RU486 facilitates or disrupts the sensitization of sexual behaviors by estradiol in the ovariectomized Long–Evans rat: Effect of timecourse

An acute injection of estradiol benzoate (EB) to the ovariectomized (OVX) rat activates low levels of lordosis, and subsequent progesterone (P) administration augments lordosis and recruits a complete pattern of sexual behavior including appetitive behaviors (e.g., hops/darts and solicitations). However, repeated injections of 5 μg or 10 μg EB (but not 2 μg EB), administered every 4 days to sexually-experienced OVX rats results in a behavioral sensitization, such that lordosis quotients (LQs) and appetitive behaviors progressively increase. We have shown that adrenal P does not play a critical role because behavioral sensitization to EB is not prevented by adrenalectomy. Here we tested whether P receptors play a role by examining the effect of chronic administration of the P receptor antagonist RU486 at a dose that reliably inhibits sexual behavior in fully primed OVX rats. Females were treated with EB (5 or 10 μg), and 5 mg RU486 dissolved in 0.4 mL vehicle (VEH; 80% sesame oil, 15% benzyl benzoate, 5% benzyl alcohol) 48 h and 5 h prior to each of 7 tests, respectively, occurring at 4-day intervals in unilevel 4-hole pacing chambers. Control animals were treated with 2, 5, or 10 μg EB + VEH. As expected, sensitization did not occur in females treated with 2 μg EB + VEH, and those females received fewer intromissions and ejaculations than all other groups. RU486 did not prevent the sensitization of LQ, moderate and high lordosis magnitudes (LM2 and LM3) or appetitive sexual behaviors on early tests, and in fact potentiated appetitive behaviors, LQ, LM2 and LM3, consistent with its facilitative actions in females treated with EB-alone, as we and others have reported previously. However, despite the initial facilitation, blocking P receptors by chronic administration of RU486 inhibited the maintenance of behavioral sensitization to EB.     

Olfactory conditioned same-sex partner preference in female rats: Role of ovarian hormones

The dopamine D2-type receptor agonist quinpirole (QNP) facilitates the development of conditioned same-sex partner preference in males during cohabitation, but not in ovariectomized (OVX) females, primed with estradiol benzoate (EB) and progesterone (P). Herein we tested the effects of QNP on OVX, EB-only primed females. Females received a systemic injection (every four days) of either saline (Saline-conditioned) or QNP (QNP-conditioned) and then cohabited for 24 h with lemon-scented stimulus females (CS +), during three trials. In test 1 (female-female) preference was QNP-free, and females chose between the CS + female and a novel female. In test 2 (male-female) they chose between the CS + female and a sexually experienced male. In test 1 Saline-conditioned females displayed more hops & darts towards the novel female, but QNP-conditioned females displayed more sexual solicitations towards the CS + female. In test 2 Saline-conditioned females displayed a clear preference for the male, whereas QNP-conditioned females displayed what we considered a bisexual preference. We discuss the effect of dopamine and ovarian hormones on the development of olfactory conditioned same-sex preference in females.     

Vaginocervical stimulation attenuates the sensitization of appetitive sexual behaviors by estradiol benzoate in the ovariectomized rat

The acute administration of estradiol benzoate (EB) to the ovariectomized (OVX) rat induces low levels of lordosis while sexually appetitive behaviors (e.g., hops, darts, solicitations) are absent, yet the repeated administration of EB results in a behavioral sensitization in which lordosis is potentiated and sexually appetitive behaviors are induced. We have shown that repeated copulation attenuates the sensitization of appetitive sexual behaviors. Here, we assessed which component of male stimulation during copulation is involved in the attenuation. On 8 occasions, sexually experienced OVX Long–Evans rats were treated with 10 μg EB and 48 h later assigned to one of six groups that differed in their experience on intermediates tests (2–7). One was given repeated access to a male (EB/Male), and another was placed in the copulation chamber alone (EB/Alone) on intermediate tests. Three groups were given one of three somatosensory stimuli by the experimenter: manual flank stimulation (FLS), clitoral stimulation (CLS), or vaginocervical stimulation (VCS). Finally, the control group was left undisturbed in the animal care facility (ACF). Sexual behaviors were measured on Tests 1 and 8. VCS received from the experimenter (VCS) or from the male during copulation (EB/Male) attenuated the magnitude of the sensitization of appetitive sexual behaviors compared with those that were not brought to the testing rooms (ACF), and the effect was most pronounced on sexual solicitations. These results suggest that VCS received during penile intromission inhibits the sensitization of sexually appetitive behaviors by repeated administration of EB. As such, repeated administration of EB may oppose those mechanisms that induce estrous termination, perhaps by sensitizing inhibitory processes within the ventromedial hypothalamus that typically prevent the display of sexual behaviors (i.e., by facilitating disinhibition).     

