Synsepalum dulcificum extracts exhibit cytotoxic activity on human colorectal cancer cells and upregulate c-fos and c-jun early apoptotic gene expression

Objective: To explore cytotoxicity of Synsepalum dulcificum(S. dulcificum) Daniell(Sapotaceae) on human colon cancer(HCT-116 and HT-29), human monocytic leukemia(THP-1) and normal(HDFn) cell lines, and its effect on the expression of early apoptotic genes, c-fos and c-jun. Methods: Leaf, stem and berry of S. dulcificum were separately extracted by using 2 solvents, 10% ethanol(EtOH) and 80% methanol(MeOH). PrestoB lue~? cell viability assay and q RT-PCR assay were conducted to examine the above objectives respectively. Results: Stem MeOH, stem EtOH, and berry EtOH extracts of S. dulcificum were cytotoxic to HCT-116 and HT-29 human colon cancer cells. For HCT-116, IC_(50) values of these 3 extracts were not significantly different(P>0.05) from that of the positive control bleomycin(IC_(50) of 33.57 μg/mL), while for HT-29, IC_(50) values of these 3 extracts were significantly lower(P<0.05) than that of bleomycin(IC_(50) of 25.24 μg/mL). None of the extracts were cytotoxic to the THP-1 monocytic leukemia cells and HDFn normal human dermal fibroblasts. For both HCT-116 and HT-29, these extracts significantly up-regulated(P<0.05) the expression of c-fos and c-jun compared to the untreated negative control. Conclusions: The results of this study suggest that cytotoxicity of stem MeOH, stem EtOH, and berry EtOH extracts of S. dulcificum on HCT-116 and HT-29 colon cancer cells is due to the induced apoptosis which is caused by the up-regulation of the expression of early apoptotic genes, c-fos and c-jun.

Antioxidant,cytotoxic,and anti-venom activity of Alstonia parvifolia Merr.Bark

Objective:To evaluate antioxidant,cytotoxic,and anti-venom capacity of crude bark extracts of Alstonia parvifolia Merr.Methods:Gas chromatography-mass spectrometry(GC-MS)and energy dispersive X-ray analyses were accomplished to characterize the chemical constituents of Alstonia parvifolia.Biochemical characterization was evaluated using an inhibitory phospholipase A_(2)(PLA_(2))assay,DPPH,and cytotoxicity assays.Using the constituents listed in the GC-MS analyses,molecular docking was conducted to inspect the binding energies between the chosen compounds and selected PLA_(2) isoforms.Results:GC-MS analyses showed that the Alstonia parvifolia crude extract consisted predominantly of acetylmarinobufogenin(14.89%),γ-sitosterol(10.44%),3-O-methyl-D-glucose(5.88%),3,5-dimethoxy-4-hydroxyphenylacetic acid(5.30%),(2α,5α)-17-methoxyaspidofracti­nin-3-one(AFM)(4.08%),and 2,3,5,6,7,8,9-heptahydro-1-phenyl-5-(p-chlorophenylimino)-1H-benzo[e][1,4]thiazepine(HPT)(1.37%).The principal elemental components of Alstonia parvifolia were Ca(4.012%)and K(1.496%),as exhibited by energy dispersive X-ray examination.Alstonia parvifolia showed significant free radical scavenging ability(IC_(50):0.287 mg/mL)and was non-cytotoxic to normal HDFn cells(IC_(50)>100μg/mL).Moreover,Alstonia parvifolia was favorably cytotoxic to MCF-7(IC_(50):4.42µg/mL),followed by H69PR,HT-29,and THP-1,with IC_(50) values of 4.94,5.07,and 6.27µg/mL,respectively.Alstonia parvifolia also displayed notable inhibition against PLA_(2) activity of Naja philippinensis Taylor venom with IC_(50) of(15.2±1.8)μg/mL.Docking and cluster analyses projected negative binding energies from AFM(−6.36 to−9.68 kcal/mol),HPT(−7.38 to−9.77 kcal/mol),and acetylmarinobufogenin(−7.22 to−9.59 kcal/mol).These calculations were for the particular interactions of Alstonia parvifolia constituents to PLA_(2) homologues where the utmost affinity was detected in HPT owing to the dipole interactions with amino acid residues.Conclusions:The bark extract of Alstonia parvifolia shows great potential as an anti-venom agent due to its low cytotoxic profile,remarkable PLA_(2) inhibition,and docking binding energies between its bioactive constituents and PLA_(2) homologues.

