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Abstract: Serotonin (5-HT) is known to play a role in the suppression of the lordosis response in males. We have previously shown that there is a sex difference in the density of 5-HT immunoreactive (5-HT-ir) fibers in the ventrolateral division of the adult ventromedial nucleus of the hypothalamus (VMNvl) and that neonatal administration of estradiol (E2) increases 5-HT-ir in the female VMNvl to male-typical levels. Here we demonstrate that postnatal administration of the ERα agonist 1,3,5-tris(4-Hydroxyphenyl)-4-propyl-1H-pyrazole (PPT), but not the ERβ agonist diarylpropionitrile (DPN), also masculinizes 5-HT-ir in the female VMNvl, suggesting a mechanistic role for ERα in this process. Sexual receptivity, as ascertained by the lordosis quotient, was unaffected by either PPT or DPN treatment but nearly abolished by estradiol benzoate (EB), a synthetic estrogen with high affinity for both ERα and ERβ. Collectively, these observations show that postnatal estrogens increase the density of 5-HT projections to the VMNvl via an ERα dependent mechanism, but that this increased inhibitory input is not sufficient to suppress the lordosis response. [Copyright &y& Elsevier]

Objective. Neuropathic pain associated with postherpetic neuralgia (PHN) and painful diabetic peripheral neuropathy (DPN) can be intractable and may not respond to commonly used treatments, such as tricyclic antidepressants (TCAs) and opioids. This long-term, open-label study was a preliminary evaluation of pregabalin for patients whose pain had been judged refractory to other treatments for neuropathic pain. Design. Patients had previously participated in double-blind, placebo-controlled, randomized trials of pregabalin in DPN and PHN. They had moderate to severe neuropathic pain despite treatment with gabapentin, a TCA, and a third medication (e.g., other anticonvulsants, opioid, selective serotonin reuptake inhibitor, tramadol). Flexible-dosage pregabalin 150–600 mg/day was taken for 3-month periods followed by 3- to 28-day pregabalin “drug holidays,” with an analysis up to 15 months (five treatment cycles). Pain intensity was measured using the visual analog scale of the Short-Form McGill Pain Questionnaire. Results. In total, 81 patients were included in this analysis. Pregabalin 150–600 mg/day was associated with clinically meaningful and sustained pain reduction during each treatment cycle. During pregabalin “drug holidays,” pain quickly returned to baseline levels; it was reduced again when pregabalin was reinstated. Conclusions. These results suggest that pregabalin may be beneficial in patients with neuropathic pain who have had an unsatisfactory response to treatment with other medications. [ABSTRACT FROM AUTHOR]

Abstract: It is well established that estrogen administration during neonatal development can advance pubertal onset and prevent the maintenance of regular estrous cycles in female rats. This treatment paradigm also eliminates the preovulatory rise of gonadotropin releasing hormone (GnRH). It remains unclear, however, through which of the two primary forms of the estrogen receptor (ERα or ERβ) this effect is mediated. It is also unclear whether endocrine disrupting compounds (EDCs) can produce similar effects. Here we compared the effect of neonatal exposure to estradiol benzoate (EB), the ERα specific agonist 1,3,5-tris(4-hydroxyphenyl)-4-propyl-1H-pyrazole (PPT), the ERβ specific agonist diarylpropionitrile (DPN) and the naturally occurring EDCs genistein (GEN) and equol (EQ) on pubertal onset, estrous cyclicity, GnRH activation, and kisspeptin content in the anteroventral periventricular (AVPV) and arcuate (ARC) nuclei. Vaginal opening was significantly advanced by EB and GEN. By 10 weeks post-puberty, irregular estrous cycles were observed in all groups except the control group. GnRH activation, as measured by the percentage of immunopositive GnRH neurons that were also immunopositive for Fos, was significantly lower in all treatment groups except the DPN group compared to the control group. GnRH activation was absent in the PPT group. These data suggest that neonatal exposure to EDCs can suppress GnRH activity in adulthood, and that ERα plays a pivotal role in this process. Kisspeptins (KISS) have recently been characterized to be potent stimulators of GnRH secretion. Therefore we quantified the density of KISS immunolabeled fibers in the AVPV and ARC. In the AVPV, KISS fiber density was significantly lower in the EB and GEN groups compared to the control group but only in the EB and PPT groups in the ARC. The data suggest that decreased stimulation of GnRH neurons by KISS could be a mechanism by which EDCs can impair female reproductive function. [Copyright &y& Elsevier]

Previous studies have shown the oestradiol modulation of brain N-methyl-d-aspartate (NMDA) receptors composed of the NR1/2B subunits. The contribution of oestrogen receptor subtypes in this oestradiol modulation of NMDA receptors and its subunits is not known. The following experiments investigated whether an oestrogenic receptor subtype is involved in the oestradiol effect on NMDA receptor specific binding and subunit mRNA levels. Ovariectomised Sprague-Dawley rats were treated 2 days after ovariectomy for 2 weeks with 17β-oestradiol, an agonist for oestrogen receptor (ER)α 4,4′,4″-(4-propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (PPT) or an agonist for ERβ 2,3-bis(4-hydroxyphenyl)-propionitrile (DPN) and compared with control vehicle-treated ovariectomised and intact rats. Uterus weights, used as a peripheral measure of oestrogenic activity, decreased after ovariectomy and increased by oestradiol and PPT but not DPN treatment. In the hippocampal CA1 oriens and CA1 radiatum, [3H]Ro 25-6981 specific binding, a NMDA/NR2B ligand, was decreased in ovariectomised compared to intact rats and this was prevented by 17β-oestradiol or PPT but not DPN treatments; a similar pattern was observed in the CA2/3 and dentate gyrus but did not reach statistical significance. In situ hybridisation of the mRNA of the NMDA/2B subunit in the hippocampus CA1, CA2/3 and dentate gyrus showed a decrease in ovariectomised rats compared to controls and this was also prevented by 17β-oestradiol and PPT but not DPN treatments. In cingulate and prefrontal cortices, ovariectomy increased [3H]Ro 25-6981 specific binding compared to intact controls, which was corrected by 17β-oestradiol treatment but neither by PPT, nor DPN. In the cortical regions, the lack of effect of the ERα or ERβ agonist whereas 17β-oestradiol was active, suggesting that the oestradiol modulation of cortical NMDA receptors requires both ERs or that this modulation does not involve ERs. In the hippocampus, the results obtained suggest an oestrogenic genomic modulation of NMDA receptors containing the NR2B subunit, implicating an ERα. [ABSTRACT FROM AUTHOR]