The inhibitory effects of corncob bedding on sexual behavior in the ovariectomized Long–Evans rat treated with estradiol benzoate are overcome by male cues

The mechanisms underlying the sensitization of sexual behaviors by repeated administration of estradiol benzoate (EB) to ovariectomized (OVX) rats are not well understood. Here we tested whether two housing conditions play a role. Sexual behavior in the female rat is dependent on the activation of ERα (estrogen receptor alpha) by estradiol. Corncob (CC) bedding has been reported to have adverse effects on the reproductive behavior and physiology of rats, and to disrupt ERα signaling in mice. In addition, some rodent behaviors are stimulated by olfactory stimuli and enhanced in the presence of estradiol. Upon arrival to the facilities OVX Long–Evans rats were housed on either Sani-Chips (SC) or CC in a room that housed only females (F) or males and females (M). Females were first given four sexual training sessions with 10 μg EB + 500 μg progesterone (P; administered 48 h and 4 h prior to training, respectively), followed by a 2-week hormone washout period. Next, 10 μg EB was administered s.c. every 4 days, 48 h prior to each of 8 test sessions in a unilevel 4-hole pacing chamber. On the final training day (i.e., when primed with EB + P), no inhibitory effects of corncob bedding were found, however a facilitation of the lordosis quality occurred in SC/F. Although all groups appear to have sensitized to the repeated administration of EB, CC/F animals displayed fewer high quality lordosis magnitudes and hop/darts, and received fewer mounts and intromissions overall. They also had a lower lordosis quotient (LQ) on tests 2–4 although this effect disappeared by test 5. These results suggest that although CC may inhibit some components of female sexual behavior when primed with EB alone, cues from sexually vigorous males can overcome that inhibition. Moreover, they suggest that male cues can facilitate mechanisms of estradiol sensitization. We recommend that quality control studies be conducted at individual institutions to assess any impact of corncob bedding on animal physiology and behavior.     

Non-ovarian aromatization is required to activate female sexual motivation in testosterone-treated ovariectomized quail

Although aromatase is expressed in both male and female brains, its functional significance in females remains poorly understood. In female quail, sexual receptivity is activated by estrogens. However it is not known whether sexual motivation is similarly estrogen-dependent and whether estrogens locally produced in the brain contribute to these behavioral responses. Four main experiments were designed to address these questions. In Experiment 1 chronic treatment of females with the anti-estrogen tamoxifen decreased their receptivity, confirming that this response is under the control of estrogens. In Experiment 2 chronic treatment with tamoxifen significantly decreased sexual motivation as treated females no longer approached a sexual partner. In Experiment 3 (a) ovariectomy (OVX) induced a significant decrease of time spent near the male and a significantly decreased receptivity compared to gonadally intact females, (b) treatment with testosterone (OVX + T) partially restored these responses and (c) this effect of T was prevented when estradiol synthesis was inhibited by the potent aromatase inhibitor Vorozole (OVX + T + VOR). Serum estradiol concentration was significantly higher in OVX + T than in OVX or OVX + T + VOR females. Together these data demonstrate that treatment of OVX females with T increases sexual motivation and that these effects are mediated at least in part by non-gonadal aromatization of the androgen. Finally, assays of aromatase activity on brain and peripheral tissues (Experiment 4) strongly suggest that brain aromatization contributes to behavioral effects observed here following T treatment but alternative sources of estrogens (e.g. liver) should also be considered.     