MutL homolog 1 methylation and microsatellite instability in sporadic colorectal tumors among Filipinos

BACKGROUND Colorectal cancer(CRC)ranks third in terms of incidence and second in mortality worldwide.In CRC,the silencing of mismatch repair genes,including the mutL homolog 1(hMLH1)has been linked to microsatellite instability(MSI),the lengthening or shortening of microsatellite repeats.Very limited data have been presented so far on the link of hMLH1 methylation and MSI in Southeast Asia populations with sporadic CRC,and on its clinical significance.AIM To investigate the significance of the MSI status and hMLH1 methylation in CRC Filipino patients.METHODS Fifty-four sporadic CRC patients with complete clinical data were included in this study.Genomic DNA from CRC tumor biopsies and their normal tissue counterparts were profiled for MSI by high resolution melting(HRM)analysis using the Bethesda Panel of Markers(BAT25,BAT26,D2S123,D5S346,and D17S250).hMLH1 methylation screening was performed using bisulfite conversion and methylation specific polymerase chain reaction.Statistical analysis was conducted to calculate their associations to clinicopathological characteristics and survival relevance(Kaplan-Meier curves and the log-rank test).RESULTS hMLH1 methylation was observed in 9%and 35%of CRC and normal samples,respectively.Higher incidence of consistently methylated hMLH1 found in both normal and CRC was noticed for relation to location of tumor(P<0.05).As for MSI status,D2S123 the most common unstable microsatellite and MSI-high(MSIH)was the most common MSI profile,counted for 46%and 50%of normal and CRC tissues,respectively.The presence of MSI-low(MSI-L)and microsatellite stable(MSS)was 43%and 11%for normal,and 31%and 19%for CRC samples.The mean month of patients’survival was shorter in patients whose normal and tumor tissues had methylated compared to those with unmethylated hMLH1 and with MSI-H compared to those with MSI-L/MSS(P<0.05).This was supported by significant difference in Kaplan-Meier with log-rank analysis.This data indicated that hMLH1 methylation and high MSI status have prognostic value.CONCLUSION This study showed the clinical significance of hMLH1 methylation and MSI status in sporadic CRC Filipino patients,especially in the normal part of the tumor.

Intestinal parasitic infections in Aetas and domesticated swine in Pampanga,Philippines

Indigenous peoples account for about 6%of the world’s population and are highly vulnerable to parasitic infections[1,2].Aetas,one of the indigenous people groups in the Philippines,rely on backyard swine production as their primary means of livelihood[3].Literature indicated that zoonoses arise from close physical contact and the cohabitation of humans with domesticated animals[4].Ascaris spp.

Soil-transmitted helminth egg contamination from soil of indigenous communities in selected barangays in Tigaon, Camarines Sur, Philippines

Objective:To provide baseline data on the prevalence and intensity of soil-transmitted helminthiasis egg contamination in the soil among indigenous communities.Methods:A total of 317 soil samples from three barangays of indigenous communities communities in Tigaon,Camarines Sur,Philippines were examined for soil-transmitted helminthiasis egg contamination using optimized sugar flotation method.Results:Of the soil samples examined,141(44.48%)were contaminated by Ascaris spp.,Toxocara spp.,and Trichuris spp.with cumulative prevalence varying across the study sites(P<0.01).Ascaris spp.was predominant in all study sites,followed by Toxocara spp.and Trichuris spp.with a prevalence of 41.96%,7.57%,and 5.36%,respectively.Interestingly,Toxocara pp.has the highest intensity of contamination,followed by Ascaris spp.and Trichuris spp.in term of geometric mean soil-transmitted helminthiasis eggs recovered per one gram soil sample(34.25,21.45,and 11.85 respectively).Each study site harbors significant amount of soiltransmitted helminthiasis eggs and zoonotic Toxocara eggs,which present high risk of soil-transmitted helminthiasis infection,particularly among children observed to play and cohabitate with animals known to be hosts of these parasites.Conclusions:The alarming rate of soil-transmitted helminthiasis and Toxocara egg contamination reported in this study suggests that additional measures should be undertaken to control soil-transmitted helminthiasis and zoonotic intestinal infections in the country.

Transmission dynamics and control strategies assessment of avian influenza A(H5N6)in the Philippines

Due to the outbreaks of Highly Pathogenic Avian Influenza A(HPAI)H5N6 in the Philippines(particularly in Pampanga and Nueva Ecija)in August 2017,there has been an increase in the need to cull the domestic birds to control the spread of the infection.However,this control method poses a negative impact on the poultry industry.In addition,the pathogenicity and transmissibility of the H5N6 in both the birds and the humans remain largely unknown which call for the necessity to develop more strategic control methods for the virus.In this study,we constructed a mathematical model for the bilinear and half-saturated incidence to compare their corresponding effect on transmission dynamics of H5N6.The simulations of half-saturated incidence model were similar to what occurred during the H5N6 outbreak(2017)in the Philippines.Instead of culling the birds,we implemented other control strategies such as non-medicinal(personal protection and poultry isolation)and medicinal(poultry vaccination)ways to prevent,reduce,and control the rate of the H5N6 virus transmission.Among the proposed control strategies,we have shown that the poultry isolation strategy is still the most effective in reducing the infected birds.