Evaluation of several routine methods for fosfomycin and mecillinam susceptibility testing of Enterobacterales urine isolates.

OBJECTIVES: Performance evaluation of routine laboratory methods to determine the susceptibility of Enterobacterales urinary isolates to fosfomycin (oral administration) and mecillinam. METHODS: We collected 347 Enterobacterales isolates from monomicrobial midstream urine samples from women with significant bacteriuria and leukocyturia. Mostly non-Escherichia coli isolates (i.e. Klebsiella spp., Citrobacter koseri, Enterobacter cloacae complex and Proteus mirabilis) were included (n = 298). Performance of VITEK®2, ETEST®, and disc diffusion to determine fosfomycin and mecillinam susceptibility was evaluated following International Organization for Standardization (ISO) 20776-2:2021 (or 20776-2:2007 for disc diffusion) in comparison with the agar dilution reference method. RESULTS: For fosfomycin testing, VITEK®2 and ETEST® were close to reaching ISO requirements (essential agreement  ≥ 90%; bias  ±30%) for C. koseri, E. coli and P. mirabilis. Categorical agreement (CA) and major error rates were acceptable for disc diffusion. Fosfomycin displayed lower activity against E. cloacae complex and Klebsiella spp., with MIC50 (minimum inhibitory concentration required to inhibit the growth of 50% of tested isolates) equal to the E. coli EUCAST breakpoint (8 mg/L). For these species, the three alternative techniques overestimated MICs and resistance, and did not meet performance criteria. For mecillinam testing of Enterobacterales isolates, apart from P. mirabilis, ETEST® nearly fulfilled ISO requirements, and CA rates were acceptable for disc diffusion. ISO criteria were reached for C. koseri and E. coli testing with VITEK®2, apart from too high rates of very major errors. For P. mirabilis, performances were unacceptable, whatever the routine method used. CONCLUSIONS: Commercially available tests may serve as alternatives to agar dilution to assess fosfomycin (oral) and mecillinam susceptibility of Enterobacterales urinary isolates, with important interspecies variabilities. Additional studies comprising more fosfomycin- and mecillinam-resistant isolates are needed to strengthen our conclusions.

Antimicrobial susceptibility profiles of anaerobic bacteria, isolated from human clinical specimens, within different European and surrounding countries. A joint ESGAI study.

OBJECTIVES: Studies on the antimicrobial susceptibility profile of anaerobic bacteria are underrepresented in the literature. Within this study we aim to give an extensive overview of the differences in antimicrobial susceptibility profiles between different European and surrounding countries. METHODS: Minimal inhibitory concentration (MIC) data of different antibiotics were collected from 10 participating laboratories, representing an equal number of countries. All MIC's were determined using Etest, according to the protocol used by the participating laboratory. Anaerobic genera represented by at least 10 clinical isolates were included in the study. RESULTS: Each country tested different antibiotics, sometimes depending on the kind of infection and/or the anaerobic species isolated. All countries tested clindamycin and metronidazole. Resistance rates differed remarkably between the different countries. Especially in Kuwait, resistance was high for all tested antibiotics. Unexpected metronidazole resistance was observed for Finegoldia magna isolates, Peptoniphilus isolates and Eggerthella lenta isolates. CONCLUSIONS: Due to the extensive differences in antimicrobial susceptibility profile of anaerobic bacteria isolated within different countries, we strongly recommend to perform this kind of study on a regular basis.

FORTE - a multipurpose high-vacuum diffractometer for tender X-ray diffraction and spectroscopy at the SIRIUS beamline of Synchrotron SOLEIL.

A new high-vacuum multipurpose diffractometer (called FORTE from the French acronyms of the project) has recently been installed at the tender/hard X-ray SIRIUS beamline of Synchrotron SOLEIL, France. The geometry chosen allows one to work either in the classical Eulerian four-circle geometry for bulk X-ray diffraction (XRD) or in the z-axis geometry for surface XRD. The diffractometer nicely fits the characteristics of the SIRIUS beamline, optimized to work in the 1.1-4.5 keV range, and allows one to perform unprecedented diffraction anomalous fine structure (DAFS) experiments in the tender X-ray region, also around non-specular reflections, covering a large reciprocal-space volume. Installation of an X-ray fluorescence detector on a dedicated flange allows simultaneous DAFS and X-ray absorption (XAS) measurements. The access to the tender X-ray region paves the way to resonant investigations around the L-edges of second-row transition elements which are constituents of functional oxide materials. It also enables access to several edges of interest for semiconductors. Finally, the control architecture based on synchronized Delta Tau units opens up exciting perspectives for improvement of the mechanical sphere of confusion.