Estradiol modulates the anorexic response to central glucagon-like peptide 1

Estrogens suppress feeding in part by enhancing the response to satiation signals. Glucagon-like peptide 1 (GLP-1) acts on receptor populations both peripherally and centrally to affect food intake. We hypothesized that modulation of the central GLP-1 system is one of the mechanisms underlying the effects of estrogens on feeding. We assessed the anorexic effect of 0, 1, and 10 μg doses of GLP-1 administered into the lateral ventricle of bilaterally ovariectomized (OVX) female rats on a cyclic regimen of either 2 μg β-estradiol-3-benzoate (EB) or oil vehicle 30 min prior to dark onset on the day following hormone treatment. Central GLP-1 treatment significantly suppressed food intake in EB-treated rats at both doses compared to vehicle, whereas only the 10 μg GLP-1 dose was effective in oil-treated rats. To follow up, we examined whether physiologic-dose cyclic estradiol treatment influences GLP-1-induced c-Fos in feeding-relevant brain areas of OVX females. GLP-1 significantly increased c-Fos expression in the area postrema (AP) and nucleus of the solitary tract (NTS), and the presence of estrogens may be required for this effect in the paraventricular nucleus of the hypothalamus (PVN). Together, these data suggest that modulation of the central GLP-1 system may be one of the mechanisms by which estrogens suppress food intake, and highlight the PVN as a region of interest for future investigation.     

Estrogen-dependent estrous behavior in rabbits is antagonized by the antiprogestin RU486

Estrogen receptor-alpha (ERα) dimerizes with unliganded progesterone receptor (PR) in target tissues to trigger genomic and non-genomic effects. In ovariectomized rats the antiprogestin RU486 or antisense nucleotides against PR antagonize estradiol-induced sexual receptivity. We determined the relevance of unliganded PR for the expression of estrogen-dependent scent-marking (chinning) and sexual receptivity by injecting RU486 to: a) ovariectomized (ovx) rabbits given estradiol benzoate (EB; 5 μg/day); b) intact rabbits. Chinning and lordosis were quantified in ovx animals before (5 days; baseline) and during hormonal treatments: EB + RU486 (20 mg/day; n = 18) or EB + vehicle (n = 18). On treatment day 4 LQ (lordosis quotient) increased in both groups, relative to baseline (mean ± se): LQ = 1 ± 5 (baseline) vs 25 ± 21 (EB + RU486) and 2 ± 6 (baseline) vs 37 ± 29 (EB + vehicle). On day 9 LQ values were: 22 ± 23 (EB + RU486) and 54 ± 39 (EB + vehicle). Chinning increased only in the EB + vehicle group (day 9 = 55 ± 46 vs baseline = 17 ± 20 marks/10 min). In intact rabbits one RU486 injection: reduced the LQ from 72 ± 7to 36 ± 8 five hrs later, increased the latency to receive first ejaculation from 45 to 98 s, and decreased the number of ejaculations received in the test from 3 to 2 but did not modify mounting latency or chinning. Results support a participation of unliganded PR for the induction (ovx) and maintenance (intact) of rabbit estrous behavior by estrogens.     

Sensitization of sexual behavior in ovariectomized rats by chronic estradiol treatment

The ovariectomized (OVX) rat treated with estradiol benzoate (EB) is used to elucidate neuroendocrine mechanisms of sexual behavior. Chronic behavioral and pharmacological manipulations can be confounded by rising baselines, since females are behaviorally more sensitive to repeated EB injections. The literature lacks a systematic examination of chronic effects of EB administered alone to the sexually experienced OVX rat. Long–Evans rats were repeatedly treated (8 tests) with s.c. injections of 2, 5, or 10 μg EB at different time intervals (4 or 8 days). Female sexual behaviors as well as receipt of mounts, intromissions and ejaculations from the male were observed in the unilevel 4-hole pacing chamber. The effects of adrenalectomy (ADX) and strain (Long–Evans vs. Wistar) were also assessed. Long–Evans OVX rats treated with 5 μg EB every 8 days showed persistently low levels of sexual behavior. Sensitization was most robust following 10 μg EB at 4-day intervals. Very few sexual behaviors were ever induced by 2 μg EB. ADX did not affect the development of behavioral sensitization by 10 μg EB. Therefore, to achieve a low steady state of sexual behaviors in sexually experienced Long-Evans OVX rats 5 μg of EB administered every 8 days is optimal, whereas a persistently high level of sexual behaviors is induced with 10 μg EB administered every 4 days. OVX Wistar rats are behaviorally more sensitive to EB. Behavioral sensitization to EB may serve as a mechanism to optimize reproductive success.     

