To examine the in vivo role(s) of type I interferons (IFNs) and to determine the role of a component of the type I IFN receptor (IFNAR1) in mediating responses to these IFNs, we generated mice with a null mutation (-/-) in the IFNAR1 gene. Despite compelling evidence for modulation of cell proliferation and differentiation by type I IFNs, there were no gross signs of abnormal fetal development or morphological changes in adult IFNAR1 -/-mice. However, abnormalities of hemopoietic cells were detected in IFNAR1 -/-mice. Elevated levels of myeloid lineage cells were detected in peripheral blood and bone marrow by staining with Mac-i and Gr-1 antibodies. Furthermore, bone marrow macrophages from IFNAR1 -/-mice showed abnormal responses to colony-stimulating factor 1 and lipopolysaccharide. IFNAR1 -/-mice were highly susceptible to viral infection: viral titers were undetected 24 hr after infection of IFNAR1 +/+ mice but were extremely high in organs of IFNAR1-/-mice, demonstrating that the type I IFN system is a major acute antiviral defence. In cell lines derived from IFNAR1 -/-mice, there was no signaling in response to IFN-a or -,B as measured by induction of 2'-5' oligoadenylate synthetase, antiviral, or antiproliferative responses. Importantly, these studies demonstrate that type I IFNs function in the development and responses of myeloid lineage cells, particularly macrophages, and that the IFNAR1 receptor component is essential for antiproliferative and antiviral responses to IFN-a and -1.

Expression of Ets2, a proto-oncogene and transcription factor, occurs in a variety of cell types. During murine development it is highly expressed in newly forming cartilage, including in the skull precursor cells and vertebral primordia. Ets2 is located on human chromosome 21 (ref. 8) and is overexpressed in Down's syndrome (trisomy 21). Here we generate transgenic mice to investigate the consequences of overexpression of Ets2. We find that mice with less than 2-fold Ets2 overexpression in particular organs develop neurocranial, viscerocranial and cervical skeletal abnormalities. These abnormalities have similarities with the skeletal anomalies found in trisomy-16 mice and humans with Down's syndrome, in which the gene dosage of Ets2 is increased. Our results indicate that Ets2 has a role in skeletal development and implicate the overexpression of Ets2 in the genesis of some skeletal abnormalities that occur in Down's syndrome.

Idiopathic Parkinson's disease involves the loss of midbrain dopaminergic neurons, resulting in the presynaptic breakdown of dopaminergic transmission in the striatum. Huntington's disease and some neurodegenerative diseases with Parkinsonian features have postsynaptic defects caused by striatal cell death. Mice were generated in which an attenuated form of the diphtheria toxin gene (tox-176) was expressed exclusively in D1 dopamine receptor (D1R)-positive cells with the aim of determining the effect of this mutation on development of the basal ganglia and on the locomotor phenotype. Transgenic mice expressing Cre, a site-specific DNA recombinase, were crossed with a second line in which a transcriptionally silenced tox-176 gene was inserted into the D1R gene locus by homologous recombination. Young doubly transgenic mutant mice expressing the tox-176 gene displayed bradykinesia, dystonia, and had falls caused by myoclonic jerks. The mutant brain had evidence of apoptosis and reactive gliosis and, consistent with the D1R expression pattern, the striatum was reduced in volume, and the Islands of Calleja were absent. In contrast, the cortex was of normal thickness. D1Rs were not detectable in mutants by in situ hybridization or ligand autoradiography, whereas D2 dopamine receptor (D2R) mRNA and protein was present in the striatum. In addition, substance P and dynorphin, neuropeptides known to be expressed in D1R-positive striatonigral projection neurons were not detectable. Enkephalin, a marker found in D2-positive striatopallidal projection neurons was expressed in the mutant brain. The mutant represents a novel neurodegenerative disease model with a dramatic extrapyramidal phenotype.