An impulsive modelling framework of fire occurrence in a size-structured model of tree-grass interactions for savanna ecosystems.

Fires and mean annual rainfall are major factors that regulate woody and grassy biomasses in savanna ecosystems. Within the savanna biome, conditions of long-lasting coexistence of trees and grasses have been often studied using continuous-time modelling of tree-grass competition. In these studies, fire is a time-continuous forcing while the relationship between woody plant size and fire-sensitivity is not systematically considered. In this paper, we propose a new mathematical framework to model tree-grass interactions that takes into account both the impulsive nature of fire occurrence and size-dependent fire sensitivity (via two classes of woody plants). We carry out a qualitative analysis that highlights ecological thresholds and bifurcation parameters that shape the dynamics of the savanna-like systems within the main ecological zones. Through a qualitative analysis, we show that the impulsive modelling of fire occurrences leads to more diverse behaviors including cases of grassland, savanna and forest tristability and a more realistic array of solutions than the analogous time-continuous fire models. Numerical simulations are carried out with respect to the three main ecological contexts (moist, mesic, semi-arid) to illustrate the theoretical results and to support a discussion about the bifurcation parameters and the advantages of the model.

About contamination by sterile females and residual male fertility on the effectiveness of the sterile insect technique. Impact on disease vector control and disease control.

The sterile insect technique (SIT) is a technique to control pests and vectors of diseases by releasing mainly sterile males. Several challenges need to be solved before large-scale field application in order to guarantee its success. In this paper we intend to focus on two important issues: residual fertility in released (sterile) males and contamination of each release by sterile females. Indeed, sterile males are never 100% sterile, that is there is always a small proportion, ɛ, of fertile males (sperm of) within the sterile males population. Among the sterile insects that are released, a certain proportion, ϵF, of them are sterile females due to imperfect mechanical sex-separation technique. This can be particularly problematic when arthropod viruses are circulating, because mosquito females, even sterile, are vectors of diseases. Various upper bound values are given in the entomological literature for ϵF and ɛ without clear explanations. In this work, we aim to show that these values are related to the biological parameters of the targeted vector, the sterile insects release rate, and the epidemiological parameters of a vector-borne disease, like Dengue. We extend results studied separately in Aronna and Dumont (2020), Dumont and Yatat-Djeumen (2022). To study the impact of both issues, we develop and study a SIT-entomological-epidemiological mathematical model, with application to Dengue. Qualitative analysis of the model is carried out to highlight threshold values that shape the overall dynamics of the system. We show that vector elimination is possible only when Nɛ<1, where N is the basic-offspring number related to the targeted wild population. To ensure the success of SIT control, we recommend that the issue of residual fertility be addressed as a priority and then that contamination by sterile females be minimized with each release.

Direct evidence of Fe(2+)-Fe3+ charge ordering in the ferrimagnetic hematite-ilmenite Fe(1.35)Ti(0.65)O(3-δ) thin films.

In this Letter we highlight direct experimental evidence of Fe(2+)-Fe3+ charge ordering at room temperature in hematite-ilmenite Fe(1.35)Ti(0.65)O(3-δ) epitaxial thin films grown by pulsed laser deposition, using aberration-corrected scanning transmission electron microscopy coupled to high-resolution energy electron-loss spectroscopy. These advanced spectromicroscopy techniques demonstrate a strong modulation of the Fe2+ valence state along the c axis. Density functional theory calculations provide crucial information on the key role of oxygen vacancies in the observed charge distributions. Their presence at significant levels leads to the localization of extra electrons onto reduced Fe2+ sites, while Ti remains solely +4. The magnetic and transport properties of these films are reviewed in the light of the present results regarding their ferrimagnetic character correlated with the Fe2+ modulation and their semiconducting behavior interpreted by an Efros-Shklovskii variable-range hopping conduction regime via Fe2+ and Fe3+ centers. The experimental evidence of only one type of mixed valence state, i.e., Fe2+ and Fe3+, in the Fe(2-x)Ti(x)O(3-δ) system will thus help to interpret further the origin of its geomagnetic properties and to illuminate fundamental issues regarding its spintronic potential.

