14-3-3ζ Regulates Immune Response through Stat3 Signaling in Oral Squamous Cell Carcinoma

Ectopic expression of 14-3-3ζ has been found in various malignancies, including lung cancer, liver cancer, head and neck squamous cell carcinoma (HNSCC), and so on. However, the effect of 14-3-3ζ in the regulation of interactions between tumor cells and the immune system has not been previously reported. In this study, we aimed to investigate whether and how 14-3-3ζ is implicated in tumor inflammation modulation and immune recognition evasion. In oral squamous cell carcinoma (OSCC) cell lines and cancer tissues, we found that 14-3-3ζ is overexpressed. In OSCC cells, 14-3-3ζ knockdown resulted in the up-regulated expression of inflammatory cytokines. In contrast, 14-3-3ζ introduction attenuated cytokine expression in human normal keratinocytes and fibroblasts stimulated with interferon-γ (IFN-γ) and lipopolysaccharide (LPS). Furthermore, supernatants from 14-3-3ζ knockdown OSCC cells dramatically altered the response of peritoneal macrophages, dendritic cells and tumor-specific T cells. Interestingly, Stat3 was found to directly interact with 14-3-3ζ and its disruption relieved the inhibition induced by 14-3-3ζ in tumor inflammation. Taken together, our studies provide evidence that 14-3-3ζ may regulate tumor inflammation and immune response through Stat3 signaling in OSCC.     

Herbicide Safener-Inducible Gene Expression in Arabidopsis thaliana

The potential use of a new chemical-inducibie gene expressionsystem in Arabidopsis thaliana has been examined. The systemis based on the maize In2-2 promoter which is activated by benzenesulfonamideherbicide safe-ners. Plants transformed with the ß-glucuronidase(gus) reporter gene under the control of the In2-2 promoterwere grown in the presence of different safeners and the inducedGUS activity pattern was studied histochemically. In the absenceof safeners, the In2-2 promoter was not active. Applicationof different safeners induced distinct gus expression patterns,including expression in the root, hy-dathodes, and the shootapical meristem. Plants maintained continuously on inducingconcentrations of the safeners were retarded in growth. Thegrowth inhibition effects of the Sa5 safener could be overcomein a sul-fonylurea-resistant background. In2-2 promoter activitycould also be induced by the sulfonylurea herbicide chlor-sulfuron.In the sulfonylurea-resistant background, which derives fromherbicide-resistant acetolactate synthase activity, inductionof the In2-2 promoter by chlorsulfuron was lower. Furthermore,branched-chain amino acids, known to inhibit acetolactate synthaseactivity, also induced In2-2 promoter activity. Our data suggesta strong correlation between In2-2 expression and inhibitionof the acetolactate synthase activity. (Received November 12, 1996; Accepted February 21, 1997)     

Incorporation of 14C-radioactivity into lipid classes at different maturity of Laminaria japonica Aresch. cultured under natural and partially controlled conditions

A study to assess which environmental or developmental factors predominate in the biosynthesis of lipids of Laminaria japonica Aresch. blades was undertaken by means of ¹⁴C-labelling technique. In experiment 1, kelp blades at different growth stages were collected in different cultural seasons. In experiment 2, kelp blades of different sizes and maturity cultured simultaneously for two months in the same sea area were collected at the same time.The following results were obtained. In experiment 1, the ¹⁴C-incorporation into whole lipids was lowest in juvenile blades collected at the end of autumn and highest in blades of middle size collected in winter. However, the highest counts were incorporated in PC among complex lipid classes from all size classes of blades in both experiments 1 and 2. In experiment 2, ¹⁴C-incorporation patterns of individual lipid classes were characteristically different depending on the sizes of blades even under the same cultural condition. Thus, the biosynthesis of lipids in this kelp seems to be affected essentially by developmental factors.Abbreviation Comp. lip. complex lipid - FA non-esterified fatty acids - Fucost fucosterol - DG diacylglycerol - DGDG digalactosyldiacylglycerol - MG monoacylglycerol - MGDG monogalactosyldiacylglycerol - NL neutral lipids - PC phosphatidylcholine - PE phosphatidylethanolamine - PG phosphatidylglycerol - PI phosphatidyl inositol - PS phosphatidylserine - SQDG sulphoquinovosyldiacyl glycerol - TG triacylglycerol     

Antennal circulatory organs in Onychophora,Myriapoda and Hexapoda: Functional morphology and evolutionary implications

