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This study is the first to demonstrate that estradiol and estrogen receptor α and β stimulation during early development sexually differentiates the electrophysiological properties of caudate-putamen medium spiny neurons, the primary output neuron of the striatal regions. Overall, this evidence provides new insight into the neuroendocrine mechanism by which caudate-putamen neuron electrophysiology is sexually differentiated and demonstrates the powerful action of early hormone exposure upon individual neuron electrophysiology.

The neuroendocrine environment in which the brain operates is both dynamic and differs by sex. How differences in neuroendocrine state affect neuron properties has been significantly neglected in neuroscience research. Behavioral data across humans and rodents indicate that natural cyclical changes in steroid sex hormone production affect sensorimotor and cognitive behaviors in both normal and pathological contexts. These behaviors are critically mediated by the caudate-putamen. In the caudate-putamen, medium spiny neurons (MSNs) are the predominant and primary output neurons. MSNs express membrane-associated estrogen receptors and demonstrate estrogen sensitivity. However, how the cyclical hormone changes across the estrous cycle may modulate caudate-putamen MSN electrophysiological properties remains unknown. Here, we performed whole-cell patch-clamp recordings on male, diestrus female, proestrus female, and estrus female caudate-putamen MSNs. Action potential, passive membrane, and miniature excitatory post-synaptic current properties were assessed. Numerous MSN electrical properties robustly differed by cycle state, including resting membrane potential, rheobase, action potential threshold, maximum evoked action potential firing rate, and inward rectification. Strikingly, when considered independent of estrous cycle phase, all but one of these properties do not significantly differ from male MSNs. These data indicate that female caudate-putamen MSNs are sensitive to the estrous cycle, and more broadly, the importance of considering neuroendocrine state in studies of neuron physiology.

In this paper, the degradation of tetracycline (TC) by peroxymonosulfate (PMS) activated with zero-valent iron (Fe0) was systematically studied through batches of experiments. First, effects of the key parameters (i.e., Fe0 dosage, PMS dosage, initial pH and co-existing ions) on TC degradation were investigated. Under the optimal conditions, high TC removal efficiency (88.5%) was achieved after 5 min treatment. Also, four control experiments were conducted to demonstrate the excellent performance of Fe0/PMS process and the synergistic effect between Fe0 and PMS. Compared with Fe2+ (the relatively common PMS activator), Fe0 was an efficient and long-lasting activator which consume less PMS and produce less dissolved iron ions. Then, the characteristics of reacted Fe0 particles were analyzed by SEM-EDS, XRD and XPS. The results shows that a few iron corrosion products generated and some of them deposited on the surface of reacted Fe0 particles. The generated iron corrosion products could promote PMS activation or adsorb TC directly. Next, the possible degradation pathway of TC was elaborated according to the intermediates detected by LC-QTOF-MS/MS and the toxicity in solution during reaction was evaluated. Finally, the common free radicals were monitored and the reaction mechanism was proposed.

Exposure to steroid sex hormones such as 17β-estradiol (estradiol) during early life potentially permanently masculinize neuron electrophysiological phenotype. In rodents, one crucial component of this developmental process occurs in males, with estradiol aromatized in the brain from testes-sourced testosterone. However, it is unknown whether most neuron electrophysiological phenotypes are altered by this early masculinization process, including medium spiny neurons (MSNs) of the rat caudate-putamen. MSNs are the predominant and primary output neurons of the caudate-putamen and exhibit increased intrinsic excitability in females compared to males. Here, we hypothesize that since perinatal estradiol exposure occurs in males, then a comparable exposure in females to estradiol or its receptor agonists would be sufficient to induce masculinization. To test this hypothesis, we injected perinatal female rats with estradiol or its receptor agonists and then later assessed MSN electrophysiology. Female and male rats on postnatal day 0 and 1 were systemically injected with either vehicle, estradiol, the estrogen receptor (ER)α agonist PPT, the ERβ agonist DPN, or the G-protein-coupled receptor 1 (GPER-1) agonist G1. On postnatal days 19 ± 2, MSN electrophysiological properties were assessed using whole cell patch clamp recordings. Estradiol exposure abolished increased intrinsic excitability in female compared to male MSNs. Exposure to either an ERα or ERβ agonist masculinized female MSN evoked action potential firing rate properties, whereas exposure to an ERβ agonist masculinized female MSN inward rectification properties. Exposure to ER agonists minimally impacted male MSN electrophysiological properties. These findings indicate that perinatal estradiol exposure masculinizes MSN electrophysiological phenotype via activation of ERα and ERβ. NEW & NOTEWORTHY This study is the first to demonstrate that estradiol and estrogen receptor α and β stimulation during early development sexually differentiates the electrophysiological properties of caudate-putamen medium spiny neurons, the primary output neuron of the striatal regions. Overall, this evidence provides new insight into the neuroendocrine mechanism by which caudate-putamen neuron electrophysiology is sexually differentiated and demonstrates the powerful action of early hormone exposure upon individual neuron electrophysiology.