Mathematical studies on the sterile insect technique for the Chikungunya disease and Aedes albopictus.

Chikungunya is an arthropod-borne disease caused by the Asian tiger mosquito, Aedes albopictus. It can be an important burden to public health and a great cause of morbidity and, sometimes, mortality. Understanding if and when disease control measures should be taken is key to curtail its spread. Dumont and Chiroleu (Math Biosc Eng 7(2):315-348, 2010) showed that the use of chemical control tools such as adulticide and larvicide, and mechanical control, which consists of reducing the breeding sites, would have been useful to control the explosive 2006 epidemic in Réunion Island. Despite this, chemical control tools cannot be of long-time use, because they can induce mosquito resistance, and are detrimental to the biodiversity. It is therefore necessary to develop and test new control tools that are more sustainable, with the same efficacy (if possible). Mathematical models of sterile insect technique (SIT) to prevent, reduce, eliminate or stop an epidemic of Chikungunya are formulated and analysed. In particular, we propose a new model that considers pulsed periodic releases, which leads to a hybrid dynamical system. This pulsed SIT model is coupled with the human population at different epidemiological states in order to assess its efficacy. Numerical simulations for the pulsed SIT, using an appropriate numerical scheme are provided. Analytical and numerical results indicate that pulsed SIT with small and frequent releases can be an alternative to chemical control tools, but only if it is used or applied early after the beginning of the epidemic or as a preventive tool.

Sterile insect technique with accidental releases of sterile females. Impact on mosquito-borne diseases control when viruses are circulating.

The sterile insect technique (SIT) is a technique to control some vectors of diseases by releasing sterile males. However, during these releases, sterilized females can be (accidentally) released and since only females are vectors of diseases, it is important to study their impact when arthropod viruses are circulating. To that aim, we develop and study an entomological-epidemiological model, considering either permanent or periodic releases. Qualitative analyses of the continuous and periodic models are conducted. We highlight a critical sterile males release rate, ΛMcrit, above which the control of wild population is always effective, using massive releases. Estimating the basic reproduction number of the epidemiological model, R02, we show that if it is above a certain threshold, R0,∗2, that depends on the basic offspring number, N, and the release rate of sterile females, the epidemiological risk can only be controlled using (very) massive releases. Otherwise, we can estimate the basic reproduction number of the SIT epidemiological model, R0,SIT2, that shapes the stability property of the (periodic) disease-free equilibrium. We show that it might be possible to take R0,SIT2 below 1 using non-massive, but large enough, releases. However, practically, it seems more efficient to consider massive releases, followed by small releases once the vector population is small enough. In addition to SIT, we also recommend mechanical control, i.e. the reduction of breeding sites, that greatly improves the efficacy of SIT, in terms of duration or size of the releases. Our results reveal that outside an epidemic period, the release of sterile females is not an issue, as long as the sterile males release rate is greater than ΛMcrit. Within an epidemic period, we show that sterile females releases do not really impact the SIT efficiency, as long as the release rate, ΛF, is lower than a critical value, ΛFcrit, that depends on the mosquito and epidemiological threshold parameters, N, and R02. To illustrate numerically our theoretical results, we consider Dengue parameters. We estimate all thresholds and also the effective reproduction number, Reff2, and highlight the importance of early permanent or periodic SIT control to prevent or mitigate the risk of a Dengue epidemic, with and without sterile females releases.

On a temporal model for the Chikungunya disease: modeling, theory and numerics.