Summary A comparative investigation of the antennal circulatory organs in representatives of the Onychophora, all subtaxa of the Myriapoda and numerous taxa of the Hexapoda (comprising a total of 54 species) revealed an unexpected diversity in structure and function.In the Onychophora, antennal vessels exist which are connected to the enlarged anterior end of the aorta dorsal to the brain.In the Chilopoda, Diplopoda and Symphyla, antennal vessels exist which originate from the dorsal vessel caudal to the brain. They extend under the optic lobes, lateral to the circumoesophageal connectives, into the antennae.In the Hexapoda, the investigations include representatives of all higher taxa, apart from the Paraneoptera and the Holometabola. Generally, antennal vessels exist. In the Diplura, they originate from the anterior end of the aorta in front of the brain. In all other insects the antennal vessels are separate from the dorsal vessel. Their proximal ends form ampullary enlargements which are attached to the frontal cuticle near the antenna bases. They communicate via valved ostia with the haemolymph sinus in front of the brain. In the Archaeognatha, Zygentoma, Odonata, certain Plecoptera and the Notoptera, no muscles are connected to these organs. In all other groups the ampullae are pulsatile as a result of associated muscles (antennal hearts). These muscles diverge widely in their attachments and act either as compressors (Dermaptera) or dilators of the ampullae (Embioptera, Blattopteroidea, Orthopteroidea, and some Plecoptera).In the Collembola and Ephemeroptera, special antennal circulatory organs are lacking. In some forms the anatomical arrangement of the inner organs, in conjunction with short diaphragms at the antenna bases, apparently leads to a channelling of haemolymph flow. This condition may be explained by the very short antennae of these insects and is considered as a convergent and apomorphic state in these taxa.The antennal vessels are supposed to be homologous within the Tracheata and to represent the lateral arteries of the antenna segment. An origin from the dorsal vessel is considered an ancestral state, which was lost in the stem lineage of the Ectognatha. Specific space constraints within the cephalic capsule are discussed as the possible reason for this loss. The evolution of pulsatile antennal circulatory organs in the Neoptera is the result of the association of muscles with the proximal ampullary ends of the antennal vessels. The attachments and innervation of these muscles indicate a derivation from precerebral pharyngeal dilators.Abbreviations Amp ampulla - Ant antenna - ant anterior - AN antennal nerve - Ao aorta - AV antennal vessel - Br brain - BrSi brain sinus - CC corpora cardiaca - CoeC circumoesophageal connectives - CM compressor muscle of ampulla - CT connective tissue - Dia diagphragm - do dorsal - DM dilator muscle of ampulla - DM1 ampullo-ampullary dilator muscle - DM2 ampullo-pharyngeal dilator muscle - DM3 ampullo-frontal dilator muscle - DM Acc accessory dilator muscle of ampulla - DV dorsal vessel - EB elastic band - FbDM fronto-buccal pharynx dilator muscle - FG frontal ganglion - FSa frontal sac - FSe frontal septum - FSi frontal sinus - Lb labium - LV lateral vessel of aorta - MA mouth-angle - Nr nervus recurrens - Oc ocellus - Oe oesophagus - OeSi oesophageal sinus - Ost ostium - Ph pharynx - Pl labial palpus - RM retractor muscle of mouth-angle - RMl lateral retractor of mouth-angle - RMm medial retractor of mouth-angle - SceSi supracerebral sinus - SD salivary duct - T tentorium     

Synthesis and cytokinin activity of (R)-(+)- and (S)-(−)-dihydrozeatins and their ribosides

(R)-(+)- and (S)-(?)-dihydrozeatins [(R)-(+)- and (S)-(?)-6-(4-hydroxy-3-methylbutylamino)purines, 1a and 1b] and their ribosides {(?)-6-[(R)-4-hydroxy-3-methylbutylamino]- and (?)-6-[(S)-4-hydroxy-3-methyl-butylamino]-9-β-D-ribofuranosylpurines, 3a and 3b} were synthesized and tested for their cytokinin activity by four bioassay systems, the growth of tobacco callus, the seed germination of lettuce, the fr. wt increase of excised radish cotyledons and the retardation of chlorophyll degradation in radish cotyledons. In tobacco callus bioassay, 1a was more active than 1b. The ribosides 3a and 3b were not less active than their corresponding aglycones 1a and 1b. In other bioassays used the activity followed the order: 1a >3a >1b >3b. In tobacco callus bioassay and lettuce seed germination, trans-zeatin [6-(4-hydroxy-3-methylbut-trans-2-enylamino)purine] showed stronger cytokinin activity than 1a.     

