Evaluation of antioxidant and anticancer properties of the seed extracts of Syzygium fruticosum Roxb. growing in Rajshahi, Bangladesh.

BACKGROUND: The use of plants and their derived substances increases day by day for the discovery of therapeutic agents owing to their versatile applications. Current research is directed towards finding naturally-occurring antioxidants having anticancer properties from plant origin since oxidants play a crucial role in developing various human diseases. The present study was designed to investigate the antioxidant and anticancer properties of Sygygium fruticosum (Roxb.) (abbreviated as SF). METHODS: The dried coarse powder of seeds of SF was exhaustively extracted with methanol and the resulting crude methanolic extract (CME) was successively fractionated with petroleum ether, chloroform and ethyl acetate to get petroleum ether (PEF), chloroform (CHF), ethyl acetate (EAF) and lastly aqueous (AQF) fraction. The antioxidant activities were determined by several assays: total antioxidant capacity assay, DPPH free radical scavenging assay, hydroxyl radical scavenging assay, ferrous reducing antioxidant capacity and lipid peroxidation inhibition assay. The in vivo anticancer activity of SF was determined on Ehrlich's Ascite cell (EAC) induced Swiss albino mice. RESULTS: All the extractives showed strong antioxidant activities related to the standard. The total antioxidant capacity (TAC) of the fractions was in the following order: EAF>AQF>CME>PEF>CHF. The TAC of EAF at 320 µg/mL was 2.60±0.005 which was significantly higher (p < 0.01) than that of standard catechin (1.37 ± 0.005). The ferrous reducing antioxidant capacity of the extracts was in the following order: EAF>AQF>CME>AA>CHF>PEF. In DPPH free radical scavenging assay, the IC50 value of EAF was 4.85 µg/mL, whereas that of BHT was 9.85 µg/mL. In hydroxyl radical scavenging assay and lipid peroxidation inhibition assay, the EAF showed the most potent inhibitory activity with IC50 of 43.3 and 68.11 µg/mL, respectively. The lipid peroxidation inhibition assay was positively correlated (p < 0 .001) with both DPPH free radical scavenging and hydroxyl radical scavenging assay. The total phenolic contents of SF were also positively correlated (p < 0 .001) with DPPH free radical scavenging, hydroxyl radical scavenging and lipid peroxidation inhibition assay. Based on antioxidant activity, EAF was selected for cytotoxic assay and it was found that EAF inhibited 67.36% (p < 0.01) cell growth at a dose of 50 mg/kg (ip) on day six of EAC cell incubation. CONCLUSIONS: Our results suggest that EAF of seeds of SF possess significant antioxidant and moderate anticancer properties. Seeds of SF may therefore be a good source for natural antioxidants and a possible pharmaceutical supplement.

Traditional uses, pharmacological activities, and phytochemical constituents of the genus Syzygium: A review.

The genus Syzygium comprises 1200-1800 species that belong to the family of Myrtaceae. Moreover, plants that are belonged to this genus are being used in the traditional system of medicine in Asian countries, especially in China, India, and Bangladesh. The aim of this review is to describe the scientific works and to provide organized information on the available traditional uses, phytochemical constituents, and pharmacological activities of mostly available species of the genus Syzygium in Bangladesh. The information related to genus Syzygium was analytically composed from the scientific databases, including PubMed, Google Scholar, Science Direct, Web of Science, Wiley Online Library, Springer, Research Gate link, published books, and conference proceedings. Bioactive compounds such as flavanone derivatives, ellagic acid derivatives and other polyphenolics, and terpenoids are reported from several species of the genus Syzygium. However, many members of the species of the genus Syzygium need further comprehensive studies regarding phytochemical constituents and mechanism-based pharmacological activities.

Alternative splicing of Mef2c promoted by Fox-1 during neural differentiation in P19 cells.

