Teixeira A.L.,Institute of Biological science
NeuroMolecular Medicine | Year: 2011
Brain-derived neurotrophic factor (BDNF) is the most widely distributed neurotrophin in the central nervous system where it plays several pivotal roles in synaptic plasticity and neuronal survival. As a consequence, BDNF became a key target in the physiopathology of several neurological and psychiatric diseases. Recent studies have reported altered levels of BDNF in the circulation, i.e. serum or plasma, of patients with Alzheimer's disease (AD), and low BDNF levels in the CSF as predictor of future cognitive decline in healthy older subjects. Altered BDNF circulating levels have also been reported in other neurodegenerative and psychiatric disorders, hampering its use as a specific biomarker for AD. Therefore, BDNF seems to be an unspecific biomarker of neuropsy-chiatric disorders marked by neurodegenerative changes. © Springer Science+Business Media, LLC 2011.
Wanner S.P.,Institute of Biological science |
Martins F.S.,Federal University of Minas Gerais |
Nicoli J.R.,Federal University of Minas Gerais |
Coimbra C.,Institute of Biological science
Journal of Nutrition | Year: 2014
Dietary supplementation with L-arginine has been shown to improve the intestinal barrier in many experimental models. This study investigated the effects of arginine supplementation on the intestinal permeability and bacterial translocation (BT) induced by prolonged physical exercise under heat stress. Under anesthesia, male Swiss mice (5-wk-old) were implanted with an abdominal sensor to record their core body temperature (Tcore). After recovering from surgery, the mice were divided into 3 groups: a non-supplemented group that was fed the standard diet formulated by the American Institute of Nutrition (AIN-93G; control), a non-supplemented group that was fed the AIN-93G diet and subjected to exertional hyperthermia (H-NS), and a group supplemented with L-arginine at 2% and subjected to exertional hyperthermia (H-Arg). After 7 d of treatment, the H-NS and H-Arg mice were forced to run on a treadmill (60 min, 8 m/min) in a warm environment (34°C). The control mice remained at 24°C. Thirty min before the exercise or control trials, the mice received a diethylenetriamine pentaacetic acid (DTPA) solution labeled with technetium-99m (99mTc-DTPA) or 99mTc-Escherichia coli by gavage to assess intestinal permeability and BT, respectively. The H-NS mice terminated the exercisewith Tcore values of;40°C, and, 4 h later, presented a 12-fold increase in the blood uptake of 99mTc-DTPA and higher bacterial contents in the blood and liver than the control mice. Although supplementation with arginine did not change the exercise-induced increase in Tcore, it prevented the increases in intestinal permeability and BT caused by exertional hyperthermia. Our results indicate that dietary L-arginine supplementation preserves the integrity of the intestinal epithelium during exercise under heat stress, acting through mechanisms that are independent of Tcore regulation. © 2014 American Society for Nutrition.
Saito V.M.,Federal University of Minas Gerais |
Rezende R.M.,Institute of Biological Science |
Teixeira A.L.,Federal University of Minas Gerais |
Teixeira A.L.,Neurology Group
Current Neuropharmacology | Year: 2012
In recent years, a growing interest has been dedicated to the study of the endocannabinoid system. The isolation of Cannabis sativa main psychotropic compound, Δ 9-tetrahydrocannabinol (THC), has led to the discovery of an atypical neurotransmission system that modulates the release of other neurotransmitters and participates in many biological processes, including the cascade of inflammatory responses. In this context, cannabinoids have been studied for their possible therapeutic properties in neuroinflammatory diseases. In this review, historic and biochemical aspects of cannabinoids are discussed, as well as their function as modulators of inflammatory processes and therapeutic perspectives for neurodegenerative disorders, particularly, multiple sclerosis. © 2012 Bentham Science Publishers.