Myelin lipids: A phylogenetic study

The lipid composition of CNS and PNS myelin was studied in rat,Xenopus, trout andTorpedo. The main difference lay in the proportion of cerebrosides, which decreased in the sequence rat > Xenopus > Torpedo > trout. In additionTorpedo CNS and PNS myelins were extremely rich in sulfatides. In some respects,Torpedo appeared closer to tetrapods than trout. Otherwise the proportion of the different lipid classes did not reveal any clear evolutionary trends.The presence of hydroxylated galactolipids in CNS myelin was investigated in several additional species. Considerable amounts were found inTorpedo, Polypterus, Protopterus, lizard, and chicken, with the highest values in rat and anurans. Only very small amounts of hydroxylated cerebrosides were detected in trout and in axolotl, while newt had none. This parameter appears therefore of doubtful usefulness for phylogenetic studies. In contrast to myelin proteins, myelin lipids are of limited value for establishing phylogenetic relationship among vertebrates.Abbreviations CH cholesterol - CNS central nervous system - FA fatty acids - GC cerebrosides - HPTLC high-performance thin-layer chromatography - PC phosphatidylcholine - PE phosphatidylethanolamine and ethanolamine plasmalogens - PI phosphatidylinositol - PLP proteolipid protein - PNS peripheral nervous system - PS phosphatidylserine - SM sphingomyelin - SU sulfatides     

Essential Metals in Cryptococcus neoformans: Acquisition and Regulation

Transition metals such as iron, zinc, and copper play key roles in numerous biological processes, including innate immunity in vertebrates. In fact, control of the availability of these essential metals is an important component of the immune response to limit the growth of pathogens during infection. In turn, pathogens must overcome the extreme limitation of essential metals and potential metal toxicity to successfully cause disease. Recent work illuminates the mechanisms by which the pathogenic fungus Cryptococcus neoformans manages iron, copper, and zinc availability in support of proliferation in vertebrate hosts. In particular, the fungus makes use of high-affinity metal transport systems and complex regulatory networks that play critical roles in the virulence of the fungus.     

Design and synthesis of chromone-nitrogen mustard derivatives and evaluation of anti-breast cancer activity

Chromone has emerged as one of the most important synthetic scaffolds for antitumor activity, which promotes the development of candidate drugs with better activity. In this study, a series of nitrogen mustard derivatives of chromone were designed and synthesised, in order to discover promising anti-breast tumour candidates. Almost all target derivatives showed antiproliferative activity against MCF-7 and MDA-MB-231 cell lines. In particular, methyl (S)-3-(4-(bis(2-chloroethyl)amino)phenyl)-2-(5-(((6-methoxy-4-oxo-4H-chromen-3-yl)methyl)amino)-5-oxopentanamido)propanoate showed the most potent antiproliferative activity with IC₅₀ values of 1.83 and 1.90 μM, respectively, and it also exhibited certain selectivity between tumour cells and normal cells. Further mechanism exploration against MDA-MB-231 cells showed that it possibly induced G2/M phase arrest and apoptosis by generating intracellular ROS and activating DNA damage. In addition, it also inhibited MDA-MB-231 cells metastasis, invasion and adhesion. Overall, methyl (S)-3-(4-(bis(2-chloroethyl)amino)phenyl)-2-(5-(((6-methoxy-4-oxo-4H-chromen-3-yl)methyl)amino)-5-oxopentanamido)propanoate showed potent antitumor activities and relatively low side effects, and deserved further investigation.     

Crystal and molecular structure of a collagen-like polypeptide (Pro-Pro-Gly)10

(Pro-Pro-Gly)₁₀ forms single crystals, providing X-ray diffraction data to 0.22 nm resolution. In the crystals, the polypeptides form triplexes that aggregate end-to-end in quasi-infinite helices with axial translation per tripeptide h = 0.287 nm and the corresponding rotation t = ?102.9 °. The structure, which may be an allomorph of collagen, has been refined by the linked-atom least-squares procedure. In addition, three water molecules per tripeptide have been detected by Fourier difference syntheses. One of them forms an intrachain hydrogen-bonded bridge O(Pro₂) - - - W - - - O(Gly). There are also interchain hydrogen bonds (Gly)NH - - - O(Pro₁) within the triplex.     

