Antimicrobial activities of the tissue extracts of Babylonia spirata Linnaeus, 1758 (Mollusca: Gastropoda) from Thazhanguda,southeast coast of India

Objective

To investigate the antimicrobial activity of the tissue extracts of Babylonia spirata (B. spirata) against nine bacterial and three fungal pathogens.

Methods

Crude extract of gastropod was tested for inhibition of bacterial and fungal growth. Antibacterial assay was carried out by disc diffusion method and in vitro antifungal activity was determined against Czapex Dox agar. The antimicrobial activity was measured accordingly based on the inhibition zone around the disc impregnated with gastropod extract. Molecular size of muscle protein was determined using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). And fourier transform infrared spectroscopy (FTIR) spectro photometry analysis was also studied.

Results

The maximum inhibition zone (12 mm) was observed against Pseudomonas aeruginosa in the crude ethanol extract of B. spirata and the minimum inhibition zone (2 mm) was noticed against Staphylococcus aureus in the crude methanol extract of B. spirata. Water extract of B. spirata showed the highest activity against Vibrio parahaemolyticus, Staphylococcus aureus and Candida albicans. Ethanol, acetone, methanol, chloroform and water extracts showed antimicrobial activity against almost all the bacteria and fungus. Compared with water extracts, ethanol and methanol extracts showed higher activity against all pathogens. The molecular weight of protein of the gastropod sample ranged from 2-110 kDa on SDS-PAGE. FTIR analysis revealed the presence of bioactive compounds signals at different ranges.

Conclusions

The research shows that the great medicinal value of the gastropod muscle of B. spirata may be due to high quality of antimicrobial compounds.     

The zoonotic implications of pentastomiasis in the royal python (python regius)

Pentastomes are worm-like endoparasites of the phylum Pentastomida found principally in the respiratory tract of reptiles, birds, and mammals. They cause a zoonotic disease known as pentastomiasis in humans and other mammals. The autopsy of a Nigerian royal python (Python regius) revealed two yellowish-white parasites in the lungs, tissue necrosis and inflammatory lesions. The parasite was confirmed to be Armillifer spp (Pentastomid); this is the first recorded case of pentastomiasis in the royal python (Python regius) in Nigeria. This report may be an alert of the possibility of on-going zoonotic transmission of pentastomiasis from snake to man, especially in the sub-urban/rural areas of Nigeria and other West African countries where people consume snake meat.     

Anticancer effect of the extracts from Polyalthia evecta against human hepatoma cell line (HepG2)

Objective

To investigate the anticancer activity of Polyalthia evecta (P. evecta) (Pierre) Finet & Gagnep against human hepatoma cell line (HepG2).

Methods

The anticancer activity was based on (a) the cytotoxicity against human hepatoma cells (HepG2) assessed using a neutral red assay and (b) apoptosis induction determined by evaluation of nuclei morphological changes after DAPI staining. Preliminary phytochemical analysis of the crude extract was assessed by HPLC analysis.

Results

The 50% ethanol-water crude leaf extract of P. evecta (EW-L) showed greater potential anticancer activity with high cytotoxicity [IC₅₀ = (62.8 ± 7.3)µg/mL] and higher selectivity in HepG2 cells than normal Vero cells [selective index (SI) = 7.9]. The SI of EW-L was higher than the positive control, melphalan (SI = 1.6) and the apoptotic cells (46.4 ± 2.6) % induced by EW-L was higher than the melphalan (41.6 ± 2.1)% (P<0.05). The HPLC chromatogram of the EW-L revealed the presence of various kinds of polyphenolics and flavonoids in it.

Conclusions

P. evecta is a potential plant with anticancer activity. The isolation of pure compounds and determination of the bioactivity of individual compounds will be further performed.     

Roles of virulence genes (PsaA and CpsA) on the invasion of Streptococcus pneumoniae into blood system

Background

Streptococcus pneumoniae (SP) is the major cause of childhood mortality worldwide, we need to understand virulence genes of SP so can better target the treatment.We investigated the expression of virulence genes PsaA and CpsA in different strains of SP interacting with monocyte cell line (THP-1) or pneumocyte cell line (A549) and the possible mechanism of SP invasion of the blood system.

Methods

A total of 23 strains of SP were collected from hospitalized patients (blood-derived and sputum-derived) in the Second Affiliated Hospital of Wenzhou Medical College. The strains and ATCC 49619 were cultured, and RNAs were extracted. THP-1 and A549 cells were stimulated by different SP and ATCC 49619 for 4 h and 8 h, respectively. Quantitative real-time PCR was used to analyze the mRNA expression of PsaA and CpsA. The data were analyzed by SPSS 17.0.