News Article | February 15, 2017
C57BL/6N mice, ICR mice and Wistar rats were purchased from SLC Japan (Shizuoka, Japan). All animals were maintained under specific pathogen-free conditions. All animal experiments were approved by the Insititutional Animal Care and Use Committee, and performed in accordance with the guidelines of the University of Tokyo and the National Institute for Physiological Sciences. Diabetes was induced in 8-week-old C57BL/6N male mice by intravenous injection of 150 mg kg−1 of STZ. Mice with nonfasting blood glucose levels over 350 mg dl−1 1 week after STZ administration were used. Embryo culture and manipulation are described11. Rodent islets conventionally are isolated by collagenase perfusion of the pancreata through the common bile duct. However, the pancreata of Pdx1mu/mu chimaeric rats could not be perfused in this way because the pancreaticobiliary junction was maldeveloped in all (Extended Data Fig. 5). Therefore, we isolated islets by digestion of minced pancreata with collagenase. Pancreata removed from interspecific chimaera were inflated by interstitial injection of Gey’s balanced salt solution (GBSS; Sigma-Aldrich). GBSS-filled pancreata were minced using scissors. Small pieces of chopped pancreata were digested with collagenase XI (Sigma-Aldrich) to release islets from exocrine tissue. After 6–8 min incubation, islets were picked up using glass micropipettes and transplanted beneath the kidney capsule of 10-week-old male mice with STZ-induced diabetes, as previously described11. To prevent acute graft rejection, 0.5 mg per g (body weight) per day of tacrolimus, was injected intraperitoneally on the day of transplantation and on each of the following 4 days, in addition to an anti-inflammatory cocktail (all components, Affymetrix) containing anti-mouse interferon-γ mAb (rat IgGκ, 16-7312, clone R4-6A2), anti-mouse tumour necrosis factor-α mAb (rat IgG1κ, 16-7322, clone MP6-XT3) and anti-mouse IL-1β (hamster IgG, 16-7012, clone B122). The mRNA of TALENs (left and right) and rat Exo1 were generated by in vitro transcription. Linearized plasmids were transcribed from T7 promoter using mMESSAGE mMACHINE T7 ULTRA Transcription Kit (Thermo Fisher Scientific) and resultant mRNAs were cleaned up by MEGAclear Kit (Thermo Fisher Scientific). 3 or 10 ng μl−1 of each mRNA was prepared by dilution in RNase- free-water and mixture of right TALEN, left TALEN and Exo1 were injected into the male pronuclei of zygotes by microinjection, as previously reported18. TALEN potential off-target sites were predicted by TALENoffer software. We chose 21 candidates (5 in exonic loci, 13 in intronic loci, 3 in intergenic loci) from TOP200 candidates19. We performed PCR amplification of genomic DNA from Pdx1+/muA, Pdx1+/muB and wild-type Wistar rats, subjecting the amplicons to Sanger sequencing. Genomic DNA was isolated from fluorescent-marker-negative cells isolated by FACS from chimaeric-rat blood samples. The TALEN target region of Pdx1 was amplified by PCR using the following primers: (forward) 5′-GCTGAGAGTCCGTGAGCTGCCCAG-3′ and (reverse) 5′-GGAACGCTTAAAGATCGTAGCAGC-3′). The PCR products were sequenced. Total RNA was isolated from duodenum of Pdx1muA/muB mice and reverse-transcribed by Superscript III reverse transcriptase (Thermo Fisher Scientific) with oligodT primer. Pdx1muA or Pdx1muB full-length cDNA were amplified by PCR using the following primers: (forward) 5′-GGCGCTGAGAGTCCGTGAGCTGC-3′ and (reverse) 5′-TTTTTTTTTTTTTTTGAAACCTCAAACAG-3′. Nonfasting blood glucose levels were determined (Medisafe-Mini glucometer; Terumo) weekly after islet transplantation. GTTs in overnight-fasted chimaera rats was conducted 0, 15, 30, 60 and 120 min after intraperitoneal injection of glucose (50% d-glucose solution, 2.5 g per kg body weight). Tail-vein blood was sampled by phlebotomy. Non-fasting serum mouse or rat c-peptide levels were analysed by enzyme-linked immunosorbent assay (ELISA) (mouse c-peptide ELISA kit, Shibayagi and Morinaga Institute of Biological