Spermatogenesis is a complex process requiring the coordinate expression of a number of testis-specific genes. One of these, Pdha-2, codes for the murine spermatogenesis-specific isoform of the E1␣ subunit of the pyruvate dehydrogenase complex. To begin to delineate the mechanisms regulating its expression in vivo, we have generated transgenic mice lines carrying Pdha-2 promoter deletion constructs. Here we report that transgenic mice harboring a construct containing only 187 bp of promoter and upstream sequences (core promoter) is sufficient for directing the testis-specific expression of a chloramphenicol acetyltransferase (CAT) reporter gene. Like the endogenous Pdha-2, the CAT gene is expressed in testis in a stage-specific manner. Our studies also show a correlation between CpG methylation within the core promoter and its capacity to regulate transcription. In NIH 3T3 cell lines stably transfected with the Pdha-2 core promoter-CAT construct, high levels of CAT reporter expression are observed, whereas the endogenous Pdha-2 gene is repressed. In these cells, the CpG dinucleotides residing within the transfected promoter are hypomethylated whereas those residing in the endogenous promoter are methylated. Furthermore, promoter activity can be abated by the in vitro methylation of its CpG dinucleotides. DNase I footprint analysis indicates that at least one site for the methylation-mediated repression may occur through the ATF/cyclic AMP response element binding element located within the core promoter. Mutations within this element reduces activity to approximately 50% of the wild-type promoter activity. These results suggest that tissue-specific gene expression may be modulated by other mechanisms in addition to specific transcription factor availability and cooperativity. We propose that methylation may be a mechanism by which repression of the testis-specific Pdha-2 gene is established in somatic tissue.

The present study aims to focus on the bioprospecting of marine macroalgae of Turbinaria species, plenteous biomass of the world ocean. Three types of solvents, i.e., H 2 O, MeOH/H 2 O (80:20, v/v) and hexane/i-PrOH (50:50, v/v), were used for extraction. Both the biological activity and the pattern of present chemicals were characterized. For the cell proliferation assay, the human embryonic kidney 293 cells, cervix/breast/pancreatic adenocarcinoma, and osteosarcoma cells were used. For the antioxidant activity determination, both intracellular assay with human embryonic kidney and cervix adenocarcinoma cells, as well as the biochemical DPPH test, were employed. To complete the information about macroalgae composition, organic compounds were characterized by the liquid chromatography coupled with high resolution tandem mass spectrometry. Attention was concentrated mainly on the lipidomic profile characterization. In spite the fact that any significant antiproliferative effect was not observed for cancer cells, both the Turbinaria species were shown to be good protectors against the oxidative stress of the noncancer cells. Most of the antioxidants were determined in the hexane/i-PrOH extract. As regards the lipids identified, most of them belonged to the triacylglycerols followed by sphingomyelins, diacylglycerols, and polar (lyso) phospholipids. Additionally to fatty acids with 14, 16 and 18 carbons, also those with odd carbon numbers were frequently present.

ABSTRAKPenelitian ini bertujuan untuk mengkaji pengaruh dosis mulsa dan sistem tanam rumput dan legum pada tanah salin terhadap pertumbuhan, produksi hijauan dan kualitas rumput benggala. Tanah salin pada percobaan ini diklasifikasikan dalam tanah sangat salin dengan kesuburan tanah yang rendah. Rancangan percobaan adalah rancangan acak kelompok dengan 3 kelompok. Perlakuan adalah penanaman rumput benggala : M1 = monokultur, tanpa mulsa, M2 = monokultur, mulsa 3 ton/ha, M3 = monokultur, mulsa 6 ton/ha, M4 = tumpangsari dengan legum turi, tanpa mulsa, M5 = tumpangsari dengan turi, mulsa 3 ton/ha, M6 = tumpangsari dengan turi, mulsa 6 ton/ha. Data yang diperoleh diolah dengan analisis ragam dilanjutkan dengan uji wilayah berganda Duncan. Kadar air tanah tertinggi ditemukan pada perlakuan tumpang sari rumput dan legum dengan mulsa 6 ton/ha. Pertumbuhan, produksi dan kualitas hijauan rumput benggala tertinggi pada perlakuan dengan mulsa. Aplikasi mulsa 3 ton/ha pada tanah salin sudah meningkatkan pertumbuhan, produksi dan kualitas hijauan rumput benggala. Tumpangsari rumput dan turi tidak mempengaruhi pertumbuhan, produksi dan kualitas hijauan rumput benggala pada tanah salin.Kata kunci : mulsa, produksi, rumput benggala, tanah salin, tumpangsari ABSTRACTThe research was conducted to evaluate the effect of mulch and mixed cropping grass -legume at saline soil on growth, forage yield and nutritional quality of guinea grass. Saline soil used in this research was classified into strongly saline soil with low soil fertility. The research was arrranged in randomized complete block design with 3 blocks. The treatments were : M1 = guinea grass monoculture, without mulch; M2 = guinea grass monoculture, 3 ton/ha mulch; M3 = guinea grass monoculture, 6 ton/ha mulch, M4 = mixed cropping grass with Sesbania grandiflora, without mulch; M5 = mixed cropping grass with Sesbania grandiflora, 3 ton/ha mulch; M6 = mixed cropping grass with Sesbania grandiflora, 6 ton/ha mulch. Data were analyzed using analysis of variance, then followed by Duncan's Multiple Range Test. The highest soil moisture content was achieved at mixed cropping grasslegume with 6 ton/ha of mulch. The effect of mulch at saline soil significantly increased plant growth, forage yield and nutritional quality of guinea grass. Application of 3 ton/ha mulch increased plant growth, forage yield and nutritional quality of guinea grass. Plant growth, forage yield and nutritional quality of guinea grass were not affected by monoculture or mixed cropping with Sesbania at saline soil.

