Annual Research & Review in Biology

Aims: This study was designed to investigate how CO2 and nitrogen availability alters growth, leaf gas exchange, primary and secondary metabolites production (total phenolics and flavonoid) in three varieties of L. pumila. namely var alata, var pumila and var lanceolata. under CO₂ enrichment (1200 µmol mol^-1) combined with four levels of nitrogen fertilization (0, 90, 180 and 270 kg N ha^-1) for 15 weeks.

Study Design: Three-month old L. pumila seedlings of var. alata, var. pumila and var. lanceolata were left for a month to acclimatize in a nursery until ready for the treatments. Carbon dioxide enrichment treatment started when the seedlings reached 4 months old where plants were exposed to 1200 µmol^-1 mol^-1 CO₂ and fertilized with four levels of nitrogen concentrations viz. 0, 90, 180 and 270 kg N Ha^-1. The fertilization with nitrogen levels were split into three applications. A control at ambient CO₂ (400 µmol^-1 mol^-1) with standard N fertilization (180 kg N ha^-1) was included to compare plant responses to high CO₂ combined with different levels of N.

Place and Duration of Study: Faculty of Agriculture Glasshouse Complex, Universiti Putra Malaysia from February to May 2011.

Methodology: The seedlings were raised in specially constructed growth houses receiving 12-h photoperiod and average photosynthetic photon flux density of 300 µmol m^-2 s^-1. Carbon dioxide at 99.8% purity was supplied from a high–pressure CO₂ cylinder and injected through a pressure regulator into fully sealed 2 m x 3 m growth houses at 2-h daily and applied continuous from 800 to 1000. Total plant biomass was taken by calculating the dry weight of root, stem and leaf per seedling. Total leaf area per plant was measured using a leaf area meter (LI-3100, Lincoln Inc, USA). The measurement of photosynthesis was obtained from a closed system of infra-red gas analyzer LICOR 6400 Portable Photosynthesis System. The response of net CO₂ exchange (A) to changing intercellular CO₂ concentration (Ci) was conducted at 27°C, 50% relative humidity and at light saturating conditions of 800 µmol m^-2s^-1. Anthrone method was used to determine the content of total available carbohydrate. Follin–Ciocalteau reagent was used to determine total phenolics and flavonoid using spectrophotometer.

Results: In all parameters measured there were no significant was observed in the three varieties. As the levels of nitrogen increased from 0 to 270 kg N ha^-1, plant growth, photosynthesis and total chlorophyll content was enhanced. However, carbohydrate, total phenolics and flavonoid was reduced with the increase in nitrogen fertilization. It was found that the high application of nitrogen can increased the photosynthetic capacity of L. pumila by having high Vc_max (RuBisCO CO₂ fixation capacity), A sat (CO2 saturation point) and low dark respiration rate in the analysis of A/Ci curves. The production of secondary metabolites was related to increased total non-structural carbohydrate (TNC) under low nitrogen condition. Photosynthesis, displayed a significant negative relationship with secondary metabolites production. Total chlorophyll content exhibited very significant negative relationships with total soluble sugar, starch and total non-structural carbohydrate.

Conclusion: This study demonstrated that enrichment with high levels of CO₂ is able to enhance the production of plant secondary metabolites. However, increased nitrogen fertilization can reduce the production of these plant secondary metabolites under CO₂ enrichment. When there were accumulation of TNC in plant leaves and reduction in photosynthesis, the production of plant secondary metabolites might be up-regulated. The increase in the production of plant secondary metabolites was indicated by reduction in chlorophyll contents.

Aims: This research aims to investigate the effectiveness of ethyl acetate extract of endophytic fungi isolated Sir G₅ from soursop leaf (Annona muricata L.) in vivo breast tumor growth using Sprague-dawley (SD) female strains induced by 7,12-dimethylbenz(α)anthracene (DMBA).

Study Design: This study was an in vivo experiment using a completely randomized design   method (RAL).

