Suppression of natural killer cell activity in mouse spleen lymphocytes by several dopamine receptor antagonists

The effects of dopaminergic receptor inhibitors such as thiothixine (D₁/D₂), fluphenazine (D₁/D₂), trifluoperazine (D₁/D₂), pimozide (D₂), flupenthixol (D₁/D₂), (+/–)-SKF 83566 (D₁), and spiperone (D₂) on splenic natural killer (NK) cell cytotoxic activities were assessed in vitro using mouse spleen lymphocytes or enriched NK cells. Both the activities of the splenic NK cell cytotoxicity and the effector-target cell conjugation were suppressed by thiothixine, fluphenazine, and trifluoperazine at concentrations from 2.64 to 14.78 M. In addition, the augmentation of the cytolytic activity of NK cells induced by interferon- or interleukin-2 was antagonized by pretreatment with these neuroleptic compounds. However, neither the splenic NK cell cytotoxicity nor the effector-target cell conjugation were affected by treatment with other neuroleptic compounds such as pimozide, flupenthixol, (+/–)-SKF 83566, and spiperone. Thus, it appears that neuroleptic compounds such as thiothixine, fluphenazine, and trifluoperazine may act through the mechanisms other than a dopaminergic pathway to affect the NK cell-target cell interaction.     

Binding site of activators of the cystic fibrosis transmembrane conductance regulator in the nucleotide binding domains

The use of substances that could activate the defective chloride channels of the mutant cystic fibrosis transmembrane conductance regulator (CFTR) has been suggested as possible therapy for cystic fibrosis. Using epithelia formed by cells stably transfected with wildtype or mutant (G551D, G1349D) CFTR, we estimated the apparent dissociation constant, K_D, of a series of CFTR activators by measuring the increase in the apical membrane current. Modification of apparent K_D of CFTR activators by mutations of the nucleotide-binding domains (NBDs) suggests that the binding site might be in these regions. The human NBD structure was predicted by homology with murine NBD1. An NBD1-NBD2 complex was constructed by overlying monomers to a bacterial ABC transporter NBD dimer in the head-to- tail conformation. Binding sites for CFTR activators were predicted by molecular docking. Comparison of theoretical binding free energy estimated in the model to free energy estimated from the apparent dissociation constants, K_D, resulted in a remarkably good correlation coefficient for one of the putative binding sites, located in the interface between NBD1 and NBD2.Received 21 September 2004; received after revision 6 December 2004; accepted 10 December 2004     

Effects of selective dopamine D1 and D2 antagonists on male rat sexual behavior

Summary The effects of selective dopamine (DA) D₁ and D₂ antagonists on male rat sexual behavior were investigated. The D₁ antagonist (+)SCH-23390, 25–100 g kg^–1 s.c. –20 min, and the D₂ antagonist raclopride, 0.1–1.6 mg kg^–1 s.c., –20 min, decreased both the number of mounts and intromissions preceding ejaculation. No statistically significant effects in the time up to ejaculation or in the time up to the first intromission were noted, whereas both compounds produced a statistically significant increase in the post-ejaculatory interval. The effect can generally be characterized as psychomotor inhibition, and no evidence was obtained for a specific role of DA D₁ or D₂ receptors in the mediation of male rat sexual behavior.The expert technical assistance of Ms Elisabeth Wallin is gratefully acknowledged. The figures were skilfully prepared by Ms Madelene Kröning at the Department of Psychology. This study received support from the Bank of Sweden tercentenary Foundation, The Swedish MRC and Wilhelm and Martina Lundgren Foundation.     

Gitterkonstanten und Raumgruppe von Tetrammincuprisulfat

Summary The space-group of Cu(NH₃)₄SO₄·H₂O isD _2h ¹⁶ orD _2h ¹³ (D _2h ⁵ , D _2h ¹ ) with a=7,07, b=12,12, c=10,66 Å ± 1%.     