The absorption properties of N-(2-hydroxyethyl) morpholine (HEM), morpholine (MP) and N-(2-aminoethyl) morpholine (AEM) for SO

The nucleus accumbens core (AcbC) is a striatal brain region essential for integrating motivated behavior and reward processing with premotor function. In humans and rodents, research has identified sex differences and sex steroid hormone sensitivity in AcbC-mediated behaviors, in disorders, and in rats in the electrophysiological properties of the AcbC output neuron type, the medium spiny neuron (MSN). It is unknown whether the sex differences detected in MSN electrophysiological properties extend to mice. Furthermore, MSNs come in distinct subtypes with subtle differences in electrophysiological properties, and it is unknown whether MSN subtype-specific electrophysiology varies by sex. To address these questions, we used male and female Drd1a-tdTomato line 6 bacterial artificial chromosome transgenic mice. We made acute brain slices of the AcbC, and performed whole cell patch-clamp recordings across MSN subtypes to comprehensively assess AcbC MSN subtype electrophysiological properties. We found that ( 1 mice MSNs did not exhibit the sex differences detected in rat MSNs, and 2) electrophysiological properties differed between MSN subtypes in both sexes, including rheobase, resting membrane potential, action potential properties, intrinsic excitability, input resistance in both the linear and rectified ranges, and miniature excitatory postsynaptic current properties. These findings significantly extend previous studies of MSN subtypes performed in males or animals of undetermined sex and indicate that the influence of sex upon AcbC MSN properties varies between rodent species. NEW & NOTEWORTHY This research provides the most comprehensive assessment of medium spiny neuron subtype electrophysiological properties to date in a critical brain region, the nucleus accumbens core. It additionally represents the first evaluation of whether mouse medium spiny neuron subtype electrophysiological properties differ by sex.

In this study, the effect of initial pH on tetracycline (TC) removal by zero-valent iron (ZVI) has been investigated thoroughly. The experimental results show that TC degradation efficiency performed effectively across the initial pH range 2.5–6.5, almost all removal efficiencies in this range reached 90% after 30 min treatment. As pH increasing to alkaline, removal efficiencies gradually decreased. Experimental results indicate that removal of TC by Fe0/air process is mainly attributed to adsorption (by Fe0 and its corrosion products), oxidation and reduction. pH significantly affected the formation of iron corrosion products, the production of hydroxyl radicals (HO ) and the variation of TC species. Adsorption is the main removal path under neutral conditions while oxidation is the prime removal way under acidic conditions. Finally, the intermediates and toxicity analysis indicates that less toxicity products generated through ring-opening reactions. The study provides new insight into the removal of TC by ZVI and support a theoretical foundation for further research.

The performance of mFe/Cu/O3 and mFe0/O3 processes was comparatively investigated through optimized the key operational parameters for degradation of p-nitrophenol (PNP). The COD removal attained by mFe/Cu/O3 process was higher than that of mFe0/O3 process under the corresponding conditions. Additionally, under the optimal conditions, the COD removal (93.6%) obtained by mFe/Cu/O3 process was more than twice of the sum (44.6%) of that by ozone (35.3%) and mFe/Cu alone (9.3%). Finally, the key active materials in mFe/Cu/O3 system, degradation pathway of PNP and reaction mechanism of mFe/Cu/O3 process were proposed according to the analysis results of intermediate products, SEM–EDS and XRD.

A novel n-n type WC/BiOCl binary photocatalyst was synthesized, and applied to sulfamethoxazole (SMX) degradation. The physical and photoelectric properties of the WC/BiOCl were characterized by various experimental and calculational methods. An intimate interfacial contact between WC and BiOCl was identified, which help to reduce the aggregation of BiOCl and more importantly, build an internal electrical field between WC and BiOCl. It is interesting to find that the WC layer(s) can help to reduce the interfacial charge transfer resistance and act as electron transfer and storage sites. Subsequently, the holes are engaged in the oxidation of SMX by WC/BiOCl. Additionally, WC/BiOCl photocatalyst has more reactive sites and absorbs more incident photons compared with pure BiOCl. Quenching and electron spin resonance (ESR) experiments indicate that O2·- is the main active species, holes and OH• are also involved. The degradation pathway of SMX and the enhancing mechanism for the photodegradation through the combination of WC and BiOCl were explored. This work provides a novel binary photocatalyst for SMX degradation.

Bisphenol A (BPA) is a high volume endocrine disrupting chemical found in a wide variety of products including plastics and epoxy resins. Human exposure is nearly ubiquitous, and higher in children than adults. Because BPA has been reported to interfere with sex steroid hormone signaling, there is concern that developmental exposure, even at levels below the current FDA No Observed Adverse Effect Level (NOAEL) of 5 mg/kg body weight (bw)/day, can disrupt brain sexual differentiation. The current studies were conducted as part of the CLARITY-BPA (Consortium Linking Academic and Regulatory Insights on BPA Toxicity) program and tested the hypothesis that perinatal BPA exposure would induce morphological changes in hormone sensitive, sexually dimorphic brain regions. Sprague-Dawley rats were randomly assigned to 5 groups: BPA (2.5, 25, or 2500 µg/kg bw/day), a reference estrogen (0.5 µg ethinylestradiol (EE2)/kg bw/day), or vehicle. Exposure occurred by gavage to the dam from gestational day 6 until parturition, and then to the offspring from birth through weaning. Unbiased stereology was used to quantify the volume of the sexually dimorphic nucleus (SDN), the anteroventral periventricular nucleus (AVPV), the posterodorsal portion of the medial amygdala (MePD), and the locus coeruleus (LC) at postnatal day 28. No appreciable effects of BPA were observed on the volume of the SDN or LC. However, AVPV volume was enlarged in both sexes, even at levels below the FDA NOAEL. Collectively, these data suggest the developing brain is vulnerable to endocrine disruption by BPA at exposure levels below previous estimates by regulatory agencies.