Mef2c protein is one of the MADS-box type transcription factors involved in muscular differentiation and synaptic formation. Previously, it has been reported that the Mef2c gene is responsible for three alternative splicing regulations. Here, we investigated the alternative splicing variants of Mef2c during neural differentiation of P19 cells and during cardio muscular differentiation of P19 clone 6 (P19CL6). We detected that two Mef2c mRNA isoforms, using exon α1 with and without the γ region at exon 10, are mainly produced in immature P19 cells. Remarkably, Mef2c isoforms containing exon ß specifically appeared in the neural cell stage. Because most transcripts contain exon ß in the neural cell stage and in the brain, this suggests that the alternative splicing of exon ß is highly regulated. Among known regulators, Fox-1 was specifically expressed in the neural cell stage in correlation with Mef2c exon ß. Fox-1 promoted exon ß inclusion in transfection experiments using Mef2cß minigene. Moreover, we found that the promotion required RNA-binding activity of Fox-1 and GCAUG sequence located in adjacent intron of exon ß. Taken together, our results suggest that Fox-1, expressed specifically in the neural cell stage, promoted Mef2c exon ß inclusion via the GCAUG.

Antidiabetic and hypolipidemic effects of different fractions of Coccinia cordifolia L. on normal and streptozotocin-induced diabetic rats.

The present study was carried out to observe the antidiabetic and hypolipidemic effects of petroleum-ether, ethyl acetate and chloroform fractions isolated from ethanolic extract of the leaves of Coccinia cordifolia Linn. (150 mg/kg body weight) on normal and streptozotocin (STZ)-induced diabetic rats for one day experiment. Single doses (150 mg/kg, i.p.) of C. cordifolia extracts were given to normal and diabetic rats. The fasting blood glucose (FBG), serum triglyceride (TG) and serum total cholesterol (TC) levels were investigated in normal and STZ-diabetic rats on 0, 1, 2, 3, 6, 10, 16, and 24th hours. In normoglycemic rats the pet-ether and ethyl acetate fractions of C. cordifolia reduced blood glucose level significantly (39.66% and 40.68% at 16th and 24th hour respectively). In the STZ-diabetic rats pet-ether and ethyl acetate fractions also reduced blood glucose level significantly (50.39% and 50% at 10th and 24th hour respectively). Ethyl acetate fraction is most effective which reduced total cholesterol level by 31.04% and 36.69% in normal and STZ-diabetic rats respectively. Ethyl acetate fraction reduced triglyceride level by 43.82% and 42.01% in normal and STZ-diabetic rats respectively. Our results indicate that pet-ether and ethyl acetate fractions of C. cordifolia have potentiality against diabetes.

Trichosanthes dioica seed lectin inhibits Ehrlich ascites carcinoma cells growth in vivo in mice by inducing G0 /G1 cell cycle arrest.

Trichosanthes dioica seed lectin (TDSL), having a molecular mass of 57 ± 2 kDa was purified in an alternative way. For the purification process, the galactose-sepharose-4B affinity column was used. The purified TDSL agglutinated human and mouse erythrocytes at the minimum concentration of 8  µg/ml. d-lactose and d-galactose were the most potent inhibitory sugars as observed. The purified lectin was a glycoprotein having 3.0% of a neutral sugar. The lectin exhibited maximum activity up to 60°C and pH range from 7.0 to 10.0 and stable up to 4.0 M urea as tested. The lectin demonstrated mild toxicity when administered against brine shrimp nauplii, and the LC50 value was calculated to be 84.0 µg/ml. Minimum agglutination of Ehrlich ascites carcinoma (EAC) cells caused by the lectin was found at the protein concentration of 1.56 µg/ml. TDSL inhibited 7, 50.2%, and 60.3% of the EAC cells growth in vivo in mice when administered with 0.75, 1.5, and 3.0 mg kg-1  day-1 (i.p.), respectively, for five consecutive days. After lectin treatment, red blood cell (RBC) and hemoglobin levels were increased significantly toward the normal compared with EAC cells-bearing control and normal mice. The tumor burden reduced to 29.5% and 67% after treatment with 1.5 and 3.0 mg kg-1  day-1 of the lectin. TDSL triggered the cell cycle arrest at the G0 /G1 phase, which was observed using flow cytometry. In conclusion, TDSL can be a candidate for the potent anticancer agents that exerts low toxicity toward brine shrimp nauplii. PRACTICAL APPLICATIONS: A 57 ± 2 kDa lectin (designated TDSL) was purified from Trichosanthes dioica seeds using a galactose-sepharose-4B affinity column. The lectin demonstrated mild toxicity and agglutinated Ehrlich ascites carcinoma (EAC) cells. The lectin inhibited 50.2% and 60.3% of the EAC cell growth in vivo in mice when administered with 1.5 and 3.0 mg kg-1  day-1 (i.p.), respectively, for five consecutive days. The lectin increased RBC and hemoglobin level toward the normal compared with lectin-treated EAC cells-bearing, EAC cells-bearing control and normal mice. The tumor burden reduced to 29.5% and 67% after treatment with 1.5 and 3.0 mg kg-1  day-1 lectin. TDSL triggered the cell cycle arrest at the G0 /G1 phase. The lectin can be a candidate for potent anticancer agents.

