Labelling of Rat Brain ß-Adrenoceptors: (3H)CGP-12177 or (125I)Iodocyanopindolol?

Abstract

Binding of (¹²⁵I) iodocyanopindolol (ICYP) and (³H) CGP-12177 to rat brain homogenates was characterized and compared. ICYP was shown to bind to both ß-adrenergic and serotonin1B (5HT_1B) receptors whereas (³H)CGP-12177 only labelled the first ones. The addition of 10 μM serotonin (5HT) prevented ICYP binding to 5HT receptors and under these experimental conditions both ligands labelled a similar total number of ß-adrenoceptors in the different rat brain regions. ICYP displayed a higher affinity for cerebellar (mainly ß₂-subtype) than for cerebral cortex ß-adrenoceptors (mainly ß-subtype) suggesting a subtype selectivity. A multiple displacement binding approach using CGP-20712A, a ß₁-subtype ligand, as competitor revealed a 2.6 fold selectivity of ICYP for the ß₂-adrenoceptor subtype. On the other hand, (³H)CGP-12177 binds only to ß-adrenoceptors and is not subtype selective in the rat brain homogenate. Considering both its high specificity and its lack of subtype selectivity (³H)CGP-12177 seems to be a more suitable ligand than ICYP to non-selectively label ß-adrenoceptors in rat brain.     

Non-Specific Binding of the Fluorescent B-Adrenergic Receptor Probe Alprenolol-NBD

Abstract

The fluorescent ß-adrenergic receptor probe alprenolol-NBD was found to exhibit a high affinity (K_d 3.2 nM) and a low capacity (10 fmol/mg protein) for the ß₂-adrenergic receptor on living Chang liver cells but also a high affinity (K_d 320 nM) for non-ß-adrenergic receptor binding sites with a very high capacity (28,000 fmol/mg protein). Calculations are presented which make clear that less than 3% of the binding of alprenolol-NBD during visualization experiments is ß-adrenergic receptor related. Furthermore, it is shown that besides the downregulation of ß-adrenergic receptors during incubation with isoproterenol, the high-affinity non-ß-receptor binding sites are also deminishing during incubation with isoproterenol. Based on our findings it is concluded that the results of Henis et al. (1) who claimed the visualization of the ß-adrenergic receptor population on Chang liver cells by alprenolol-NBD must be interpreted as an almost completely non-specific fluorescence.     

Restoration of ß-Globin Gene Expression in Mammalian Cells by Antisense Oligonucleotides That Modify the Aberrant Splicing Patierns of Thalassemic Pre-mRNAs

Abstract

Antisense 2′-methoxy-oligoribonucleotides targeted to aberrant splice sites in two thalassemic human ß-globin pre-mRNAs, IVS2-654 and IVS2-705, expressed in HeLa cells efficiently restore correct splicing with the use of Lipofectamine, Cellfectin and DMRIE-C, in effect reactivating a defective ß-globin gene.     

The hydrophobic amino acid cluster at the cytoplasmic end of transmembrane helix III modulates the coupling of the ß1-adrenergic receptor to Gs

Abstract

A cluster of hydrophobic amino acids at the cytoplasmic end of trans-membranal helix III (TM-III) is a common feature among class-A of G protein-coupled receptors (GPCR). We mutagenized alanine 159^3.53 to glutamic acid and isoleucine160^3.54 to arginine (A159E/I160R) in TM-III of the human ß₁-adrenergic receptor (ß₁-AR) to disrupt the function of the hydrophobic cluster. Structurally, the combined mutations of A159E/I160R caused an almost 90° tilt in the rotation of Arg156^3.50 in the E/DRY motif of TM-III and displaced Tyr166^3.60 in intracellular loop 2. The A159E/I160R ß₁-AR was uncoupled from G_s as determined by cyclic AMP/adenylyl cyclase assays and by FRET-based proximity measurements between the ß₁-AR and G_sα. Isoproterenol induced ß-arrestin trafficking in cells expressing both the wild-type ß₁-AR and the A159E/I160R ß₁-AR. Isoproterenol markedly increased the phosphorylation of ERK_1/2 in cells expressing the WT ß₁-AR and this effect was dependent on the activation of the G_s-cyclic AMP-dependent protein kinase?→?Rap?→?B-raf axis. However, in cells bearing the A159E/I160R ß₁-AR, isoproterenol failed to increase the phosphorylation of ERK_1/2. These results indicate that mutations in the G_sα-binding pocket of the GPCR interfered with receptor coupling to G_s and with its downstream signaling cascades.     