Reunion Island faced two episodes of Chikungunya, a vector-borne disease, in 2005 and in 2006. The latter was of unprecedented magnitude: one third of the population was infected. Until the severe episode of 2006, our knowledge of Chikungunya was very limited. The principal aim of our study is to propose a model, including human and mosquito compartments, that is associated to the time course of the first epidemic of Chikungunya. By computing the basic reproduction number R(0), we show there exists a disease-free equilibrium that is locally asymptotically stable if the basic reproduction number is less than 1. Moreover, we give a necessary condition for global asymptotic stability of the disease-free equilibrium. Then, we propose a numerical scheme that is qualitatively stable and present several simulations as well as numerical estimates of the basic reproduction number for some cities of Reunion Island. For the episode of 2005, R(0) was less than one, which partly explains why no outbreak appeared. Using recent entomological results, we investigate links between the episode of 2005 and the outbreak of 2006. Finally, our work shows that R(0) varied from place to place on the island, indicating that quick and focused interventions, like the destruction of breeding sites, may be effective for controlling the disease.

Investigation of pre-XDR Beijing Mycobacterium tuberculosis transmission to a healthcare worker in France, 2016.

A case of occupational contamination of a healthcare worker by a pre-extensively drug-resistant (pre-XDR) Beijing strain of Mycobacterium tuberculosis at the University Hospital of Montpellier, France is reported. The index case was identified using genetic fingerprinting of isolates. This report underscores the risk of healthcare-associated contamination by pre-XDR tuberculosis (TB) in low-incidence countries and the importance of molecular tools for TB care. It also calls for increased vigilance in the management of multi-drug-resistant/XDR TB patients.

Human behaviors: A threat to mosquito control?

In this work, we consider a simple theoretical model that enables us to take into account private human decisions that may interfere with public mosquito control. The model reflects the trade-off between perceived costs and observed efficacy. Our theoretical results emphasize that households may reduce their protective behavior in response to mechanical elimination techniques piloted by a public agent, leading to an increase in the total number of mosquitoes in the surrounding environment and generating a barrier for vector-borne diseases control. Our study is sufficiently generic to be applied to different arboviral diseases. It also shows that vector-control models and strategies have to take into account individual behaviors.

Genome Sequences of Multiresistant Staphylococcus capitis Pulsotype NRCS-A and Methicillin-Susceptible S. capitis Pulsotype NRCS-C.

Here, we report the draft genome sequences of methicillin-susceptible Staphylococcus captis pulsotype NCRS-C (CR02 strain) and multiresistant Staphylococcus captis pulsotype NCRS-A (CR07 strain).

The molecular pharmacology of CGRP and related peptide receptor subtypes.

Calcitonin gene-related peptides (alpha and beta isoforms), better known as CGRPalpha and CGRPbeta, were isolated twenty years ago. In fact, these were the first peptides to be characterized using a molecular cloning strategy, which is not the traditional approach of biochemical extraction and purification. Paradoxically, progress in the characterization of CGRP receptor subtypes has been extremely slow as a result of difficulties in their cloning and the lack of selective receptor subtype agonists and antagonists. However, exciting progress has been made overthe pasttwo years and is briefly reviewed here.

Evidence that the inhibition of luteinizing hormone secretion exerted by central administration of neuropeptide Y (NPY) in the rat is predominantly mediated by the NPY-Y5 receptor subtype.