The absorption properties of N-(2-hydroxyethyl) morpholine (HEM), morpholine (MP) and N-(2-aminoethyl) morpholine (AEM) for SO2 were studied using sulfolane (SUL) as solvent in this work. Among these solvent combinations, HEM/SUL shows the best cyclic absorption performance, and the capacity of HEM-SUL-40 (40 wt% of HEM and 60 wt% of SUL) to absorb 8580 mg/m3 SO2 (the remainder is N2) is 192.18 mg/g at 293.15 K. The absorption capacity of the second cycle is 97.5% of the first absorption cycle, which is higher than 70% of the Cansolv amine solution in a commercial application with similar experimental conditions. However, MP/SUL is difficult to desorb at high temperature, and the absorption capacity of AEM/SUL is much lower than HEM/SUL and MP/SUL. According to the FTIR, 1H NMR and 13C NMR, all three cyclic amines have charge transfer effects with SO2. The structure of HEM/SUL can be recovered after heating, but MP cannot be recovered. ΔrGm° in the reaction against HEM with SO2 increases significantly with increasing temperature. The ΔrGm° of HEM-SO2 and MP-SO2 at 353.15 K is −12.56 kJ/mol and −16.29 kJ/mol, respectively, which further explains the easy desorption of HEM-SO2 and the difficult desorption of MP-SO2 at high temperature.

A coagulation-flocculation as pre-treatment combined with mFe/Cu/O 3 (CF-mFe/Cu/O 3 ) process was developed to degrade the pollutants in automobile coating wastewater (ACW). In coagulation-flocculation (CF) process, high turbidity removal efficiency (97.1%) and low COD removal efficiency (10.5%) were obtained under the optimal conditions using Al 2 (SO 4 ) 3 ·18H 2 O and CaO. The effluent of CF process (ECF) was further disposed by mFe/Cu/O 3 process, and its key operating parameters were optimized by batch experiments. Optimally, COD removal efficiency of ECF obtained by the mFe/Cu/O 3 process (i.e., 87.6% after 30 min treatment) was much higher than those of mFe/Cu alone (8.3%), ozone alone (46.6%), and mFe/Cu/air (6.1%), which confirms the superiority of the mFe/Cu/O 3 process. In addition, the analysis results of UV–vis, excitation-emission matrix (EEM) fluorescence spectra and GC/MS further confirm that the phenol pollutants of ECF had been effectively decomposed or transformed after CF-mFe/Cu/O 3 process treatment. Meanwhile, B/C ratio of ACW increased from 0.19 to 0.56, which suggests the biodegradability was improved significantly. Finally, the operating cost of CF-mFe/Cu/O 3 process was about 1.83 USD t −1 for ACW treatment. Therefore, the combined process is a promising treatment technology for the coating wastewater from automobile manufacturing.

A micro-size Fe0/O3 process (mFe0/O3) was setup to degrade p-nitrophenol (PNP) in aqueous solution, and its key operational parameters (i.e., initial pH, ozone flow rate, and Fe0 dosage) were optimized by the semibatch experiments, respectively. Under the optimal conditions, COD removal efficiency (89.5%) obtained by the mFe0/O3 process was about two times of the sum (44.8%) of COD removal obtained by Fe0 alone and O3 alone. The results suggest that the synergetic effect between O3 and Fe0 in the mFe0/O3 process played a vital role in the degradation of PNP. In addition, treatment efficiency of the mFe0/O3 process also was much higher than some of the present technologies (e.g., Fe0/air process, MnO2/O3 and Al2O3/O3), which confirm the superiority of the mFe0/O3 process. Furthermore, the degradation mechanism and pathway of PNP was proposed according to the analysis results of UV–vis, FTIR, IC and GC–MS. According to the present literature and analysis data of SEM, EDS and XRD analysis, it can be concluded that the high-efficient mFe0/O3 process was mainly resulted from the combination of homogeneous or heterogeneous catalytic ozonation, Fenton-like reaction, adsorption and precipitate. Therefore, the mFe0/O3 process was a promising technology for the refractory industrial wastewater.

