Conventional mammalian models of neurodegeneration are often limited by futile axonogenesis with minimal functional recuperation of severed neurons. The emergence of zebrafish, a non-mammalian model with excellent neuroregenerative properties, may address these limitations. This study aimed to establish an adult zebrafish-based, neurotoxin-induced Parkinson's disease (PD) model and subsequently validate the regenerative capability of dopaminergic neurons (DpN). The DpN of adult male zebrafish (Danio rerio) were lesioned by microinjecting 6-hydroxydopamine (6-OHDA) neurotoxin (6.25, 12.5, 18.75, 25, 37.5, 50 and 100 mg/kg) into the ventral diencephalon (Dn). This was facilitated by an optimised protocol that utilised 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanineperchlorate (DiI) dye to precisely identify the injection site. Immunostaining was utilised to identify the number of tyrosine hydroxylase immunoreactive (TH-ir) DpN in brain regions of interest (i.e. olfactory bulb, telencephalon, preoptic area, posterior tuberculum and hypothalamus). Open tank video recordings were performed for locomotor studies. The Dn was accessed by setting the injection angle of the microinjection capillary to 60° and injection depth to 1200 μm (from the exposed brain surface). 6-OHDA (25 mg/kg) successfully ablated >85% of the Dn DpN (preoptic area, posterior tuberculum and hypothalamus) whilst maintaining a 100% survival. Locomotor analysis of 5-min recordings revealed that 6-OHDA-lesioned adult zebrafish were significantly (p < 0.0001) reduced in speed (cm/s) and distance travelled (cm). Lesioned zebrafish showed full recovery of Dn DpN 30 days post-lesion. This study had successfully developed a stable 6-OHDA-induced PD zebrafish model using a straightforward and reproducible approach. Thus, this developed teleost model poses exceptional potentials to study DpN regeneration.

BackgroundMedicinal plants are important source of drugs with pharmacological activities. Therefore, there is always rising demands to discover more therapeutic agents from various species. Orthosiphon stamineus, Gynura procumbens and Ficus deltoidea are high valued medicinal plants of Malaysia contain rich source of phenolic and flavonoid compounds. The aims of the present study were to evaluate anti-oxidant, antimicrobial and anti-proliferative effects on A549, HeGP2 and MCF7 cell lines of four different extracts of Orthosiphon stamineus, Gynura procumbens and Ficus deltoidea.MethodologyThe leaves of all selected plants were extracted with methanol, chloroform, ethyl acetate and butanol separately with simple cold maceration. Antioxidant activity of all crude extracts were quantitatively measured against DPPH and Ferric Reducing Assay. Antimicrobial evaluation was done by Microdilution and MTT assay and antipoliferative activity of all extracts of selected plant were evaluated against A549, HePG2 and MCF7 cell lines.ResultsResults showed that methanol extract exhibited highest percentage free radical scavenging activity of almost all extracts of selected plants. Antimicrobials results showed chloroform and methanol extracts of O. stamineus extract were the two most active extracts against resistant MRSA but not S. aureus. Only methanol extract of G. procumbens showed antimicrobial activity against the tested pathogens. Chloroform and methanol extracts of F. deltoidea elicited antimicrobial activity against S. aureus but not MRSA. Antiproliferative activity against three tested cell lines results showed that ethyl acetate extract of O. stamineus showed good effect whereas methanol extract of F. deltoidea and G. procumbens exhibited good antiproliferative activity.ConclusionsThe results of the present investigation demonstrated significant variations in the antioxidant, antimicrobial and antiproliferative effects of different solvent extracts. These data could be helpful in isolation of pure potent compounds with good biological activities from the extracts of plants.

A series of benzimidazole derivatives (1-20) was synthesized and evaluated for its in vitro antimicrobial, antitubercular and anticancer activities. Compound 10 was found to be the most active antibacterial agent. The compounds active in in vitro evaluation against M. tuberculosis were further assessed for their in vivo activity in mice and for their capacity to inhibit the vital mycobacterial enzymes viz., isocitrate lyase, pantothenate synthetase and chorismate mutase. The dose of the compounds in antitubercular evaluation that proved fatal and highly toxic to mice was 5.67 mg/kg while lethal dose varied from 1.82 mg/kg to 3.23 mg/kg body weight of the mice. A dose of 1.34 mg/kg was found to be safe for each of the compounds. All compounds inhibited the mycobacterial enzymes but to a lesser extent than streptomycin sulphate used as positive control. Compound 19, exhibiting inhibition of 67.56%, 53.45%, and 47.56% against isocitrate lyase, pantothenate synthetase and chorismate mutase, respectively is the most potent antitubercular compound among the synthesized benzimidazole derivatives. Further, compound 19 also emerged as a potent anticancer agent (IC 50 = 0.0013 mM) than 5-flourouracil against breast cancer cell line (MCF 7).

There is growing interest in the use of probiotic lactic acid bacteria (LAB) for prevention of hypercholesterolaemia. This study assessed the cholesterol lowering ability of Pediococcus acidilactici LAB4 and Lactobacillus plantarum LAB12 in growth media. Both LAB yielded >98% (39.2 μg/ml) cholesterol lowering in growth media. Nile Red staining indicated direct assimilation of cholesterol by the LAB. The LAB were then explored for their prophylactic (pre-treatment of HT29 cells with LAB prior to cholesterol exposure) and biotherapeutic (treatment of HT29 cells with LAB after exposure to cholesterol) use against short and prolonged exposure of HT29 cells to cholesterol, respectively. For HT29 cells pre-treated with LAB, cholesterol lowering was accompanied by down-regulation of ATP-binding cassette family transporter-type A1 (ABCA1), cluster of differentiation 36 (CD36) and scavenger receptor class B member 1 (SCARB1). HT29 cells treated with LAB after prolonged exposure to cholesterol source, on the other hand, was associated with up-regulation of ABCA1, restoration of CD36 to basal level and down-regulation of Neimann-Pick C1-Like 1 (NPC1L1). The present findings implied the potential use of LAB4 and LAB12 as part of the strategies in prevention and management of hypercholesterolaemia.

