribution of genotypes AA, AG, GG was 5%, 42%, 52% for mothers and 8%, 42%, 50% for newborns. The allele frequencies of A and G were 0.275and 0.725 for mothers, 0.125 and 0.875 for newborns. The commonest allele and genotype for the H19 1737 A/G polymorphism were G and GG respectively for Sri Lankan population. Birth weight, crown heel length and head circumferences were not affected by maternal or newborn ...
Human lymphatic filariasis (HLF) is a neglected tropical disease which threatens nearly 1.4 billi... more Human lymphatic filariasis (HLF) is a neglected tropical disease which threatens nearly 1.4 billion people in 73 countries worldwide. Wuchereria bancrofti is the major causative agent of HLF and it closely resembles cattle filarial parasite Setaria digitata. Due to difficulties in procuring W. bancrofti parasite material, S. digitata cDNA library has been constructed to identify novel drug targets against HLF and many of the cDNA sequences are yet to be assigned structure and function. In this study, a 549 bp long cDNA (sdrbp) has been sequenced and characterized in silico. The shortest ORF of 249 bp from the isolated cDNA encodes a polypeptide of 82 amino acids and shows an amino acid identity of 54% with the RRM domain of human cleavage stimulation factor-64 kDa subunit (CstF-64). Structure of the protein (sdRBP) obtained by homology modelling using RRM of CstF-64 as template adopts classical RRM topology (β1α1β2β3α2β4). sdRBP model built was validated by superimposition tools and...
Sequence variants of human mitochondrial DNA (mt DNA) have been implicated in a variety of disord... more Sequence variants of human mitochondrial DNA (mt DNA) have been implicated in a variety of disorders and conditions. Massive parallel sequencing is becoming increasingly popular due to its efficiency and cost-effectiveness. In relation to acquiring significant sequence information like levels of heteroplasmy in mt DNA, it offers a marked improvement compared to previous methods used. Here we describe a variant calling pipeline for human mitochondrial DNA using Next Generation Sequencing (NGS) data obtained by enriching the sample only for mitochondria prior to sequencing.
Gedunin is one of the major compounds found in the neem tree(Azadirachta indica). In the present ... more Gedunin is one of the major compounds found in the neem tree(Azadirachta indica). In the present study, antiproliferative potential of gedunin was evaluated in human embryonal carcinoma cells (NTERA-2, a cancer stem cell model) and peripheral blood mononuclear cells (PBMCs), using Sulforhodamine (SRB) and WST-1 assays, respectively. The effects of gedunin on expression of heat shock protein 90 (HSP90), its cochaperone Cdc37, and HSP client proteins (AKT, ErbB2, and HSF1) were evaluated by real-time PCR. Effects of gedunin on apoptosis were evaluated by (a) apoptosis associated morphological changes, (b) caspase 3/7 expression, (c) DNA fragmentation, (d) TUNEL assay, and (e) real-time PCR of apoptosis related genes (Bax,p53,andsurvivin). Gedunin showed a promising antiproliferative effect in NTERA-2 cells with IC50values of 14.59, 8.49, and 6.55 μg/mL at 24, 48, and 72 h after incubations, respectively, while exerting a minimal effect on PBMCs. Expression of HSP90, its client protein...
Garcinol (GA), a polyisoprenylated benzophenone derivative, is one of the major phytochemicals fo... more Garcinol (GA), a polyisoprenylated benzophenone derivative, is one of the major phytochemicals found in a number of fruits of Garcinia. This study aimed to develop a new nanodelivery system comprising GA, hyaluronic acid (HA), and poly(lactic-co-glycolic acid) (PLGA) to actively target breast cancer stem cells (bCSCs) grown under hypoxic conditions. HA has been reported to show higher affinity for the cluster-determinant 44 receptor (CD44), while PLGA is an FDA-approved biodegradable polymer used in clinical applications. Nanoparticles (NPs) were prepared by emulsion solvent diffusion technique using DMAB as the emulsifier. Following preparation, particle size, surface charge, and polydispersity index of NPs were characterized. Antiproliferative effects of NPs were assessed by the WST-1 cell proliferation assay. Apoptotic effects of NPs were evaluated by the caspase-3/7 assay. Effects of prepared NPs on the expression of genes related to hypoxia-inducing factors (HIF-1α and HIF-2α) ...
Additional file 1: Table S1. Optimised concentration, incubation time of the primary antibodies a... more Additional file 1: Table S1. Optimised concentration, incubation time of the primary antibodies and antigen retrieval buffer
Common genetic variations of imprinted genes that are maternally expressed could elucidate the ma... more Common genetic variations of imprinted genes that are maternally expressed could elucidate the maternal-specific inheritance of low birth weight. Human H19 is an imprinted growth regulatory gene located at chromosome 11p15.5 exclusively expressed from the maternal allele coding for a regulatory RNA implicated in the regulation of insulin-like growth factor II. We have previously described H19 1737A/G genotype frequencies in 24 healthy mother-baby pairs. PCR based RFLP method was used for genotyping. Here we report data for 167 mother-baby pairs including the previous subjects. Distribution of genotypes AA, AG, GG was 5%, 42%, 52% for mothers and 8%, 42%, 50% for newborns. The allele frequencies of A and G were 0.275and 0.725 for mothers, 0.125 and 0.875 for newborns. The commonest allele and genotype for the H19 1737 A/G polymorphism were G and GG respectively for Sri Lankan population. Birth weight, crown heel length and head circumferences were not affected by maternal or newborn ...
Human lymphatic filariasis (HLF) is a neglected tropical disease which threatens nearly 1.4 billi... more Human lymphatic filariasis (HLF) is a neglected tropical disease which threatens nearly 1.4 billion people in 73 countries worldwide. Wuchereria bancrofti is the major causative agent of HLF and it closely resembles cattle filarial parasite Setaria digitata. Due to difficulties in procuring W. bancrofti parasite material, S. digitata cDNA library has been constructed to identify novel drug targets against HLF and many of the cDNA sequences are yet to be assigned structure and function. In this study, a 549 bp long cDNA (sdrbp) has been sequenced and characterized in silico. The shortest ORF of 249 bp from the isolated cDNA encodes a polypeptide of 82 amino acids and shows an amino acid identity of 54% with the RRM domain of human cleavage stimulation factor-64 kDa subunit (CstF-64). Structure of the protein (sdRBP) obtained by homology modelling using RRM of CstF-64 as template adopts classical RRM topology (β1α1β2β3α2β4). sdRBP model built was validated by superimposition tools and...
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Papers by Prof. Kamani Tennekoon