Steroid sex hormones and genetic sex regulate the phenotypes of motivated behaviors and relevant disorders. Most studies seeking to elucidate the underlying neuroendocrine mechanisms have focused on how 17β-estradiol modulates the role of dopamine in striatal brain regions, which express membrane-associated estrogen receptors. Dopamine action is an important component of striatal function, but excitatory synaptic neurotransmission has also emerged as a key striatal substrate and target of estradiol action. Here, we focus on excitatory synaptic input onto medium spiny neurons (MSNs) in the striatal region nucleus accumbens core (AcbC). In adult AcbC, miniature excitatory postsynaptic current (mEPSC) frequency is increased in female compared with male MSNs. We tested whether increased mEPSC frequency in female MSNs exists before puberty, whether this increased excitability is due to the absence of estradiol or testosterone during the early developmental critical period, and whether it is accompanied by stable neuron intrinsic membrane properties. We found that mEPSC frequency is increased in female compared with male MSNs before puberty. Increased mEPSC frequency in female MSNs is abolished after neonatal estradiol or testosterone exposure. MSN intrinsic membrane properties did not differ by sex. These data indicate that neonatal masculinization via estradiol and/or testosterone action is sufficient for down-regulating excitatory synaptic input onto MSNs. We conclude that excitatory synaptic input onto AcbC MSNs is organized long before adulthood via steroid sex hormone action, providing new insight into a mechanism by which sex differences in motivated behavior and other AbcC functions may be generated or compromised.

Manganese-doped bismuth oxychloride (Mn-BiOCl) photocatalysts were facilely synthesized for the photodegradation of metronidazole (MTZ), and the as-prepared Mn-BiOCl was systematically characterized. The results show that the introduction of Mn2+ to BiOCl increases its MTZ removal efficiency through increasing its specific surface area, broadening its light absorption region, and facilitating photoinduced carrier separation. The enhancement of the optical properties is ascribed to the generation of an intermediate Mn 3p-O 2d energy level in the forbidden gap of Mn-BiOCl based on the density functional theory calculations. Mn-BiOCl also exhibited favorable stability and recyclability after four cycles. Moreover, the results of the quenching experiments indicate that superoxide radicals (O2 −), hydroxyl radicals (OH ) and holes (h+) participated in MTZ degradation over Mn-BiOCl, and the electron spin resonance (ESR) test demonstrated that more OH was generated in the Mn-BiOCl process than the BiOCl process. This was explained by the H2O adsorption energy and bond parameter on the photocatalyst’s surface. Finally, the photodegradation pathway of MTZ was elaborated based on the intermediate analysis, and a mechanism for the promotion of MTZ photodegradation through Mn-doping BiOCl was proposed.

The caudate-putamen is a striatal brain region essential for sensorimotor behaviors, habit learning, and other cognitive and premotor functions. The output and predominant neuron of the caudate-putamen is the medium spiny neuron (MSN). MSNs present discrete cellular subtypes that show differences in neurochemistry, dopamine receptor expression, efferent targets, gene expression, functional roles, and most importantly for this study, electrophysiological properties. MSN subtypes include the striatonigral and the striatopallidal groups. Most studies identify the striatopallidal MSN subtype as being more excitable than the striatonigral MSN subtype. However, there is some divergence between studies regarding the exact differences in electrophysiological properties. Furthermore, MSN subtype electrophysiological properties have not been reported disaggregated by biological sex. We addressed these questions using prepubertal male and female Drd1a-tdTomato line 6 BAC transgenic mice, an important transgenic line that has not yet received extensive electrophysiological analysis. We made acute caudate-putamen brain slices and assessed a robust battery of 16 relevant electrophysiological properties using whole-cell patch-clamp recording, including intrinsic membrane, action potential, and miniature EPSC (mEPSC) properties. We found that: (1) MSN subtypes exhibited multiple differential electrophysiological properties in both sexes, including rheobase, action potential threshold and width, input resistance in both the linear and rectified ranges, and mEPSC amplitude; (2) select electrophysiological properties showed interactions between MSN subtype and sex. These findings provide a comprehensive evaluation of mouse caudate-putamen MSN subtype electrophysiological properties across females and males, both confirming and extending previous studies.

In this paper, the Fe/Cu bimetallic particles and ozone were combined to decompose or transform the toxic and refractory pollutants in dinitrodiazophenol (DDNP) wastewater. Firstly, operational parameters including theoretical Cu mass loading (TMLCu), Fe/Cu dosage, initial pH, O3 flow rate and treatment time were optimized respectively. The maximum COD removal efficiency (85.3%) and color removal efficiency (95.0%) were obtained under the optimal conditions (i.e., theoretical Cu mass loading (TMLCu) = 0.02 g Cu per g Fe, Fe/Cu dosage = 20 g L−1, initial pH = 5.0, O3 flow rate = 0.3 L min−1, treatment time = 15 min). Then, in order to confirm the superiority of the Fe/Cu/O3 process, three control experiment systems including Fe/Cu, O3 and Fe0/O3 processes were set up under the same conditions. In addition, the UV-vis and FTIR results confirm the main pollutants in DDNP wastewater were oxidized and generated intermediates, which revealed the superiority of Fe/Cu/O3 process. Thus, the DDNP wastewater could be treated by Fe/Cu/O3 process in a promising way.

Endocrine disrupting compounds (EDCs) are hypothesized to promote obesity and early puberty but their interactive effects with hormonally active diets are poorly understood. Here we assessed individual and combinatorial effects of soy diet or the isoflavone genistein (GEN; administered as the aglycone genistin GIN) with bisphenol A (BPA) on body weight, ingestive behavior and female puberal onset in Wistar rats. Soy-fed dams gained less weight during pregnancy and, although they consumed more than dams on a soy-free diet during lactation, did not become heavier. Their offspring (both sexes), however, became significantly heavier (more pronounced in males) pre-weaning. Soy also enhanced food intake and accelerated female pubertal onset in the offspring. Notably, pubertal onset was also advanced in females placed on soy diet at weaning. Males exposed to BPA plus soy diet, but not BPA alone, had lighter testes. BPA had no independent effects.