Science; rat c-peptide ELISA kit, MERCODIA AB). Serum was isolated from 10-week-old Pdx1muA/muB + mPSCs chimaeras, C57BL/6N mice and Wistar rats. Serum was obtained from STZ-treated diabetic mice transplanted with mouseR islets 260 or 372 days after transplantation. SGE2 (EGFP-expressing mES cells) were derived from blastocysts generated from mating C57BL/6N female mice with C57BL/6N-Tg male mice (CAG-EGFP) (SLC Japan). mRHT (mES cells) were derived from blastocysts generated from mating male and female H2B-tdTomato knock-in mice with human histone H2B and tdTomato fusion gene in the mouse ROSA locus (T.K., unpublished data). Wlv3i-1 (rES cells) and GT3.2 (miPSCs) have been previously described11, 31. Maintenance of mPSCs and rPSCs has been previously described32, 33. Briefly, mPSCs were cultured on mitomycin-C-treated mouse embryonic fibroblasts in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, 0.1 mM 2-mercaptoethanol, 0.1 mM nonessential amino acids, 1 mM sodium pyruvate, and 1,000 units per ml of mouse leukaemia inhibitory factor (all Thermo Fisher Scientific) and 1% l-glutamine-penicillin-streptomycin (Sigma-Aldrich). rPSCs were cultured on mitomycin-C-treated mouse embryonic fibroblasts in N2B27 medium supplemented with 1 μM mitogen-activated protein kinase inhibitor PD0325901 and 3 μM glycogen synthase kinase inhibitor CHIR99021 (both Axon Groeningen). All PSC lines were authenticated by chimaera formation. These cell lines were not contaminated with mycoplasma. Isolated pancreata and islets were fixed in 4% paraformaldehyde in phosphate-buffered saline solution (PBS). Paraffin-embedded sections were incubated with blocking buffer (Active Motif) for 1 h at room temperature. The sections were incubated with primary antibodies, diluted in blocking buffer for 1 h at room temperature, and washed three times with PBS. They were then incubated with secondary antibodies for 1 h at room temperature. Primary antibodies used were guinea pig anti-insulin (Abcam; ab7842), rabbit anti-glucagon (Nichirei Bioscience, 422271), rabbit anti-somatostatin (Nichirei Bioscience 422651), rabbit anti-cytokeratin 19 (Abcam; ab52625, clone EP1580Y), mouse anti-amylase (SantaCruz; SC-46657, clone G-10) and goat anti-GFP (Abcam; ab6673), with Alexa-488-, Alexa-546-, and Alexa-633-conjugated secondary antibodies (Thermo Fisher Scientific). After antibody treatment, sections were mounted with Vectashield (Vector Laboratories), a mounting medium containing DAPI (Thermo Fisher Scientific) for nuclear counterstaining, and sections were observed under fluorescence microscopy. Three to five sections per slide were imaged and processed using Image J. For detection of lymphoid infiltration, DAB immunohistochemistry was performed with rabbit anti-CD3 (Abcam; ab5690) and rabbit anti-CD11b (Bioss Inc.; bs-1014R). Islets or small pieces of kidney that included transplanted islets were dispersed into single cells with collagenase type1A (Sigma-Aldrich). Dispersed cells stained with phycoerythrin (PE)-conjugated anti-mouse CD31 (Thermo Fisher Scientific; A16201, clone 390) or allophycocyanin (APC)-conjugated anti-rat CD31 (Thermo Fisher Scientific; 50-0310-82, clone TLD-3A12) were subjected to FACS CantoII analysing (BD Biosciences). Data were collected for all of the dispersed cells and analysed. The experiments were not randomized and the investigators were not blinded to allocation during experiments and outcome assessment. Sample size was estimated on the basis of previous publications. Statistical significance was calculated by F-test and Student’s t-test (compare two groups) and the similarity to the Mendelian ratio was analysed by chi-square test (with Excel and Graphpad Prism software). P < 0.05 was considered to be statistically significant. Data are presented as mean ± s.d. Immunohistochemistry and flow-cytometry studies were repeated three times independently with similar results. All relevant data that are included with this study are available from corresponding auther upon reasonable request.
