LC-MS Phytochemical Profiles of Phenolic Compounds and Antimicrobial, Antioxidant, and Antiplasmodial Activities of Ethanol Extracts of Pycnostachys erici-rosenii R.E.Fr and Leucas martinicensis (Jack.) R.Br (Lamiaceae)

This study has been conducted to analyze the chemical composition and antimicrobial, antioxidant and antiplasmodial properties of ethanol extracts of Pycnostachys erici-rosenii R.E.Fr and Leucas martinicensis (Jack.) R.Br that are not yet well known. The HPLC-DAD-MS-ESI+ method was used to investigate the chemical profile, the disc diffusion and microdilution method for the antimicrobial activities, the DPPH and FRAP assays for antioxidant property and SYBR Green I-based growth inhibition assay on Pf 3D7 (CQ-sensitive) strain for antiplasmodial properties. The results indicated significant amounts of hydroxybenzoic acid, flavonol and hydroxycinnamic acid in the two species. The total polyphenols analysis gave 209.0 ± 16.1 mg GAE/100g DW for P. erici-rosenii and 175.3 ± 1.6 mg GAE/100g DW for L. martinicensis while for total flavonoids analysis 33.4 ± 1.8 mg TE/100g DW was found for P. erici-rosenii and 39.6 ± 0.6 mg TE/100g DW for L. martinicensis. Moreover, antimicrobial tests revealed 0.125 mg/mL on E. faecalis ATCC25922 for ethanol extracts P. erici-rosenii and L. martinicensis. The antioxidant assays revealed IC50 56.17 μg/mL and 78.43 μg/mL for DPPH assay, and 112.03 μM TE/100mL and 90.46 μM TE/100mL) for FRAP assay for ethanol extracts of P. erici-rosenii and L. martinicensis respectively. Finally, for Antiplasmodial activity, IC50 was 4.07 μg/mL for P. erici-rosenii and 12.29 μg/mL for L. martinicensis. Therefore, the antimicrobial, the antioxidant, and the antiplasmodial results suggest that the ethanolic extracts of the aerial parts of P. erici-rosenii and L. martinicensis contain compounds with antimicrobial, antioxidant, and antiplasmodial properties, which can be searched in new drug discovery for treatment of infectious diseases.

Synergistic growth inhibitory effects of Phyllanthus emblica and Terminalia bellerica extracts with conventional cytotoxic agents:Doxorubicin and cisplatin against human hepatocellular carcinoma and lung cancer cells

AIM: To examine the growth inhibitory effects of Phyllanthus emblica (P. emblica) and Terminalia bellerica (T. bellerica) extracts on human hepatocellular carcinoma (HepG2), and lung carcinoma (A549) cells and their synergistic effect with doxorubicin or cisplatin. METHODS: HepG2 and A549 cells were treated with P. emblica and T. bellerica extracts either alone or in combination with doxorubicin or cisplatin and effects on cell growth were determined using the sulforhodamine B (SRB) assay. The isobologram and combination index (CI) method of Chou-Talalay were used to evaluate interactions between plant extracts and drugs. RESULTS: P. emblica and T. bellerica extracts demonstrated growth inhibitory activity, with a certain degree of selectivity against the two cancer cell lines tested. Synergistic effects (CI < 1) for P. emblica /doxorubicin or cisplatin at different dose levels were demonstrated in A549 and HepG2 cells. The T. bellerica/ cisplatin or doxorubicin also showed synergistic effects in A549 and HepG2 cells. In some instances, the combinations resulted in antagonistic effects. The dose reduction level was different and specific to each combination and cell line. CONCLUSION: The growth inhibitory activity of doxorubicin or cisplatin, as a single agent, may be modified by combinations of P. emblica or T. bellerica extracts and be synergistically enhanced in some cases. Depending on the combination ratio, the doses for each drug for a given degree of effect in the combination may be reduced. The mechanisms involved in this interaction between chemotherapeutic drugs and plant extracts remain unclear and should be further evaluated.

