Adrenotensin: An ADM gene product with the opposite effects of ADM

The purpose of the present study was to investigate the effects of putative products of the ADM gene, other than ADM including, prodepin, proADM_45–92 and proADM_153–185 on cat pulmonary arterial (PA) rings with or without precontraction with U46619. Addition of proADM_153–185 (3×10⁻¹⁰−10⁻⁶M) increased tension in a concentration-dependent manner in cat PA rings without precontraction. When vessels were precontracted with U46619, ADM(3×10⁻¹⁰−10⁻⁶M) produced a concentration-dependent vasorelaxant response, whereas proADMus-iss produced a weak concentration-dependent contractile response. Prodepin and proADM_45–92 up to 10⁻⁶M had no activity on PA rings. Since proADM_{1531̄85}, similar to ADM, would be expected to be released in free form following endopeptidase-induced cleavage, the present data suggest proADM undergoes proteolytic processing to release peptides with divergent vascular effects.Thus, the present data also suggest that proADM_153–185 may represent a novel product of the ADM gene and term this putative new substance “adrenotensin”.     

Mass and molecular weight of isolated nuclear rings

Nuclear rings are cell structures found at the nuclear cortex wedged between the nuclear envelope and the chromatin fiber network. In previous publications we have dealt with their morphology, relationships with the nuclear membranes, chromatin fibers and cytoskeletal filaments; and more recently, with their measurements at high electron microscope resolution. In this article we have calculated the mass and molecular weight of 336 isolated nuclear rings from human circulating lymphocytes using a photometric procedure and polystyrene latex spheres as the standard for weight calibration. Our results show a range of mass of 0.4–35.5 × 10⁻¹⁶g (equivalent to 0.2–21.2 × 10⁸ Da with a positively skewed distribution (median: 3.3 × 10⁻¹⁶g or 2.0 × 10⁸ Da). Mass and volume of nuclear rings were highly correlated. In addition, it was possible to calculate the area, the whole mass and the mass per unit area of the nuclear envelope present in the center of the nuclear rings. The mass of this area also shows a lognormal distribution (median of mass/unit area: 37.3 × 10⁻⁸ pg/nm² or 1.9 × 10⁵ Da/nm²). We discuss the significance of this results as parameters for the characterization of the nuclear rings and their possible implications for a new interpretation of nuclear cortex architecture, nucleocytoplasmic traffic and macromolecule segregation between the two main cell compartments.     

Comparative nuclear magnetic resonance studies of diffusional water permeability of red blood cells from different species. V—Rabbit (Oryctolagus Cuniculus)

• 1.1. The diffusional water permeability (P_d) of rabbit red blood cell (RBC) membrane has been monitored by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition with p-chloromercuribenzene sulfonate (PCMBS).
• 2.2. The values of P_d were around 6.3 × 10⁻³ cm/sec at 15°C, 7.0 × 10⁻³cm/sec at 20°C, 8.0 × 10⁻³ cm/sec at 25°C, 9.1 × 10⁻³ cm/sec at 30°C and10.7 × 10⁻³ cm/sec at 37°C.
• 3.3. Systematic studies on the effects of PCMBS on water diffusion indicated that the maximal inhibition was reached in 15 min at 37°C with 0.5 mM PCMBS.
• 4.4. The values of maximal inhibition were around 71–74% at all temperatures.
• 5.5. The basal permeability to water was estimated as 1.6 × 10⁻³cm/sec at 15°C, 2.0 × 10⁻³cm/sec at 20°C, 2.4 × 10⁻³cm/sec at 25°C, 2.6 × 10⁻³cm/sec at 30°C, and 3.1× 10^{−3 cm/secat 37°C}.
• 6.6. The activation energy of water diffusion was around 18 kJ/mol and increased to 27 kcal/mol after incubation with PCMBS in conditions of maximal inhibition of water diffusion.
• 7.7. The membrane polypeptide electrophoretic pattern of rabbit RBCs has been compared with its human counterpart.
• 8.8. The rabbit membrane contained a higher amount of spectrin (bands 1 and 2), while the band 6 (glyceraldehyde-3-phosphate dehydrogenase) was markedly less intense.
• 9.9. Considerable differences in the electrophoretic patterns of the two sources of RBC membranes appeared in the bands migrating in the band 4.5 region and in front of band 7, where some polypeptides were apparent in higher amounts in the rabbit RBC membrane.