Previous studies have shown that high dose 17β-estradiol (10-5 M) has a G2/M blocking effect in SW-13 human adrenal carcinoma cultures and strongly enhances apoptosis. To examine the differential effects of estrogen α and β-receptors in this system, we incubated SW-13 cells with specific α- and β-estrogen receptor agonists, PPT [4,4′,4′′-(propyl-[1H]-pyrazole-1,3,5-triyl) trisphenol] and DPN [2,3-bis (4-hydroxyphenyl) propionitrile], respectively (each at 10-5 M). Flow cytometry was used to analyze the percentages of cells in various phases of the cell cycle [sub-G1 (apoptosis), G1, S, and G2/M] in each experimental condition. Exposure to 17 β-estradiol for 48 hours increased apoptosis more than 5-fold (from 3.6�0.5 to 20�2.2% of cells; p<0.01). The α-estrogen agonist PPT had a similar effect, increasing apoptosis to 22�1.7% (p<0.01), but the β-agonist DPN caused no change (3.6�0.5 vs. 3.9�0.8%). While estrogen and the α-estrogen agonist decrease apoptosis in this system, both of these compounds decreased the percentage of cells in G1 (from 59�1.4% for control to 34�2.3% for estrogen and 40�2.0% for PPT; p<0.01 for both agents relative to control); the β-agonist again had no effect. Estrogen was also found to block the cell cycle in G2/M, increasing it from 15�0.4 to 21�1.0% of cells (p<0.01), but neither the α- nor β-estrogen agonists had any effect at this point in the cell cycle, indicating that the influence of estrogen was not likely to be either α- or β-receptor mediated. There was no apparent effect of any of these agents on DNA synthesis, as indicated by unchanged percentages of cells in S phase. These studies suggest that induction of apoptosis by estrogen in SW-13 human adrenal cortical carcinoma cultures is mediated by the α-receptor, but the G2/M blocking effect of estrogen is not likely to be related to either α or β mechanisms. [ABSTRACT FROM AUTHOR]

Abstract: Endocrine active compounds (EACs) have been shown to influence a number of reproductive endpoints but less is known about how they might affect other hormone dependent behaviors including anxiety and aggression. Recent evidence suggests that these effects may be mediated through the beta form of the estrogen receptor (ERβ). Using male Long Evans rats, we sought to determine how neonatal exposure to EACs affects anxiety and aggression in adulthood. Anxiety was assessed using the elevated plus maze and aggression was assessed 8weeks later using the resident intruder test. To gain insight into which ER subtype (ERα vs ERβ) might be mediating these effects we used agonists specific for ERα (1,3,5-tris(4-Hydroxyphenyl)-4-propyl-1H-pyrazole (PPT)) or ERβ (Diarylpropionitrile (DPN)) as additional treatment groups. For these experiments the synthetic EAC bisphenol-A (BPA) and the phytoestrogen metabolite equol (EQ) were used. Male neonates were injected with either 0.05 ml sesame oil (control), 50 μg estradiol benzoate (EB), 1 mg/kg DPN, 1 mg/kg PPT, 50 μg/kg BPA, or 10 mg/kg EQ daily for 4 days beginning on the day of birth (PND 0). Compared to the oil treated controls, significantly fewer open arm entries were made by the males neonatally treated with DPN, EQ, or BPA. The DPN and EQ treated males were also more aggressive compared to the controls. These findings suggest that neonatal exposure to EACs with agonistic activity on ERβ may influence affective behavior in adulthood, including anxiety and aggression. [Copyright &y& Elsevier]

Abstract: Estrogen has been shown to attenuate the inflammatory response following injury or lipopolysaccharide treatment in several organ systems. Estrogen''s actions are transduced through two estrogen receptor sub-types, estrogen receptor (ER) -alpha and estrogen receptor-beta, whose actions may be overlapping or independent of each other. The present study examined the effects of ERα- and ERβ-specific ligands in regulating the inflammatory response in primary astrocyte cultures. Pre-treatment with 17β-estradiol (ERα/ERβ agonist), HPTE (ERα agonist/ERβ antagonist) and DPN (ERβ agonist) led to attenuation of IL-1β, TNFα, and MMP-9 in astrocyte media derived from young adult (3-4 mos.) and reproductive senescent female (9-11 mos., acyclic) astrocyte cultures, while pretreatment with PPT (ERα agonist) attenuated IL-1β (but not MMP-9) in both young and senescent-derived astrocyte cultures. Our previous work determined that 17β-estradiol was unable to attenuate the LPS-induced increase in IL-1β in olfactory bulb primary microglial cultures derived from either young adult or reproductive senescent females. In young adult-derived microglial cultures, the LPS-induced increase in IL-1β was not attenuated by pre-treatment with 17β-estradiol, PPT or HPTE. Interestingly, the ERβ agonist, DPN significantly decreased IL-1β following LPS treatment in young adult-derived microglia. Thus while both microglia and astrocytes synthesize and release inflammatory mediators, the present data shows that compounds which bind ERβ are more effective in attenuating proinflammatory cytokines in both cell types and may therefore be a more effective agent for future therapeutic use. [Copyright &y& Elsevier]