Anti-insulin resistant effect of ferulic acid on high fat diet-induced obese mice

Objective: To evaluate the insulin sensitivity action of ferulic acid(FA) in skeletal muscle and hypothalamus of high-fat diet(HFD)-induced obese mice. Methods: Obese mouse model was induced by HFD(45 kcal% lard fat) for 16 weeks. After 8 weeks of HFD feeding, these obese mice were orally treated with FA at doses of 25 and 50 mg/kg/day for 8 weeks. At the end of all treatments, the epididymal fat, pancreas, skeletal muscle and hypothalamus were removed for biochemical parameter and protein expression examinations. Results: FA treatment significantly decreased leptin level in fat tissue and insulin level in pancreas(P < 0.05). Interestingly, obese mice treated with FA increased the protein expressions of insulin receptor substrate-1, phosphatidylinositol 3-kinase, and phosphorylated-protein kinase B in both muscle and brain(P < 0.05). The phosphorylations of adenosine monophosphate-activated protein kinase and acetyl-CoA carboxylase in muscle, and leptin receptor protein in hypothalamus were also increased(P < 0.05). The pancreatic islets histology showed smaller size in obese mice treated with FA compared to untreated obese mice. Conclusions: These findings indicate the beneficial effect of FA in improving insulin resistance in HFD-induced obese mice. These effects are probably mediated via modulating the insulin receptor substrate/phosphatidylinositol 3-kinase/protein kinase B or adenosine monophosphate-activated protein kinase pathways.

Hexadecanoic acid-enriched extract of Halymenia durvillei induces apoptotic and autophagic death of human triple-negative breast cancer cells by upregulating ER stress

Objective:To investigate the effect of the hexane solvent fraction of Halymenia durvillei(HDHE)on triple-negative breast cancer.Methods:The phytochemical profile of HDHE was investigated by GC-MS.The cytotoxicity of HDHE against MDA-MB-231 cells was determined.The apoptotic and autophagic effects of HDHE were analyzed.The expression of molecular markers controlling apoptosis,autophagy,DNA damage,and endoplasmic reticulum(ER)stress was determined.Results:HDHE contains a mixture of fatty acids,mainly hexadecanoic acid.HDHE at a cytotoxic concentration induced apoptotic death of MDA-MB-231 cells through mitochondrial membrane dysfunction,and induction of apoptosis markers,and increased the expression of proteins related to DNA damage response.HDHE also induced the expression of LC-3,a marker of autophagic cell death at a cytotoxic concentration.Moreover,HDHE modulated the expression of ER stress genes.Conclusions:The hexadecanoic acid-enriched extract of Halymenia durvillei promotes apoptosis and autophagy of human triple-negative breast cancer cells.This extract may be further explored as an anticancer agent for triple-negative breast cancer.

Renoprotective effect of umbelliferone in high-fat diet/streptozotocin-induced type 2 diabetic rats

Objective:To evaluate the renoprotective effect of umbelliferone in high-fat diet/streptozotocin-induced type 2 diabetic rats.Methods:We established a streptozotocin-induced type 2 diabetic model in male Wistar rats.The rats were fed with high-fat diet(45 kcal%lard fat)and injected with 35 mg/kg streptozotocin.Diabetic rats were treated with umbelliferone for 8 weeks.At the end of the experimental period,the serum and kidney were used for measuring biochemical parameters,protein expression and histological analysis.Results:After 8-week treatment,umbelliferone decreased fasting plasma glucose,concentrations of malondialdehyde and monocyte chemoattractant protein-1 in the plasma and tissues.It also significantly reduced serum creatinine,blood urea nitrogen,serum advanced glycation end products,as well as kidney weight in type 2 diabetic rats(P<0.05).Moreover,umbelliferone reduced the 24-h urine albumin,but increased 24-h urine creatinine excretion(P<0.05).In renal protein expression,umbelliferone decreased the levels of transforming growth factor-β1 and fibronectin while increasing the levels of superoxide dismutase and catalase(P<0.05).Renal histological examination revealed an enlarged glomerular size in diabetic rats,which was smaller in umbelliferone-treated diabetic rats.Conclusions:Umbelliferone alleviates renal dysfunction in diabetes via decreasing hyperglycemia,oxidative stress,inflammation and glycation.