The human Rad9/Rad1/Hus1 damage sensor clamp interacts with DNA polymerase beta and increases its DNA substrate utilisation efficiency: implications for DNA repair

In eukaryotic cells, checkpoints are activated in response to DNA damage. This requires the action of DNA damage sensors such as the Rad family proteins. The three human proteins Rad9, Rad1 and Hus1 form a heterotrimeric complex (called the 9-1-1 complex) that is recruited onto DNA upon damage. DNA damage also triggers the recruitment of DNA repair proteins at the lesion, including specialized DNA polymerases. In this work, we showed that the 9-1-1 complex can physically interact with DNA polymerase β in vitro. Functional analysis revealed that the 9-1-1 complex had a stimulatory effect on DNA polymerase β activity. However, the presence of 9-1-1 complex neither affected DNA polymerase λ, another X family DNA polymerase, nor the two replicative DNA polymerases α and δ. DNA polymerase β stimulation resulted from an increase in its affinity for the primer–template and the interaction with the 9-1-1 complex stimulated deoxyribonucleotides misincorporation by DNA polymerase β. In addition, the 9-1-1 complex enhanced DNA strand displacement synthesis by DNA polymerase β on a 1 nt gap DNA substrate. Our data raise the possibility that the 9-1-1 complex might attract DNA polymerase β to DNA damage sites, thus connecting directly checkpoints and DNA repair.     

Sympathetic, sensory, and nonneuronal contributions to the cutaneous vasoconstrictor response to local cooling

Previous work indicates that sympathetic nerves participate in the vascular responses to direct cooling of the skin in humans. We evaluated this hypothesis further in a four-part series by measuring changes in cutaneous vascular conductance (CVC) from forearm skin locally cooled from 34 to 29 degrees C for 30 min. In part 1, bretylium tosylate reversed the initial vasoconstriction (-14 +/- 6.6% control CVC, first 5 min) to one of vasodilation (+19.7 +/- 7.7%) but did not affect the response at 30 min (-30.6 +/- 9% control, -38.9 +/- 6.9% bretylium; both P < 0.05, P > 0.05 between treatments). In part 2, yohimbine and propranolol (YP) also reversed the initial vasoconstriction (-14.3 +/- 4.2% control) to vasodilation (+26.3 +/- 12.1% YP), without a significant effect on the 30-min response (-26.7 +/- 6.1% YP, -43.2 +/- 6.5% control; both P < 0.05, P > 0.05 between sites). In part 3, the NPY Y1 receptor antagonist BIBP 3226 had no significant effect on either phase of vasoconstriction (P > 0.05 between sites both times). In part 4, sensory nerve blockade by anesthetic cream (Emla) also reversed the initial vasoconstriction (-20.1 +/- 6.4% control) to one of vasodilation (+213.4 +/- 87.0% Emla), whereas the final levels did not differ significantly (-37.7 +/- 10.1% control, -37.2 +/- 8.7% Emla; both P < 0.05, P > 0.05 between treatments). These results indicate that local cooling causes cold-sensitive afferents to activate sympathetic nerves to release norepinephrine, leading to a local cutaneous vasoconstriction that masks a nonneurogenic vasodilation. Later, a vasoconstriction develops with or without functional sensory or sympathetic nerves.     

Nutritional n-3 polyunsaturated fatty acids deficiency alters cannabinoid receptor signaling pathway in the brain and associated anxiety-like behavior in mice

N-3 polyunsaturated fatty acids (PUFAs) cannot be synthesized de novo in mammals and need to be provided by dietary means. In the brain, the main n-3 PUFA is docosahexaenoic acid (DHA), which is a key component of neuronal membranes. A low dietary level of DHA has been associated with increased risk of developing neuropsychiatric diseases; however, the mechanisms involved remain to be determined. In this study, we found that long-term exposure to an n-3 deficient diet decreases the level of DHA in the brain and impairs the cannabinoid receptor signaling pathway in mood-controlling structures. In n-3 deficient mice, the effect of the cannabinoid agonist WIN55,212-2 in an anxiety-like behavior test was abolished. In addition, the cannabinoid receptor signaling pathways were altered in the prefrontal cortex and the hypothalamus. Consequently, our data suggest that behavioral changes linked to an n-3 dietary deficiency are due to an alteration in the endocannabinoid system in specific brain areas.     