Anxiety-like behaviors in adulthood are altered in male but not female rats exposed to low dosages of polychlorinated biphenyls in utero

Exposure to polychlorinated biphenyls (PCBs), a class of endocrine-disrupting chemicals, can result in altered reproductive behavior in adulthood, especially when exposure occurs during critical periods of brain sexual differentiation in the fetus. Whether PCBs alter other sexually dimorphic behaviors such as those involved in anxiety is poorly understood. To address this, pregnant rat dams were injected twice, on gestational days 16 and 18, with the weakly estrogenic PCB mixture Aroclor 1221 (A1221) at one of two low dosages (0.5 mg/kg or 1.0 mg/kg, hereafter 1.0 and 0.5), estradiol benzoate (EB; 50 μg/kg) as a positive estrogenic control, or the vehicle (3% DMSO in sesame oil). We also conducted a comprehensive assessment of developmental milestones of the F1 male and female offspring. There were no effects of treatment on sex ratio at birth and age at eye opening. Puberty, assessed by vaginal opening in females and preputial separation in males, was not affected in females but was advanced in males treated with A1221 (1.0). Males and females treated with A1221 (both dosages) were heavier in early adulthood relative to controls. The earliest manifestation of this effect developed in males prior to puberty and in females slightly later, during puberty. Anxiety-like behaviors were tested using the light:dark box and elevated plus maze tests in adulthood. In females, anxiety behaviors were unaffected by treatment. Males treated with A1221 (1.0) showed reduced indices of anxiety and increased activity in the light:dark box but not the elevated plus maze. EB failed to replicate the phenotype produced by A1221 for any of the developmental and behavioral endpoints. Collectively, these results indicate that PCBs increase body weight in both sexes, but their effects on anxiety-like behaviors are specific to males. Furthermore, differences between the results of A1221 and EB suggest that the PCBs are likely acting through mechanisms distinct from their estrogenic activity.     

Impact of 1.8-GHz radiofrequency radiation (RFR) on DNA damage and repair induced by doxorubicin in human B-cell lymphoblastoid cells

In the present in vitro study, a comet assay was used to determine whether 1.8-GHz radiofrequency radiation (RFR, SAR of 2 W/kg) can influence DNA repair in human B-cell lymphoblastoid cells exposed to doxorubicin (DOX) at the doses of 0 μg/ml, 0.05 μg/ml, 0.075 μg/ml, 0.10 μg/ml, 0.15 μg/ml and 0.20 μg/ml. The combinative exposures to RFR with DOX were divided into five categories. DNA damage was detected at 0 h, 6 h, 12 h, 18 h and 24 h after exposure to DOX via the comet assay, and the percent of DNA in the tail (% tail DNA) served as the indicator of DNA damage. The results demonstrated that (1) RFR could not directly induce DNA damage of human B-cell lymphoblastoid cells; (2) DOX could significantly induce DNA damage of human B-cell lymphoblastoid cells with the dose–effect relationship, and there were special repair characteristics of DNA damage induced by DOX; (3) E–E–E type (exposure to RFR for 2 h, then simultaneous exposure to RFR and DOX, and exposure to RFR for 6 h, 12 h, 18 h and 24 h after exposure to DOX) combinative exposure could obviously influence DNA repair at 6 h and 12 h after exposure to DOX for four DOX doses (0.075 μg/ml, 0.10 μg/ml, 0.15 μg/ml and 0.20 μg/ml) in human B-cell lymphoblastoid cells.     