Anti-lipogenic effect of Senna alata leaf extract in high-fat diet-induced obese mice

Objective: To examine the effect of Senna alata(S. alata) leaf extracts on the regulation of lipid metabolism in high-fat diet-induced obese mice.Methods: The obesity condition was induced in the male ICR mice by feeding them with high-fat diet(45 kcal% fat) for 12 weeks. At the 7th week of diet feeding, the obese mice were treated with the water extract of S. alata leaf at 250 and 500 mg/kg/day, respectively, that continued for six weeks. At the end of the treatment period, the biochemical parameters were determined. The hepatic histology and the gene and protein expressions were also examined.Results: In comparison with the obese control mice, the mice treated with S. alata showed a significant reduction in the elevated blood glucose levels and a decrease in the serum insulin and leptin levels. A reduction in the serum total cholesterol, triglyceride,non-esterified fatty acid, and hepatic triglyceride levels were also observed. The histological examination of the obese mice treated with S. alata showed a reduced lipid accumulation in the liver tissue. Hepatic lipogenic gene expression showed that S. alata decreased the activity of sterol regulatory element binding protein 1c, fatty acid synthase,and acetyl-Co A carboxylase. S. alata could suppress hepatic peroxisome proliferatoractivated receptor gamma(PPARg) protein. Moreover, the protein expression of PPARa in liver tissue was clearly increased by S. alata treatment.Conclusion: The treatment with S. alata could decrease several parameters of impaired lipid metabolism in the obese mice by downregulating sterol regulatory element binding protein 1c and PPARg and upregulating PPARa. This study is the first report on the role of S. alata leaf extract in alleviating the abnormal lipid metabolism in obese conditions.

Effects of Solanum torvum fruit water extract on hyperlipidemia and sex hormones in high-fat fed male rats

Objective:To study the effect of water extract of Solanum torvum(S.torvum) on blood lipid and sex hormone levels in high-fat diet(HFD) fed male rats.Methods:Male Wistar rats were maintained on a standard diet or HFD for 10 weeks.During the last 4 weeks,the standard diet groups received distilled water or S.torvum(400 mg/kg) and the HFD groups received distilled water or S.torvum(100,200 and400 mg/kg).Body weight,lipid profiles,sex hormone,internal organs weight and liver histopathology were all measured.Moreover,kidney function was evaluated using blood urea nitrogen and creatinine levels,and liver function by the levels of aspartate aminotransferase and alanine aminotransferase.Results:The result showed that rats in the HFD control group had increased body weight and hyperlipidemia,but had decreased levels of both testosterone and estradiol.When receiving the S.torvum extract at a dose of 100 mg/kg,treated rats had significantly increased sex hormone levels of both types,and decreased total cholesterol levels,and at a dose of 200 mg/kg,treated rats had significantly decreased levels of triglyceride.Long term administration of the S.torvum did not produce any toxic signs in livers and kidneys.Pathological examinations of livers showed lipid accumulation in the HFD group,but the treatment of S.torvum slightly reduced lipid deposition in liver tissue.Conclusions:S.torvum extract can reverse the level of sex hormones to their normal level and reduce serum cholesterol in HFD-induced obese male rats.Furthermore,the long term oral administration of S.torvum extract is harmless.

Genetic Structure of the Red-spotted Tokay Gecko, Gekko gecko(Linnaeus, 1758)(Squamata: Gekkonidae) from Mainland Southeast Asia

This study was performed to explore the genetic diversity and genetic structure of red-spotted tokay geckos(Gekko gecko) from 23 different geographical areas in Thailand, Lao PDR and Cambodia. The mitochondrial tRNAGln/tRNA-Met/partial NADH dehydrogenase subunit 2 from 166 specimens was amplified and sequenced. A total of 54 different haplotypes were found. Highly significant genetic differences occurred between populations from different localities. The haplotype network revealed six major haplogroups(G1 to G6) belonging to different clades(clade A–E). Clade D and clade E were newly observed in this study. Haplogroup G4(clade D) was a sympatric population with haplogroup G1(clade B). The populations from northern Thailand were divided into two distinct haplogroups separated by mountain range. Genetic structure and genetic differentiation of the tokay in Southeast Asia was related to the geographical region sampled, spatial distance and natural barriers. Our results indicate that red-spotted tokay geckos from mainland Southeast Asia are cryptically diverse. Morphological comparisons, in addition to an intensive genetic investigation covering the whole species range, are needed to clarify the systematic and population structure of this species group.