Abstract: Clinical and experimental studies show a modulatory role of estrogens in the brain and suggest their beneficial action in mental and neurodegenerative diseases. The estrogen receptors ERα and ERβ are present in the brain and their targeting could bring selectivity and reduced risk of cancer. Implication of ERs in the effect of estradiol on dopamine, opiate and glutamate neurotransmission is reviewed. The ERα agonist, PPT, is shown as estradiol to modulate hippocampal NMDA receptors and AMPA receptors in cortex and striatum of ovariectomized rats whereas the ERβ agonist DPN is inactive. Striatal DPN activity suggests implication of ERβ in estradiol modulation of D2 receptors and transporters in ovariectomized rats and is supported by the lack of effect of estradiol in ERβ knockout (ERKOβ) mice. Both ERα and ERβ agonists modulate striatal preproenkephalin (PPE) gene expression in ovariectomized rats. In male mice PPT protects against MPTP toxicity to striatal dopamine; this implicates Akt/GSK3β signaling and the apoptotic regulators Bcl2 and Bad. This suggests a role for ERα in striatal dopamine neuroprotection. ERKOα mice are more susceptible to MPTP toxicity and not protected by estradiol; differences in ERKOβ mice are subtler. These results suggest therapeutic potential for the brain of ER specific agonists. [Copyright &y& Elsevier]

Abstract: Selective estrogen receptor (ER) agonists can indicate which receptor subtypes are implicated in neuroprotection. This study investigated the contribution of ERβ, using the selective agonist diarylpropiolnitrile (DPN), in a rat model of stroke. Lister Hooded rats were ovariectomized and implanted with mini-pumps containing either DPN (8 mg kg−1 day−1) (n =7) or vehicle (n =5). Sensorimotor function was assessed using a neurological score and the spontaneous forelimb use asymmetry (cylinder) test. One week later the animals received a middle cerebral artery occlusion (MCAO), and T2-weighted MRI at 48 h post-MCAO quantified ischemic damage. Functional recovery was tested for 7 days post-MCAO and brains processed for histological verification of infarct size. The MRI images revealed no significant differences in hemispheric lesion volumes between vehicle- and DPN-treated groups (35.6±3.5% and 30.8±1.7%, respectively [mean±SEM]; Student''s unpaired t-test df =10, t =−1.357, p =0.453); this was confirmed histologically at 7 days. MCAO induced significant decline in neurological score performance (from 22 to 11 at 2 h post-MCAO) in the vehicle-treated animals, which was not significantly influenced by DPN. MCAO also induced significant changes in forelimb use in the cylinder test (10% reduction in contralateral, 20% reduction in both, and 30% increase in ipsilateral forelimb use) but this response was not significantly different between groups [F(1,1)=2.929, p =0.118, repeated-measures ANOVA]. In conclusion, pretreatment with the ERβ agonist DPN did not influence infarct size or sensorimotor function in rats exposed to MCAO. [Copyright &y& Elsevier]

Abstract: The estrogen-dependent process of vitellogenesis is a key function on oviparous fish reproduction and it has been widely used as an indicator of xenoestrogen exposure. The two estrogen receptor (ER) subtypes, ERα and ERβ, are often co-expressed in the liver of fish. The relative contribution of each ER subtype to modulate vitellogenin production by hepatocytes was studied using selected compounds known to preferentially interact with specific ER subtypes: propyl-pyrazole-triol (PPT) an ERα selective agonist, methyl-piperidino-pyrazole (MPP) an ERα selective antagonist, and diarylpropionitrile (DPN) an ERβ selective agonist. First, the relative binding affinity of the test compounds to estradiol for rainbow trout hepatic nuclear ER was determined using a competitive ligand binding assay. All the test ligands achieved complete displacement of specific [3H]-estradiol binding from the nuclear ER extract. This indicates that the test ligands have the potential to modify the ER function in the rainbow trout liver. Secondly, the ability of the test compounds to induce or inhibit vitellogenin production by primary cultures of rainbow trout hepatocytes was studied. Estradiol and DPN were the only compounds that induced a dose-dependent increase on vitellogenin synthesis. The lack of vitellogenin induction by PPT indicates that ERα could not have a role on this reproductive process whereas the ability of DPN to induce vitellogenin production supports the participation of ERβ. In addition, this hypothesis is reinforced by the results obtained from MPP plus estradiol. On one hand, the absence of suppressive activity of MPP in the estradiol-induced vitellogenin production does not support the participation of ERα. On the other hand, once blocked ERα with MPP, the only manifestation of agonist activity of estradiol would be achieved via ERβ. In conclusion, the present results indicate that vitellogenin production is mainly mediated through ERβ, implying, furthermore that compounds which only exhibit ERα selectivity are not detected by vitellogenin bioassay. [Copyright &y& Elsevier]