Results

The mRNA level of PsaA and CpsA were all significantly increased in clinical SP strains when compared to ATCC49619 after tedTHP-1 and A549 cells were stimulated. Clinical SPs showed higher virulence compared with ATCC49619.

Conclusions

The expression of CpsA is the basis of the pathogenicity of SP. The expression of virulence gene PsaA may be helpful to the invasion of SP to the blood system.     

Isolation and efficacy of entomopathogenic fungus (Metarhizium anisopliae) for the control of Aedes albopictus Skuse larvae: suspected dengue vector in Pakistan

Objective

To isolate the entomopathogenic fungus Metarhizium anisopliae (M. anisopliae) in the local environment, and evaluate its efficacy against the suspected dengue vector Aedes albopictus in Pakistan.

Methods

According to the standard procedure, M. anisopliae was isolated from the dead mosquitoes which were collected from the field or dead after the collection. Bioassay was performed to determine its efficacy.

Results

The results indicated that M. anisopliae had larvicidal effect with LC₅₀ value 1.09×10⁵ and LC₉₀ value 1.90×10¹³ while it took 45.41 h to kill 50% of tested population.

Conclusions

Taking long time to kill 50% population when compare with the synthetic insecticides, is the only drawback for the use of entomopathogenic fungus but these bio-pesticides are safe for the use.     

Effect of a commercial air ionizer on dust mites Dermatophagoides pteronyssinus and Dermatophagoides farinae (Acari: Pyroglyphidae) in the laboratory

Objective

To investigate the short and long term efficacy of a commercial air ionizer in killing Dermatophagoides pteronyssinus (D. pteronyssinus) and Dermatophagoides farinae (D. farinae) mites.

Methods

The effect of a commercial ionizer on D. pteronyssinus and D. farinae was evaluated in the laboratory, using a specially designed test. Mortality was assessed after 6, 16 and 24 hours for direct exposure and after 24, 36, 48, 60 and 72 hours for exposure in simulated mattress. New batches of mites were used for each exposure time.

Results

LT₅₀ for direct exposure of ionizer was 10 hours for D. pteronyssinus and 18 hours for D. farinae. The LT₅₀ for exposure in simulated mattress was 132 hours or 5.5 days for D. pteronyssinus and 72 hours or 3 days for D. farinae. LT₉₅ for direct exposure of ionizer was 36 hours for D. pteronyssinus and D. farinae. Meanwhile, the LT₉₅ for exposure in simulated mattress was 956 hours or 39.8 days for D. pteronyssinus and 403 hours or 16.8 days for D. farinae.

Conclusions

This study demonstrates the increasing mite mortalities with increasing exposure time of a commercial ionizer and suggests that negative ions produced by an ionizer kill dust mites and can be used to reduce natural mite populations on exposed surfaces such as floors, clothes, curtains, etc. However, there is reduced efficacy on mites inside stuffed materials as in mattresses and furniture.     

Morphological study of the asymmetrical buccal cavity of the flatfish common solea (Solea solea) and its relation to the type of feeding

Objective

To investigate the surface architecture of the asymmetrical buccal cavity of Solea solea which are considered one of the most important predators in benthic communities.

Methods

Adult Solea solea were obtained from Mediterranean Sea near Damietta. The heads were removed and processed for scanning electron microscopy. Its buccal cavity is asymmetrical and divided into roof and floor and the tongue for histological studies.

Results

The buccal cavity roof is formed from upper jaw, velum and the palate. The upper jaw has several wing like processes with teeth arranged in several rows which may help in cutting and pushing the food to the entrance of the digestive canal while the floor is formed from the lower jaw and the tongue. The tongue is divided into apex, body and root. There is a gradual decrease of goblet cells in the tongue from anterior to posterior. These goblet cells function in protection of the epithelium.

Conclusions

Teeth in the floor of the buccal cavity and taste buds can be considered adaptive changes of the oral cavity related to the feeding habits and was a source to identify new and better methods of nutrition in aquaculture of Solea solea.     

Plant profile,phytochemistry and pharmacology of Cordia dichotoma (Indian cherry): A review

More than half of the world''s population relies on the traditional medicine and major role of the traditional medicine including the use of plant extract and their active constituents. Among them, Cordia dichotoma Forst., a small to moderate size plant of family Boragenaceae, commonly called bhokar, lasura, gonda, Indian cherry and shlesmataka. Plant parts such as leaves, fruit, bark and seed have been reported for possessing antidiabetic, antiulcer, anti-inflammatory, immune-modulator and analgesic activity. Screening of fruit, leaves and seed shows the presence of pyrrolizidine alkaloids, coumarins, flavonoids, saponins, terpenes and sterols. Present review focuses on details of geographical distribution, physicochemical parameters, phytoconstituents and pharmacological properties of Cordia dichotoma reported so far.     