Participation of ACTH1–10 and ACTH4–10 on the melatonin modulation of benzodiazepine receptors in rat cerebral cortex

In this study, we examined the effect of intracerebroventricular (i.c.v) injection of melatonin and/or ACTH_1–10 and ACTH_4–10 on [³H]flunitrazepam binding sites in the cerebral cortex of hypophysectomized rats. Hypophysectomy increased the B_max (maximum number of binding sites) of benzodiazepine (BNZ) receptors for at least 7 days after surgery, without changing K_D (dissociation constant). The i.c.v. injection of melatonin to hypophysectomized rats significantly increased B_max, whereas the same doses of melatonin were ineffective in sham-operated animals. In both cases, K_D values were unchanged. The i.c.v injection of ACTH_1–10 to hypophysectomized animals significantly increased B_max, an effect that was enhanced by simultaneous i.c.v. injection of ACTH_1–10+melatonin, reaching higher values of B_max than the i.c.v. injection of these hormones individually. No significant changes in K_D values were found after ACTH_1–10 and/or melatonin administration. However, the i.c.v. injection of ACTH_4–10 to hypophysectomized rats did not change B_max, although it significantly increased K_D values, indicating a decrease in the BNZ binding affinity. Melatonin injection counteracted this effect of ACTH_4–10, returning K_D to the control value. Moreover, although the lower dose of i.c.v. melatonin used, 10 ng, was unable to modify B_max of BNZ binding in the ACTH_4–10-injected group, the higher dose, 20 ng, significantly increased B_max. The results suggest that these ACTH-derived peptides can modulate the effect of melatonin on brain benzodiazepine receptors.     

The action of various vitamin D3 metabolites on calcium and phosphorus metabolism in chick embryo calvariae

Summary Chick embryos from vitamin D-deficient hens given physiological doses of 1,25-dihydroxyvitamin D₃ or 24,25-dihydroxyvitamin D₃ or both become severely hypocalcemic, hyperphosphatemic and fail to hatch as compared to those derived from hens given 25-hydroxyvitamin D₃ or 24,25-difluoro-25-hydroxyvitamin D₃. Calvariae from the former contain less mineral and on incubation in vitro produce significantly lower calcium and higher phosphate concentration in the medium than do the calvariae derived from the embryos of hens supported on 25-hydroxyvitamin D₃ or 24,24-difluoro-25-hydroxyvitamin D₃.     

Radioimmunological determination of prostaglandin D2 synthesis in human thrombocytes

Summary A specific radioimmunoassay for prostaglandin D₂ was developed. Using the radioimmunoassay, prostaglandin D₂ synthesis by human thrombocytes was measured. While the cyclooxygenase inhibitor indomethacin inhibits formation of prostaglandin D₂, increased formation of prostaglandin D₂ was observed in the presence of the thromboxane synthetase inhibitor imidazole.This work was supported by the Deutsche Forschungsgemeinschaft.     

Distribution of the 1,25 dihydroxy-vitamin D3 receptor in the bursa of Fabricius of chicken

The vitamin D₃ metabolite 1,25(OH)₂D₃ is probably involved in B lymphocyte ontogeny. We therefore determined the distribution of the 1,25(OH)₂D₃ receptor in the bursa of Fabricius and spleen cells of 7-day-old chicks, by immunohistochemistry using a monoclonal antibody against the chick intestinal cell 1,25(OH)₂D₃ receptor. The bursa cells of young (7-day-old) chicks contained large amounts of receptor while the spleen cells did not. The bursa cells of older (35-day-old) chicks contained fewer receptors, but the number of receptors in the spleen increased.     

Deletion mapping of a new gustatory mutant inDrosophila melanogaster

Summary A gustatory mutant ofDrosophila melanogaster insensitive to the taste of salt has been isolated. Genetic crosses and a deletion mapping analysis show that this mutation, designatedgust-M ₁ is located in the 93C_3–6-93D_6–7 region of the third chromosome.gust-M ₁ is also insensitive to the taste of quinine sulfate. The behavior of this mutant may be explained by assuming thatgust-M ₁ could be a mutation perturbing functions in the central nervous system affecting the responses to both compounds.     

Demonstration of vitamin D3 metabolism inmytilus edulis

Summary Radiolabeled vitamin D₃ was converted into several polar metabolites upon incubation with tissue homogenates from the common musselMytilus edulis. Chromatographic analysis indicated that the metabolites have chromatographic mobilities different from those of known standards. The results suggest that vitamin D₃ is metabolized in mussels via pathways that differ from the vertebrate systems.Acknowledgments. The authors thank Dr Pirjo Rantamäki for the mussels and F. Hoffmann-La Roche & Co. for supplying the vitamin D₃ metabolite standards. This work was supported by a grant from the Magnus Ehrnrooth Foundation to T. K.     