Phytochemical Analysis and Cholinesterase Inhibitory and Antioxidant Activities of Enhydra fluctuans Relevant in the Management of Alzheimer's Disease.

Enhydra fluctuans, a popular vegetable in Bangladesh, is used in folk medicine to treat diseases of the nervous system. The objective of this study was to investigate the phytochemical profile and cholinesterase inhibitory and antioxidant potential of the extracts of E. fluctuans. Among the four tested extracts, the chloroform extract was found to exert the highest inhibition against both the acetylcholinesterase and butyrylcholinesterase enzymes with the IC50 (concentration required for 50% inhibition) values of 83.90 µg/mL and 48.14 µg/mL, respectively. Likewise, the chloroform extract showed the highest radical scavenging activity and reducing power. In DPPH radical scavenging assay, the IC50 value was found to be 113.27 µg/mL, and in reducing power assay, the absorbance was found to be 1.916 at a concentration of 50 µg/mL. Phytochemical analyses revealed that the chloroform extract contained 19.16 mg gallic acid equivalent (GAE)/g extract of phenolics and 41.84 mg catechin equivalent (CE)/g extract of flavonoids, which appeared to be the highest among the extracts. A significant correlation was observed between phenolic content and butyrylcholinesterase inhibition and antioxidant activity, while a moderate correlation was seen between flavonoid content and cholinesterase inhibition and antioxidant activity. These findings suggest that E. fluctuans is a natural source of cholinesterase inhibitors and antioxidants, which could be utilized as functional foods for Alzheimer's disease management.

Antioxidative role of palm grass rhizome ameliorates anxiety and depression in experimental rodents and computer-aided model.

Palm grass (Curculigo recurvata) is an ethnomedicinally important herb reported to have significant medicinal values. The present study aimed to evaluate the antidepressant and anxiolytic activities of a methanol extract of C. recurvata rhizome (Me-RCR) through different approaches. The antidepressant and anxiolytic properties of Me-RCR were assessed by using elevated plus maze (EPM), hole-board (HBT), tail suspension (TST), and forced swimming (FST) tests in Swiss Albino mice. The in-depth antioxidative potential of Me-RCR was also evaluated through DPPH radical scavenging activity, ferric-reducing power capacity, total phenolic, flavonoid, flavonol, and antioxidant content analysis. Computational investigations were performed using computer-aided methods for screening the anxiolytic, antidepressant, and antioxidative activities of the selected lead molecules. Treatment with Me-RCR (200 and 400 mg/kg, b.w.) notably increased the number of open arm entries and the time spent in the EPM test. In the HBT, Me-RCR exhibited significant anxiolytic activity at a dose of 200 mg/kg, whereas similar activity was observed at 400 mg/kg in the EPM test. Me-RCR significantly decreased the immobility time in a dose-dependent manner in both TST and FST. The IC50 for DPPH and reducing power capacity assay were found to be 18.56 and 193 µg/mL, respectively. Promising outcomes were noted for the determination of total phenolics, flavonoids, flavonols, and antioxidant capacity. In the case of computer-aided studies, nyasicoside showed promising binding energy for antidepressant and anxiolytic activities, whereas isocurculigine demonstrated promising effects as an antioxidant. Overall, these findings suggest that Me-RCR could be a favourable therapeutic candidate for the treatment of mental and psychiatric disorders, as well as a good source of antioxidants.

Retinoic acid treatment and cell aggregation independently regulate alternative splicing in P19 cells during neural differentiation.