Place and Duration of Study: This research was conducted from July 2016 to February 2017 in Microbiology Laboratory of Biology Research Center, Indonesian Institute of Sciences (LIPI), Cibinong-Bogor, and Research Laboratory of Biochemistry Department of Bogor Agricultural University.

Methodology: Twenty-four female rats were divided into six groups; normal group (administered per os (p.o) with aquadest 2 ml for twenty two weeks), negative control (-) DMBA (administered intraperitoneal (i.p.) with 20 mg/kg body weight (bw) of 7,12-dimethylbenz(α)anthracene), positive control (+) Doxo (administered i.p with 20 mg/kg bw of 7,12-dimethylbenz(α)anthracene and p.o. with 2 µg/20 0 g bw of Doxorubicin), DI (administered i.p. with 20 mg/kg bw of 7,12-dimethylbenz(α)anthracene and p.o. with extract of 20 mg/kg bw isolate Sir G₅), DII (administered i.p. with 20 mg/kg bw of 7,12- dimethylbenz(α) anthracene and p.o. with 40 mg/kg bw extract of isolate Sir G₅) and DIII (administered i.p. with 20 mg/kg b.w.of 7,12- dimethylbenz(α)anthracene and with 120 mg/kg bw of extract of isolate Sir G₅).

Results: The result showed that body weight rats was not significantly different at each group (P>0.05). The quantity and volume of tumor samples from isolat Sir G₅ extract treatment groups was significantly lower (P<0.05) than negative control DMBA and positive control doxo.

Conclusion: Overall, it can be concluded that the ethyl acetate extract of endophytic fungi isolated Sir G5 from soursop leaf (Annona muricata L.) can inhibit the growth of rat breast tumor particulary on the treatment dose DI (20 mg/Kg bw).

Background: Feeding preferences of Odontotermes obesus on five different commercial wood species were studied under choice and no choice trials both in laboratory and field conditions. Feeding preferences as a quantitative parameter of wood consumption by O. Obesus were recognized as follows: Cupressus sempervirens linn >Ficus microphylapers > Ficus banghalensis linn > Ficus relgiosa linn > phylanthus embicallium. Toxicity and LC₅₀ value of fipronil, Imidacloprid indicate that Imidacloprid is more toxic than fipronil based on LC₅₀ values.

Results: The wooden species used under lab no choice bioassays were follows as, Cupressus sempervirens linn, Ficus microphylapers, Ficus banghalensis, Ficus relgiosa linn  and Phylanthus emblicallium. Survival rate, Weight loss and mean consumption were calculated. Mean wood mass loss (mg) in five different wood pairs when exposed in combination to Odontotermes obesus for CS/FM (Cupressus sempervirens linn and Ficus microphylapers), FM/FR (Ficus microphylapers and Ficus relgiosa lin), FR/FB (Ficus relgiosa linn and Ficus banghalensis linn), FB/PE (Ficus banghalensis linn and Phylanthus emblicallium), PE/CS (Phylanthus emblicallium and Cupressus sempervirens linn) in 2 week choice under field conditions. Maximinm consumption (mg) was recorded pair of PE/CS (Phylanthus emblicallium Vs. Cupressus sempervirens linn). Cupressus sempervirens linn was found to be the most palatable wood species whereas Phylanthus emblicallium proved the most resistant one. Minimum feeding was recorded in Ficus banghalensis linn (2.9 mg) and maximum in Cupressus sempervirens linn (9.3 mg). Whereas under no choice field conditions. Minimum feeding was recorded in Phylanthus emblicallium (4.8 mg) and maximum on Cupressus sempervirens linn (11.8 mg). Feeding preferences as a quantitative parameter of wood consumption by O. Obesus were recognized as follows: Cupressus sempervirens linn >Ficus microphylapers > Ficus banghalensis linn > Ficus relgiosa linn > phylanthus embicallium. Toxicity and LC₅₀ value of fipronil, Imidacloprid indicate that Imidacloprid is more toxic than fipronil based on LC₅₀ values.