Association of Blastocystis hominis with colorectal cancer:A systematic review of in vitro and in vivo evidences

BACKGROUND Recently,there have been several findings that showed intestinal colonisation of Blastocystis hominis(Blastocystis)as a risk factor to the worsening of colorectal cancer(CRC).However,studies have shown controversial results in the pathogenicity of Blastocystis.AIM To review systematically the evidence available on the association between CRC and Blastocystis and the prevalence of Blastocystis in CRC patients and to investigate cytopathic and immunological effects of Blastocystis in in vitro and in vivo studies.METHODS PRISMA guidelines were utilised in conducting this systematic review.Original articles published before February 2,2020 were included.PubMed,Science Direct,Scopus and Google scholar databases were searched.Manual searching was carried out to find articles missed during the online search.RESULTS Out of 12 studies selected for this systematic review,seven studies confirmed the prevalence of Blastocystis and found it to be between 2%-28%in CRC patients,whereby subtype 1 and subtype 3 were predominantly seen.A total of four studies employing in vitro human colorectal carcinoma cell line study models showed significant cytopathic and immunological effects of Blastocystis.In addition,one in vivo experimental animal model study showed that there was a significant effect of infection with Blastocystis on exacerbation of colorectal carcinogenesis.CONCLUSION Blastocystis is a commonly identified microorganism in CRC patients.These studies have provided supportive data that Blastocystis could exacerbate existing CRC via alteration in host immune response and increased oxidative damage.Future studies of CRC and Blastocystis should attempt to determine the various stages of CRC that are most likely to be associated with Blastocystis and its relationship with other intestinal bacteria.

Thai pigmented rice bran extracts inhibit production of superoxide, nitric oxide radicals and inducible nitric oxide synthase in cellular models

Objective:To study the inhibitory effect of rice bran extracts of Thai black Kam Muang and red Hawm Dawk Mali Deang on oxidative stress factors including superoxide(O2·-),nitric oxide(NO·),and inducible nitric oxide synthase(iNOS).Methods:Bran extracts(40%ethanol)of Kam Muang and Hawm Dawk Mali Deang were obtained and evaluated for in vitro 2-2′-azino-di-(3-ethylbenzthiazoline sulfonate)(ABTS)and NO·scavenging activity.Their inhibitory effects on cellular O2·-and NO·were measured in phorbol 12-myristate 13-acetate-stimulated neutrophil-like HL-60 cells and lipopolysaccharidestimulated RAW264.7 macrophages,respectively,and their viability was monitored using the MTT assay.The effect on iNOS expression was also assessed by the Western blotting assay.Total contents of phenolics,flavonoids,and subtypes were also determined.Results:Hawm Dawk Mali Deang exhibited about 3.5-fold greater cellular O2·-inhibitory activity than Kam Muang[EC50 values of(23.57±4.54)and(81.98±1.45)μg/mL,respectively]in phorbol 12-myristate 13-acetate-stimulated HL-60 cells.Hawm Dawk Mali Deang exhibited about 2-fold higher in vitro ABTS·+and NO·scavenging activity than Kam Muang,but it exerted cellular NO·inhibitory activity of only about 26%(undetermined EC50 value)in lipopolysaccharide-stimulated RAW264.7 cells.Conversely,Kam Muang exerted potent cellular NO·inhibitory activity[EC50 value:(281.13±59.18)μg/mL]and dose-dependently decreased iNOS levels.No cytotoxicity of both extracts was detected in both cell types.As for corresponding contents,Hawm Dawk Mali Deang contained higher contents of phenolics and flavonoids than Kam Muang.Moreover,Kam Muang and Hawm Dawk Mali Deang had a high content of total anthocyanins[(14.73±0.52)mg C3GE/g of extract]and total proanthocyanidins[(115.13±1.47)mg CE/g of extract],respectively.Conclusions:Based on these data,bran extracts of Thai black Kam Muang and red rice Hawm Dawk Mali Deang can help lower oxidative stress and inflammation attributed partly to O2·-and NO·.