To induce neural differentiation of P19 cells, two different treatments, RA (retinoic acid) and cell aggregation, are required. However, there has been no report that RA treatment alone or cell aggregation alone could control alternative splicing regulation in P19 cells. Therefore, we focused on alternative splicing effects by neural induction (RA treatment and/or cell aggregation) in P19 cells. We analysed the splicing patterns of several genes, including 5-HT3R-A (5-hydroxytryptamine receptor), Actn1 (actinin alpha1), CUGBP2 (CUG-binding protein) and PTB (polypyrimidine track-binding protein), which showed different responses during the early neural induction of P19 cells. We show here that RA treatment alone changes the alternative splice mechanism of 5-HT3R-A. Cell aggregation alone controls alternative splicing regulation of Actn1. Both treatments (RA and cell aggregation) compensate and regulate the alternative splicing mechanism of CUGBP2. However, PTB is independent of RA and cell aggregation. Taken together, our results suggest that RA treatment and cell aggregation independently regulate the alternative splicing mechanism in the early stage of P19 cells during neural differentiation.

Curculigo recurvata W.T.Aiton exhibits anti-nociceptive and anti-diarrheal effects in Albino mice and an in silico model.

BACKGROUND: Curculigo recurvata (C. recurvata) is an enthnomedicinally important herb reported to have significant medicinal values. The present study aimed to explore the in vivo and in silico anti-nociceptive and anti-diarrheal effects of a C. recurvate rhizome methanol extract (Me-RCR). METHODS: The analgesic effects of Me-RCR were assessed using acetic acid-induced writhing and the formalin-induced flicking test. The drugs were administered intraperitoneally (IP) at doses of 200 and 400 mg/kg body weight (bw). Anti-diarrheal activity was evaluated by assessing intestinal motility, hypersecretion, and fecal score in mice at oral doses of 200 and 400 mg/kg·bw. Computer facilitated analyses for anti-nociceptive and anti-diarrheal activities of three isolated compounds from C. recurvata were undertaken to identify the best-fit phytoconstituents. RESULTS: The Me-RCR showed significant (P < .05) peripheral anti-nociception at the highest dose. The extract inhibited both early and late phases of nociception in the formalin-induced writhing test. In the castor oil-induced diarrhoea model, the extract significantly (P < .05) prolonged the onset time of diarrhoea, inhibited percentage of diarrhoea, and decreased both the volume and weight of intestinal contents. Rates of intestinal fluid accumulation inhibition were (33.61 ± 1.00)% and (46.44 ± 0.89)% at Me-RCR doses of 200 and 400 mg/kg·bw, respectively. Moreover, a significant (P < .05) reduction in gastrointestinal motility was observed. An absorption, distribution, metabolism, excretion and/or toxicity (ADME/T) test showed that the selected compounds yielded promising results, satisfying Lipinski's rule of five for predicting drug-like potential. Notably, of the three phytoconstituents curculigine and isocurculigine possessed the highest affinity for the COX-1 and COX-2. Isocurculigine was also identified as the most effective anti-diarrheal compound in the computer-facilitated model. CONCLUSION: An extract of the plant C. recurvata showed potential analgesic and anti-diarrheal activity due to the presence of one or more active secondary metabolite(s).

Expression analysis of Fgf8a &Fgf8b in early stage of P19 cells during neural differentiation.

Fgf8 is a member of the fibroblast growth factor (FGF) family that plays an important role in early neural development. Cellular aggregation and retinoic acid (RA) are needed for mouse embryonic carcinoma (EC) P19 cell neural differentiation. We have examined the Fgf8 gene in P19 cells during neural differentiation and identified 2 alternatively spliced Fgf8 isoforms, Fgf8a and Fgf8b, among the 8 known splicing isoforms in mammals. The expression of Fgf8a and Fgf8b mRNAs transiently and rapidly increased in the early stage of P19 cells during RA-induced neural differentiation, followed by a decline in expression. The relative amount of Fgf8b was clearly higher than that of Fgf8a at different time-points measured within 24h after RA treatment. Increased Fgf8b mRNA expression was cellular-aggregation dependent. The results demonstrated that cellular-aggregation-induced Fgf8b, but not Fgf8a, may play a pivotal role in early neural differentiation of P19 cells.

Oral glucose tolerance test (OGTT) in normal control and glucose induced hyperglycemic rats with Coccinia cordifolia l. and Catharanthus roseus L.