Methodische Hinweise zur Gewinnung von Pansensaft

Abstract

erarbeitet von der RGW-Arbeitsgruppe zum Thema 9.2. ?Die Verwendung nichteiweißartiger Stickstoffsubstanzen in der Fütterung der Wiederkäuer“ anläßlich der methodischen Konferenz vom 6. bis 11. Februar 1969 in Berlin, Deutsche Demokratische Republik.

(Beteiligte Länder: Volksrepublik Bulgarien, Ungarische Volksrepublik, Deutsche Demokratische Republik, Volksrepublik Polen, Sozialistische Rumänische Republik, Union der Sozialistischen Sowjetrepubliken, Tschechoslowakische Sozialistische Republik).     

Methodische Empfehlung zur Bestimmung von einigen Stoffwechselprodukten im Pansensaft und Blut

Abstract

erarbeitet von der RGW-Arbeitsgruppe zum Thema 9.2 ?Die Verwendung nichteiweißartiger Stickstoffsubstanzen in der Fütterung der Wiederkäuer“ anläßlich der methodischen Konferenz vom 6. bis 11. Februar 1969 in Berlin, Deutsche Demokratische Republik.

(Beteiligte Länder: Volksrepublik Bulgarien, Ungarische Volksrepublik, Deutsche Demokratische Republik, Volksrepublik Polen, Sozialistische Rumänische Republik, Union der Sozialistischen Sowjetrepubliken, Tschechoslowakische Föderative Sozialistische Republik).     

Binding of 125I-Cyanopindolol to Beta-1-Adreno-Ceptors in a High and Low Affinity State

Abstract

Investigation of binding properties of (+), (?) and (±)¹²⁵Iodocyanopindolol (ICYP) to ß₁-adrenoceptors of guinea pig left ventricle membranes revealed that these radioligands bind to receptors in a high and low affinity state which is not influenced by guanylnucleotides. The contribution of the (+)enantiomer to the binding of the racemic ligand at low receptor concentrations can be neglected since the dissociation time courses of (±) and (?)ICYP are identical. The existence of two affinity states of ß-adrenoceptors interacting with the antagonist ICYP was evident from 1: biphasic dissociation kinetics and 2: from curvilinear Scatchard plots.     

Zinc status is independently related to the bone mineral density,fracture risk assessment tool result,and bone fracture history: Results from a U.S. nationally representative survey

PurposePrevious reports have identified the important role of zinc in bone health. Although the risk of zinc deficiency is still a concern in the U.S., there has never been an in-depth study of the association between zinc status and bone health in a sample representing the country.MethodsWe included 2,895 subjects (aged ≥ 40 years) from National Health and Nutrition Examination Survey (NHANES) 2013–2014 to explore the relationship among three biomarkers of zinc (serum, food, and total intake), the bone mineral density (BMD) of the total spine and femur, the FRAX® scores, and the previous history of bone fractures.ResultsWe showed a one-unit increase in the ln-serum zinc level was associated with an increase in the total spine BMD (ß = 0.068; S.E. = 0.028; P = 0.030) and total femur BMD (ß = 0.061; S.E. = 0.017; P = 0.003), while a one-unit increase in the ln-food zinc intake amount was correlated with an increase in the total femur BMD in the participants (ß = 0.023; S.E. = 0.009; P = 0.021). The ln-total zinc intake amount was correlated with an increase in the total femur BMD in women (ß = 0.016; S.E. = 0.007; P = 0.041). We also found food zinc intake was negatively correlated with the FRAX® score, while increased levels of all three zinc biomarkers were associated with a decreased incidence of previous bone fractures.ConclusionsIn this representative survey of American adults above 40 years old, higher zinc status was associated with higher total spine and femoral BMD, lower FRAX® scores, and lower incidence of previous fractures. If this finding is causal, increased zinc intake remains an important issue for Americans.     

Synthesis of Homo-C-D4T and Homo-C-Thymidine

Abstract

ß-C-Vinyl glycoside of D-arabinofuranose was converted to the β-C-hydroxymethyl glycoside 12 which was condensed with N-3-benzoyl thymine under Mitsunobu conditions to furnish homo-C-analog 14 of the 2′,3′-dideoxy-2′,3′-didehydrothymidine (D4T). Synthesis of the homo-C-thymidine 18 was accomplished by the same procedure from compound 15.     

Intensive Markstammkohlfütterung und ihr Einfluß auf Verdaulichkeit,Blutbild sowie Blut-, Milch- und Harnbestandteile von Kühen,Ochsen und Schafen

Abstract

The effect of energy deposition and the utilization of feedstuffs and rations were compared between growing and adult pigs. The good agreement in the data leads to the conclusion, that the energy deposition effect of feedstuffs and rations in growing pigs can be estimated with an equation based on measurements in adult pigs. Exceptions are feedstuffs which are digested bacterially to a relatively high part in the large intestine, as e. g. sugar beet pulp, raw potatoes or similar feed.