Positional Distribution of Fatty Acids in Glycerolipids of the Marine Red Alga, Porphyra yezoensis

The positional distribution of fatty acids in glycerolipidsfrom thalli of Porphyra yezoensis was studied by enzymatic hydrolysis.In monogalactosyl diacylglycerol, icosapentaenoic acid was amajor fatty acid at both the sn-1 and sn-2 positions of theglycerol moiety, whereas palmitic acid was a minor componentat both positions. In digalactosyl diacylglycerol and sulfoquinovosyldiacylglycerol, icosapentaenoic and palmitic acids were almostexclusively distributed at the sn-1 and sn-2 positions, respectively.In phosphatidylglycerol, palmitic and trans--13-hexadecenoicacid were exclusively located at the sn-2 position. In phosphatidylcholine,icosapentaenoic acid occurred in both the sn-l and sn-2 positions,whereas palmitic acid was confined to the sn-1 position. Itis suggested that monogalactosyl diacylglycerol in P. yezoensissynthesized in both the cytoplasmic and chloroplastic pathways,while the diacylglycerol moieties of the other chloroplast lipidsare virtually all derived from the chloroplastic pathway. (Received March 7, 1986; Accepted April 6, 1987)     

Design,Synthesis, and Evaluation of Chalcone Derivatives as Multifunctional Agents against Alzheimer's Disease

Fifteen chalcone derivatives 3a – 3o were synthesized, and evaluated as multifunctional agents against Alzheimer's disease. In vitro studies revealed that these compounds inhibited self-induced Aβ_1-42 aggregation effectively ranged from 45.9–94.5 % at 20 μM, and acted as potential antioxidants. Their structure-activity relationships were summarized. In particular, (2E)-3-[4-(dimethylamino)phenyl]-1-(pyridin-2-yl)prop-2-en-1-one ( 3g ) exhibited an excellent inhibitory activity of 94.5 % at 20 μM, and it could disassemble the self-induced Aβ_1-42 aggregation fibrils with ratio of 57.1 % at 20 μM concentration. In addition, compound 3g displayed good chelating ability for Cu²⁺, and could effectively inhibit and disaggregate Cu²⁺-induced Aβ aggregation. Moreover, compound 3g exerted low cytotoxicity, significantly reversed Aβ_1-42-induced SH-SY5Y cell damage. More importantly, compound 3g remarkably ameliorated scopolamine-induced memory impairment in mice. In summary, all the results revealed compound 3g was a potential multifunctional agent for AD therapy.     

Fibulin-4 and fibulin-5 in elastogenesis and beyond: Insights from mouse and human studies

The fibulin family of extracellular matrix/matricellular proteins is composed of long fibulins (fibulin-1, -2, -6) and short fibulins (fibulin-3, -4, -5, -7) and is involved in protein–protein interaction with the components of basement membrane and extracellular matrix proteins. Fibulin-1, -2, -3, -4, and -5 bind the monomeric form of elastin (tropoelastin) in vitro and fibulin-2, -3, -4, and -5 are shown to be involved in various aspects of elastic fiber development in vivo. In particular, fibulin-4 and -5 are critical molecules for elastic fiber assembly and play a non-redundant role during elastic fiber formation. Despite manifestation of systemic elastic fiber defects in all elastogenic tissues, fibulin-5 null (Fbln5^−/−) mice have a normal lifespan. In contrast, fibulin-4 null (Fbln4^−/−) mice die during the perinatal period due to rupture of aortic aneurysms, indicating differential functions of fibulin-4 and fibulin-5 in normal development. In this review, we will update biochemical characterization of fibulin-4 and fibulin-5 and discuss their roles in elastogenesis and outside of elastogenesis based on knowledge obtained from loss-of-function studies in mouse and in human patients with FBLN4 or FBLN5 mutations. Finally, we will evaluate therapeutic options for matrix-related diseases.     