Vasoprotective effects of rice bran water extract on rats fed with high-fat diet

Objective:To elucidate the protective effects of rice bran water extract on the expression of endothelial nitric oxide synthase(e NOS),nuclear factor-kappa B(NF-k B),and a cluster of differentiation 36(CD36) in the vasculature of high-fat diet-fed rats.Methods:Male Sprague-Dawley rats were divided into three groups.Group I served as control,Group II was treated with high-fat diet,and Group III was treated with high-fat diet and rice bran water extract at 2 205 mg/kg/day.After four weeks,the metabolic parameters,malondialdehyde as a marker of oxidative stress,and histological features of the aorta were evaluated.The levels of transcripts and proteins in aorta were determined by real-time PCR and Western blot analysis,respectively.Results:In comparison with the Group II,rice bran water extract administration resulted in a significant reduction in body weight,visceral fat tissue weights,blood glucose levels,and serum total-cholesterol and free fatty acid levels in Group III.Serum triglyceride levels tended to decrease in the Group III.Also,rice bran water extract administration obviously decreased malondialdehyde levels in both serum and aorta.Interestingly,rice bran water extract treatment demonstrated a significant up-regulation of e NOS expression and down-regulation of NF-k B p65 and CD36 expressions.Nonetheless,all groups showed normal histology of aorta.Conclusions:Rice bran water extract exhibited vasoprotective effects in the high-fat diet-induced obesity condition by modulating the expression of e NOS,NF-kB,and CD36 and metabolic parameters.

Multilocus enzyme electrophoresis analysis of Echinostoma revolutum and Echinostoma malayanum(Trematoda:Echinostomatidae) isolated from Khon Kaen Province,Thailand

Objective:To explore the genetic variation and differentiation of 2 echinostomes from genus Echinostoma,i.e.Echinostoma revolutum(E.revolution) and Echinostoma malayanum(E. malayanum) from Khon Kaen Province,Thailand.Methods:These parasites were compared at 22 enzymes encoding a presumptive 30 loci by using multilocus enzyme electrophoresis (MEE) technique.Results:Twenty-two loci can be used as diagnostic markers to differentiate these 2 species.E.revolutum and E.malayanum had fixed genetic differences at 70%of loci, whereas both species had fixed genetic differences from the liver fluke,Opisthorchis viverrini at 91%of loci.Intraspecific variation within a population of E.revolutum was observed at 5 polymorphic loci.Conclusions:MEE is a powerful technique to investigate genetic variation and differentiation of E.revolutum and E.malayanum.

Effect of Cymbopogon citratus Stapf extract on lipid profile and antioxidant status in streptozotocin-induced diabetic rats

OBJECTIVE To investigate the effects of Cymbopogon citratus Stapf water extract on lipid profile and antioxidant status in streptozotocin-induced diabetic rats.METHODS Diabetes was induced by injection of streptozotocin(STZ,50mg·kg-1)intraperitoneally.Diabetic rats were divided into 5groups,consisting of 6rats.GroupⅠ,reserved as diabetic control,was administered distilled water and groupⅡ,reserved as positive control,was administered glibenclamide(10mg·kg-1·d-1)throughout the duration of the experiment.Those in groupⅢ,ⅣandⅤ were administered 250,500 and 1000mg·kg-1·d-1 of the extract,respectively for 28 d.RESULTS Treatment with 500 and 1000mg·kg-1·d-1 of the extract resulted in reduction of serum AST,ALT,serum cholesterol,triglycerides and LDL,whereas HDL was found to be increased compared with diabetic control rats(P<0.05).Moreover,increased serum activities of superoxide dismutases and catalase were found in diabetic rats treated with the extract whereas serum thiobarbituric acid reactive substance was decreased,in comparison with diabetic control rat(P<0.05).CONCLUSION Cymbopogon citratus Stapf water extract provides a benefit effect on serum lipid and antioxidant effect in diabetic rats.Thus,the extract may lower cardiovascular disease risk and others complications related to hyperlipidemia and oxidative stress in diabetic patients.

Effects of Brassica oleracea extract on impaired glucose and lipid homeostasis in highfat diet-induced obese mice

Objective: To examine the effect of Brassica oleracea extract(BO) on impaired glucose and lipid homeostasis in high-fat diet(HFD)-induced obese mice. Methods: Obesity of ICR mice was induced by feeding a HFD(45 kcal% lard fat) for 16 weeks. During the last 8 weeks of study period, obese mice were additionally administered with BO(100 and 200 mg/kg/day). The metabolic parameters were determined. The gene expressions of hepatic lipogenesis were also studied. Results: After 8 weeks of treatment, BO(100 and 200 mg/kg) significantly reduced hyperglycemia and improved insulin sensitivity(P < 0.05). The serum lipid(total cholesterol, triglyceride, and non-esterified fatty acid) and hepatic triglyceride and nonesterified fatty acid were decreased(P < 0.05). The levels of insulin and leptin in serum were also decreased(P < 0.05). Moreover, the expressions of hepatic lipogenic genes including sterol regulatory element-binding protein 1 c, fatty acid synthase, and acetyl-CoA carboxylase were decreased by BO treatment(P < 0.05). Conclusions: These results suggest that BO is a new therapeutic agent for improving the homeostasis of glucose and lipid in HFD-induced obese mice probably by suppression of lipogenic genes in liver tissue.