Household survey of container-breeding mosquitoes and climatic factors influencing the prevalence of Aedes aegypti (Diptera: Culicidae) in Makkah City,Saudi Arabia

Objective

To investigate the prevalence of container breeding mosquitoes with emphasis on the seasonality and larval habitats of Aedes aegypti (Ae. aegypti) in Makkah City, adjoining an environmental monitoring and dengue incidence.

Methods

Monthly visits were performed between April 2008 and March 2009 to randomly selected houses. During each visit, mosquito larvae were collected from indoors and outdoors containers by either dipping or pipetting. Mosquitoes were morphologically identified. Data on temperature, relative humidity, rain/precipitations during the survey period was retrieved from governmental sources and analyzed.

Results

The city was warmer in dry season (DS) than wet season (WS). No rain occurred at all during DS and even precipitations did fall, wetting events were much greater during WS. Larval survey revealed the co-breeding of Aedes, Culex and Anopheles in a variety of artificial containers in and around homes. 32 109 larvae representing 1st , 2nd, 3rd, and 4th stages were collected from 22 618 container habitats. Culicines was far the commonest and Aedes genus was as numerous as the Culex population. Ae. aegypti larval abundance exhibited marked temporal variations, overall, being usually more abundant during WS. Ten types of artificial containers were found with developing larvae. 70% of these habitats were located indoors. 71.42% of indoor containers were permanent and 28.58% was semi-permanent during WS. Cement tanks was the only container type permanent during DS. Ae. aegypti larval indices (CI, HI, BI) recorded were greater during WS.

Conclusions

Taken together, these results indicate a high risk of dengue transmission in the holy city.     

Synergistic anticancer effect of the extracts from Polyalthia evecta caused apoptosis in human hepatoma (HepG2) cells

Objective

To evaluate the anticancer activity of the extract fraction of Polyalthia evecta (P. evecta) (Pierre) Finet & Gagnep and the synergistic anticancer effect of the extracts from P. evecta by using the ATR/FT-IR spectroscopy.

Methods

The 50% ethanol-water crude leaf extract of P. evecta (EW-L) was prepared and was further fractionated to isolate various fractions. The anticancer activity was investigated from cytotoxicity against HepG2 using a neutral red assay and apoptosis induction by evaluation of nuclei morphological changes after DAPI staining. Synergistic anticancer effects of the extracts from P. evecta were performed using the ATR/FT-IR spectroscopy.

Results

The result showed that the EW-L showed higher cytotoxicity and apoptosis induction in HepG2 cells than its fractionated extracts. The hexane extract exhibited higher cytotoxicity and apoptosis induction than the water extracts, but less than the EW-L. The combined water and hexane extracts apparently increased cytotoxicity and apoptosis induction. The %apoptotic cells induced by the extract mixture were increased about 2-fold compared to the single hexane extract.

Conclusions

The polar extract fraction is necessary for the anticancer activity of the non-polar extract fraction. The ATR/FT-IR spectra illustrates the physical interaction among the constituents in the extract mixture and reveals the presence of polyphenolic constituents in the EW-L, which might play a role for the synergistic anticancer effect.     

Comparison of protein patterns between Plasmodium falciparum mutant clone T9/94-M1-1(b3) induced by pyrimethamine and the original parent clone T9/94

Objective

To compare the protein patterns from the extracts of the mutant clone T9/94-M1-1(b3) induced by pyrimethamine, and the original parent clone T9/94 following separation of parasite extracts by two-dimensional electrophoresis (2-DE).

Methods

Proteins were solubilized and separated according to their charges and sizes. The separated protein spots were then detected by silver staining and analyzed for protein density by the powerful image analysis software.

Results

Differentially expressed protein patterns (up- or down-regulation) were separated from the extracts from the two clones. A total of 223 and 134 protein spots were detected from the extracts of T9/94 and T9/94-M1-1(b3) clones, respectively. Marked reduction in density of protein expression was observed with the extract from the mutant (resistant) clone compared with the parent (sensitive) clone. A total of 25 protein spots showed at least two-fold difference in density, some of which exhibited as high as ten-fold difference.

Conclusions

These proteins may be the molecular targets of resistance of Plasmodium falciparum to pyrimethamine. Further study to identify the chemical structures of these proteins by mass spectrometry is required.     

The microstructural effects of aqueous extract of Garcinia kola (Linn) on the hippocampus and cerebellum of malnourished mice

Objective

To assess the neuroprotective effects of aqueous extract of Garcinia kola on neurotoxin administered malnourished mice adopting histological procedure.