Diffusion of d-glucose measured in the cytosol of a single astrocyte

Astrocytes interact with neurons and endothelial cells and may mediate exchange of metabolites between capillaries and nerve terminals. In the present study, we investigated intracellular glucose diffusion in purified astrocytes after local glucose uptake. We used a fluorescence resonance energy transfer (FRET)-based nano sensor to monitor the time dependence of the intracellular glucose concentration at specific positions within the cell. We observed a delay in onset and kinetics in regions away from the glucose uptake compared with the region where we locally super-fused astrocytes with the d-glucose-rich solution. We propose a mathematical model of glucose diffusion in astrocytes. The analysis showed that after gradual uptake of glucose, the locally increased intracellular glucose concentration is rapidly spread throughout the cytosol with an apparent diffusion coefficient (D _app) of (2.38 ± 0.41) × 10^?10 m² s^?1 (at 22–24 °C). Considering that the diffusion coefficient of d-glucose in water is D = 6.7 × 10^?10 m² s^?1 (at 24 °C), D _app determined in astrocytes indicates that the cytosolic tortuosity, which hinders glucose molecules, is approximately three times higher than in aqueous solution. We conclude that the value of D _app for glucose measured in purified rat astrocytes is consistent with the view that cytosolic diffusion may allow glucose and glucose metabolites to traverse from the endothelial cells at the blood–brain barrier to neurons and neighboring astrocytes.     

Simple integrated rate equations for reversible bimolecular reactions

Summary If the complete rate equations for reversible, one-step, bimolecular reactions are written withP _e–P as the concentration variable (whereP _e is the equilibrium, andP is the instantaneous, product concentration), the 3 possible stoichiometries can be reduced to a single straightforward differential equation. This can be solved very economically. For each stoichiometry, weret is time,k ₁ is the forward rate constant,K _e is the equilibrium constant, and P isP–P _o. The termsP _c–P _o andD+P _c–P _o are the physically possible and physically impossible roots of the quadratic equation forP _e–P _o in terms of the initial concentrations andK _c.D is the discriminant in this equation. All 3 quantities can be calculated if the equilibrium constant is known. A plot oft against ln{[1–P/(D+P _c–P _o)]/[1–P/(P _c–P _o)]} should be a straight line for any second order reaction. For each stoichiometry,P _c–P _o approachesA _o, the initial concentration of the first reactant, as the equilibrium constant increases. When a second reactant is present,D+P _e–P _o approachesB _o. The limiting equation is then that of an irreversible bimolecular reaction. For AP+Q,D approaches –K _e as the equilibrium constant becomes large, and the value of the second logarithmic term in the integrated equation approaches zero. The limiting equation is that of an irreversible, unimolecular reaction.Acknowledgments. I thank Dr. Athel Cornish-Bowden for many helpful discussions. This work was partially supported by a grant from Utah State University.     

The monamycins,a new family of cyclodepsipeptide antibiotics

Zusammenfassung Die Analyse von Monamycin, einem Antibiotika-Komplex, führte zur Identifizierung der 15 Cyclohexadepsipeptidkomponenten. Für Monamycin D₁ wurde Strukturformel V abgeleitet.     

Chemical structure of polymyxin D1

Zusammenfassung Es wird über die Strukturaufklärung von Polymyxin D₁, einem Antibioticum ausBacillus polymyxa ATCC 10401, berichtet, dem die Formel der Figur zukommt.     

Arthritogenicity and protective effect against adjuvant arthritis of wax DS fractions (glycolipids without a nitrogen-containing moiety) ofMycobacterium tuberculosis var.hominis andbovis

Résumé Les cires D_S des Mycobactéries (souches humaines et bovines), glycolipides dépourvus de la partie azotée présente dans les cires D_P, ne provoquent pas d'arthrite chez le rat; contrairement aux cires D_P acétylées, elles ont un effet protecteur vis-à-vis d'une cire D_P arthrogène seulement si elles sont injectées 40 jours avant cette dernière.     