Saxitoxin monitoring in three species of Florida puffer fish

Beginning in April 2002, three species of Florida puffer fish from around the state of Florida, USA were monitored for the presence of saxitoxin (STX). In total, 873 southern (Sphoeroides nephelus), 171 checkered (S. testudineus), and 53 bandtail (S. spengleri) puffer fish were collected between 2002 and 2006 from eight regions: Jacksonville, the Indian River Lagoon, Tequesta, the Florida Keys, Charlotte Harbor, Tampa Bay, Cedar Key, and Apalachicola. Emphasis was placed on collecting specimens from the Indian River Lagoon (IRL), where recreational harvesting of puffer fish led to 28 cases of saxitoxin puffer fish poisoning (SPFP) between January 2002 and May 2004. Southern puffer fish from the northern IRL routinely contained the highest concentrations of STX, with average levels in the skin of 1787 μg STXequiv./100 g tissue. Elevated concentrations were also found in the muscle (1102 μg STXequiv./100 g), gut contents (539 μg STXequiv./100 g), gonads (654 μg STXequiv./100 g), and liver (214 μg STXequiv./100 g). Lower, yet significant (above the action limit of 80 μg STXequiv./100 g tissue), concentrations of STX were also detected in the skin (599 μg STXequiv./100 g), muscle (233 μg STXequiv./100 g), gut contents (197 μg STXequiv./100 g), and gonads (239 μg STXequiv./100 g) of southern puffer fish from Tequesta in the southern IRL, as well as in the gonads (122 μg STXequiv./100 g) of Jacksonville southern puffer fish and the skin (265 μg STXequiv./100 g) of Tampa Bay southern puffer fish. STX concentrations above the action limit were also found in the skin of bandtail puffer fish from the IRL (620 μg STXequiv./100 g), Tequesta (374 μg STXequiv./100 g), and the Florida Keys (230 μg STXequiv./100 g). Checkered puffer fish collected from the IRL, Tequesta, and the Florida Keys on average were nontoxic, containing STX levels below the action limit in all tissues.     

Effect of follicular wave synchronization on in vitro embryo production in heifers

Aiming to achieve the ideal time of ovum pick-up (OPU) for in vitro embryo production (IVP) in crossbred heifers, two Latin square design studies investigated the effect of ovarian follicular wave synchronization with estradiol benzoate (EB) and progestins. For each experiment, crossbred heifers stage of estrous cycle was synchronized either with a norgestomet ear implant (Experiment 1) or a progesterone intravaginal device (Experiment 2) for 7 d, followed by the administration of 150 μg d-cloprostenol. On Day 7, all follicles >3 mm in diameter were aspirated and implants/devices were replaced by new ones. Afterwards, implant/device replacement was conducted every 14 d. Each experiment had three treatment groups. In Experiment 1 (n = 12), heifers in Group 2X had their follicles aspirated twice a week and those in Groups 1X and 1X-EB were submitted to OPU once a week for a period of 28 d. Heifers from Group 1X-EB also received 2 mg EB i.m. immediately after each OPU session. In Experiment 2 (n = 11), animals from Group 0EB did not receive EB while heifers in Groups 2EB and 5EB received 2 and 5 mg of EB respectively, immediately after OPU. The OPU sessions were performed once weekly for 28 d. Therefore, in both experiments, four OPU sessions were performed in heifers aspirated once a week and in Experiment 1, eight OPU sessions were done in heifers aspirated twice a week. Additionally, during the 7-d period following follicular aspiration, ovarian ultrasonography examinations were conducted to measure diameter of the largest follicle and blood samples were collected for FSH quantification by RIA. In Experiment 1, all viable oocytes recovered were in vitro matured and fertilized. Results indicated that while progestin and EB altered follicular wave patterns, this treatment did not prevent establishment of follicular dominance on the ovaries of heifers during OPU at 7-d intervals. Furthermore, the proposed stage of follicular wave synchronization strategies did not improve the number and quality of the recovered oocytes, or the number of in vitro produced embryos.     

Embryo production in superstimulated llamas pre-treated to inhibit follicular growth