Sex differences in neuron electrophysiological properties were traditionally associated with brain regions directly involved in reproduction in adult, postpubertal animals. There is growing acknowledgement that sex differences can exist in other developmental periods and brain regions as well. This includes the dorsal striatum (caudate/putamen), which shows robust sex differences in gene expression, neuromodulator action (including dopamine and 17β-estradiol), and relevant sensorimotor behaviors and pathologies such as the responsiveness to drugs of abuse. Here we examine whether these sex differences extend to striatal neuron electrophysiology. We test the hypothesis that passive and active medium spiny neuron (MSN) electrophysiological properties in prepubertal rat dorsal striatum differ by sex. We made whole cell recordings from male and females MSNs from acute brain slices. The slope of the evoked firing rate to current injection curve was increased in MSNs recorded from females compared with males. The initial action potential firing rate was increased in MSNs recorded from females compared with males. Action potential after-hyperpolarization peak was decreased, and threshold was hyperpolarized in MSNs recorded from females compared with males. No sex differences in passive electrophysiological properties or miniature excitatory synaptic currents were detected. These findings indicate that MSN excitability is increased in prepubertal females compared with males, providing a new mechanism that potentially contributes to generating sex differences in striatal-mediated processes. Broadly, these findings demonstrate that sex differences in neuron electrophysiological properties can exist prepuberty in brain regions not directly related to reproduction.

Perinatal life is a critical window for sexually dimorphic brain organization, and profoundly influenced by steroid hormones. Exposure to endocrine-disrupting compounds may disrupt this process, resulting in compromised reproductive physiology and behavior. To test the hypothesis that neonatal bisphenol A (BPA) exposure can alter sex-specific postnatalEsr2(Erβ) expression in brain regions fundamental to sociosexual behavior, we mappedEsr2mRNA levels in the principal nucleus of the bed nucleus of the stria terminalis (BNSTp), paraventricular nucleus (PVN), anterior portion of the medial amygdaloid nucleus (MeA), super optic nucleus, suprachiasmatic nucleus, and lateral habenula across postnatal days (PNDs) 0–19. Next, rat pups of both sexes were subcutaneously injected with 10 μg estradiol benzoate (EB), 50 μg/kg BPA (LBPA), or 50 mg/kg BPA (HBPA) over the first 3 days of life andEsr2levels were quantified in each region of interest (ROI) on PNDs 4 and 10. EB exposure decreasedEsr2signal in most female ROIs and in the male PVN. In the BNSTp,Esr2expression decreased in LBPA males and HBPA females on PND 10, thereby reversing the sex difference in expression. In the PVN,Esr2mRNA levels were elevated in LBPA females, also resulting in a reversal of sexually dimorphic expression. In the MeA, BPA decreasedEsr2expression on PND 4. Collectively, these data demonstrate that region- and sex-specificEsr2expression is vulnerable to neonatal BPA exposure in regions of the developing brain critical to sociosexual behavior in rat.

The caudate-putamen is a striatal brain region essential for sensorimotor behaviors, habit learning, and other cognitive and premotor functions. The output and predominant neuron of the caudate-putamen is the medium spiny neuron (MSN). MSNs present discrete cellular subtypes that show differences in neurochemistry, dopamine receptor expression, efferent targets, gene expression, functional roles, and most importantly for this study, electrophysiological properties. MSN subtypes include the striatonigral and the striatopallidal groups. Most studies identify the striatopallidal MSN subtype as being more excitable than the striatonigral MSN subtype. However, there is some divergence between studies regarding the exact differences in electrophysiological properties. Furthermore, MSN subtype electrophysiological properties have not been reported disaggregated by biological sex. We addressed these questions using prepubertal male and female Drd1a-tdTomato line 6 BAC transgenic mice, an important transgenic line that has not yet received extensive electrophysiological analysis. We made acute caudate-putamen brain slices and assessed a robust battery of 16 relevant electrophysiological properties using whole-cell patch-clamp recording, including intrinsic membrane, action potential, and miniature EPSC (mEPSC) properties. We found that: (1) MSN subtypes exhibited multiple differential electrophysiological properties in both sexes, including rheobase, action potential threshold and width, input resistance in both the linear and rectified ranges, and mEPSC amplitude; (2) select electrophysiological properties showed interactions between MSN subtype and sex. These findings provide a comprehensive evaluation of mouse caudate-putamen MSN subtype electrophysiological properties across females and males, both confirming and extending previous studies.