The effects of phenoxybenzamine on the aortic pressure-diameter relationship in dogs

Summary The normalized diameter (D/D_13.3 where D_13.3 equals D at 13.3 kPa under control conditions) was measured at selected pressure levels under different hemodynamic conditions. Hemorrhage caused the normalized diameter to decrease (–3.3%) when compared to control values at a given pressure. Volume expansion anda-blockade with phenoxybenzamine caused D/D_13.3 to increase (+3.3% and +8.5% respectively).This work was supported in part by PHS grant HL-23239 and a grant from the Central Ohio Heart Chapter of the American Heart Association. To whom reprint request should be addressed.     

Structure and dynamics of rotary V<Subscript>1</Subscript> motor

Rotary ATPases are unique rotary molecular motors that function as energy conversion machines. Among all known rotary ATPases, F₁-ATPase is the best characterized rotary molecular motor. There are many high-resolution crystal structures and the rotation dynamics have been investigated in detail by extensive single-molecule studies. In contrast, knowledge on the structure and rotation dynamics of V₁-ATPase, another rotary ATPase, has been limited. However, recent high-resolution structural studies and single-molecule studies on V₁-ATPase have provided new insights on how the catalytic sites in this molecular motor change its conformation during rotation driven by ATP hydrolysis. In this review, we summarize recent information on the structural features and rotary dynamics of V₁-ATPase revealed from structural and single-molecule approaches and discuss the possible chemomechanical coupling scheme of V₁-ATPase with a focus on differences between rotary molecular motors.     

Influence of heavy water (D2O) on the multiplication of Adeno and Mengo virus

Zusammenfassung Der Einfluss von Deuteriumoxyd (50%) auf die Vermehrung von Mengo-Virus (kleiner RNS-Virus) und Adeno-7-Virus (ein DNS-Virus) in L-Zellen wurde untersucht. Nach 8 h im H₂O-Medium ist der Titer vom Mengo-Virus signifikant erhöht. 24 h nach Infektion sind die Titer von Mengo-Virus in H₂O und D₂O enthaltenden Medien identisch. Der Adenovirustiter in H₂O enthaltendem Medium ist 24 und 48 h nach Infektion signifikant höher als im Medium mit D₂O.     

Structural variations in wheat HKT1;5 underpin differences in Na<Superscript>+</Superscript> transport capacity

An important trait associated with the salt tolerance of wheat is the exclusion of sodium ions (Na⁺) from the shoot. We have previously shown that the sodium transporters TmHKT1;5-A and TaHKT1;5-D, from Triticum monoccocum (Tm) and Triticum aestivum (Ta), are encoded by genes underlying the major shoot Na⁺-exclusion loci Nax1 and Kna1, respectively. Here, using heterologous expression, we show that the affinity (K _m) for the Na⁺ transport of TmHKT1;5-A, at 2.66 mM, is higher than that of TaHKT1;5-D at 7.50 mM. Through 3D structural modelling, we identify residues D⁴⁷¹/a gap and D⁴⁷⁴/G⁴⁷³ that contribute to this property. We identify four additional mutations in amino acid residues that inhibit the transport activity of TmHKT1;5-A, which are predicted to be the result of an occlusion of the pore. We propose that the underlying transport properties of TmHKT1;5-A and TaHKT1;5-D contribute to their unique ability to improve Na⁺ exclusion in wheat that leads to an improved salinity tolerance in the field.     

Trichinella spiralis: Acceleration and inhibition of cyst calcification in rats

Summary Daily administration of vitamin D₃ (75,000 IU/kg b.wt) for 7 days acceleratedTrichinella spiralis cyst calcification in rats with a 14-week-old infection. When disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) was administered (50 mg/kg b.wt) from 2 days before until 2 days after vitamin D₃ treatment, cyst calcification was inhibited. Thus, the ability to inhibitT. spiralis calcification has been demonstrated for the first time.Supported in part by EKU faculty research grant No. 03-05.Much appreciation goes to Dr Gordon S. Hassings and the Proctor and Gamble Company, Miami Valley Laboratories, Cincinnati, Ohio USA for providing the EHDP used in this study.