     

Auxin Synthesis in Crown Gall Tumor Tissue: A Comparison of Three Putative Precursors

Axenic crown gall tumor callus (from Vinca rosea L.) which is known to synthesize its own auxin is able to convert exogenous ¹⁴C-indole or tryptamine to indoleacetic acid [5.4 and 10 × 10⁻⁶μmol × h⁻¹× (g fr wt)⁻¹ respectively], but little or no ³H-tryptophan is converted [less than 6.4 × 10⁻⁸×μmol × h⁻¹× (g fr wt)⁻¹].     

Effects of VA-045, a novel apovincaminic acid derivative,on isolated blood vessels: Cerebroarterial selectivity

We investigated the effects of VA-045, an apovincaminic acid derivative, on isolated blood vessels. VA-045 (10⁻⁷−10⁻⁵M) and vinpocetine (10⁻⁷−10⁻⁵M) inhibited the 64 mM KCl-induced and 10⁻⁶M norepinephrine (NE)-induced contraction of rat aortic strips. VA-045 (10⁻⁷−10⁻⁴M) and vinpocetine (10⁻⁷−10⁻⁴M) inhibited the activity of cyclic AMP and cyclic GMP phosphodiesterase in porcine coronary artery. VA-045 (3×10⁻⁹−3×10⁻⁶M) relaxed the 64 mM KCl-induced contraction of the canine basilar artery without affecting the peripheral arteries. These results indicate that VA-045 selectively dilates canine cerebral artery, and that it may be a useful agent for the treatment of cerebrovascular diseases such as stroke.     

FeS2 nanosheets–luminol–O2 chemiluminescence method for determination of venlafaxine hydrochloride,imipramine hydrochloride,and cefazolin sodium

This study introduces a novel chemiluminescence (CL) approach utilizing FeS₂ nanosheets (NSs) catalyzed luminol–O₂ CL reaction for the measurement of three pharmaceuticals, namely venlafaxine hydrochloride (VFX), imipramine hydrochloride (IPM), and cefazolin sodium (CEF). The CL method involved the phenomenon of quenching induced by the pharmaceuticals in the CL reaction. To achieve the most quenching efficacy of the pharmaceuticals in the CL reaction, the concentrations of reactants comprising luminol, NaOH, and FeS₂ NSs were optimized accordingly. The calibration curves demonstrated exceptional linearity within the concentration range spanning from 4.00 × 10⁻⁷ to 1.00 × 10⁻³ mol L⁻¹, 1.00 × 10⁻⁷ to 1.00 × 10⁻⁴ mol L⁻¹, and 4.00 × 10⁻⁶ to 2.00 × 10⁻⁴ mol L⁻¹ with detection limits (3σ) of 3.54 × 10⁻⁷, 1.08 × 10⁻⁸, and 2.63 × 10⁻⁶ mol L⁻¹ for VFX, IPM, and CEF, respectively. This study synthesized FeS₂ NSs using a facile hydrothermal approach, and then the synthesized FeS₂ NSs were subjected to a comprehensive characterization using a range of spectroscopic methods. The proposed CL method was effective in measuring the aforementioned pharmaceuticals in pharmaceutical formulations as well as different water samples. The mechanism of the CL system has been elucidated.     

Histological and pharmacological investigations of the two symmetrically situated giant neurons,RPeNLN and LPeNLN,identified on the anterior surface of the pedal ganglia of an african giant snail (achatina fulica férussac)

• 1.1. Morphological and pharmacological investigations were made of two giant neurons, RPeNLN (right pedal nerve large neuron) and LPeNLN (left pedal nerve large neuron), situated symmetrically on the anterior surface of the pedal ganglia of an African giant snail (Achatina fulica Férussac).
• 2.]2. The two neurons (about 250–300 μm in diameter) were the largest ones identified in the ganglia of the snail species. The axonal pathways of the two neurons were symmetrical; of their four main axonal branches, the three main branches innervated the ipsilateral pedal nerves, whereas the last main branch projected to the contralateral pedal nerves.
• 3.]3. The pharmacological features of the two neurons were very similar. Both were inhibited markedly by dopamine [minimum effective concentrations (MECs): 3 × 10⁻⁶-10⁻⁵M], dl-octopamine (MECs: 2 × 10⁻⁶-2 × 10⁻⁵M), 5-hydroxytryptamine (MEC: 3 × 10⁻⁶M), GABA (MEC: 3 × 10⁻⁵ M), l-homocysteic acid (MECs: 3 × 10⁻⁵-10-10⁻⁴M) and erythro-β-hydroxy-l-ghitanuc acid (MEC: 3× 10⁻⁵M). Acetylcholine showed varied effects, either excitatory or inhibitory, on the two neurons examined. No substances were found to have any marked excitatory effects on the neurons.