Es wurden Vergleiche zur Energieansatzwirkung und Energieverwertung von Futtermitteln und Rationen zwischen wachsenden und adulten Schweinen durchgeführt. Die gute Übereinstimmung in den Ergebnissen führt zu der Schlußfolgerung, daß die Energieansatzwirkung von Futtermitteln und Rationen beim wachsenden Schwein mit einer Gleichung geschätzt werden kann, die auf Messungen am ausgewachsenen Schwein basiert. Ausnahmen sind Futtermittel, die zu einem größeren Anteil bakteriell im Dickdarm verdaut werden, wie z.B. Trockenschnitzel, rohe Kartoffeln oder ähnliche Futtermittel.     

miR-223在结肠癌组织中的表达及促进结肠癌细胞迁移侵袭的机制研究

目的:探讨微小RNA-223 (mi R-223)在结肠癌组织中的表达及对结肠癌HT-29细胞侵袭、迁移能力的影响及机制。方法:检测mi R-223在结肠癌组织与癌旁组织中的表达。通过脂质体转染法将mi R-223模拟物(mi R-223 mimics,mi R-223 mimics组)及microRNA无关序列(mi R-223 NC,NC组)转染入结肠癌HT-29细胞。采用Real-time PCR检测转染后细胞中mi R-223和TWIST的表达,Western blot检测TWIST的蛋白表达,Tranwell检测细胞的迁移与侵袭能力。双荧光素酶报告基因检测mi R-223对TWIST基因启动子活性的影响。采用Transwell迁移与侵袭实验检测mi R-223 mimic及Twist si RNA共转染后人结肠癌细胞系HT-29迁移与侵袭能力的变化。结果:与癌旁结肠组织比较,mi R-223在结肠癌组织中呈现明显高表达(P0.05);与空白对照组和mi R-223 NC组比较,转染mi R-223 mimics后的HT-29细胞中的mi R-223表达显著增加(P0.05)。与阴性对照组和空载转染组相比较,mi R-223 mimics转染组穿透的细胞数目明显增加(P0.05),且mi R-223 mimics转染组的细胞侵袭能力显著增强(P0.05)。与mi R-223 NC组和空白对照组比较,转染mi R-223 mimics的HT-29细胞的TWIST基因m RNA和蛋白表达均显著增加(P0.05)。双荧光素酶检验结果显示TWIST为mi R-223的下游靶基因。共转染TWIST si RNA和mi R-223 mimics的结肠癌HT-29细胞的迁移与侵袭能力较单独转染mi R-223 mimics的HT-29细胞显著减弱(P0.05)。结论:mi R-223可能通过上调下游靶基因TWIST水平促进结肠癌HT-29细胞的迁移与侵袭。     

Binary and ternary complexes of FLNa-Ig21 with cytosolic tails of αMß2 integrin reveal dual role of filamin mediated regulation

BackgroundCytoskeletal protein filamin A is critical for the outside-in signaling of integrins. Although molecular mechanisms of filamin-integrin interactions are not fully understood. Mostly, the membrane distal (MD) part of the cytosolic tail (CT) of β subunit of integrin is known to interact with filamin A domain 21 (FLNa-Ig2). However, binary and ternary complexes of full-length CTs of leucocyte specific ß2 integrins with FLNa-Ig21 are yet to be elucidated.MethodsBinding interactions of the CTs of integrin αMß2 with FLNa-Ig21 are extensively investigated by NMR, ITC, cell-based functional assays and computational docking.ResultsThe αM CT demonstrates interactions with FLNa-Ig21 forming a binary complex. Filamin/αM interface is mediated by sidechain-sidechain interactions among non-polar and aromatic residues involving MP helix of αM and the canonical CD face of FLNa-Ig21. Functional assays delineated an interfacial residue Y1137 of αM CT is critical for in-cell binding to FLNa-Ig2. In addition, full-length ß2 CT occupies two distinct binding sites in complex with FLNa-Ig21. A ternary complex of FLNa-Ig21 with CTs has been characterized. In the ternary complex, αM CT moves away to a distal site of FLNa-Ig21 with fewer interactions.ConclusionOur findings demonstrate a plausible dual role of filamin in integrin regulation. The molecular interactions of the ternary complex are critical for the resting state of integrins whereas stable FLNa-Ig21/αM CT binary complex perhaps be required for the activated state.General significanceFilamin binding to both α and β CTs of other integrins could be essential in regulating bidirectional signaling mechanisms.     