A number of studies have indicated that neuropeptide Y (NPY) is a central regulator of the gonadotropic axis, and the Y1 receptor was initially suggested to be implicated. As at least five different NPY receptor subtypes have now been characterized, the aim of the present study was to reinvestigate the pharmacological profile of the receptor(s) mediating the inhibitory action of NPY on LH secretion by using a panel of NPY analogs with different selectivity toward the five NPY receptor subtypes. When given intracerebroventricularly (icv) to castrated rats, a bolus injection of native NPY (0.7-2.3 nmol) dose-dependently decreased plasma LH. Peptide YY (PYY; 2.3 nmol) was as potent as NPY, suggesting that the Y3 receptor is not implicated. Confirming previous data, the mixed Y1, Y4, and Y5 agonist [Leu31,Pro34]NPY (0.7-2.3 nmol) inhibited LH release with potency and efficacy equal to those of NPY. Neither the selective Y2 agonist C2-NPY (2.3 nmol) nor the selective Y4 agonist rat pancreatic polypeptide affected plasma LH, excluding Y2 and Y4 subtypes for the action of NPY on LH secretion. The mixed Y4-Y5 agonist human pancreatic polypeptide (0.7-7 nmol) as well as the mixed Y2-Y5 agonist PYY3-36 (0.7-7 nmol) that displayed very low affinity for the Y1 receptor, thus practically representing selective Y5 agonists in this system, decreased plasma LH with potency and efficacy similar to those of NPY, indicating that the Y5 receptor is mainly involved in this inhibitory action of NPY on LH secretion. [D-Trp32]NPY, a selective, but weak, Y5 agonist, also inhibited plasma LH at a dose of 7 nmol. Furthermore, the inhibitory action of NPY (0.7 nmol) on LH secretion could be fully prevented, in a dose-dependent manner (6-100 microg, icv), by a nonpeptidic Y5 receptor antagonist. This antagonist (60 microg, icv) also inhibited the stimulatory action of NPY (0.7 nmol) on food intake. The selectivity of PYY3-36, human PP, [D-Trp32]NPY, and the Y5 antagonist for the Y5 receptor subtype was further confirmed by their ability to inhibit the specific [125I][Leu31,Pro34]PYY binding to rat brain membrane homogenates in the presence of the Y1 receptor antagonist BIBP3226, a binding assay system that was described as being highly specific for Y5-like receptors. With the exception of [D-Trp32]NPY, all analogs able to inhibit LH secretion were also able to stimulate food intake. Taken together, these results indicate that the Y5 receptor is involved in the negative control by NPY of the gonadotropic axis.

Species differences in the expression and distribution of the neuropeptide Y Y1, Y2, Y4, and Y5 receptors in rodents, guinea pig, and primates brains.

The respective distribution of neuropeptide Y Y1, Y2, Y4, and Y5 receptor subtypes was investigated in rodents (rat and mouse), guinea pig, and primates (marmoset and vervet monkeys and human) brains, representing three orders of mammals. [125I] [Leu31,Pro34]PYY (total Y1-like; Y1, Y4, and Y5) and [125I]PYY3-36 (total Y2-like; Y2 and possibly Y5) binding sites were discretely but similarly distributed in the rat and mouse brain, each having its unique pattern. In contrast, surprisingly low levels of [125I]PYY3-36 binding sites were found in the guinea pig brain including in the hippocampal formation. [125I][Leu31,Pro34]PYY/BIBP3226-insensitive binding sites (Y5-like) were found in different areas of the rat and guinea pig brains. The primate brains also revealed a different distribution binding profile for these various NPY receptor subtypes. Although the human and vervet brains contained very low amounts of [125I][Leu31,Pro34]PYY sites (Y1-like) in most brain regions, except for the dentate gyrus of the hippocampus, the marmoset brain contains significant amounts of both [125I][Leu31,Pro34]PYY (Y1-like) and [125I]PYY3-36 (Y2-like) binding sites. Additionally, [125I][Leu31,Pro34]PYY/BIBP3226-insensitive binding sites were not clearly detected in the vervet and human brains. On the other hand, Y5-like binding sites were observed in few regions of the marmoset brain. Finally, [125I]hPP (Y4/Y5-like) were very discretely distributed in the rat brain, being concentrated in the paraventricular nucleus of the hypothalamus and the interpeduncular nucleus. The marmoset brain is apparently not enriched with specific [125I]hPP sites. Taken together, these data show that significant species differences exist in the level of expression and distribution of various NPY receptor subtypes in the mammalian brain.

Blood pressure-independent effect of angiotensin AT1 receptor blockade on renal endothelin-1 production in hypertensive uremic rats.