Hurtado A.Q.,Integrated Services for the Development of Aquaculture and Fisheries |
Montano M.N.E.,University of the Philippines |
Martinez-Goss M.R.,Institute of Biological science
Journal of Applied Phycology | Year: 2013
An abundance of marine algae along the Philippines' long, irregular coastline (36,289 km) has made it inevitable for Filipinos to exploit algae for food, feeds, medicine, or other purposes. Among the most popular of these algae are the carrageenophytes, which include four genera, six species, and 21 morphotypes/varieties/cultivars under the Family Solieriaceae. However, commercial production did not begin until 1973. The development of carrageenophyte farming for commercial purposes evolved from simple fixed-bottom monoline farming by coastal farmers, who refined the technology themselves, with the help and guidance of local and international scientists, which has led to a commercially viable industry with a maximum estimated production of 97,000-102,820 dry, metric tons in 2004. Farm gate revenues for that year were estimated at US$ 82.45-87.4 million (four croppings year-1), whose main recipients were the local seaweed farmers. However, in 2008, production started to decline, which was brought primarily by the deteriorating quality of propagules and the perennial occurrence of "ice-ice" and harmful endophytes caused by environmental stresses due to unfavorable weather conditions, and secondarily, the peace and order problem in the major producing areas like Zamboanga, Maguindanao, Basilan, and Sulu in Mindanao and insufficient government support. This paper aims to assess the present situation and suggest how to possibly reverse the situation to its usual productive periods. © 2012 Springer Science+Business Media Dordrecht.
Romero T.R.L.,Institute of Biological science |
Pacheco D.D.F.,Institute of Biological science |
Duarte I.D.G.,Institute of Biological science
Life Sciences | Year: 2013
Aims: Recently, we demonstrated that peripheral antinociception induced by δ opioid receptor is dependent of Ca2 +-activated Cl - channels (CaCCs). Because opioid and cannabinoid receptors share some common mechanisms of action, our objective was to identify a possible relationship between CaCCs and the endocannabinoid system. Main methods: To induce hyperalgesia, rat paws were treated with intraplantar prostaglandin E2 (PGE2, 2 μg). Nociceptive thresholds to pressure (grams) were measured using an algesimetric apparatus 3 h following injection. Probabilities were calculated using ANOVA/Bonferroni's test, and values that were less than 5% were considered to be statistically significant. Key findings: Administration of the cannabinoid agonist CB1 anandamide (12.5, 25 and 50 μg/paw) and the cannabinoid agonist CB2 PEA (5, 10 and 20 μg/paw) decreased the PGE2-induced hyperalgesia in a dose-dependent manner. The possibility of the higher doses of anandamide (50 μg) and PEA (20 μg) having a central or systemic effect was excluded because the administration of the drug into the contralateral paw did not elicit antinociception in the right paw. As expected, the antinociceptive effects induced by anandamide and PEA were blocked by the CB1 and CB 2 receptor antagonists AM251 and AM630, respectively. The peripheral antinociception was induced by anandamide but not PEA and was dose-dependently inhibited by the CaCC blocker niflumic acid (8, 16 and 32 μg). Significance: These results provide the first evidence for the involvement of CaCCs in the peripheral antinociception induced by activation of the CB1 cannabinoid receptor. © 2012 Elsevier Inc.
Moghadamtousi S.Z.,Institute of Biological science |
Kamarudin M.N.A.,Institute of Biological science |
Chan C.K.,Institute of Biological science |
Goh B.H.,Institute of Biological science |
Kadir H.A.,Institute of Biological science
American Journal of Chinese Medicine | Year: 2014
Loranthus parasiticus Merr (L. parasiticus) is a member of Loranthaceae family and is an important medicinal plant with a long history of Chinese traditional use. L. parasiticus, also known as Sang Ji Sheng (in Chinese), benalu teh (in Malay) and baso-kisei (in Japanese), is a semiparasitic plant, which is mostly distributed in the southern and southwestern regions of China. This review aims to provide a comprehensive overview of the ethnomedicinal use, phytochemistry and pharmacological activity of L. parasiticus and to highlight the needs for further investigation and greater global development of the plant's medicinal properties. To date, pharmacological studies have demonstrated significant biological activities, which support the traditional use of the plant as a neuroprotective, tranquilizing, anticancer, immunomodulatory, antiviral, diuretic and hypotensive agent. In addition, studies have identified antioxidative, antimutagenic, antiviral, antihepatotoxic and antinephrotoxic activity. The key bioactive constituents in L. parasiticus include coriaria lactone comprised of sesquiterpene lactones: coriamyrtin, tutin, corianin, and coriatin. In addition, two proanthocyanidins, namely, AC trimer and (+)-catechin, have been recently discovered as novel to L. parasiticus. L. parasiticus usefulness as a medicinal plant with current widespread traditional use warrants further research, clinical trials and product development to fully exploit its medicinal value. © 2014 World Scientific Publishing Company & Institute for Advanced Research in Asian Science and Medicine.