     

Determination of pioglitazone hydrochloride by flow injection chemiluminescence tris(2,2′-bipyridyl)ruthenium(II)–silver(III) complex system

A novel flow injection-chemiluminescence (FI–CL) approach is proposed for the assay of pioglitazone hydrochloride (PG-HCl) based on its enhancing influence on the tris(2,2′-bipyridyl)ruthenium(II)–silver(III) complex (Ru(bipy)₃²⁺-DPA) CL system in sulfuric acid medium. The possible CL reaction mechanism is discussed with CL and ultraviolet (UV) spectra. The optimum experimental conditions were found as: Ru(bipy)₃²⁺, 5.0 × 10⁻⁵ M; sulfuric acid, 1.0 × 10⁻³ M; diperiodatoargentate(III) (DPA), 1.0 × 10⁻⁴ M; potassium hydroxide, 1.0 × 10⁻³ M; flow rate 4.0 ml min⁻¹ for each flow stream and sample loop volume, 180 μl. The CL intensity of PG-HCl was linear in the range of 1.0 × 10⁻³ to 5.0 mg L⁻¹ (R² = 0.9998, n = 10) with limit of detection [LOD, signal-to-noise ratio (S/N) = 3] of 2.2 × 10⁻⁴ mg L⁻¹, limit of quantification (LOQ, S/N = 10) of 6.7 × 10⁻⁴ mg L⁻¹, relative standard deviation (RSD) of 1.0 to 3.3% and sampling rate of 106 h⁻¹. The methodology was satisfactorily used to quantify PG-HCl in pharmaceutical tablets with recoveries ranging from 93.17 to 102.77 and RSD from 1.9 to 2.8%.     

Protein kinase C β1 overexpression augments phorbol ester-induced increase in endothelial permeability

We studied the postulated involvement of the protein kinase C β₁ (PKCβ₁) isoform in the regulation of endothelial permeability using human dermal microvascular endothelial cell line (HMEC-1). We overexpressed the recombinant PKCβ₁ gene via retroviral-mediated transduction in these cells. PKCβ₁ gene transfer was stable, and PKCβ₁ protein production was persistent for at least 1 month posttransduction. Addition of 2 × 10⁻⁹ M and 2 × 10⁻⁸ M phorbol 12-myristate 13-acetate (PMA) to the control (nontransduced) HMEC-1 cells increased the transendothelial ¹²⁵I-albumin clearance rate (an index of endothelial permeability) from 2.5 ± 0.2 × 10⁻² μl/min to 5.4 ± 1.2 × 10⁻² μl/min and 16.8 ± 3.1 × 10⁻² μl/min, respectively. However, addition of 2 × 10⁻⁹ M PMA to PKCβ₁-overexpressing HMEC-1 cells produced a maximal increase in the transendothelial ¹²⁵I-albumin clearance rate of 15.9 ± 2.0 × 10⁻² μl/min. Challenge of these cells with 2 × 10 ⁻⁸ M PMA did not further augment the increase in permeability. Activation with PMA was associated with the translocation of the PKCβ₁ from the cytosol to the membrane. These data show that PKCβ₁ overexpression augments the increase in endothelial permeability in response to PKC activation, suggesting an important function for the PKCβ₁ isoform in the regulation of endothelial barrier. © 1996 Wiley-Liss, Inc.     

Investigation on silent bacterial infections in specimens from pregnant women affected by spontaneous miscarriage