Abstract: Our previous analyses indicated that both estrogen receptor (ER) subtypes, ERα and ERβ, contribute to estrogen neuroprotection [Zhao, L., Wu, T.-W., Brinton, R.D., 2004. Estrogen receptor subtypes alpha and beta contribute to neuroprotection and increased Bcl-2 expression in primary hippocampal neurons. Brain Res. 1010, 22–34]. In the present study, we sought to determine the underlying mechanisms by which ERα and ERβ promote neuronal function, with a focus on neuroprotection, and whether these mechanisms are consistent with a classical nuclear or membrane ER-mediated response. Results of these analyses demonstrated that both the ERα-selective agonist, PPT (100 pM), and the ERβ-selective agonist, DPN (100 pM), were effective in dynamically but differentially regulating intracellular calcium (Ca2+) signaling in hippocampal neurons. Consistent with the direct measurement of neuroprotective outcomes [Zhao, L., Wu, T.-W., Brinton, R.D., 2004. Estrogen receptor subtypes alpha and beta contribute to neuroprotection and increased Bcl-2 expression in primary hippocampal neurons. Brain Res. 1010, 22–34], PPT and DPN exerted comparable efficacy in attenuating excitotoxic glutamate (200 μM)-induced intracellular Ca2+ rise. In contrast, DPN was more efficacious than PPT in potentiating a physiological concentration of glutamate (25 μM)-induced intracellular Ca2+ rise in these neurons. Further analyses revealed that both PPT and DPN increased ERK phosphorylation, however, the temporal profile and magnitude of response were unique to each molecule. The presence of the L-type Ca2+ channel inhibitor, nifedipine (10 μM), partially inhibited 17β-estradiol- and PPT-induced increase in phosphorylated ERK expression, whereas it induced a complete inhibition of DPN-induced increase in ERK phosphorylation. Additional neuroprotective experiments demonstrated that the MAPK inhibitor, PD 98059 (5 μM), partially blocked 17β-estradiol-induced promotion of neuronal survival against excitotoxic glutamate (200 μM)-induced neurotoxicity, whereas it completely blocked both PPT- and DPN-induced neuroprotection. The presence of the nuclear ER antagonist, ICI 182,780 (1 μM), not only failed to block all 3 molecule-induced neuroprotection, but coadministration of ICI 182,780 and each single molecule exerted a comparable or even greater neuroprotection. Taken together, as an expansion of our previous analyses, these data indicate that both ERα and ERβ contribute to neuronal mechanisms leading to estrogen promotion of neuronal function but with unique signaling profiles. Activation of ERβ and induction of intracellular Ca2+ influx via the L-type channels appears to be more closely associated with estrogen promotion of memory mechanisms. However, ERα and ERβ play an equivalently important role in mediating estrogen neuroprotection, and, which is dependent upon the activation of the MAPK signaling. Further, the present analyses suggest that separate from a classical nuclear ER-mediated response, estrogen promotes neuronal survival likely through a non-nuclear cytoplasm or membrane-associated ER-mediated rapid signaling cascade. [Copyright &y& Elsevier]

Abstract: The specific roles of estrogen receptor (ER) subtypes α and β in mediating estrogen’s influences on lupus autoimmunity are unknown. Herein we found that ovariectomized NZB/NZW F1 mice treated with propyl pyrazole triol (ERα-selective agonist) had significantly shorter survival, earlier development of albuminuria, higher serum concentrations of total IgG and prolactin, increased serum levels of anti-DNA IgG3, IgG2a and IgG2b and decreased anti-DNA IgG1 level compared to vehicle controls. In contrast, diarylpropionitrile (ERβ-selective agonist) administration significantly decreased serum anti-DNA IgG2b level but did not significantly affect serum levels of other anti-DNA IgG subclasses, serum total IgG or prolactin concentration, mortality or the occurrence of albuminuria. These findings suggest that ERα activation plays the predominant and immunostimulatory role in estrogen-mediated modulation of lupus while ERβ activation appears to have a slightly immunosuppressive effect on this disease. ERα activation coincidentally increased serum prolactin concentrations and may accelerate lupus disease activity also through this mechanism. [Copyright &y& Elsevier]

Abstract: The kinetics of ruthenium(III)-catalysed oxidation of paracetamol by diperiodatonickelate(IV) (DPN) in aqueous alkaline medium at a constant ionic strength of 1.0moldm−3 was studied spectrophotometrically using a rapid kinetic accessory. The reaction exhibits 1:2 stoichiometry (DPN:paracetamol). The reaction shows first order dependence on [DPN] and [ruthenium(III)] and (apparent) less than unit order in both [paracetamol] and [alkali] under the experimental conditions. However, the order in [paracetamol] and [alkali] changes from first order to zero order as the concentrations change from lower to higher concentrations. Addition of periodate has a retarding effect on the reaction. The effects of added products, ionic strength and dielectric constant of the reaction medium have been investigated. The main products were identified by spot test, IR and NMR. A mechanism involving the monoperiodatonickelate(IV) (MPN) as the reactive species of the oxidant has been proposed. The active species of ruthenium(III) is understood as [Ru(H2O)5OH]2+. The reaction constants involved in the different steps of mechanism are calculated. The activation parameters with respect to the slow step of the mechanism are computed and discussed and thermodynamic quantities are also calculated. [Copyright &y& Elsevier]

Abstract: Estrogens have multiple influences on immune functions. Estrogen receptors (ERs) have two distinct subtypes – α and β. To explore the specific roles of each ER subtype in estrogen-mediated immunomodulation, we investigated the effects of ER subtype-selective agonists on immune functions in ovariectomized Balb/c mice. Treatment with ERα-selective agonist propyl pyrazole triol (PPT) caused thymic atrophy and significant changes in thymic CD4/CD8 phenotypic profile. In contrast, ERβ-selective agonist diarylpropionitrile (DPN) alone had no effect on thymic weight, cellularity or CD4/CD8 phenotype expression. When coadministered with PPT, DPN partially antagonized PPT-evoked decrease in thymic cellularity and also partially attenuated PPT-induced shifts in thymic T-cell phenotype. These results indicate that ERα plays a predominant role in estrogen-induced thymic atrophy and ERβ activation may partially down-regulate ERα-mediated effects on thymic cellularity and T-cell phenotype expression. In addition, PPT administration induced a reduction in the percentage of mature B cells in the spleen, and enhanced IFN-γ production but suppressed IL-6 production from in vitro Con A-stimulated splenocytes as estradiol (E2) did, whereas DPN treatment had no effects either alone or with PPT, suggesting ERα mediates these estrogen actions. Treatment with PPT or DPN did not augment anti-DNP antibody production after DNP-KLH immunization as E2 did, implying that not merely one ER signaling pathway is involved in mediating estrogen''s effects on specific humoral immune responses. Our study further indicates ER subtype-selective agonists provide a novel approach to explore each ER subtype-mediated immunomodulation. [Copyright &y& Elsevier]