OBJECTIVE: We recently reported that treatment of uremic rats with reduced renal mass with the angiotensin II (Ang II) subtype 1 receptor (AT1) antagonist losartan reduces endothelin-1 (ET-1) levels in blood vessels and in glomeruli. Although this suggests an important role for Ang II in the modulation of ET-1 production, the concomitant decrease in blood pressure may also be involved. The present study was designed to investigate whether the modulation of ET-1 production in uremic rats is related to tissue-specific effects of AT1 receptor blockade or to the antihypertensive effect of losartan. DESIGN: One week after renal mass reduction, uremic rats were treated with the conventional triple therapy (TRx) [reserpine (5 mg/l), hydralazine (80 mg/l) and hydrochlorothiazide (25 mg/l)] or losartan (20 mg/kg per day) for 6 weeks. Immunoreactive-ET-1 (ir-ET-1) levels in plasma and urine, as well as in vascular and renal tissues were measured by a specific radioimmunoassay after sample extraction and purification. RESULTS: Before treatment, systolic blood pressure was significantly higher in uremic animals compared to sham-operated controls (165+/-4 versus 123+/-2 mmHg, respectively; P < 0.01). Treatment with the TRx or with losartan normalized systolic blood pressure in uremic rats, whereas it was further increased in untreated uremic animals. At week 6, serum creatinine, proteinuria and urinary ET-1 and transforming growth factor-beta1 (TGF-beta1) excretion, as well as vascular and glomerular ET-1 content were increased in uremic rats compared to the controls (P < 0.01). Treatment of uremic rats with the TRx or with losartan reduced ET-1 content in the thoracic aorta and the mesenteric arterial bed (P < 0.01). However, losartan, but not the TRx, significantly attenuated the rise of serum creatinine, proteinuria and urinary ET-1 and TGF-beta1 excretion, as well as ET-1 content in glomeruli of uremic rats. Compared with the controls, renal preproET-1 mRNA expression was also significantly higher in uremic rats. Treatment of uremic rats with losartan prevented renal preproET-1 mRNA overexpression, indicating that changes in glomerular ET-1 content and urinary ET-1 excretion were related to modulation of renal ET-1 production. CONCLUSIONS: These findings indicate that the effect of losartan on ET-1 production in peripheral blood vessels may be mediated, in part, by the reduction of blood pressure. In contrast, the reduction of renal ET-1 production is mediated by tissue-specific effects of AT1 receptor blockade, and may contribute to the renal protective effects of losartan.

Structure-activity study of hCGRP8-37, a calcitonin gene-related peptide receptor antagonist.

A structure-activity study was carried out to determine the importance of the N-terminal amino acids of hCGRP8-37 in binding and antagonistic activity to CGRP receptors. Therefore, fragments of hCGRP8-37 as well as analogs obtained by the replacement of residues 9-12 by L-alanine were synthesized by solid-phase peptide synthesis, using BOP as a coupling reagent. The affinities of the peptides to CGRP receptors were evaluated in the rat brain, guinea pig atrium, and guinea pig vas deferens membrane preparations. Their antagonistic activities were measured in the guinea pig atria and rat vas deferens bioassays. The pharmacological characterization showed that arginine-11 and leucine-12 play a crucial role for the affinity of hCGRP8-37. Interestingly, it was observed that [Ala11]hCGRP8-37 was able to potentiate the twitch response of the electrically stimulated rat vas deferens. On the other hand, the substantial antagonistic potencies of analogs [Ala9]-, [Ala10]-, and [Ala12]hCGRP8-37, as compared to those of the fragments hCGRP10-37, hCGRP11-37, and hCGRP12-37, suggest that the side chains of Thr-9, His-10, and Leu-12 assume mainly a structural role. Accordingly, the conformational characterization of these peptides by circular dichroism spectroscopy revealed that the residues 9-12 are important for the integrity of the amphiphilic alpha-helix of hCGRP8-37.

Characterization of neuropeptide Y receptor subtypes in the normal human brain, including the hypothalamus.