Ribeiro F.M.,Federal University of Minas Gerais |
Camargos E.R.S.,Institute of Biological science |
De Souza L.C.,Federal University of Minas Gerais |
Teixeira A.L.,Federal University of Minas Gerais
Revista Brasileira de Psiquiatria | Year: 2013
The prevalence of neurodegenerative diseases, such as Alzheimer's disease (AD) and Parkinson's disease (PD), increases with age, and the number of affected patients is expected to increase worldwide in the next decades. Accurately understanding the etiopathogenic mechanisms of these diseases is a crucial step for developing disease-modifying drugs able to preclude their emergence or at least slow their progression. Animal models contribute to increase the knowledge on the pathophysiology of neurodegenerative diseases. These models reproduce different aspects of a given disease, as well as the histopathological lesions and its main symptoms. The purpose of this review is to present the main animal models for AD, PD, and Huntington's disease. © 2013 Associação Brasileira de Psiquiatria.
Ismail S.,Institute of Biological science |
Dadrasnia A.,Institute of Biological science
PLoS ONE | Year: 2015
Environmental contamination by petroleum hydrocarbons, mainly crude oil waste from refineries, is becoming prevalent worldwide. This study investigates the bioremediation of water contaminated with crude oil waste. Bacillus salamalaya 139SI, a bacterium isolated from a private farm soil in the Kuala Selangor in Malaysia, was found to be a potential degrader of crude oil waste. When a microbial population of 108 CFU ml-1 was used, the 139SI strain degraded 79% and 88% of the total petroleum hydrocarbons after 42 days of incubation in mineral salt media containing 2% and 1% of crude oil waste, respectively, under optimum conditions. In the uninoculated medium containing 1% crude oil waste, 6% was degraded. Relative to the control, the degradation was significantly greater when a bacteria count of 99 × 108 CFU ml-1 was added to the treatments polluted with 1% oil. Thus, this isolated strain is useful for enhancing the biotreatment of oil in wastewater. © 2015 Ismail, Dadrasnia.
Rothan H.A.,University of Malaya |
Mohamed Z.,Institute of Biological Science |
Suhaeb A.M.,University Hospital |
Rahman N.A.,University of Malaya |
Yusof R.,University of Malaya
OMICS A Journal of Integrative Biology | Year: 2013
Dengue virus infects millions of people worldwide, and there is no vaccine or anti-dengue therapeutic available. Antimicrobial peptides have been shown to possess effective antiviral activity against various viruses. One of the main limitations of developing these peptides as potent antiviral drugs is the high cost of production. In this study, high yield production of biologically active plectasin peptide was inexpensively achieved by producing tandem plectasin peptides as inclusion bodies in E. coli. Antiviral activity of the recombinant peptide towards dengue serotype-2 NS2B-NS3 protease (DENV2 NS2B-NS3pro) was assessed as a target to inhibit dengue virus replication in Vero cells. Single units of recombinant plectasin were collected after applying consecutive steps of refolding, cleaving by Factor Xa, and nickel column purification to obtain recombinant proteins of high purity. The maximal nontoxic dose (MNTD) of the recombinant peptide against Vero cells was 20 lM (100 lg/mL). The reaction velocity of DENV2 NS2B-NS3pro decreased significantly after increasing concentrations of recombinant plectasin were applied to the reaction mixture. Plectasin peptide noncompetitively inhibited DENV2 NS2BNS3pro at Ki value of 5.03 - 0.98 lM. The percentage of viral inhibition was more than 80% at the MNTD value of plectasin. In this study, biologically active recombinant plectasin which was able to inhibit dengue protease and viral replication in Vero cells was successfully produced in E. coli in a time-and cost-effective method. These findings are potentially important in the development of potent therapeutics against dengue infection. © Mary Ann Liebert, Inc.