The aim of this study is to investigate the hypoglycemic effects of petroleum ether, chloroform and ethyl acetate fractions isolated from ethanolic extracts of Coccinia cordifolia and Catharanthus roseus on normal control and orally glucose-induced hyperglycemic rats. Single doses (150 mg/kg) of different fractions of C. cordifolia and C. roseus extracts were intraperitonelly administered. The serum blood glucose level was obtained by pricking the tail vein using glucometer at time 0, 30, 60, 90, 150 and 270 minutes. In the orally glucose induced hyperglycemic rats, chloroform-coccinia (CHCl3-CC) fraction showed maximum reduction of blood glucose level by 21.94% on 60 minute of the experiment. On the other hand maximum reduction (p<0.05) of 17.92% was observed for petroleum ether-catharanthus (PET-CR) on 30 minute of the experiment. Metformin HCl was used as standard drug. Our results indicate that the CHCl3-CC fraction is relatively more potent than other fractions of C. cordifolia. Similarly the PET-CR is found to be better than other fractions of catharanthus. Phytochemical screening test results showed that chloroform fraction of C. cordifolia contain saponins and flavonoids compounds, which are known to be hypoglycemic. On the other hand petroleum ether fraction of C. roseus contains tannins, flavonoids and alkaloid compounds produced varying degree of blood sugar reduction. On the pharmacological point of view C. cordifolia and C. roseus appears to be a valuable plant, which can be useful, at least as an adjunct, in the therapy of diabetes.

Antimicrobial and cytotoxic activities of Achyranthes ferruginea.

Chloroform extract (CE) of Achyranthes ferruginea and N-trans-feruloyl-4-methyldopamine (1) showed remarkable antimicrobial activities against a wide range of bacteria and fungi. Both crude extract (CE) and compound 1 showed significant cytotoxicity of LC(50) at 16.21 microg/ml and 11.70 microg/ml, respectively.

Antibacterial and cytotoxic compounds from the bark of Cananga odorata.

O-Methylmoschatoline, liriodenine and 3,4-dihydroxybenzoic acid isolated from the barks of Cananga odorata showed antibacterial activities against a number of Gram (+) and Gram (-) bacteria. The compounds also showed antifungal and cytotoxic activities.

Momordica charantia seed lectin: toxicity, bacterial agglutination and antitumor properties.

In last three decades, several studies were carried out on the D-galactose-specific lectin of Momordica charantia seeds (MCL). In the present study, in vitro growth inhibition (8-23 %) at different concentrations (6-24 µg/ml) of MCL was observed against Ehrlich ascites carcinoma (EAC) cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. MCL also showed 28, 45, and 75 % growth inhibitions against EAC cells when administered 1.2, 2.0, and 2.8 mg/kg/day (i.p.), respectively for five consequent days in vivo in mice. After lectin treatment, the level of red blood cell and hemoglobin was increased significantly with the decrease of white blood cell and maintained the normal level when compared with EAC-bearing control and normal mice without EAC cells. Although MCL caused cell cycle arrest at G0/G1 phase of EAC cells, any irregular shape or apoptotic morphological alterations in the lectin-treated EAC cells was not observed by an optical and fluorescence microscope. Lectin showed toxicity against brine shrimp nauplii with an LC50 value of 49.7 µg/ml. Four out of seven pathogenic bacteria were agglutinated by MCL in the absence of inhibitory sugar D-lactose/D-galactose. In conclusion, MCL showed strong cytotoxic effect and therefore can be used as a potent anticancer chemotherapeutic agent.

Effects of the phosphodiesterase IV inhibitor rolipram on Th1 and Th2 immune responses in mice.

The present study was designed to investigate the effect of the phosphodiesterase IV inhibitor rolipram on Th1 and Th2 immune responses in mice. Mice were immunized subcutaneously at the base of the tail with ovalbumin (OVA) emulsified with complete Freund's adjuvant (day 0) and were treated daily with oral administration of various doses of rolipram from days 0 to 20. On day 21, production of anti-OVA IgG and proliferative responses to the antigen were determined. Anti-OVA IgG2a and interferon-gamma (IFN-gamma), as indicators of Th1 responses, and anti-OVA IgG1 and interleukin-10 (IL-10), as indicators of Th2 responses, were also measured. The results showed that treatment with rolipram failed to affect the production of OVA-specific IgG but decreased the proliferation of spleen cells to the antigen. Its inhibitory effect on these immune responses was correlated with a marked decrease in IFN-gamma but not IL-10 production, although neither anti-OVA IgG2a nor IgG1 production was affected by rolipram. These results suggest that rolipram may preferentially inhibit Th1 responses more effectively than Th2 responses. Administration of rolipram resulted in suppression of antigen (OVA)-induced arthritis in mice. The suppression of joint inflammation by rolipram was associated with the inhibition of the OVA-specific proliferative responses of spleen cells and IFN-gamma secretion. These results indicate that rolipram may be effective in regulating Th1-mediated diseases such as rheumatoid arthritis.