Occurrence of Chlamydia spp.in wild birds in Thailand

Objective: To determine the occurrence of Chlamydia spp.in wild birds in Thailand.Methods: Cloacal and tracheal swabs of 313 wild birds from 11 orders, 27 families, and 51 species were tested to determine the occurrence of Chlamydia infection.The outer membrane protein A(ompA) gene was amplified from positive samples to construct a phylogenetic tree.Results: At the time of sample collection, none of the birds showed clinical signs of any disease.Of 313 wild birds, two Asian openbill stork(Anastomus oscitans) were positive for Chlamydia spp., representing 0.64%(2/313) and 4.9%(2/41) occurrence for birds overall and for the Asian openbill stork, respectively.Phylogram analysis based on deduced amino acid of the ompA gene showed that Chlamydia spp.in Asian openbill storks was closely related to that in wildfowl(Pica pica and Cygnus olor) from Poland in a different branch with a 95% bootstrap value and had a shorter evolutionary distance to Chlamydia abortus.Conclusions: Asymptomatic Asian openbill storks could be a potential source of Chlamydia infection in domestic animals, poultry, and humans who share their habitat.

Improvement of lipid profile and atherogenic index in rat treated with Cymbopogon citratus Stapf water extract

OBJECTIVE To investigate the effects of Cymbopogon citratus Stapf water extract(CCS)on lipid profile,atherogenic index and antioxidant enzymes in Sprague-Dawley rats.METHODS Rats aged eight weeks(250±20)g were orally administered with CCS at 250 and 500mg·kg-1·d-1 for one month.RESULTS Administration of CCS extract to rats caused significant decrease of serum total cholesterol,triglycerides and LDL levels,whereas,the HDL level was increased compared with untreated rats(P<0.05).Moreover,the CCS extract showed a significant decreased atherogenic index in comparison with untreated rats(P<0.05).Furthermore,serum activities of superoxide dismutases and catalase were also improved in rats treated with CCS.This was consistent with decrease of serum thiobarbituric acid reactive substance.CONCLUSION The present study provides evidence that CCS water extract provides a benefit effect on serum lipid and atherogenic index and exhibits antioxidant effect in vivo.

Genetic variation and phylogenetic relationship of Hypoderaeum conoideum(Bloch, 1782) Dietz, 1909(Trematoda: Echinostomatidae) inferred from nuclear and mitochondrial DNA sequences

Objective:To explore genetic variations of Hypoderaeum conoideum collected from domestic ducks from 12 different localities in Thailand and Lao PDR,as well as their phylogenetic relationship with American and European isolates.Methods:The nucleotide sequences of their nuclear ribosomal DNA(ITS),mitochondrial cytochrome c oxidase subunit 1(CO1),and NADH dehydrogenase subunit 1(ND1)were used to analyze genetic diversity indices.Results:We found relatively high levels of nucleotide polymorphism in ND1(4.02%),whereas moderate and low levels were observed in CO1(2.11%)and ITS(0.96%),respectively.Based on these polymorphisms,the 20 ND1,12 CO1,and 18 ITS haplotypes were classified,and several common haplotypes were observed in all samples.At least three major lineages,namely American,European and Asian lineages,have been classified by phylogenetic analyses based on ND1 sequences.Conclusions:Our report demonstrates that the ND1 gene is the most suitable genetic marker to explore genetic variation and phylogenetic relationship of Hypoderaeum conoideum.However,a combination of all loci for ND1,CO1 and ITS would be of great value toward further genetic investigation of this endemic worldwide parasite.Thus,comprehensive molecular genetic analyses of Hypoderaeum conoideum from its worldwide distribution is needed to further understanding of the evolutionary and systematic relationships of this parasite.