Bisphenol A (BPA) exposure is ubiquitous, and in laboratory animals, early-life BPA exposure has been shown to alter sex-specific neural organization, neuroendocrine physiology, and behavior. The specific mechanisms underlying these brain-related outcomes, however, remain largely unknown, constraining the capacity to ascertain the potential human relevance of neural effects observed in animal models. In the perinatal rat brain, estrogen is masculinizing, suggesting that BPA-induced perturbation of estrogen receptor (ESR) expression may underpin later in-life neuroendocrine effects. We hypothesized that prenatal BPA exposure alters sex-specific ESR1 (ERα) and ESR2 (ERβ) expression in postnatal limbic nuclei. Sprague Dawley rats were mated and gavaged on gestational days (GDs) 6–21 with vehicle, 2.5 or 25 μg/kg bw/day BPA, or 5 or 10 μg/kg bw/day ethinyl estradiol. An additional group was restrained but not gavaged (naïve control). Offspring were sacrificed the day after birth to quantify ESR gene expression throughout the hypothalamus and amygdala by in situ hybridization. Relative to the vehicle group, significant effects of BPA were observed on ESR1 and ESR2 expression throughout the mediobasal hypothalamus and amygdala in both sexes. Significant differences in ESR expression were also observed in the mediobasal hypothalamus and amygdala of the naïve control group compared with the vehicle group, highlighting the potential for gavage to influence gene expression in the developing brain. These results indicate that ESR expression in the neonatal brain of both sexes can be altered by low-dose prenatal BPA exposure.

The rodent amygdaloid complex is composed of numerous subnuclei important for the sex-specific regulation of sociosexual behavior. Although estrogen receptors (ERs) are critical for organizing functional and cytoarchitectural sex differences in these subnuclei, a detailed developmental profile of ER expression in the amygdaloid complex is not available. Moreover, the kisspeptin gene (Kiss1) was recently identified in the adult amygdala, but it remains unknown if it is expressed during development. To fill these data gaps, rat brains (5-7/group) were assessed on postnatal days (PNDs) 0, 2, 4, 7, and 19 for ER alpha (ERα; Esr1), beta (ERβ; Esr2), and Kiss1 expression using in situ hybridization. Expression was quantified in the posterodorsal portion of the medial amygdala posterodorsal (MePD), lateral (PLCo), and medial (PMCo) components of the posterior cortical nucleus, and the amygdalohippocampal area (AHi). ERα expression was high throughout the amygdala at birth, but sexually dimorphic only in the AHi. ERα expression in the MePD and the PLCo showed a U-shaped expression pattern over time. In the PMCo, ERα expression decreased from PND 2 and remained low through PND 19. Sexually dimorphic expression of ERβ in the MePD was observed on PND 0, with higher levels in females, but reversed by PND 4 due to declining levels in females. No Kiss1 signal was observed in the postnatal amygdala, suggesting that expression arises after puberty. These data reveal that ER expression is region-specific within the neonatal amygdala. These differences likely contribute to sex differences in sociosexual behavior across the lifespan.

Developmental exposure to Bisphenol A (BPA), a component of polycarbonate and epoxy resins, has been purported to adversely impact reproductive function in female rodents. Because neonatal life is a critical window for the sexual dimorphic organization of the hypothalamic–pituitary–gonadal (HPG) axis, interference with this process could underlie compromised adult reproductive physiology. The goal of the present study was to determine if neonatal BPA exposure interferes with sex specific gene expression of estrogen receptor alpha (ERα), ER beta (ERβ) and kisspeptin (Kiss1) in the anterior and mediobasal hypothalamus. Long Evans (LE) neonatal rats were exposed to vehicle, 10 μg estradiol benzoate (EB), 50 mg/kg BPA or 50 μg/kg BPA by subcutaneous injection daily from postnatal day 0 (PND 0) to PND 2. Gene expression was assessed by in situ hybridization on PNDs 4 and 10. Within the anterior hypothalamus ERα expression was augmented by BPA in PND 4 females, then fell to male-typical levels by PND 10. ERβ expression was not altered by BPA on PND 4, but significantly decreased or eliminated in both sexes by PND 10. Kiss1 expression was diminished by BPA in the anterior hypothalamus, especially in females. There were no significant impacts of BPA in the mediobasal hypothalamus. Collectively, BPA effects did not mirror those of EB. The results show that neonatal hypothalamic ER and Kiss1 expression is sensitive to BPA exposure. This disruption may alter sexually dimorphic hypothalamic organization and underlie adult reproductive deficiencies. Additionally, the discordant effects of EB and BPA indicate that BPA likely disrupts hypothalamic organization by a mechanism other than simply acting as an estrogen mimic.

Release of gonadotropins in adult rodents is sex specific and dependent upon kisspeptin (Kiss1) neurons. This crucial pathway within the hypothalamic-pituitary-gonadal (HPG) axis is profoundly influenced by neonatal estrogens, which induce a male-like phenotype. Classically, estrogen activity is mediated via the estrogen receptors a and β (ERα and ERβ), but the relative roles each plays in organizing the sex-specific ontogeny of kisspeptin signaling pathways remain unresolved. Thus, the present study used in situ hybridization histochemistry (ISHH) to map the temporal and sexually dimorphic neonatal mRNA expression profiles of ERα, ERβ, and Kiss1 in the anterioventral periventricular nucleus (AVPV), medial preoptic area (MPOA), ventromedial nucleus (VMN), and arcuate nucleus (ARC), all regions critical for kisspeptin regulation of gonadotropin secretion. In general, females had higher levels of ERα, in all regions examined, a sex difference that persisted until postnatal day (PND) 19 except in the ARC. Males had significantly more ERβ expression in the AVPV at birth, but this sex difference was lost and then re-emerged on PND 19, with females having more than males. VMN ERβ levels were higher in females until PND 19. Kiss1 was not detectable until PND 11 in the anterior hypo-thalamus, but expression levels were equivalent at birth in the ARC. By PND 2, ARC ERα and Kiss1 levels were abundant, sexually dimorphic (higher in females), and, respectively, showed a U- and a bell-shaped pattern with age. Sex differences in ARC Kiss1 expression provide evidence that Kiss1 may play a role in the sexual dimorphic organization of the neonatal brain. These detailed profiles of neonatal Kiss1 and ERs mRNA levels will help elucidate the relative roles each plays in the sex-specific, estrogen-dependent organization of gonadotropin signaling pathways.