Abstract: Estradiol was previously reported to increase striatal D2 receptor density. The following experiments investigated the contribution of each estrogen receptor in estradiol modulation of D2 receptors. Ovariectomized Sprague–Dawley rats were treated for 2 weeks with an agonist for ERα, 4,4′,4″-(4-propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (PPT), an agonist for ERβ, 2,3-bis(4-hydroxyphenyl)-propionitrile (DPN) and compared to estradiol treatment. Ovariectomy decreased D2 agonist and antagonist striatal binding sites, specific binding was measured using [3H]quinpirole and [3H]spiperone. Estradiol prevented this decrease, while DPN but not PPT mimicked the estradiol increase of D2 receptor specific binding. In the nucleus accumbens, ovariectomy decreased [3H]quinpirole specific binding in the core and left the shell unchanged. Similarly, estradiol and DPN but not PPT prevented this decrease. Neither ovariectomy nor treatments affected [3H]spiperone specific binding in this area. In the olfactory tubercle, neither ovariectomy nor treatments changed D2 receptor binding. Finally, both ovariectomy and treatments did not affect D2L, D2S mRNA and D2L/D2S ratios measured by semi-quantitative RT-PCR. The present results show, for the first time, that an ERβ agonist treatment modulates D2 receptors and suggest that ERβ is involved in the estradiol modulation of D2 receptors. [Copyright &y& Elsevier]

Abstract: We describe a method of resistless photolithography using laser for the fabrication of microscopic markers and electrodes. A single shot of laser (355 nm, 100 mJ) is used to induce local surface melting and thus transfer a pattern from the mask (TEM grid) on to the surface of silicon. With a silicon substrate pre-coated with a layer of phosphorus, the laser pulse selectively produces doped regions that are highly conducting. The electrodes and markers thus obtained are robust and can withstand harsh chemical treatments. The utility of the marker for dip-pen nanolithography is illustrated by performing gold colloid nanopatterning. [Copyright &y& Elsevier]

Dip-pen nanolithography has been successfully demonstrated with hydrosols of Pd, Au, magnetic γ-Fe2O3 and luminescent Eu3+ doped LaPO4 on various substrates such as silicon and mica. The magnetic nature of γ-Fe2O3 patterns has been studied by MFM measurements. The patterns were independently imaged and characterized by employing LEEM and XPEEM. [ABSTRACT FROM AUTHOR]

Ca2+ uptake by mitochondria is a key element in the control of cellular Ca2+ homeostasis and Ca2+-dependent phenomena. It has been known for many years that this Ca2+ uptake is mediated by the mitochondrial Ca2+ uniporter, a specific Ca2+ channel of the inner mitochondrial membrane. We have shown previously that this channel is strongly activated by a series of natural phytoestrogenic flavonoids. We show here that several agonists and antagonists of estrogen receptors (ERs) also modulate the activity of the uniporter.The specific α-ER agonist 4,4′,4″-(4-propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (PPT) was the strongest activator, increasing the rate of mitochondrial Ca2+ uptake in permeabilized HeLa cells by 10-fold at 2 μM. Consistently, PPT largely increased the histamine-induced mitochondrial [Ca2+] peak and reduced the cytosolic one.Diethylstilbestrol and 17-β-estradiol (but not 17-α-estradiol) were active at pharmacological concentrations while the β-estrogen-receptor agonist 2,3-bis(4-hydroxyphenyl)-propionitrile (DPN) was little effective.The ER modulators tamoxifen and 4-hydroxy-tamoxifen inhibited mitochondrial Ca2+ uptake (IC50 2.5±1.5 and 2.5±1.4 μM, mean±s.d., respectively) both in the presence and in the absence of PPT, but raloxifene and the pure estrogen antagonist ICI 182,780 produced no effect.Activation by PPT was immediate and inhibition by tamoxifen or 4-hydroxy-tamoxifen required only 5 min to reach maximum.Tamoxifen did not modify mitochondrial membrane potential and PPT induced a slow mitochondrial depolarization at higher concentrations than those required to activate mitochondrial Ca2+ uptake.These results suggest that some kind of ER or related protein located in mitochondria controls the activity of the Ca2+ uniporter by a nongenomic mechanism. This novel mechanism of action of estrogen agonists and antagonists can provide a new interpretation for several previously reported effects of these compounds.British Journal of Pharmacology (2005) 145, 862–871. doi:10.1038/sj.bjp.0706265; published online 23 May 2005 [ABSTRACT FROM AUTHOR]

Abstract: To investigate the role of the estrogen receptor (ER) in mediating neuroprotection, the neuroprotective profiles of selective ER agonists for ERα and ERβ, propylpyrazole triol (PPT) and 2,3-bis(4-hydroxyphenyl) proprionitrile (DPN), respectively, were compared to that of 17β-estradiol and 17α-estradiol in primary neuron cultures challenged by β-amyloid toxicity. All compounds were found to be neuroprotective in an ER-dependent manner. However, protein kinase C (PKC) inhibition completely blocked the protective effects of 17β-estradiol and 17α-estradiol and significantly attenuated PPT but not DPN neuroprotection. These data indicate that estrogen-mediated neuroprotection likely involves a variety of mechanisms and that protection due to PKC activation is more likely due to ERα compared to ERβ. [Copyright &y& Elsevier]