Miscarriage is one of the main complications occurring in pregnancy. The association between adverse pregnancy outcomes and silent bacterial infections has been poorly investigated. Ureaplasma parvum and urealiticum, Mycoplasma genitalium and hominis and Chlamydia trachomatis DNA sequences have been investigated by polymerase chain reaction (PCR) methods in chorionic villi tissues and peripheral blood mononuclear cells (PBMCs) from females with spontaneous abortion (SA, n = 100) and females who underwent voluntary interruption of pregnancy (VI, n = 100). U. parvum DNA was detected in 14% and 15% of SA and VI, respectively, with a mean of bacterial DNA load of 1.3 × 10⁻¹ copy/cell in SA and 2.8 × 10 ⁻³ copy/cell in VI; U. urealiticum DNA was detected in 3% and 2% of SA and VI specimens, respectively, with a mean DNA load of 3.3 × 10⁻³ copy/cell in SA and 1.6 × 10⁻³ copy/cell in VI; M. hominis DNA was detected in 5% of SA specimens with a DNA load of 1.3 × 10⁻⁴ copy/cell and in 6% of VI specimens with a DNA load of 1.4 × 10⁻⁴ copy/cell; C. trachomatis DNA was detected in 3% of SA specimens with a DNA load of 1.5 × 10⁻⁴ copy/cell and in 4% of VI specimens with a mean DNA load of 1.4 × 10⁻⁴ copy/cell. In PBMCs from the SA and VI groups, Ureaplasma spp, Mycoplasma spp and C. trachomatis DNAs were detected with a prevalence of 1%–3%. Bacteria were investigated, for the first time, by quantitative real-time PCR (qPCR) in chorionic villi tissues and PBMCs from women affected by SA and VI. These data may help to understand the role and our knowledge of the silent infections in SA.     

A novel LncRNA-based prognostic score reveals TP53-dependent subtype of lung adenocarcinoma with poor survival

The prognostic signatures play an essential role in the era of personalised therapy for cancer patients including lung adenocarcinoma (LUAD). Long noncoding RNA (LncRNA), a relatively novel class of RNA, has shown to play a crucial role in all the areas of cancer biology. Here, we developed and validated a robust LncRNA-based prognostic signature for LUAD patients using three different cohorts. In the discovery cohort, four LncRNAs were identified with 10% false discovery rate and a hazard ratio of >10 using univariate Cox regression analysis. A risk score, generated from the four LncRNAs’ expression, was found to be a significant predictor of survival in the discovery and validation cohort (p = 9.97 × 10 ⁻⁸ and 1.41 × 10 ⁻³, respectively). Further optimisation of four LncRNAs signature in the validation cohort, generated a three LncRNAs prognostic score (LPS), which was found to be an independent predictor of survival in both the cohorts ( p = 1.00 × 10 ⁻⁶ and 7.27 × 10 ⁻⁴, respectively). The LPS also significantly divided survival in clinically important subsets, including Stage I ( p = 9.00 × 10 ⁻⁴ and 4.40 × 10 ⁻², respectively), KRAS wild-type (WT), KRAS mutant ( p = 4.00 × 10 ⁻³ and 4.30 × 10 ⁻², respectively) and EGFR WT ( p = 2.00 × 10 ⁻⁴). In multivariate analysis LPS outperformed, eight previous prognosticators. Further, individual members of LPS showed a significant correlation with survival in microarray data sets. Mutation analysis showed that high-LPS patients have a higher mutation rate and inactivation of the TP53 pathway. In summary, we identified and validated a novel LncRNA signature LPS for LUAD.     

A comparison of the 8-hydroxydeoxyguanosine,chromosome aberrations and micronucleus techniques for the assessment of the genotoxicity of mercury compounds in human blood lymphocytes

We compared the mechanism of action of micronuclei (MN), unstable chromosome aberrations, and 8-hydroxydeoxyguanosine (8-OHdG) levels to evaluate the genotoxicity of methyl mercuric chloride (CH₃HgCl) and mercuric chloride (HgCl₂) in human peripheral lymphocytes. The chromosome aberrations in human peripheral lymphocytes exposed to various concentrations of CH₃HgCl or HgCl₂ increased in a concentration-dependent manner and were significantly higher than the control when the cells were incubated with 1 × 10⁻⁵ M (HgCl₂) or 2 × 10⁻⁶ M (CH₃HgCl). The increase in the incidence of micronucleated lymphocytes was significant among the exposed groups, being 2 × 10⁻⁵ M (HgCl₂) and 5 × 10⁻⁶ M (CH₃HgCl) compared with the control. CH₃HgCl was about 4-fold more potent than HgCl₂. We determined the 8-OHdG levels in human peripheral blood mononuclear cells(PBMC) and found that they were significantly higher in the exposed groups at 1 × 10⁻⁵ M (HgCl₂) and 5 × 10⁻⁶ M (CH₃HgCl) compared with the control. A detectable (p < 0.05) increase in the level of 8-OHdG was induced by CH₃HgCl at a concentration that was about 50% of the amount of HgCl₂ required to produce a similar response. The data confirmed the value of the MN and/or chromosome aberration assays for assessing of HgCl₂- and/or CH₃HgCl-induced genotoxicity, and indicated that they are about the same concentration as the 8-OHdG assay. The presence of genotoxic effects in peripheral blood lymphocytes exposed to the mercuric compounds indicated by the chromosome aberrations and the MN assays could be partly due either to the disturbance of the spindle mechanism, or to the elevated level of 8-OHdG brought by the generation of reactive oxygen species.     