Methods

The study was carried out using thirty-two adult malnourished mice which were randomly assigned into four groups (n=8): A, B, C and D. Group A served as control, while the other groups served as the experimental groups. Animals in group A were fed malnourished diet ad libitum and given water liberally. Animals in group B were administered with 3-Nitropropionic acid (3-NP) (neurotoxin) only at 20 mg/kg body weight, group C were given only Garcinia kola extracts, and group D were pre-treated with Garcinia kola extracts at 200 mg/kg for seven days prior to administration of neurotoxin at 20 mg/kg body weight. After three days of neurotoxins administration in the relevant groups, the brains were excised and fixed in formal calcium for histological processing.

Results

The study showed that hippocampal and cerebellar neurons of animals in group B exhibited some cellular degeneration and blood vessel blockage, which were not seen in groups A, C and D. Cresyl violet staining was least intense in group B than in groups A, C and D. Despite the fact that animals in group D has equal administration of 3-Nitropropionic acid concentration, there were no traces of neural degeneration as it was evidenced in group B.

Conclusions

It is concluded that Garcinia kola has protective effects on the neurons of the hippocampus and cerebellum of malnourished mice.     

白色念珠菌ITS2的实时荧光定量PCR快速检测方法的建立及评价

目的 建立一种快速鉴定白色念珠菌的实时荧光定量PCR检测方法。方法 在白色念珠菌核糖体RNA编码基因（rDNA）中的内转录间隔区2（ITS2）上设计特异性引物和TaqMan-MGB探针,通过实时荧光定量PCR分析系统建立白色念珠菌的检测方法,对其敏感度、重复性和特异性进行方法学评价,并在模拟白色念珠菌血症的血标本中初步探讨其应用价值。结果 所建方法对白色念珠菌纯菌液的准确检测下限为10¹CFU/ml,相应的批内、批间变异系数（CV）分别为1.57%和2.20%。对135株白色念珠菌临床分离株、其他非白念菌株、细菌临床分离株和血液标本进行检测,结果显示该方法特异性为100%。另外,对于模拟白色念珠菌血症的血标本,该方法的最低检测下限为10²CFU/ml。结论 所建方法具有特异性和敏感度高、重复性好的特性,可适用于临床上白色念珠菌引起的血流感染的快速鉴定。     

Isolation and structural elucidation of cytotoxic compounds from the root bark of Diospyros quercina (Baill.) endemic to Madagascar

Objective

To isolate and characterize the cytotoxic compounds from Diospyros quercina (Baill.) G.E. Schatz & Lowry (Ebenaceae).

Methods

An ethno-botanical survey was conducted in the south of Madagascar from July to August 2010. Bio-guided fractionation assay was carried out on the root bark of Diospyros quercina, using cytotoxicity bioassay on murine P388 leukemia cell lines as model. The structures of the cytotoxic compounds were elucidated by 1D and 2D NMR spectroscopy and mass spectrometry.

Results

Biological experiments resulted in the isolation of three bioactive pure compounds (named TR-21, TR-22, and TR-23) which exhibited very good in vitro cytotoxic activities with the IC₅₀ values of (0.017 5±0.0060) µg/mL, (0.089±0.005) µg/mL and (1.027±0.070) µg/mL respectively. Thus, they support the claims of traditional healers and suggest the possible correlation between the chemical composition of this plant and its wide use in Malagasy folk medicine to treat cancer.

Conclusions

The ability of isolated compounds in this study to inhibit cell growth may represent a rational explanation for the use of Diospyros quercina root bark in treating cancer by Malagasy traditional healers. Further studies are, therefore, necessary to evaluate the in vivo anti-neoplastic activity of these cytotoxic compounds as effective anticancer drugs.     

1, 25-二羟基维生素D3通过下调葡萄糖转运蛋白-1表达抑制膀胱癌细胞增殖

目的：研究 1,25-二羟基维生素 D₃(1,25-(OH)₂-VitD₃)对膀胱癌细胞增殖的影响,并探讨其作用机制。方法：体外培养膀胱癌细胞 T24,MTT 法检测不同浓度 1,25-(OH)₂-VitD₃对细胞活力的影响,并筛选 1,25-(OH)₂-VitD₃干预细胞的最佳时间及半数抑制浓度。将细胞分为 4 组：对照组、葡萄糖转运蛋白-1(glucose transporter-1,GLUT1)抑制组、1,25-(OH)₂-VitD₃组、1,25-(OH)₂-VitD₃+GLUT1 抑制组。采用 Hoechst 33258 染色观察各组细胞形态变化;试剂盒检测细胞内葡萄糖摄取、三磷酸腺苷(adenosine triphosphate,ATP)产生及乳酸形成水平;Real-time PCR 及 Western blot 检测 GLUT1、己糖激...