Llamas are monotocous and the length of their gestation period varies between 342 and 350 days. Thus the average number of offspring any female can produce throughout her reproductive life is very limited to spread a desired genome. The multiple ovulation and embryo transfer (MOET) technique allows an alternative to this limitation and reduces the generation interval. The objective of this study was to evaluate embryo recovery in superstimulated llamas which had previously been hormone-treated to inhibit follicular growth. A total of 50 female llamas were monitored daily via rectal palpation and ultrasound and divided according to their ovarian follicular growth into four phases. The females in each phase were then randomly divided into two groups: A (n = 20) received a single dose of 1 mg of estradiol benzoate (EB) on the first day of the treatment + 100 mg of progesterone (P4) i.m. for 5 days with 5 animals per phase and B (n = 20) received 1 mg EB at onset + 150 mg P4 i.m. for a period of 5 days with 5 animals per phase. Group C (n = 10) or control did not receive any prior hormonal treatment and the females were in follicular phase I. All groups were monitored daily and, in the presence of ovarian follicles smaller than the dominant size at the end of treatment, all were superstimulated with 1000 IU eCG. For plasma progesterone concentration recording, daily blood samples were collected from days ?1 to 5 in the treated females in Group A and B. No significant differences were observed regarding the inhibition of follicle growth and in the plasma progesterone concentrations between Group A and B. The ovarian response to superstimulation was 56.2%, 71.4% and 90%, with the average number of dominant follicles produced per female being 4.4 ± 0.9; 4.8 ± 0.7 and 4.6 ± 0.6 in Groups A, B and C, respectively. The embryo recovery rate was 77.7%; 90% and 66.7% and the average number of embryos recovered per female was 2.9 ± 0.9; 2.6 ± 0.9 and 2.4 ± 0.8 for Groups A, B and C, respectively. In Groups A and B, the static follicular phase (III) seemed to be ideal for initiating the assisted reproductive technique of MOET. Although prior administration of P4 + EB seems to have no effect on the number of females that responded to the superstimulation treatments, the number of embryos recovered showed a tendency to be higher when ovarian follicle growth inhibition was performed beforehand.     

Plasma gossypol dynamics in white-tailed deer: Implications for whole cottonseed as a supplemental feed

Whole cottonseed (WCS) is a potential supplemental feed for white-tailed deer (Odocoileus virginianus) in rangeland conditions because of its high digestible energy and protein content, moderate fiber content, and resistance to degradation in moist conditions. WCS also contains the polyphenolic secondary metabolite gossypol, which reduces palatability to non-target monogastric species but may be of concern for deer nutrition. Plasma gossypol stabilization when fed a constant dry matter intake, plasma gossypol depletion after WCS was removed from the diet, and the relationship between WCS consumption and plasma gossypol concentration was studied in 10 mature male (N = 5) and female (N = 5) captive white-tailed deer. Consumption of WCS by 73 free-ranging white-tailed deer (59 males, 14 females) was estimated using results of the captive study. Plasma gossypol concentrations declined exponentially and averaged 0.74 μg/mL 35 days after WCS was removed from the diet. Plasma gossypol concentration was linearly related to WCS consumption (P < 0.001), with females having 0.35 μg/mL greater (P = 0.04) plasma gossypol than males for a given rate of dry matter consumption. All female and 93% of male white-tailed deer captured in WCS supplemented pastures had detectable plasma gossypol. Female averaged 1.88 μg/mL of plasma gossypol and males averaged 4.84 μg/mL of plasma gossypol. Based on the captive deer data, these plasma values suggest an average WCS consumption of ∼2.6 g/kg BW/day for female free-ranging deer and ∼5.6 g/kg BW/day for male free-ranging deer. Inferentially, a large proportion of free-ranging white-tailed deer in rangeland conditions will consume WCS, with females consuming 125 g WCS/day and males consuming 428 g WCS/day. That plasma gossypol levels decrease rapidly after cottonseed is removed from the diet suggests that the long withdrawal periods often used prior to breeding season may not be needed. However, although 93% of gossypol was eliminated from the animals after a five-week withdrawal period, a small amount of gossypol can still be detected. While our preliminary data on these animals suggests that these levels are not detriment to animal health or reproduction, ranch managers may want to take a conservative approach to the feeding of WCS until these questions are answered.     

Changes of substance P during fracture healing in ovariectomized mice

Neuropeptides may play an important role in the healing process of osteoporotic fractures. The objective of this study was to determine the role of substance P during osteoporotic fracture healing.One hundred ninety-two mice were randomized into ovariectomy (OVX) and control (CON) group (n = 96, respectively). Femoral shaft fracture was created 3 weeks after OVX. Bone mineral density (BMD), micro-CT (µCT) analysis of fracture callus formation and mineralization, µCT analysis of fracture site neovascularization and biomechanical property as well as substance P levels were evaluated 1, 2, 4, and 8 weeks after fracture and compared with CON group.Following OVX-induced bone loss, fracture healing in OVX mice was significantly poorer than that in CON mice, with a significant decrease of substance P at the fracture site at all time points and with the level at early stage (1 and 2 weeks) higher than later stage (4 and 8 weeks). Impaired angiogenesis was also noted in OVX mice. No significant change of substance P level in serum was found between different groups or time points.In conclusion, fracture healing is inferior in OVX-induced bone loss and associated with a significant decrease of substance P. Substance P may play an important role during osteoporotic fracture healing.     