Unraveling 14-3-3 Proteins in C4 Panicoids with Emphasis on Model Plant Setaria italica Reveals Phosphorylation-Dependent Subcellular Localization of RS Splicing Factor

14-3-3 proteins are a large multigenic family of regulatory proteins ubiquitously found in eukaryotes. In plants, 14-3-3 proteins are reported to play significant role in both development and response to stress stimuli. Therefore, considering their importance, genome-wide analyses have been performed in many plants including Arabidopsis, rice and soybean. But, till date, no comprehensive investigation has been conducted in any C₄ panicoid crops. In view of this, the present study was performed to identify 8, 5 and 26 potential 14-3-3 gene family members in foxtail millet (Si14-3-3), sorghum (Sb14-3-3) and maize (Zm14-3-3), respectively. In silico characterization revealed large variations in their gene structures; segmental and tandem duplications have played a major role in expansion of these genes in foxtail millet and maize. Gene ontology annotation showed the participation of 14-3-3 proteins in diverse biological processes and molecular functions, and in silico expression profiling indicated their higher expression in all the investigated tissues. Comparative mapping was performed to derive the orthologous relationships between 14-3-3 genes of foxtail millet and other Poaceae members, which showed a higher, as well as similar percentage of orthology among these crops. Expression profiling of Si14-3-3 genes during different time-points of abiotic stress and hormonal treatments showed a differential expression pattern of these genes, and sub-cellular localization studies revealed the site of action of Si14-3-3 proteins within the cells. Further downstream characterization indicated the interaction of Si14-3-3 with a nucleocytoplasmic shuttling phosphoprotein (SiRSZ21A) in a phosphorylation-dependent manner, and this demonstrates that Si14-3-3 might regulate the splicing events by binding with phosphorylated SiRSZ21A. Taken together, the present study is a comprehensive analysis of 14-3-3 gene family members in foxtail millet, sorghum and maize, which provides interesting information on their gene structure, protein domains, phylogenetic and evolutionary relationships, and expression patterns during abiotic stresses and hormonal treatments, which could be useful in choosing candidate members for further functional characterization. In addition, demonstration of interaction between Si14-3-3 and SiRSZ21A provides novel clues on the involvement of 14-3-3 proteins in the splicing events.     

Improved structural preservation in freeze-substituted sporidia ofUstilago avenae—a comparison with low-temperature embedding

Summary In the present study the cellular fine structure of freeze-substituted sporidia of the phytopathogenic fungusUstilago avenae is investigated by means of thin-section electron microscopy. A conventional embedding method using Spurr's low viscosity resin is compared with the recently developed methacrylate mixtures Lowicryl^® K 4 M and HM 20 resin. Generally, freeze-substitution yields improved preservation of fine structural details of the fungus compared to previously applied conventional fixation methods. Using double fixation during freeze-substitution prior to conventional embedding the fungal membrane system (plasmalemma, endoplasmic reticulum), organelles (mitochondria, nucleus etc.) and other cytoplasmic features (ribosomes, cytoskeleton) appear well resolved and smoothly contoured. Aldehyde fixed and Lowicryl embedded sporidia ofU. avenae resemble these double fixed fungal specimens fairly closely. The complete low-temperature preparation produces visualization of distinct cellular details although contrast reversal of cellular membranes (er, mitochondria etc.) is sometimes observed. In particular, fine structure resolution is enhanced in Lowicryl HM 20 embedded fungal cells. This is due also to significant improvement in staining of the cellular membranes, cytoskeleton (microfilaments and microtubules) and Golgi apparatus-like areas, using tannic acid. In case of the fungusU. avenae, freeze-substitution in combination with mild glutaraldehyde fixation and final low-temperature embedding in HM 20 resin prove suitable for improved preservation of cellular ultrastructure.Abbreviations cw cell wall - cy cytoplasm - FS freeze-substitution - FS-A GA/OsO₄ freeze-substitution and Spurr's LV-embedding - FS-B GA freeze-substitution and Lowicryl K 4 M LT-embedding - FS-C GA freeze-substitution and Lowicryl HM 20 LT-embedding - go Golgi apparatus-like body - GA glutaraldehyde - g glycogen deposit - l lipid droplet - LT low temperature - Lowicryl LT-embedding Lowicryl low-temperature embedding - Lowicryl LT-resin Lowicryl low-temperature resin - MeOH methanol - mf microfilament - mt microtubule - m mitochondrion - mvb multivesicular body - ne nuclear envelope - np nuclear pore - npl nucleoplasm - nu nucleolus - n nucleus - OsO₄ osmium tetroxide - pl plasmalemma - pr polyribosomes - Pb-citrate Reynolds' lead citrate - r ribosome - RT room temperature - rer rough endoplasmic reticulum - Spurr's LV-embedding Spurr's low viscosity embedding - Spurr's LV-resin Spurr's low viscosity resin - t tonoplast - Uac uranyl acetate - v vacuole     