Effect of Thunbergia laurifolia water extracts on hepatic insulin resistance in high-fat diet-induced obese mice

Objective:To examine the effect of water extract of Thunbergia laurifolia on hepatic insulin resistance in high-fat diet-induced obese mice.Methods:High-fat diet with 45 kcal%lard fat was used for obesity induction in ICR mice.The mice were fed with high-fat diet for 16 weeks,and during the last 8 weeks,they were treated with 200 mg/kg/day of water extracts from Thunbergia laurifolia leaf,stem and flower.Serum biochemistry,liver histology,and protein expression were examined after the treatment.Results:Extracts from all of the three parts of Thunbergia laurifolia significantly alleviated hyperglycemia,hyperlipidemia,hyperinsulinemia,and hyperleptinemia.The stem and flower extracts improved glucose tolerance.All of the extracts significantly reduced serum TNFαand monocyte chemoattractant protein-1 levels.Liver weight,triglyceride levels,and lipid accumulation were also decreased.Moreover,hepatic glucose-6-phosphatase level was significantly decreased,while the levels of PPARα,phosphorylated AMPK,and phosphorylated Akt were significantly increased with treatment of Thunbergia laurifolia extracts.Conclusions:Thunbergia laurifolia extracts can ameliorate hepatic insulin resistance in high-fat diet-induced obese mice by improving glucose and lipid homeostasis,which may be associated with stimulating phosphorylation of AMPK and Akt pathways.

Mosquito larva distribution and natural Wolbachia infection in campus areas of Nakhon Ratchasima,Thailand

Objective:To determine the prevalence of mosquito larvae in campus areas and the infection rate of endosymbiotic bacteria,Wolbachia in mosquito larvae.Method:The mosquito larvae samples were collected in residential areas and academic buildings of Suranaree University of Technology located in Northeastern Thailand during 2017-2018.Mosquito species identification was performed using GLOBE mosquito protocols and Rattanarithikul&Panthusiri’s keys.The gene encoding for the surface protein of Wolbachia was amplified by PCR and confirmed by DNA sequencing.Results:Armigeres sp.is the highest proportion of mosquito larvae followed by Culex spp.,Aedes albopictus,Aedes aegypti,and Toxorynchites spp.,respectively.Aedes aegypti have breeding sites mostly in the containers found indoors,whereas the main breeding sites of Aedes albopictus were found in both outdoors and indoors.The House Index and Breteau Index for Aedes spp.was more than 5%and 20%,respectively,in both areas,indicating that these areas are dengue sensitive.The highest proportion of Wolbachia infection was found in the larvae of Culex spp.(86.21%),followed by Aedes albopictus(69.23%)and rarely detected in Aedes aegypti(9.09%).Conclusion:The present study reported the first natural infection of Wolbachia in mosquito larvae in Thailand.Our result suggested that the mosquito species containing higher proportion of Wolbachia are less likely to be vectors for dengue.Therefore,Wolbachia transfection in mosquito larvae could be applied as a biocontrol for dengue and other mosquito-borne disease prevention.

Genetic Differentiation of the Forest Crested Lizards,Calotes emma alticristatus Schmidt,1925 and C.emma emma Gray,1845(Squamata:Agamidae)Relating to Climate Zones in Thailand

In Thailand,the forest crested lizard,Calotes emma consists of two subspecies,C.emma alticristatus Schmidt,1925 and C.emma emma Gray,1845.This study was performed to determine the genetic diversity and differentiation of C.emma from 16 different localities throughout Thailand.A total of 116 samples were analyzed using the mitochondrial cytochrome c oxidase subunit 1(CO1).Of these,65 and 51 of C.e.alticristatus and C.e.emma were classified into 23(N1-N23)and 21(S1-S21)haplotypes,respectively.There was no shared haplotype between subspecies or between different populations within each subspecies.These haplotypes were classified into four(north-A to north-D)and three(south-A to south-C)haplogroups of C.e aliristaus and C.e.emma,respectively.Phylogenetic analyses retrieved four lineages(classified asⅠtoⅣ).LineagesⅠandⅡcontained the four haplogroups of C.e.alicristatus,whereas lineageⅡandⅣcontained three haplogroups of C.e.emma.These two subspecies live separately in different climate zones,ie.C.e.alticristatus is found in an equatorial winter dry climate,whereas C.e.emma inha bits areas with an equa torial monsoonal clima te.