Estimation of total phenol and in vitro antioxidant activity of Albizia procera leaves.

BACKGROUND: Research on natural products has gained a wide popularity due to the potential of discovering active compounds. The antioxidant properties contained in plants have been proposed as one of the mechanisms for the observed beneficial effect. Therefore, the present study investigated the antioxidant activity and total phenolic contents of various solvent extracts of Albizia procera leaves. METHODS: Antioxidant activity of the methanol extract and its derived fractions petroleum ether (APP), carbon tetrachloride (APC), dichloromethane (APD), ethyl acetate (APE), and residual aqueous fraction (APA) of the leaves of Albizia procera was performed by in vitro chemical analyses. Total phenolic content of the APM and other five fractions were also determined. APM and its derived fractions were also subjected to preliminary phytochemical screening test for various constituents. RESULTS: Phytochemical screening revealed the presence of saponins, steroids, tannins, glycosides and flavonoids in the extracts. Amongst the extracts, APE showed the highest total phenolic content (449.18 ± 18.41mg of gallic acid equivalent/g of extract). In DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging test, the IC(50) value of APM, APP, APC, APD, APE and APA was 43.43, 63.60, 166.18, 41.15, 11.79, and 63.06 µg/mL, respectively. Therefore, among the APM and its derived fractions, APE showed the highest antioxidant activity which is comparable to that of standard ascorbic acid (AA) (IC(50) 10.12 µg/mL). The total antioxidant capacity was found to be varied in different fractions. The reducing activity on ferrous ion was ranked as APE > APD > APM > APA > APC. CONCLUSION: The above evidences suggest that APE of A. procera leaf is a potential source of natural antioxidant and can be used to prevent diseases associated with free radicals.

A comparative study on the antioxidant activity of methanolic extracts from different parts of Morus alba L. (Moraceae).

BACKGROUND: Antioxidants play an important role to protect damage caused by oxidative stress (OS). Plants having phenolic contents are reported to possess antioxidant properties. The present study was designed to investigate the antioxidant properties and phenolic contents (total phenols, flavonoids, flavonols and proanthrocyanidins) of methanolic extracts from Morus alba (locally named as Tut and commonly known as white mulberry) stem barks (TSB), root bark (TRB), leaves (TL) and fruits (TF) to make a statistical correlation between phenolic contents and antioxidant potential. METHODS: The antioxidant activities and phenolic contents of methanolic extractives were evaluated by in vitro standard method using spectrophotometer. The antioxidant activities were determined by total antioxidant capacity, DPPH (1,1-diphenyl-2-picrylhydrazine) radical scavenging assay, hydroxyl radical scavenging assay, ferrous reducing antioxidant capacity and lipid peroxidation inhibition assay methods. RESULTS: Among the extracts, TSB showed the highest antioxidant activity followed by TRB, TF and TL. Based on DPPH and hydroxyl radical scavenging activity, the TSB extract was the most effective one with IC50 37.75 and 58.90 µg/mL, followed by TRB, TF and TL with IC50 40.20 and 102.03; 175.01 and 114.63 and 220.23 and 234.63 µg/mL, respectively. The TSB extract had the most potent inhibitory activity against lipid peroxidation with IC50 145.31 µg/mL. In addition, the reducing capacity on ferrous ion was in the following order: TSB > TRB > TL > TF. The content of phenolics, flavonoids, flavonols and proanthocyanidins of TSB was found to be higher than other extractives. CONCLUSION: The results indicate high correlation and regression (p-value <0 .001) between phenolic contents and antioxidant potentials of the extracts, hence the Tut plant could serve as effective free radical inhibitor or scavenger which may be a good candidate for pharmaceutical plant-based products. However, further exploration is necessary for effective use in both modern and traditional system of medicines.

Comprehensive analysis of alternative splicing and functionality in neuronal differentiation of P19 cells.