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This study assessed the cholesterol lowering effect of Pediococcus acidilactici LAB4 and Lactobacillus plantarum LAB12 using adult zebrafish. Animals were fed with a high cholesterol diet (HCD) with/without LAB for seven weeks. Serum and liver cholesterol was quantified using colorimetric and dye staining methods. Expressions of npc1l1 and abca1 in the liver and intestine and appa in the brain were quantified using RT-PCR. Serum and liver cholesterol was significantly lowered in LAB4- and LAB12-fed zebrafish (≤64% and ≤71%, respectively), with reduced liver cholesterol deposition. The cholesterol lowering effect was accompanied by down-regulation of npc1l1 in intestines (≤28.7%), up-regulation of abca1 in the liver (≥30.5%) and down-regulation of appa in the brain (≤24.5%). A moderately strong positive Pearson correlation (r = 0.617, p < 0.01) was found between appa and serum cholesterol. LAB-fed zebrafish exhibited improved spatial learning and memory. LAB4 and LAB12 can be potentially used in preventing hypercholesterolaemia and Alzheimer's diseases.

Background: Triazole is an important heterocyclic moiety that occupies a unique position in heterocyclic chemistry, due to its large number of biological activities. It exists in two isomeric forms i.e. 1,2,4-triazole and 1,2,3-triazole and used as core molecule for the design and synthesis of many medicinal compounds. 1,2,4-Triazole possess broad spectrum of therapeutically interesting drug candidates such as analgesic, antiseptic, antimicrobial, antioxidant, anti urease , anti-inflammatory, diuretics, anticancer, anticonvulsant, antidiabetic, antimigrain agents. Methods: The structure of all synthesized compounds were characterized by physicochemical properties and spectral means (IR and NMR). The synthesized compounds were evaluated for their in vitro antimicrobial activity against Gram-positive ( B. subtilis ), Gram-negative ( P. aeruginosa and E. coli ) bacterial and fungal ( C. albicans and A. niger ) strains by tube dilution method using ciprofloxacin, amoxicillin and fluconazole as standards. In-vitro antioxidant and anti- urease screening was done by DPPH assay and indophenol method, respectively. The in-vitro anticancer evaluation was carried out against MCF-7 and HCT116 cancer cell lines using 5-FU and cisplatin as standards. Results, discussion and conclusion: The biological screening results reveal that the compounds T 5 (MIC BS, EC = 24.7µM, MIC PA, CA = 12.3 µM) and T 17 (MIC AN = 27.1µM) exhibited potent antimicrobial activity as comparable to standards ciprofloxacin, amoxicillin (MIC Cipro = 18.1µM, MIC Amo = 17.1µM) and fluconazole (MIC Flu = 20.4µM), respectively. The antioxidant evaluation showed that compounds T 2 (IC 50 = 34.83 µg/ml) and T 3 (IC 50 = 34.38 µg/ml) showed significant antioxidant activity and comparable to ascorbic acid (IC 50 = 35.44 µg/ml). Compounds T 3 (IC 50 = 54.01µg/ml) was the most potent urease inhibitor amongst the synthesized compounds and compared to standard thiourea (IC 50 = 54.25 µg/ml). The most potent anticancer activity showed by compounds T 2 (IC 50 = 3.84 μM) and T 7 (IC 50 = 3.25 μM) against HCT 116 cell lines as compared to standard 5-FU (IC 50 = 25.36 μM).

Probiotics are increasingly recognized for their cholesterol removal ability. Their vulnerability to harsh environmental factors during gastrointestinal transit and industrial processing remains a major challenge. This study addressed these issues by immobilizing Lactobacillus plantarum LAB12 within polymeric matrix comprised of sodium alginate (NaAlg), xanthan gum (XG) and chitosan (Ch) coating, with supplementation of b-cyclodextrin (b-CD), another potential cholesterol reducing agent. LAB12 encapsulated in Alg-XG-b-CD-Ch exhibited excellent survivability (95%) at pH 1.8 with facilitated release (>9 log CFU/g) at pH 6.8. While minimizing the loss of viable microencapsulated LAB12 at 75 and 908C, the microcapsules yielded viable counts >7 log CFU/g over a 4-week storage at 48C. Cholesterol lowering effects of b-CD supplemented beads and LAB12 against total cholesterol from egg yolk were also studied. These findings implied the potential use of Alg-XG-b-CD-Ch as protective micro-transport for probiotics and additional means of cholesterol reduction. PRACTICAL APPLICATIONSThe Alg-XG-b-CD-Ch beads yielded in this study exhibited characteristics of an ideal probiotic vehicle whereby they enhanced protection of free lactic acid bacteria (LAB) against harsh environments, facilitated targeted-cell release and conferred health benefits to the host upon consumption. The technical applications of this formulation include its use as heat-and pH-stable polymeric beads that carry L. plantarum LAB12 (the LAB model). The beads are acid tolerant (pH 1.8) under gastric conditions with the capability of targeted release at the large intestine (pH 6.8). In addition, it is also heat resistant, enabling microorganisms to survive at high temperature of up to 908C (essential during product manufacturing). The bound b-CD within the formulation is also able to enhance the cholesterol reducing ability of the LAB. Alg-XG-b-CD-Ch have successfully widen the applications of probiotics especially in health, food and agro industries.