Sex differences are widespread across vertebrate nervous systems. Such differences are sometimes reflected in the neural substrate via neuroanatomical differences in brain region volume. One brain region that displays sex differences in its associated functions and pathologies is the striatum, including the caudate-putamen (dorsal striatum), nucleus accumbens core and shell (ventral striatum). The extent to which these differences can be attributed to alterations in volume is unclear. We thus tested whether the volumes of the caudate-putamen, nucleus accumbens core, and nucleus accumbens shell differed by region, sex, and hemisphere in adult Sprague–Dawley rats. As a positive control for detecting sex differences in brain region volume, we measured the sexually dimorphic nucleus of the medial preoptic area (SDN-POA). As expected, SDN-POA volume was larger in males than in females. No sex differences were detected in the volumes of the caudate-putamen, nucleus accumbens core or shell. Nucleus accumbens core volume was larger in the right than left hemisphere across males and females. These findings complement previous reports of lateralized nucleus accumbens volume in humans, and suggest that this may possibly be driven via hemispheric differences in nucleus accumbens core volume. In contrast, striatal sex differences seem to be mediated by factors other than striatal region volume. This conclusion is presented within the context of a detailed review of studies addressing sex differences and similarities in striatal neuroanatomy.

Concerns have been raised regarding the long-term impacts of early life exposure to the ubiquitous environmental contaminant bisphenol A (BPA) on brain organization. Because BPA has been reported to affect estrogen signaling, and steroid hormones play a critical role in brain sexual differentiation, there is also concern that BPA exposure could alter neural sex differences. Here, we examine the impact of subchronic exposure from gestation to adulthood to oral doses of BPA below the current no-observed-adverse-effect level (NOAEL) of 5 mg/kg body weight (bw)/day on estrogen receptor (ESR) expression in sexually dimorphic brain regions of prepubertal and adult female rats. The dams were gavaged daily with vehicle (0.3% carboxymethylcellulose), 2.5, 25, 260, or 2700 μg BPA/kg bw/day, or 0.5 or 5.0 μg ethinyl estradiol (EE)/kg bw/day from gestational day 6 until labor began. Offspring were then gavaged directly from the day after birth until the day before scheduled sacrifice on postnatal days 21 or 90. Using in situ hybridization, one or more BPA doses produced significant decreases in Esr1 expression in the juvenile female rat anteroventral periventricular nucleus (AVPV) of the hypothalamus and significant decreases in Esr2 expression in the adult female rat AVPV and medial preoptic area (MPOA), relative to vehicle controls. BPA did not simply reproduce EE effects, indicating that BPA is not acting solely as an estrogen mimic. The possible consequences of long-term changes in hypothalamic ESR expression resulting from subchronic low dose BPA exposure on neuroendocrine effects are discussed and being addressed in ongoing, related work.