Estrogens modulate the expression of preproenkephalin (PPE) in the hypothalamus but little is known for other brain regions. The present study investigated the effect of hormonal withdrawal and replacement therapy on PPE expression in the striatum, nucleus accumbens and cortex. Ovariectomized Sprague–Dawley rats were treated for 2 weeks with estradiol, a specific ligand for estrogen receptor α (ERα), 4,4′,4″-(4-propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (PPT) and estrogen receptor β (ERβ) 2,3-bis(4-hydroxyphenyl)-propionitrile (DPN), or the selective estrogen receptor modulators (SERMs) tamoxifen and raloxifene. Brain PPE mRNA levels, measured by in situ hybridization, were high in the striatum and nucleus accumbens compared to the low expression in the cortex. Ovariectomy decreased uterine weights compared to intact uterus, which was corrected by estradiol and PPT. Tamoxifen and raloxifene partially stimulated uterine weights while DPN left it unchanged. In the anterior, median and posterior striatum and in the core and shell of the nucleus accumbens, ovariectomy decreased PPE mRNA levels compared to intact rats, this was corrected by estradiol treatment except for the posterior striatum. PPT, DPN, tamoxifen and raloxifene reproduced the estradiol effect. In the prefrontal and cingulate cortices, neither ovariectomy nor treatments changed PPE mRNA levels. These results show for the first time that estradiol increases PPE mRNA in the striatum and nucleus accumbens. This effect is observed also with estrogen receptor agonists for the ERα and ERβ as well as with SERMs. [Copyright &y& Elsevier]

It has been demonstrated in our previous studies that Calbindin-D9k (CaBP-9k) is a potent biomarker for screening estrogen-like chemicals in the rat model. Although treatments with 17beta-estradiol (E2) and endocrine disrupting compounds resulted in the up-regulation of uterine CaBP-9k, the mechanism of CaBP-9k induction by these compounds through two subtypes of estrogen receptors (ERα and ERβ) is unclear. Thus, in the present study, immature rats were treated with propyl pyrazole triol (PPT, an ERα-selective ligand), diarylpropionitrile (DPN, an ERβ-selective ligand), E2, or dimethyl sulfoxide (DMSO, a vehicle control) for three days in order to clarify which subtype of ER is involved in the uterine CaBP-9k induction. Following injection with these ER ligands, uterine CaBP-9k expression was analyzed by Northern blot and immunoblot assays. Uterine CaBP-9k expression is mainly mediated by PPT in a dose- and time-dependent manner in immature rats, whereas no significant alteration of the uterine CaBP-9k gene was observed after DPN treatment. In addition, an estrogenicity of PPT in inducing CaBP-9k expression was completely blocked by the anti-estrogen ICI 182,780, implying that uterine CaBP-9k is solely induced by ERα. A single treatment with PPT rapidly increased the protein levels of ERα and PR, an E2-mediated gene, in these tissues. Taken together, these results indicate that uterine CaBP-9k is induced by E2 and endocrine disrupting chemicals via the ERα pathway, but not ERβ, in the uterus of immature rats. [ABSTRACT FROM AUTHOR]

The diversity of the physical and consequently the functional behaviour of starches, isolated from different rice varieties, is related to their specific structures. The latter are directly related to the starch biosynthetic pathway. To fully take advantage of the different functionalities of starches from different rice varieties and to design tailor-made starches, it is important to gain insight into biosynthesis–structure–physical behaviour–functionality relations. In a first part of this review, starch composition is described with a focus on rice starch. Secondly, current knowledge on starch biosynthesis is discussed. This more specifically includes (i) the function of the rice biosynthetic enzymes (i.e. adenosine diphosphate glucose pyrophosphorylases, synthases, branching and debranching enzymes), (ii) the effect of mutations on rice starch structure and, (iii) models for amylose and amylopectin synthesis. Thirdly, starch structure [i.e. from granule (2–100 μm), to growth ring (120–500 nm), blocklet (20–500 nm), amorphous and crystalline lamellae (9 nm), and amylopectin and amylose chain levels (0.1–1.0 nm)] is dealt with. Finally, relations between rice starch structural aspects [i.e. amylopectin (core) chain length distributions] and physical behaviour (i.e. gelatinisation and amylopectin retrogradation as measured by differential scanning calorimetry) are studied. [Copyright &y& Elsevier]

Estrogen receptor (ER) mediated neuroprotection has been demonstrated in both in vitro and in vivo model systems. However, the relative contribution by either ER subtype, ERα or ERβ, to estrogen-induced neuroprotection remains unresolved. To address this question, we investigated the impact of selective ER agonists for either ERα, PPT, or ERβ, DPN, to prevent neurodegeneration in cultured hippocampal neurons exposed to excitotoxic glutamate. Using three indicators of neuronal viability and survival, we demonstrated that both the ERα selective agonist PPT and the ERβ selective agonist DPN protected hippocampal neurons against glutamate-induced cell death in a dose-dependent manner, with the maximal response occurring at 100 pM. Further analyses showed that both PPT and DPN enhanced Bcl-2 expression in hippocampal neurons, with an efficacy comparable to their neuroprotective capacity. Collectively, the present data indicate that activation of either ERα or ERβ can promote neuroprotection in hippocampal neurons, suggesting that both receptor subtypes could be involved in estrogen neuroprotection. As ERβ is highly expressed in the brain and has little or no expression in the breast or uterus, discovery and design of ERβ selective molecules could provide a strategy for activating the beneficial effects of estrogen in the brain without activating untoward effects of estrogen in reproductive organs. [Copyright &y& Elsevier]

We report the results of fabricating micrometer and submicrometer-scale patterns of cytochrome c on gold surfaces. We used direct micro-contact printing (μCP) and indirect dip-pen nanolithography (DPN) for fabricating cytochrome c arrays. The protein dots were formed in diameters of 2 μm by μCP and of ∼200 nm by DPN, respectively. We analyzed the pattern size and height of protein arrays with atomic force microscopy (AFM). We expect that these methods will be potentially useful for developing small-scale biosensors and protein chip microarrays. [Copyright &y& Elsevier]