BACKGROUND: Alternative splicing, which produces multiple mRNAs from a single gene, occurs in most human genes and contributes to protein diversity. Many alternative isoforms are expressed in a spatio-temporal manner, and function in diverse processes, including in the neural system. METHODOLOGY/PRINCIPAL FINDINGS: The purpose of the present study was to comprehensively investigate neural-splicing using P19 cells. GeneChip Exon Array analysis was performed using total RNAs purified from cells during neuronal cell differentiation. To efficiently and readily extract the alternative exon candidates, 9 filtering conditions were prepared, yielding 262 candidate exons (236 genes). Semiquantitative RT-PCR results in 30 randomly selected candidates suggested that 87% of the candidates were differentially alternatively spliced in neuronal cells compared to undifferentiated cells. Gene ontology and pathway analyses suggested that many of the candidate genes were associated with neural events. Together with 66 genes whose functions in neural cells or organs were reported previously, 47 candidate genes were found to be linked to 189 events in the gene-level profile of neural differentiation. By text-mining for the alternative isoform, distinct functions of the isoforms of 9 candidate genes indicated by the result of Exon Array were confirmed. CONCLUSIONS/SIGNIFICANCE: Alternative exons were successfully extracted. Results from the informatics analyses suggested that neural events were primarily governed by genes whose expression was increased and whose transcripts were differentially alternatively spliced in the neuronal cells. In addition to known functions in neural cells or organs, the uninvestigated alternative splicing events of 11 genes among 47 candidate genes suggested that cell cycle events are also potentially important. These genes may help researchers to differentiate the roles of alternative splicing in cell differentiation and cell proliferation.

Difference in preventive effects between the phosphodiesterase iv inhibitor rolipram and anti-arthritic drugs on antigen-induced arthritis in mice.

The efficacy of the phosphodiesterase (PDE) IV inhibitor rolipram on antigen-induced arthritis (AIA) in mice was evaluated in comparison with clinically used anti-arthritic drugs. To induce AIA, DBA/1 mice were immunized with ovalbumin (OVA) emulsified with CFA (day 0) followed by intra-articular injection of OVA on day 21. Rolipram and clinically used anti-arthritic drugs including indomethacin (IND), dexamethasone (DEX), methotrexate (MTX), auranofin (AUR), and D-penicillamine (D-PA) were orally administered daily from days 0 to 20. On day 22, anti-OVA IgG in serum, proliferative responses of spleen cells to the OVA, and anti-OVA IgG2a and interferon (IFN)-gamma as indicators of Th1 responses, as well as anti-OVA IgG1 and interleukin (IL)-10 as those of Th2 reactions, were measured. Treatment with rolipram was followed by inhibition of the early phase of AIA associated with downregulation of both OVA-specific splenocyte proliferation and decreases of IFN-gamma released from the spleen cells but no decreases of the amount of IL-10, or levels of anti-OVA IgG, IgG2a, and IgG1. All clinically used anti-arthritic drugs were more effective in suppressing the late phase of AIA compared with the early phase of joint inflammation. The suppression of AIA by clinically used anti-arthritic drugs was associated with down-regulation of not only Th1 but also Th2 responses. These results suggest that PDE IV inhibitors such as rolipram may exert their suppressive effects on AIA with relatively selective downregulation of antigen-specific Th1 responses compared with anti-arthritic drugs.

Toxicological studies of two compounds isolated from Loranthus globosus Roxb.

The sub-acute toxicities of two compounds 3,4-dimethoxycinnamyl alcohol (1) and 3,4,5-trimethoxycinnamyl alcohol (2) isolated from the plant Loranthus globosus Roxb were studied on long Evan's rats. The studies included the gross general observation such as changes in body weight, haematological profiles [total count of Red Blood Cells (RBC) and White Blood Cells (WBC), differential count of WBC, platelet count and Haemoglobin (Hb)%], biochemical parameters of blood [Serum Glutamate Oxaloacetate Transaminase (SGOT), Serum Glutamate Pyruvate Transaminase (SGPT), Serum Alkaline Phosphatase (SALP), urea and creatinine) and histopathology of the liver, kidney, heart and lung of both control and experimental groups of rats. The changes in haematological and biochemical parameters were statistically not significant after the administration of compounds 1 and 2 in a dose of 300 microg/rat/day for consecutive 14 days. No abnormality was found in the histopathology of the liver, kidney, heart and lung in the experimental groups of rats following same dose when compared with control group. This preliminary study suggests that the isolated compounds may be used safely for clinical trial.