MDCK and Vero cell lines have been used as substrates for influenza virus replication. However, Vero cells produced lower influenza virus titer yield compared to MDCK. Influenza virus needs molecules for internalisation of the virus into the host cell, such as influenza virus receptor and clathrin. Human influenza receptor is usually a membrane protein containing Sia(α2,6) Gal, which is added into the protein in the golgi apparatus by α2,6 sialyltransferase (SIAT1). Light clathrin A (LCA), light clathrin B (LCB) and heavy clathrin (HC) are the main components needed for virus endocytosis. Therefore, it is necessary to compare the expression of SIAT1 and clathrin in Vero and MDCK cells. This study is reporting the expression of SIAT1 and clathrin observed in both cells with respect to the levels of (1) RNA by using RT-PCR, (2) protein by using dot blot analysis and confocal microscope. The results showed that Vero and MDCK cells expressed both SIAT1 and clathrin proteins, and the expression of SIAT1 in MDCK was higher compared to Vero cells. On the other hand, the expressions of LCA, LCB and HC protein in MDCK cells were not significantly different to Vero cells. This result showed that the inability of Vero cells to internalize H1N1 influenza virus was possibly due to the lack of transmembrane protein receptor which contained Sia(α2,6) Gal.

Abstract.A neurodegenerative accumulation of α-synuclein effect of rotenone on the increase synuclein in the midbrain of Wistar rats. Thirty 250 g) were divided into three groups: the Blank group, the Solvent group (intraperitoneally injected with 1 ml/kg body weight of sunflower seed oil), and the Rotenone group (intraperitoneally injected with 2.5 mg/kg body weight of rotenone) for 9, 19, and 28 days. The rats were for the 9-day treatment, day 20 for the 19 for the 28-day treatment (2 rats/gr midbrains were isolated and extracted. Glutathione assay and α tests were performed. The results showed that the average oxidative stress index was highest in the Blank group (0.95 ± 0.24, 0.63 ± respectively). Meanwhile, groups (0.09 ± 0.03, 0.15 ± 0.03, and 0.13 ± 0.02 ng/mg tissue, respectively). Correlation analysis showed that the oxidative stress index proportional to the concentration of α midbrain of Wistar rats treated with rotenone indicated oxidative stress and led to an accumulation of α-synuclein protein. Here, we report our findings on the on the increase of oxidative stress and accumulation of α-synuclein in the midbrain of Wistar rats. Thirty six male rats (8-9 weeks, 200-were divided into three groups: the Blank group, the Solvent-rotenone group (intraperitoneally injected with 1 ml/kg body weight of sunflower seed tenone group (intraperitoneally injected with 2.5 mg/kg body weight of rotenone) for 9, 19, and 28 days. The rats were decapitated on day 10 day treatment, day 20 for the 19-day treatment, day 30, 40, 50 and 60 day treatment (2 rats/group or 6 rats/day of observation time). The midbrains were isolated and extracted. Glutathione assay and α-synuclein ELISA tests were performed. The results showed that the average oxidative stress index was highest in the Blank group (0.95 ± 0.24, 0.63 ± 0.23, 0.81 ± 0.27, Meanwhile, the concentration of α-Synuclein had decreased in all (0.09 ± 0.03, 0.15 ± 0.03, and 0.13 ± 0.02 ng/mg tissue, respectively). Correlation analysis showed that the oxidative stress index was inversely ortional to the concentration of α-synuclein. Our conclusion is that the midbrain of Wistar rats treated with rotenone indicated oxidative stress and led synuclein protein. Keyword