Resolution des VIII. Internationalen Pflanzenschutzkongresses

Abstract

In Anbetracht der weitreichenden Bedeutung des VIII. Internationalen Pflanzenschutzkongresses, der in der Zeit vom 22. bis 26. August 1975 in Moskau stattfand, hält es die Redaktion für erforderlich, die anläßlich des Kongresses verabschiedete Resolution im Wortlaut wiederzugeben.     

外周血miR-223和血清BNP水平在慢性心力衰竭患者中的诊断价值

目的:探究慢性心力衰竭(chronic heart failure,CHF)患者外周血microRNA-223(miR-223)和血清B型利钠肽(B-type natriuretic peptide,BNP)水平及其诊断价值。方法:选取CHF患者65例(CHF组),其中美国纽约心脏病协会(New York Heart Association,NYHA)心功能分级Ⅱ级24例,Ⅲ级22例,Ⅳ级19例。另取40例同期在体检中心进行健康体检者(Control组),采用实时荧光定量PCR检测外周血中miR-223水平,ELISA检测血清BNP含量,ROC曲线评价miR-223及BNP对CHF的诊断价值。结果:CHF组左心室短轴缩短率及左心室射血分数显著低于Control组(P0.01);左心室舒张末期内径显著高于Control组(P0.01)。CHF组不同NYHA心功能分级患者miR-223和BNP水平均高于Control组,且miR-223和BNP水平随NYHA心功能分级逐渐递增,组间两两比较均显示统计学差异(P0.05)。miR-223诊断CHF的曲线下面积(area under the curve,AUC)为0.7375,临界值为43.51时灵敏度为92.82%,特异度为89.44%;BNP诊断CHF的AUC为0.7925,临界值为128 ng/L时灵敏度为92.93%,特异度为92.58%。结论:CHF患者外周血miR-223和血清BNP水平高于健康对照人群,其对CHF的诊断具有一定的临床参考价值。     

Chemische Untersuchungen über unterschiedlich vorbehandelte Federmehle

Abstract

Dr. Carlheinrich Engelmann: Ernährung und Fütterung des Geflügels. Erschienen im Neumann-Verlag Radebeul/Berlin, 162 Seiten, 17 × 24 cm Halbleinen, MDN 6,80. Reviewed by H. Bergner

Dr. Martin Pankrath: Wasserbedarf landwirtschaftlicher Nutztiere — Mengennormen. - Erschienen im S. Hirzel Verlag, Leipzig C 1, Schuhmachergäßchen 1–3. 1. Auflage 1963, 99 Seiten mit 25 Diagrammen, 21 Tabellen und 2 Tafeln im Text. 8°, Hln., MDN 10,70. Reviewed by H. Bergner

Physiologische und wirtschaftliche Leistungsgrenzen in der Tierzucht 14 Vorträge eines internationalen Symposions im Max-Planck-Institut für Tierzucht und Tierernährung in Mariensee vom 14. bis 17. Oktober 1963.: Herausgegeben von M. Witt. 315 Seiten mit 8 Kunstdrucktafeln. Englische Summaries. Steif broschiert, Preis DM 16,-. Bestellung über Institut. Reviewed by H. Bergner     

Genomic sequence and organization of rabbitTcrb constant region genes

We previously reported that some rabbits have three different copies of T-cell receptor b (Tcrb) constant region genes unlike man, mice, and rats who generally have two copies. Two of theseCß genes were found on an ≈ 14 kilobase (kb) and one on an ≈ 6 kbEco RI fragment. The gene on the 6 kb fragment is of ß2 type. A previously described portion of the 14 kb fragment appeared to have sequences characteristic ofCß1. We have now shown that the 6 kb fragment is adjacent to and 3′ of the 14 kb fragment. Furthermore, the second linked sequence ofCß gene present on the 14 kb fragment resembles to a large extent theCß2 gene present on the 6 kb fragment. Moreover, this secondCß gene has a 5′ cluster ofJß sequences resemblingJß2 of other species. However, exon 4 and the 3′ unranslated region (3′UT) are of the ß1 type. Mapping studies using southern analyses of both genomic DNA and the 14 kb clone have identified another cluster ofJß2 sequences 5′ of the third tandemCß2 gene present on the 6 kbEco RI fragment. Thus, the second gene on the 14 kb fragment appears to be a chimeric genomicTcrb gene that may have arisen by an unequal crossing-over event analogous to that which may have deletedCß1,Dß2, andJß2 in NZW mice.     