14-3-3γ Induces Oncogenic Transformation by Stimulating MAP Kinase and PI3K Signaling

The 14-3-3 proteins are a set of highly conserved scaffolding proteins that have been implicated in the regulation of a variety of important cellular processes such as the cell cycle, apoptosis and mitogenic signaling. Recent evidence indicates that the expression of some of the family members is elevated in human cancers suggesting that they may play a role in tumorigenesis. In the present study, the oncogenic potential of 14-3-3γ was shown by focus formation and tumor formation in SCID mice using 14-3-3γ transfected NIH3T3 mouse fibroblast cells. In contrast, 14-3-3σ, a putative tumor suppressor, inhibited NIH3T3 transformation by H-ras and c-myc. We also report that activation of both MAP kinase and PI3K signaling pathways are essential for transformation by 14-3-3γ. In addition, we found that 14-3-3γ interacts with phosphatidylinositol 3-kinase (PI3K) and TSC2 proteins indicating that it could stimulate PI3K signaling by acting at two points in the signaling pathway. Overall, our studies establish 14-3-3γ as an oncogene and implicate MAPK and PI3K signaling as important for 14-3-3γ induced transformation.     

Distribution of catecholaminergic and serotoninergic systems in forebrain and midbrain of the newt,Triturus alpestris (Urodela)

Summary Mapping of monoaminergic systems in the brain of the newt Triturus alpestris was achieved with antisera against (1) thyrosine hydroxylase (TH), (2) formaldehyde-conjugated dopamine (DA), and (3) formaldehyde-conjugated serotonin (5-HT). In the telencephalon, the striatum was densely innervated by a large number of 5-HT-, DA-and TH-immunoreactive (IR) fibers; IR fibers were more scattered in the amygdala, the medial and lateral forebrain bundles, and the anterior commissure. In the anterior and medial diencephalon, TH-IR perikarya contacting the cerebrospinal fluid (CSF-C perikarya) were located in the preoptic recess organ (PRO), the organum vasculosum laminae terminalis and the suprachiasmatic nucleus. Numerous TH-IR perikarya, not contacting the CSF, were present in the posterior preoptic nucleus and the ventral thalamus. At this level, DA-IR CSF-C neurons were only located in the PRO. In the posterior diencephalon, large populations of 5-HT-IR and DA-IR CSF-C perikarya were found in the paraventricular organ (PVO) and the nucleus infundibularis dorsalis (NID); the dorsal part of the NID additionally presented TH-IR CSF-C perikarya. Most regions of the diencephalon showed an intense monoaminergic innervation. In addition, numerous TH-IR, DA-IR and 5-HT-IR fibers, orginating from the anterior and posterior hypothalamic nuclei, extended ventrally and reached the median eminence and the pars intermedia of the pituitary gland. In the midbrain, TH-IR perikarya were located dorsally in the pretectal area. Ventrally, a large group of TH-IR cell bodies and some weakly stained DA-IR and 5-HT-IR neurons were observed in the posterior tuberculum. No dopaminergic system equivalent to the substantia nigra was revealed. The possible significance of the differences in the distribution of TH-IR and DA-IR neurons is discussed, with special reference to the CSF-C neurons.Abbreviations AM amygdala - CAnt commissura anterior - CH commissura hippocampi - CP commissura posterior - Ctm commissura tecti mesencephali - DH dorsal hypothalamus - DTh dorsal thalamus - FLM fasciculus longitudinalis medialis - Fsol fasciculus solitarius - H habenula - LFB lateral forebrain bundle - ME median eminence - MFB medial forebrain bundle - NID nucleus infundibularis dorsalis - nIP neuropil of nucleus interpeduncularis - NPOP nucleus preopticus posterior - NS nucleus septi - OVLT organum vasculosum laminae terminalis - PD pars distalis - Pdo dorsal pallium - PHi primordium hippocampi - PI pars intermedia - Pl lateral pallium - PN pars nervosa - PRO preoptic recess organ - Ptec pretectal area - PVO paraventricular organ - Ra nucleus raphe - Rm nucleus reticularis medius - SCO subcommisural organ - ST striatum; strm stria medullaris thalami - strt stria terminalis thalami - TM tegmentum mesencephali - TO tectum opticum - TP tuberculum posterius - trch tractus cortico-habenularis - trmp tractus mamillopeduncularis - VH ventral hypothalamus - Vm nucleus motorius nervi trigemini - VTh ventral thalamus - II optic nerve