Conclusions: When resistant tree species are planted in combination with those of susceptible in natural environment then the termite infestation rate could be minimised to a greater extent. Natural durability is an area of increasing attention that can help in reducing the application of industrial chemicals in the surroundings. A broader diversity of trees is needed in our urban landscapes to guard against the possibility of large-scale devastation by both native and introduced insect including pest termites. Tree diversity in forests is an important driver of ecological processes including herbivory.

Aims: This study aimed to investigate the in vitro toxicity activity of ethyl acetate extracts of endophytic fungi isolated from soursop leaves (Annona muricata L.) against colon cancer and analyse the bioactive compounds from the fungal extract with best anticancer activity using Gas Chromatograph-Mass Spectrometer (GC-MS).

Study Design: The experimental design used was a completely randomized factorial design with two factors, the type and concentration of endophytic fungi extract.

Place and Duration of Study: Industrial Microbiology Laboratory of Biology Research Center, Indonesian Institute of Sciences (LIPI) Cibinong-Bogor, Laboratory of Microbiology and Immunology of Center for Animal and Primate Studies of Bogor Agricultural University and Center of Forensic Laboratory, conducted from March 2016 to March 2017.

Methodology: Cytotoxic property was determined using 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay method. Entophytes were extracted using ethyl acetate solvent. Extract of ten isolates (at concentration of 100 µg/mL) were examined for the inhibition effect to WiDr cancer cells. Five isolates that showed significantly high inhibition effect (P<0.05) were further selected for IC₅₀ determination against WiDr cell lines at extract concentration of 25, 50, 100, 200 and 400 µg/mL. Three isolates that significantly showed high activity (P<0.05) were chosen, and then measured for their toxicity (IC₅₀ value) to normal cell (Chang cell lines) at concentration of 25, 50, 100, 200 and 400 µg/mL. The bioactive compound of the isolate which had the lowest toxicity to normal cell was then analysed using GC-MS.

Results: Fungal extracts of ten isolates were examined for their cytotoxicity to WiDr cancer cells with the result showed that isolate of Sir-CA1, Sir-CA2, Sir-G2, Sir-G4 and Sir-SM2 had high inhibitions. From the five fungal extracts, isolate of Sir-CA1, Sir-G2 and Sir-SM2 showed high cytotoxicity to WiDr cancer cell. Among the three isolats, fungal extract of isolate from Sukabumi (Sir-SM2) showed the lowest toxicity to normal cell was analysed on secondary metabolite compounds using GC-MS. The fungal extract contained ester group, alkaloid, saturated fatty acid, unsaturated fatty acid, terpene, terpenoid and aromatic compounds.

Conclusion: Ethyl acetate extracts of endophytic fungi Sir-SM2 isolated from soursop leaves (Annona muricata L.) had high cytotoxic effect on colon cancer cell (IC₅₀=20.80 µg/mL) and the lowest toxicity to normal cell compared with other fungal extracts (IC₅₀=63.69 µg/mL). The analysis of bioactive compounds with GC-MS showed that fungal extract of isolate Sir- SM2 contained compounds such as alkaloid, saturated fatty acid, unsaturated fatty acid, terpene and terpenoid that had function as anticancer.

Background: In Egypt, the prevalence of chronic hepatitis C (CHC) infection is 14% of whole population and about 80% of the patients with hepatocellular carcinoma have underling hepatitis C.

Aim: To explore the diagnostic value of serum miR-122 as non-invasive diagnostic markers of (HCV)- (HCC).

Methodology: The study included 102 healthy controls (group I), in addition to 228 patients, they were divided into 2 groups: 120 CHC patients (group II), and 108 hepatitis C patients with HCC (group III). All samples underwent full clinical assessment and laboratory investigations additionally to the detection of the concentration of blood serum miR-122 expression by RT-PCR. Selected biomarkers were assessed, evaluated and correlated with degree of liver damage.

Results: Revealed that miR-122 had the highest efficiency in prediction of liver cell damage. Also, miR-122 was strongly correlated with vascular endothelial growth factor (VEGF) and alpha fetoprotein (α-FP) in HCC patients.

Conclusions: Plasma miR-122 can be considered as a potential biomarker, for detection of HCC combined with VEGF and α -FP.