Evidence that ICI 118, 551 is a potent, highly Beta 2-selective adrenoceptor antagonist and can be used to characterize Beta-adrenoceptor populations in tissues

pA₂ values for a new ß-adrenoceptor antagonist, ICI 118, 551 (erythro-DL-1(7-methylindan-4-yloxy)-3-isopropylaminobutan-2-o1), have been obtained on intrinsic tone tracheal chain preparations and on spontaneously beating atrial preparations from guinea-pigs. Two different agonists, fenoterol (ß₂-selective) and noradrenaline (ß₁-selective) were used. The slopes of the Schild plots were not significantly different from 1.0. The antagonist was very potent (pA₂ on trachea, fenoterol as agonist, was 8.69) and also highly ß₂-selective (pA₂ on atria, noradrenaline as agonist, was only 6.96). On guinea-pig trachea (which contains ß₁- and ß₂-adrenoceptors) the potency was 44 times higher when fenoterol was the agonist than when noradrenaline was the agonist. On guinea-pig atria (which contains only ß₁-adrenoceptors) the pA₂ value was the same whichever agonist was used. Thus ICI 118, 551 has been shown to be potent and the most ß₂-selective antagonist so far studied in our laboratory. In experiments carried out with selective agonists ICI 118, 551 distinguished clearly between tissues with a mixed ß-adrenoceptor population (different pA₂ values) and those with a homogeneous population (single pA₂ value). Therefore, ICI 118, 551 is a valuable addition to the group of ß₂-selective adrenoceptor antagonists.     

Exposure of preterm neonates to toxic metals during their stay in the Neonatal Intensive Care Unit and its impact on neurodevelopment at 2 months of age

BackgroundPremature neonates might be exposed to toxic metals during their stay in the neonatal intensive care unit (NICU), which could adversely affect neurodevelopment; however, limited evidence is available. The present study was therefore designed to assess the exposure to mercury, lead, cadmium, arsenic, and manganese of preterm neonates who received total parenteral nutrition (TPN) and/or red blood cell (RBC) transfusions during their NICU stay and the risk of neurodevelopment delay at the age of 2 months.MethodsWe recruited 33 preterm neonates who required TPN during their NICU admission. Blood samples were collected for metal analysis at two different time points (admission and before discharge). Metals in the daily TPN received by preterm neonates were analyzed. Neurodevelopment was assessed using the Ages and Stages Questionnaire Edition 3 (ASQ-3).ResultsAll samples of TPN had metal contamination: 96% exceeded the critical arsenic limit (0.3 μg/kg body weight/day); daily manganese intake from TPN for preterm neonates exceeded the recommended dose (1 µg/kg body weight) as it was added intentionally to TPN solutions, raising potential safety concerns. All samples of RBC transfusions exceeded the estimated intravenous reference dose for lead (0.19 µg/kg body weight). Levels of mercury, lead and manganese in preterm neonates at discharge decreased 0.867 µg/L (95% CI, 0.76, 0.988), 0.831 (95%CI, 0.779, 0.886) and 0.847 µg/L (95% CI, 0.775, 0.926), respectively. A decrease in ASQ-3-problem solving scores was associated with higher levels of blood lead in preterm neonates taken at admission (ß = −0.405, 95%CI = −0.655, −0.014), and with plasma manganese (ß = −0.562, 95%CI = −0.995, −0.172). We also observed an association between decreased personal social domain scores with higher blood lead levels of preterm neonates before discharge (ß = −0.537, 95%CI = −0.905, −0.045).ConclusionOur findings provide evidence to suggest negative impacts on the neurodevelopment at 2 months of preterm infants exposed to certain metals, possibly related to TPN intake and/or blood transfusions received during their NICU stay. Preterm neonates may be exposed to levels of metals in utero.     

Subcomponents C1q of the first component of guinea pig and mouse complement: Comparative study of their asparagine-linked sugar chains

Guinea pig and mouse C1q, subcomponents of the first component of complement, contained six asparagine-linked sugar chains on the C-terminal non-collagenous globular regions of each molecule. After N-acetylation and successive NaB³H₄-reduction of asparagine-linked sugar chains liberated by hydrazinolysis, their structure was analysed by sequential exoglycosidase digestion in combination with sugar composition analyses. The sugar chains of C1q molecules of both animals were very similar and composed of the biantennary complex type sugar chains with the following outer chains in various combinations: (± NeuNAcα → )Galß1 → GlcNAcß1 → and Galß1 → Galß1 → GlcNAcß1 →. These chain moieties were found to be linked to a common core structure of Manα1 → (Manα1 → )Manß1 → GlcNAcß1 → (Fucα1 → )GlcNAc.