Gender difference in blood cadmium concentration in the general population: Can it be explained by iron deficiency?

IntroductionGender differences in blood cadmium concentrations and the effect of iron deficiency on blood cadmium levels were analyzed in a representative sample of Koreans assessed in the Korean National Health and Nutritional Examination Survey (KNHANES) 2008–2011.MethodsA rolling sampling design was used to perform a complex, stratified, multistage probability cluster survey of a representative sample of the non-institutionalized civilian population in South Korea. Serum ferritin was categorized as low (<15.0 μg/L), low normal (15.0–<30.0 μg/L for females and 15.0–<50.0 μg/L for males), and normal (≥30.0 μg/L for females and ≥50.0 μg/L for males), and its association with blood cadmium levels was assessed after adjustment for various demographic and lifestyle factors.ResultsThe geometric mean (GM) of the blood cadmium level was significantly higher in females than in males, and significantly higher in older individuals for both genders. After controlling for covariates, multiple regression analysis with interaction terms showed that blood cadmium was correlated with serum ferritin levels only in pre-menopausal females.DiscussionIron deficiency is associated with blood cadmium levels in a representative sample of pre-menopausal females, as evaluated in KNHANES. Gender differences in blood cadmium concentration may not be due solely to an iron deficiency-associated increase in blood cadmium.     

Effects of developmental exposure to bisphenol A on spatial navigational learning and memory in rats: A CLARITY-BPA study

Bisphenol A (BPA) is a ubiquitous industrial chemical used in the production of a wide variety of items. Previous studies suggest BPA exposure may result in neuro-disruptive effects; however, data are inconsistent across animal and human studies. As part of the Consortium Linking Academic and Regulatory Insights on BPA Toxicity (CLARITY-BPA), we sought to determine whether female and male rats developmentally exposed to BPA demonstrated later spatial navigational learning and memory deficits. Pregnant NCTR Sprague–Dawley rats were orally dosed from gestational day 6 to parturition, and offspring were directly orally dosed until weaning (postnatal day 21). Treatment groups included a vehicle control, three BPA doses (2.5 μg/kg body weight (bw)/day—[2.5], 25 μg/kg bw/day—[25], and 2500 μg/kg bw/day—[2500]) and a 0.5 μg/kg/day ethinyl estradiol (EE)-reference estrogen dose. At adulthood, 1/sex/litter was tested for seven days in the Barnes maze. The 2500 BPA group sniffed more incorrect holes on day 7 than those in the control, 2.5 BPA, and EE groups. The 2500 BPA females were less likely than control females to locate the escape box in the allotted time (p value = 0.04). Although 2.5 BPA females exhibited a prolonged latency, the effect did not reach significance (p value = 0.06), whereas 2.5 BPA males showed improved latency compared to control males (p value = 0.04), although the significance of this result is uncertain. No differences in serum testosterone concentration were detected in any male or female treatment groups. Current findings suggest developmental exposure of rats to BPA may disrupt aspects of spatial navigational learning and memory.     

Antibiotic resistance profiling and phenotyping of Aeromonas species isolated from aquatic sources

This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13), bivalve (n = 10), sea cucumber (n = 16) and sea water (n = 14) and resistance to 12 antibiotics – Tetracycline (30 μg), Kanamycin (30 μg), Oxytetracycline (30 μg), Ampicillin (10 μg), Streptomycin (10 μg), Gentamicin (10 μg), Sulphamethoxazole (25 μg), Nalixidic acid (30 μg), Trimethoprim (1.25 μg), Novobiocin (5 μg), Penicilin (10 μg) and Chloramphenicol (10 μg) was tested. The results obtained from this study reveal multi drug resistance pattern among the isolates. All the isolates were completely resistant to Ampicillin, Novobiocin, Sulphamethoxazole and Trimethoprim, respectively but susceptible to Tetracycline (100%), Kanamycin (5.7%), Gentamicin (5.7%) and Oxytetracycline (24.5%). Antibiotics phenotyping of the bacteria revealed 21 different phenotypes among the isolates.     