Most cultured cell lines require addition of serum to the medium to maintain their proliferative capacity. For studies examining the cellular effects of estrogens serum is charcoal-stripped to remove steroids. Nonetheless, addition of the selective estrogen receptor modulator (SERM) 4-hydroxytamoxifen (4-OHT) inhibits the basal transcriptional activity of estrogen receptors alpha or beta (ERα or ERβ) in transfected cells. We tested the hypothesis that elimination of serum from the culture medium will block 4-OHT’s repression of basal activity. Chinese hamster ovary (CHO-K1) cells adapted to serum-free medium exhibited estrogen responsiveness that was identical with that of the cells grown in serum-containing media. 4-OHT-suppressed basal transcription of an estrogen response element (ERE)-reporter in ERα-transfected cells even in the absence of serum, indicating that the 4-OHT suppressive activity is not mediated by blocking ER interaction with serum estrogens. We speculate that 4-OHT-ER recruits co-repressors to suppress basal transcription. We discovered that CHO-K1 cells express ERα and ERβ mRNA. However only ERβ protein was expressed and use of ERβ-selective 2,3-bis(4-hydroxy-phenyl)propionitrile (DPN) and ERα-selective 4-propyl-1,3,5-tris(4-hydroxy-phenyl)pyrazole) (PPT) revealed that only ERβ was transcriptionally active. In conclusion, growing CHO-K1 in serum-free medium does not impact the estrogen responsiveness and this cell line expresses functional ERβ. [Copyright &y& Elsevier]

Increasing evidence suggested that inhibiting the apoptosis of Schwann cells (SCs) and promoting nerve growth factor (NGF) expression in sciatic nerves play key roles in preventing the onset of diabetic peripheral neuropathy (DPN). Curcumin, a primary bioactive substance in turmeric with multiple characteristics, has been shown to have many therapeutic effects in a variety of diseases. However, curcumin is poorly studied in the DPN models. We aimed to explore the therapeutic benefits and underlying mechanism of curcumin in high fat/sugar diets joint streptozotocin (STZ)-induced DPN rat models. Sprague-Dawley (SD) rats were divided into five groups (6 rats per group), control group, DPN group, Curcumin groups (50, 100, and 150 mg/kg). Curcumin was administered intragastrically once per day for 4 continuous weeks. Body weight (BW) and fasting blood glucose (FBG) were monitored in all groups. The mechanical withdraw threshold (MWT) was measured. We also assessed neuropathic change by testing nerve conductance velocity (NCV) in sciatic nerves. TEM was applied to observe the sciatic nerves ultrastructure. The SCs apoptosis in sciatic nerves was stained using TUNEL kit. NGF contents in sciatic nerves and serum were detected using western blotting and ELISA analysis. The results showed curcumin had no obvious effect on the BW and FBG change. Curcumin (100 and 150 mg/kg) attenuated the MWT, NCV, and sciatic nerves ultrastructure in DPN rats. Curcumin (50, 100 and 150 mg/kg) reduced SCs apoptosis in sciatic nerves. In addition, curcumin at 150 mg/kg had the best efficacy in increasing protein expression of NGF in sciatic nerves and serum NGF level. Our work demonstrated that curcumin has neuroprotective effects for the treatment of DPN., (© 2022 Wiley-VHCA AG, Zurich, Switzerland.)

A decoupled plasma nitridation (DPN) with post nitridation annealing (PNA) treatment method was introduced to improve the performances of MOS devices with high-k (HK)-last/gate-last integration scheme and chemical oxide interface layer (IL). By introducing N to form HfSiON, it was found that DPN + PNA treatments could provide smaller equivalent oxide thickness (EOT) for both nMOS and pMOS devices. It was also found that we could achieve the best overall device performance for the HK-last/gate-last integration scheme with a chemical oxide IL by introducing nitrogen gas with low percentage content during DPN followed by high temperature PNA. [ABSTRACT FROM AUTHOR]

A principal process during the remanufacturing of worn-out or malfunctioning products is disassembly that enables the dumping, cleaning, repair or replacement of components as desired. Products subjected to disassembly exhibit uncertainty in the product structure and component conditions. Hence the termination goal (the level of disassembly) is subject to change, and the disassembly plan must be adapted. This paper proposes a mathematically sound disassembly Petri net (DPN) for the modeling and adaptive planning of disassembly processes. The planning algorithm guarantees the optimal remanufacturing value when each node's utility function (benefit of a subassembly or part and cost of a disassembly operation) is fixed. It can be modified to deal with the cases when the disassembly operation success rates vary with the product condition and reliability of resources performing the operations. The proposed methodology and algorithms are demonstrated through the sample disassembly of a telephone. [ABSTRACT FROM PUBLISHER]

Estrogens play a key role in the sexual differentiation of the brain and behavior. While early estrogen actions exert masculinizing effects on the brain of male rodents, a diametrically opposite effect is observed in birds where estrogens demasculinize the brain of females. Yet, the two vertebrate classes express similar sex differences in the brain and behavior. Although ERα is thought to play a major role in these processes in rodents, the role of ERβ is still controversial. In birds, the identity of the estrogen receptor(s) underlying the demasculinization of the female brain remains unclear. The aim of the present study was thus to determine in Japanese quail the effects of specific agonists of ERα (propylpyrazole triol, PPT) and ERβ (diarylpropionitrile, DPN) administered at the beginning of the sensitive period (embryonic day 7, E7) on the sexual differentiation of male sexual behavior and on the density of vasotocin-immunoreactive (VT-ir) fibers, a known marker of the organizational action of estrogens on the quail brain. We demonstrate that estradiol benzoate and the ERβ agonist (DPN) demasculinize male sexual behavior and decrease the density of VT-ir fibers in the medial preoptic nucleus and the bed nucleus of the stria terminalis, while PPT has no effect on these measures. These results clearly indicate that ERβ, but not ERα, is involved in the estrogen-induced sexual differentiation of brain and sexual behavior in quail. • Quail eggs were injected at embryonic day 7 with an ERα or ERβ agonist. • The ERβ agonist DPN demasculinized copulatory behavior of adult males. • DPN reduced the density of sexually dimorphic VT-ir fibers in the preoptic area. • The ERα agonist PPT did not have any of these effects. • Quail demascuilinization is mediated by ERβ with no major contribution of ERα. [ABSTRACT FROM AUTHOR]