The interaction of cefotaxime with the serum albumin of several mammalian species

1. The interaction of cefotaxime with the serum albumin of several mammalian species; horses, swine, sheep, dogs and rabbits, was studied comparatively. The technique of ultrafiltration and spectrophotometric determination of the free antibiotic in the filtrate was used.2. Binding percentages, which vary according to the species studied, were found to be higher in swine and rabbit albumins (between 92 and 81%) and lower for sheep, dog and horse albumins (between 67 and 52%).3. The number of binding sites is usually close to 2; in the case of the horse it is 2.43. The apparent binding constants are: swine, 1.61 × 10⁴ M⁻¹; rabbit, 1.19 × 10⁴ M⁻¹; sheep, 2.33 × 10³ M⁻¹; dog, 2.00 × 10³ M⁻¹; horse, 1.42 × 10³ M⁻¹. The Scatchard model was used for data analysis.4. Possible consequences of this interaction regarding clinical use of cefotaxime on different species are discussed.     

A genome-wide association study identifies a new variant associated with word reading fluency in Chinese children

Reading disability exhibited defects in different cognitive domains, including word reading fluency, word reading accuracy, phonological awareness, rapid automatized naming and morphological awareness. To identify the genetic basis of Chinese reading disability, we conducted a genome-wide association study (GWAS) of the cognitive traits related to Chinese reading disability in 2284 unrelated Chinese children. Among the traits analyzed in the present GWAS, we detected one genome-wide significant association (p < 5 × 10⁻⁸) on word reading fluency for one SNP on 4p16.2, within EVC genes (rs6446395, p = 7.33 × 10⁻¹⁰). Rs6446395 also showed significant association with Chinese character reading accuracy (p = 2.95 × 10⁻⁴), phonological awareness (p = 7.11 × 10⁻³) and rapid automatized naming (p = 4.71 × 10⁻³), implying multiple effects of this variant. The eQTL data showed that rs6446395 affected EVC expression in the cerebellum. Gene-based analyses identified a gene (PRDM10) to be associated with word reading fluency at the genome-wide level. Our study discovered a new candidate susceptibility variant for reading ability and provided new insights into the genetics of developmental dyslexia in Chinese children.     

Kinetic investigation of the base hydrolysis of the chloropentaamminecobalt(III) ion in micellar sodium dodecyl sulfate solution

The kinetics of the base hydrolysis of the complex ion chloropentaamminecobalt(III) have been studied by conventional spectrophotometry at 25.0 °C in water and in the presence of the anionic surfactant sodium dodecyl sulfate (SDS) over the SDS concen- tration range from 1.0 × 10⁻³ to 7.5 × 10⁻² mol dm⁻³. The hydrolysis rate is strongly inhibited by the surfactant, fitting a model in which the cobalt- (III) complex is distributed between water and the micellar pseudo-phase with a binding constant equal to 3.7 × 10³ dm³ mol⁻¹. The effects of different added electrolytes on the critical micelle concentra- tion of the surfactant and on the hydrolysis rate have also been investigated and discussed.     

Newly synthesized Fe-doped CDs for colorimetric and fluorometric nanozyme-based levodopa sensing

A simple single-pot hydrothermal method was used to fabricate a Fe, N, and S co-doped carbon dots (Fe-CDs) nanozyme using ferric chloride and sunset yellow as precursors. The fabricated Fe-CDs exhibited intense green fluorescence at 460 nm with excitation-independent properties and a high quantum yield of 40.23%. This nanozyme mimics peroxidase by catalyzing the oxidation of tetramethylbenzidine (TMB) by H₂O₂ to yield a blue-coloured TMB_ox product at 652 nm. Dual detection methods were established for determining levodopa (l -dopa) by taking advantage of the high nanozyme activity and the distinct fluorescence aspect. Both determination methods are based on the oxidation of l -dopa by H₂O₂ in the presence of Fe-CDs and fading of the blue colour of the TMB_ox. The colorimetric method monitors the amount of colour fading of TMB_ox. In the fluorometric method, the formed blue TMB_ox absorbs the emission light of the Fe-CDs; when l -dopa is present, this effect decreases and the intensity of the emission light increases. The nanozyme-based detection procedures exhibit good linearity in the ranges 2.17 × 10⁻³ to 34.78 × 10⁻³ mM [limit of detection (LOD) = 0.84 × 10⁻³ mM] and 0.85 × 10³ to 16.95 × 10³ nM (LOD = 0.102 × 10³ nM) for colorimetric and fluorometric methods, respectively.     