The effect of iron and zinc supplementation and discontinuation of this practice on iron and zinc level in tissues in rats fed deficient diets

The effect of iron and iron/zinc supplementation on their levels in tissues of rats fed initially one of the three following regimen: C – control AIN-93 diet, D – iron deficient diet and R – diet with 50% reduction of all vitamins and minerals was investigated. The study was conducted on 6-week male Wistar rats, in 3 stages: (1) 4-week adaptation to the diets (C, D or R); (2) 4-week supplementation with the same regimen enriched with 10-times more iron (CSFe, DSFe, RSFe) or iron/zinc (CSFeZn, DSFeZn, RSFeZn); (3) 2-week post-supplementation period (the same diets as the stage I). Iron and zinc content in serum, the initial segment of intestine, liver and kidney were measured using FAAS method. After supplementation period (stage II) the content of iron in the intestine, liver and kidney in groups of rats fed DSFe and DSFeZn-diet were significantly higher (all p-values ≤ 0.05) than in rats fed D-diet (intestine: DSFe = 50.1 ± 9.0 μg/g wet weight, DSFeZn = 43.0 ± 9.9 μg/g vs. D = 16.5 ± 2.1 μg/g; liver: DSFe = 149 ± 30 μg/g, DSFeZn = 152 ± 25 μg/g vs. D = 56 ± 13 μg/g; kidney: DSFe = 74.0 ± 8.1 μg/g, DSFeZn = 72.7 ± 6.6 μg/g vs. D = 59.3 ± 9.5 μg/g). The same significant associations (all p-values ≤ 0.05) were observed in R rats in the intestine and liver (intestine: RSFe = 60.8 ± 6.6 μg/g, RSFeZn = 54.8 ± 6.6 μg/g vs. R = 31.5 ± 8.2 μg/g; liver: RSFe = 161 ± 10 μg/g, RSFeZn = 166 ± 21 μg/g vs. R = 136 ± 24 μg/g). After post-supplementation period the statistically significant differences between supplemented and non-supplemented rats fed D- and R-diets were still observed. There was not found the effect of applied treatments on zinc status. In conclusion, iron or iron/zinc supplementation increased similarly iron level in tissues of rats fed D-diet or R-diet with prolonged effect after supplementation discontinuation.     

Iron levels in the human brain: A post-mortem study of anatomical region differences and age-related changes

The link between brain iron homeostasis and neurodegenerative disease has been the subject of extensive research. There is increasing evidence of iron accumulation during ageing, and altered iron levels in some specific brain regions in neurodegenerative disease patients have been reported.Using graphite furnace atomic absorption spectrometry after microwave-assisted acid digestion of the samples, iron levels were determined in 14 different areas of the human brain [frontal cortex, superior and middle temporal, caudate nucleus, putamen, globus pallidus, cingulated gyrus, hippocampus, inferior parietal lobule, visual cortex of the occipital lobe, midbrain, pons (locus coeruleus), medulla and cerebellum (dentate nucleus)] of n = 42 adult individuals (71 ± 12 years old, range: 53–101 years old) with no known history or evidence of neurodegenerative, neurological or psychiatric disorders.It was found that the iron distribution in the adult human brain is quite heterogeneous. The highest levels were found in the putamen (mean ± SD, range: 855 ± 295 μg/g, 304–1628 μg/g) and globus pallidus (739 ± 390 μg/g, 225–1870 μg/g), and the lowest levels were observed in the pons (98 ± 43 μg/g, 11–253 μg/g) and medulla (56 ± 25 μg/g, 13–115 μg/g).Globally, iron levels proved to be age-related. The positive correlation between iron levels and age was most significant in the basal ganglia (caudate nucleus, putamen and globus pallidus).Compared with the age-matched control group, altered iron levels were observed in specific brain areas of one Parkinson's disease patient (the basal ganglia) and two Alzheimer's disease patients (the hippocampus).