Intestinal stem cells (ISCs) are able to generate gut-specific enterocytes, as well as neural-like enteroendocrine cells. It is unclear how the tissue identity of the ISC lineage is regulated to confer cell-lineage fidelity. Here, we show that, in adult Drosophila midgut, loss of the transcriptional repressor Tramtrack in ISCs causes a self-renewal program switch to neural stem cell (NSC)-like, and that switch drives neuroendocrine tumor development. In Tramtrack-depleted ISCs, the ectopically expressed Deadpan acts as a major self-renewal factor for cell propagation, and Sequoia acts as a differentiation factor for the neuroendocrine phenotype. In addition, the expression of Sequoia renders NSC-specific self-renewal genes responsive to Notch in ISCs, thus inverting the differentiation-promoting function of Notch into a self-renewal role as in normal NSCs. These results suggest an active maintenance mechanism for the gut identity of ISCs, whose disruption may lead to an improper acquisition of NSC-like traits and tumorigenesis. • Depletion of ttk69 in fly ISCs causes a genome-wide program switch to NSC-like • The ectopically expressed NSC factor Dpn drives NET tumorigenesis • The ectopically expressed pan-neural factor Seq generates a neuroendocrine phenotype • Seq also converts the differentiation-promoting role of Notch into a self-renewal role Li et al. show that the loss of a transcriptional repressor, Tramtrack, in adult Drosophila intestinal stem cells causes a self-renewal program switch to neural-stem-cell-like, which drives neuroendocrine tumor development. [ABSTRACT FROM AUTHOR]

Automatic skin lesion segmentation in dermoscopy images is challenging due to the diversity of skin lesion characteristics, low contrast between normal skin and lesions, and the existence of many artefacts in the images. To meet these challenges, we propose a novel segmentation topology called FC-DPN, which is built upon a fully convolutional network (FCN) and dual path network (DPN). The DPN inherits the advantages of residual and densely connected paths, enabling effective feature re-usage and re-exploitation. We replace dense blocks in fully convolutional DenseNets (FC-DenseNets) with two kinds of sub-DPN blocks, namely, sub-DPN projection blocks and sub-DPN processing blocks. This framework enables FC-DPN to acquire more representative and discriminative features for more accurate segmentation. Many images in the original ISBI 2017 Skin Lesion Challenge test dataset are given the incorrect or inaccurate ground truths, and these ground truths have been revised. The revised test dataset is called the modified ISBI 2017 Skin Lesion Challenge test dataset. The proposed method achieves an average Dice coefficient of 88.13% and a Jaccard index of 80.02% on the modified ISBI 2017 Skin Lesion Challenge test dataset and 90.26% and 83.51%, respectively, on the PH2 dataset. Extensive experimental results on the two datasets demonstrate that the proposed method exhibits better performance than FC-DenseNets and other well-established segmentation algorithms., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Neuropathies of the peripheral and autonomic nervous systems affect up to half of all people with diabetes mellitus, and are major risk factors for foot ulceration, amputation and cardiovascular dysfunction. Peripheral neuropathies manifest with either painful or painless symptoms, but many patients experience both. Once diagnosed appropriately, painful diabetic neuropathy management presents a unique challenge for physicians and should be considered as a syndrome, clinically distinct from diabetic peripheral neuropathy. The aetiology is multifactorial: metabolic changes in diabetes may directly affect neural tissue and neurodegenerative changes are precipitated by compromised nerve vascular supply. Metabolic changes include the elevated polyol pathway activity, the increased oxidative stress, the formation of advanced glycation and lipoxidation end products, and various pro-inflammatory changes. These mechanisms work in combination and interact in a mutually facilitatory fashion. This review focuses on the current therapies for the management of peripheral and cardiovascular autonomic neuropathy and of painful neuropathy as a distinct entity, based on the current knowledge of diabetic neuropathy. Moreover, the role of ACE inhibition has been explored in the treatment of Cardiovascular Autonomic Neuropathy., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Diabetic peripheral neuropathy (DPN) is a complication of diabetes connected with morbidity and mortality. DPN presents deterioration of peripheral nerves with pain, feebleness, and loss of sensation. Particular medications might display their remedial potential by controlling neuroinflammation. Palmitoylethanolamide (PEA) is an autacoid local injury antagonist distinguished for its neuroprotective, analgesic, and anti-inflammatory properties in numerous experimental models of neuroinflammation. Based on these findings, the goal of this work was to better test the neuroprotective effects of a formulation of micronized PEA (PEA-m) and the probable mechanism of action in a mouse model of DPN induced by streptozotocin (STZ) injection. Diabetic and control animals received PEA-m (10 mg/kg) by oral gavage daily starting 2 wk from STZ injection. After 16 wk, the animals were euthanized, and blood, urine, spinal cord, and sciatic nerve tissues were collected. Our results demonstrated that after diabetes induction, PEA-m was able to reduce mechanical, thermal hyperalgesia, and motor alterations as well as reduce mast cell activation and nerve growth factor expression. In addition, PEA-m decreased neural histologic damage, oxidative and nitrosative stress, cytokine release, angiogenesis, and apoptosis. Moreover, spinal microglia activation (IBA-1), phospho-P38 MAPK, and nuclear factor NF-κB inflammatory pathways were also inhibited. The protective effects of PEA-m could be correlated at least in part to peroxisome proliferator-activated receptor-α activation. In summary, we demonstrated that PEA-m represents a new therapeutic strategy for neuroinflammation pain associated with mixed neuropathies. [ABSTRACT FROM AUTHOR]