Evidence for natural transformation of Bacillus subtilis in foodstuffs

The effect of foodstuffs on the natural transformation of Bacillus subtilis was investigated. As examples of complex food matrices milk with various fat contents as well as chocolate milk were used. The frequencies of transformation varied with the fat content and ranged between 3.8×10⁻⁴ and 1.4×10⁻³. Highest frequencies of about 3×10⁻³ were observed in chocolate milk with 1.5% fat. Development of competence was observed in chocolate milk, resulting in maximal transformation frequencies upon incubation for 10–12 h at 37°C.     

Increased frequency of 6-thioguanine-resistant lymphocytes in peripheral blood of workers employed in cyclophosphamide production

The frequency of 6-thioguanine-resistant peripheral blood lymphocytes has been determined by autoradiography in a control population and a population of cyclophosphamide-exposed individuals. The mean variant frequency in a non-exposed population was found to be 2.76 ± 1.48 × 10⁻⁵. Subpopulations of smokers and non-smokers revealed statistically significantt differences in the variant frequencies, i.e. 3.52 ± 1.55 × 10⁻⁵ and 2.07 ± 1.05 × 10⁻⁵ respectively. In 20 out of a total of 23 individuals employed in cyclophosphamide synthesis and manufacturing, the variant frequency of 6-thioguanine-resistant lymphocytes was found to be higher than the maximum individual frequency found in the control population. The mean variant frequency in the cyclophosphamide-exposed population was 13.64 ± 13.56 × 10⁻⁵, a statistically significant increase as compared to the mean control frequency. There was no correlation between variant frequency and duration of employment suggesting that this test reflects the actual exposure and not a cumulative effect.     

Study of different nitrogen sources on glucose uptake and production of melanin precursors and fungal mass of Fonsecaea pedrosoi cultured in tricyclazole

The influence of tricyclazole on the production of phenolic precursors, glucose uptake and production of fungal mass of Fonsecaea pedrosoi was assessed in the presence of phenylalanine, sodium nitrate and tryptophan. Photocolorimetry was used for the quantitation of glucose and total phenol, and the dry weight method for biomass. The glucose uptake with phenylalanine was 99.85×10³ mg/ml; with tryptophan, 99.88×10³ mg/ml and with sodium nitrate, 99.90×10³ mg/ml. Production of biomass: with tryptophan, 2.7×10⁻¹ mg; with sodium nitrate, 3.0×10⁻¹ mg and with phenylalanine, 3.4×10⁻¹ mg. Tricyclazole induced higher phenol accumulation, lower glucose uptake, inhibition of melanin deposition on the cell and higher production and higher biomass production in relation to glucose uptake for all nitrogen sources tested under these conditions. A negative correlation between biomass production and glucose uptake was observed in the presence of secondary metabolism.     

Characterization of azadirachtin binding to Sf9 nuclei in vitro

[22,23-³H₂]dihydroazadirachtin was incorporated by Sf9 cells in culture and was bound specifically to the nuclear fraction. The observed association constant of the binding of the radioligand to a purified nuclear fraction was determined to be 0.037 ± 0.008 min ¹ using a one-phase exponential association equation, and binding appeared to be to a single population of sites. The binding was essentially irreversible, and the dissociation constant was estimated to be 0.00065 ± 0.00013 min ¹. An association rate constant of 7.3 × 10⁶ M ¹ min ¹ was calculated from these data. Binding was saturable, and the receptor number and affinity were determined as B_max = 23.87 ± 1.15 pmol/mg protein, K_d = 18.1 ± 2.1 nM. The order of potency of semisynthetic azadirachtin analogues for competition for the binding site was as follows (IC₃₀ in parentheses): azadirachtin (1.55 × 10⁻⁸ M) > dihydroazadirachtin (3.16 × 10⁻⁸ M) > dansyl dihydroazadirachtin (7.40 × 10⁻⁸ M) > DNP-azadirachtin (7.50 × 10⁻⁸ M) > biotin dihydroazadirachtin (1.27 × 10⁻⁷ M) ≫ 11-methoxy 22,23-dihydroazadirachtin (6.67 × 10⁻⁷ M). Arch. Insect Biochem. Physiol. 34:461–473, 1997. © 1997 Wiley-Liss, Inc.