The aim of the present study was to investigate the existence and distribution of neuropeptide Y receptor subtypes in various regions of the normal human brain using the peptide YY derivative receptor probes, [125I][Leu31,Pro34]polypeptide YY/Y1 and [125I]polypeptide YY(3-36)/Y2, in addition to the non-selective ligand [125I]polypeptide YY. Membrane binding assays performed with post mortem frontal cortex homogenates revealed that [125I]polypeptide YY and [125I]polypeptide YY(3-36) bound in a time- and protein concentration-dependent manner. Very low amounts of specific [125I][Leu31,Pro34]polypeptide YY binding could be detected even in the presence of high amounts of protein, contrasting with results obtained with [125I]polypeptide YY and [125I]polypeptide YY(3-36), a preferential Y2 receptor probe. Analysis of saturation isotherms revealed that [125I]polypeptide YY(3-36) bound to a single class of high-affinity sites (0.5-2 nM). Significantly higher binding capacities were evident for [125I]polypeptide YY(3-36) as compared to [125I][Leu31,Pro34]polypeptide YY, suggesting that the human frontal cortex, in contrast to the rat, is mostly enriched with Y2 receptors. Ligand selectivity profile confirmed the hypothesis that polypeptide YY(3-36), neuropeptide Y and polypeptide YY but not the [Leu31,Pro34] derivatives are potent competitors of [125I]polypeptide YY and [125I]polypeptide YY(3-36) binding sites. Autoradiographic studies demonstrated further that cortical areas, as well as most other regions of the human brain, are particularly enriched with Y2/[125I]polypeptide YY(3-36) sites, while only low to very low amounts of Y1 binding were detected except in the dentate gyrus of the hippocampal formation. In the human hypothalamus, a preponderance of Y2 binding sites was also noted. Taken together, these results clearly establish that the distribution of the Y1 and Y2 receptor subtypes in human is different from the rodent brain, the Y2 subtype being most abundant in the human brain.

Decreased levels of neuropeptide Y(5) receptor binding sites in two experimental models of epilepsy.

It has been suggested that the anticonvulsant effects of neuropeptide Y (NPY) could be mediated by the activation of Y(2) and/or Y(5) receptors. NPY Y(1) receptor levels are known to decrease and Y(2) to increase in rat models of epilepsy. By using an autoradiographic approach, we investigated whether epilepsy models (kainic acid and kindling) are also associated with changes in Y(5) receptors. Compared with naive controls, [125I][Leu(31), Pro(34)]PYY/BIBP3226-insensitive (Y(5)) binding sites in the hippocampus (strata oriens and radiatum of CA3 and CA1) and in the neocortex (superficial layers) were unchanged in sham-stimulated rats, but reduced by approximately 50% in kindled rats (seven days after the last stimulus evokes seizure), and further reduced (to approximately -90%) 1h after a kindled seizure. Additionally, Y(5) receptor binding sites in the hippocampus and in the neocortex were unchanged 6h after kainic acid injection, but were highly reduced at 12 and 24h. No changes in Y(5) binding levels were found in the dentate gyrus and the pyramidal cell layer of the hippocampus. The present data suggest that changes in Y(5) receptor levels occur in epilepsy models. These changes may play a role in seizure expression and/or in the maintenance of kindling hyperexcitability.

[(125)I]-GR231118: a high affinity radioligand to investigate neuropeptide Y Y(1) and Y(4) receptors.

GR231118 (also known as 1229U91 and GW1229), a purported Y(1) antagonist and Y(4) agonist was radiolabelled using the chloramine T method. [(125)I]-GR231118 binding reached equilibrium within 10 min at room temperature and remained stable for at least 4 h. Saturation binding experiments showed that [(125)I]-GR231118 binds with very high affinity (K(d) of 0.09 - 0.24 nM) in transfected HEK293 cells with the rat Y(1) and Y(4) receptor cDNA and in rat brain membrane homogenates. No specific binding sites could be detected in HEK293 cells transfected with the rat Y(2) or Y(5) receptor cDNA demonstrating the absence of significant affinity of GR231118 for these two receptor classes. Competition binding experiments revealed that specific [(125)I]-GR231118 binding in rat brain homogenates is most similar to that observed in HEK293 cells transfected with the rat Y(1), but not rat Y(4), receptor cDNA. Autoradiographic studies demonstrated that [(125)I]-GR231118 binding sites were fully inhibited by the Y(1) antagonist BIBO3304 in most areas of the rat brain. Interestingly, high percentage of [(125)I]-GR231118/BIBO3304-insensitive binding sites were detected in few areas. These [(125)I]-GR231118/BIBO3304-insensitive binding sites likely represent labelling to the Y(4) receptor subtype. In summary, [(125)I]-GR231118 is a new radiolabelled probe to investigate the Y(1) and Y(4) receptors; its major advantage being its high affinity. Using highly selective Y(1) antagonists such as BIBO3304 or BIBP3226 it is possible to block the binding of [(125)I]-GR231118 to the Y(1) receptor allowing for the characterization and visualization of the purported Y(4) subtype. British Journal of Pharmacology (2000) 129, 37 - 46