The Border Gateway Protocol (BGP) is the single inter-domain routing protocol that enables network operators within each autonomous system (AS) to influence routing deci- sions by independently setting local policies on route filtering and selection. This independence leads to fragile networking and makes analysis of policy configurations very complex. To aid the systematic and efficient study of the policy configuration space, this paper presents network reduction, a scalability technique for policy-based routing systems. In network reduction, we provide two types of reduction rules that transform policy configurations by merging duplicate and complementary router configurations to simplify analysis. We show that the reductions are sound, dual of each other and are locally complete. The reductions are also computationally attractive, requiring only local configuration information and modification. We have developed a prototype of network reduction and demonstrated that it is applicable on various BGP systems and enables significant savings in analysis time. In addition to making possible safety analysis on large networks that would otherwise not complete within reasonable time, network reduction is also a useful tool for discovering possible redundancies in BGP systems. I. INTRODUCTION The Internet today runs on a complex routing protocol called the Border Gateway Protocol (BGP). It enables autonomous systems (ASes) worldwide to achieve global connectivity, subject to each system's local policy (which paths are allowed, and the route preference used to select best paths). The convergence behavior of the global Internet depends on how each AS configures its policy. Prior work has shown that policy misconfigurations can lead to rapid oscillation between routing states, slowing or even preventing convergence (11). This happens when the conflicting local policies cannot be reconciled: there is no solution to the routing problem. Other configurations support a unique stable solution, which normal protocol execution is bound to reach. We refer to such configurations as 'safe'. While abstract formal models of BGP (8), (5), (7) allow researchers to explore how local policies affect BGP stability, the membership problem for this safe subset is NP-hard, and real network configurations are very large, drastically limiting the feasibility of the safety test. In this paper, we present a novel network reduction tech- nique that enables networking researchers to study and analyze large BGP systems in a sound and automatic fashion. Central to network reduction is two forms of rewriting rules that transform policy configurations into smaller and simpler forms while preserving safety property. These rewriting rules are directed at known patterns in real networks, which exhibit considerable structural redundancy. Once a configuration is reduced, safety analysis can be performed directly to check for possible misconfigurations. To evaluate the effectiveness of the reduction technique for scaling up safety analysis, we use an automated analyzer (20) based on the Maude rewriting logic engine (12). In the Maude- based analyzer, a BGP system is encoded as a transition system driven by concurrent rewriting rules. Safety analysis is then performed by simulating execution runs on the transition system, as well as exhaustively exploring all execution runs for possible divergence. Our experimental results show that network reduction enables us to perform safety analysis effi- ciently, often completing the analysis on large networks that would otherwise not be possible to study within reasonable time. Prior work (19) demonstrates that a limited form of rewrit- ing rule based on merging identical router-level configurations can significantly improve convergence analysis time of BGP instances. This paper introduced a new form of rewriting rule, based on a new unified model (EPD). Using EPD model, we proved that the two forms of rules are dual. Moreover, we proved that they form a complete set of reduction rules that require only local rewrites; this and other properties of the reduction rules, have provided a deeper understanding of the redundancies presented in BGP systems, and established net- work reduction a sound and effective tool for scaling up formal analysis. Specifically, we make the following contributions: Formal model for reduction. We propose an abstract model for modeling Internet topology and policies. This abstract model, which we call the Extended Path Digraph (EPD), extends prior models (7), (16), and provides a basis for re- ducing instances prior to analysis. EPD enables the unification of configuration specification and analysis within a common model, resulting in simpler reductions. We further provide a tool developed using Maude for automatically extracting EPD from existing network topologies and policies. Network reduction. We present two reduction rules that trans- form EPD policy configurations to simplify analysis. These two reduction rules merge duplicate or complementary router- level configurations into one. We show that these reductions are sound and mutually dual, and establish a confluence result

Arylhydrocarbon receptor (Ahr) activation by 2,3,7,8-tetrachlordibenzo-p-dioxin (TCDD) interferes with female reproductive functions, but there is little information on the specific targets of TCDD in the hypothalamic-pituitary-gonadal (HPG) axis. In these studies, we found that TCDD upregulated known AhR target genes, cytochrome p450 1a1 (Cyp1a1), Cyp1a2 and Cyp1b1 in the rat pituitary gland. Moreover, 75% of pituitary lactotropes and 45% of gonadotropes contained Ahr mRNA, and most Ahr-containing cells were estrogen receptor 1 (Esr1)-positive. TCDD abrogated estradiol (E(2))-induced prolactin (Prl) expression in vivo and in vitro; conversely, E(2) blocked TCDD upregulation of luteinizing hormone beta (Lhb) and glycoprotein hormone alpha polypeptide (Cga) expression. TCDD had no effect on levels of Ahr mRNA, but upregulated Esr1 mRNA. E(2) independently repressed Ahr and Esr1 expression and blocked TCDD upregulation of Esr1. Thus, complex interactions between Ahr and Esr alter Prl and luteinizing hormone (LH) synthesis by direct actions in lactotropes and gonadotropes. These findings provide important insights into how TCDD disrupts female reproductive functions.

EGCG [(-)-epigallocatechin-3-gallate] has shown its antitumor ability and perhaps a potential regimen for cancer patients. The goal of this study was to investigate the effect of EGCG on human papilloma virus (HPV) positive cervical cancer cell lines. EGCG inhibited the growth of CaSki (HPV16 positive) and HeLa (HPV18 positive) cells in a time- and concentration-dependent manner. Cell cycle arrest and apoptosis were observed in two cell lines after EGCG exposure. More importantly, we focused on EGCG regulation ability on pivotal genes involved in cervical cancer: viral oncogenes E6/E7, estrogen receptor (ER) and aromatase. Our results suggested that EGCG may be suitable for prevention and treatment of cervical cancer.

Sex differences exist in how the brain regulates motivated behavior and reward, both in normal and pathological contexts. Investigations into the underlying neural mechanisms have targeted the striatal brain regions, including the dorsal striatum and nucleus accumbens core and shell. These investigations yield accumulating evidence of sexually different electrophysiological properties, excitatory synaptic input, and sensitivity to neuromodulator/hormone action in select striatal regions both before and after puberty. It is unknown whether the electrical properties of neurons in the nucleus accumbens shell differ by sex, and whether sex differences in excitatory synaptic input are present before puberty. To test the hypothesis that these properties differ by sex, we performed whole-cell patch-clamp recordings on male and female medium spiny neurons (MSNs) in acute brain slices obtained from prepubertal rat nucleus accumbens shell. We analyzed passive and active electrophysiological properties, and miniature EPSCs (mEPSCs). No sex differences were detected; this includes those properties, such as intrinsic excitability, action potential afterhyperpolarization, threshold, and mEPSC frequency, that have been found to differ by sex in other striatal regions and/or developmental periods. These findings indicate that, unlike other striatal brain regions, the electrophysiological properties of nucleus accumbens shell MSNs do not differ by sex. Overall, it appears that sex differences in striatal function, including motivated behavior and reward, are likely mediated by other factors and striatal regions.