Evaluation of trace element and mineral status and related to levels of amino acid in children with phenylketonuria

The aim of the present study was to examine trace elements (Zn, Cu, Mn, Se, Fe, Co, Cr, Ni, Cd, Pb), minerals (Ca, Mg, K), amino acids status in children with phenylketonuria and also whether they were correlated with each other in phenylketonuric patients.It has been found out that the HPA group was significantly lower than the control group with regards to Zn, Se, K, Ca, Mg and Zn/Cr levels (p < 0.001, p < 0.01, p < 0.001, p < 0.01, p < 0.01 and p < 0.001 respectively). In the patients with HPA, significantly strong positive correlations were observed between magnesium and calcium (r = 0.791; p = 0.001), also, indicates negative significant correlation between the concentrations of magnesium and phenylalanine (r = −0.591; p = 0.026).The results of this study showed that, in the HPA group, phenylalanine-Mg relationship found, the presence of disease will in the evaluation of phenylalanine and other amino acids, together with the value of magnesium is required to consider.     

双酚 A对成年 C57BL/6J小鼠糖、脂代谢的影响

目的：研究双酚A（BPA）长期暴露对C57BL/6J小鼠血糖、血脂的影响。方法：8周龄雄性小鼠随机分为正常对照组、溶剂对照组、低剂量BPA组、高剂量BPA组,分别给予蒸馏水、玉米油、0.5mg·kg^-1·d^-1和50mg·kg^-1·d^-1BPA,90d后,进行口服糖耐量（OGTT）测定,血TC、TG、HDL-C、LDL-C含量测定,肝脏病理切片及肝脏脂肪酸合成酶（FAS）、乙酰辅酶A羧化酶（ACC）、硬脂酰CoA脱饱和酶1（SCD1）和葡萄糖激酶（GCK）表达水平的测定。结果：与正常对照组和溶剂对照组相比,低剂量BPA组小鼠糖耐量存在明显异常（P〈0.05）;血脂各指标出现异常,血TC、TG、LDL-C升高,HDL-C下降（P均〈0.05）;高剂量BPA组TC、TG和HDL-C水平差异无统计学意义（P均〉0.05）,而LDL-C水平升高（P〈0.05）。低剂量BPA组肝脏FAS、ACC、SCD1、GCK蛋白表达升高,肝脏出现明显的脂质沉积;而高剂量BPA组仅ACC和GCK蛋白水平表达略有升高,其余蛋白无明显变化。结论：长期低剂量BPA接触会引起机体糖、脂代谢紊乱。     

Exposure of neonatal rats to the endocrine disrupter Bisphenol A affects ontogenic expression pattern of testicular steroid receptors and their coregulators

In light of the adverse reports of Bisphenol A (BPA) on reproduction and considering the pivotal role played by the steroid receptors (SRs) and their coregulators in male reproduction, it was of interest to decipher the influence that BPA may have on their expression pattern during critical ’windows‘ of development. Male rats were injected with 2.4 µg per pup per day of BPA from postnatal days (PND) 1–5 and controls received vehicle. During development, the testicular expression pattern of SRs (AR, ERβ and ERα), coactivators (SRC‐1, SRC‐2 and SRC‐3) and corepressors (NCoR and SMRT) in BPA‐exposed rats were compared. A significant decrease in the expression of SRs was seen in the BPA group. SRC‐1 showed a significant decrease, whereas SRC‐2 and SRC‐3 showed a significant increase in the protein expression whereas corepressor expression remained unaltered in the BPA‐exposed groups. Such impairments in the expression pattern can be a putative mechanism of adversities on fertility as a result of BPA exposure. Copyright © 2013 John Wiley & Sons, Ltd.     

氨基酸对利用红色糖多孢菌发酵生产红霉素的产量及组分影响研究

研究红色糖多孢菌发酵生产红霉素过程中几种氨基酸对产量及组分的影响。通过摇瓶实验证实了0h时向合成培养基中分别添加数种氨基酸，对红霉素的产量与组分都有着重要的影响。本文重点研究甘氨酸在红色糖多孢菌发酵中对产量和组分的影响，试验结果表明，0h添加0．05％的甘氨酸产量明显提高（72．2％），红霉素A组分相对百分含量提高9．7％，红霉素C组分减少78．7％。通过50L FUS-50（A）发酵罐进行验证，红霉素产量达到8939u／ml，比对照提高12．2％，EMA：EMC从5．1：1提高到8.2：1。     

软骨藻酸对H4细胞氨基酸释放和Ca2+浓度的影响

目的　研究软骨藻酸 (domoicacid)对H4细胞的兴奋性、抑制性氨基酸释放和胞内游离Ca2 ( [Ca2 ]i)浓度的影响。方法　应用高效液相色谱 (HPLC)和荧光分光光度计检测 0、0 0 64、0 64和 6 4μmol/L软骨藻酸作用于细胞 2h后的兴奋性、抑制性氨基酸释放和 [Ca2 ]i浓度。结果　天冬氨酸和谷氨酸 :中、高剂量组均高于对照组 ,差异有显著性(P <0 0 5 ) ;甘氨酸 :仅高剂量组高于对照组 ,差异有显著性 (P <0 0 5 ) ;γ 氨基丁酸 :低、中和高剂量组均低于对照组 ,差异有显著性 (P <0 0 5 ) ;兴奋性氨基酸 /抑制性氨基酸 :低、中和高剂量组均高于对照组 ,差异有显著性 (P <0 0 5 )。胞内[Ca2 ]i :低、中和高剂量组分别为 ( 2 0 8 65± 11 0 1)、( 3 42 3 1± 15 0 8)和 ( 5 81 3 6± 17 2 4)nmol/L ,高于对照组 [( 14 3 2 5±11 97)nmo1/L] ,差异有显著性 (P <0 0 1)。兴奋性氨基酸与 [Ca2 ]i浓度有显著的正相关性 (r =0 93 2 0P <0 0 1)。结论　软骨藻酸能引起H4细胞兴奋性和抑制性氨基酸释放和胞内 [Ca2 ]i变化 ,这种变化可能是软骨藻酸兴奋性神经毒性的重要机制     

The effect of amino acids on intestinal smooth muscle related to their content in blood and tissue

The concentration of amino acids in the blood of the guinea pig is about 0.1 mM, while the intracellular concentration in the gastrointestinal smooth muscle cells is about ten times higher. Most amino acids, applied exogenously to isolated smooth muscles, exert effects in concentrations resembling their intracellular concentration. The results show that the inhibiting amino acids do not function as a neurotransmitter in the non-adrenergic inhibitory nervous system. The effective amino acids may possess modifying properties with respect to the contractile state of the muscle.     

Mutagenic effect of Bisphenol A on adult rat male germ cells and their fertility

The present study investigates the effects of Bisphenol A on the induction of dominant lethal mutation and male reproductive functions. The male rats were gavaged with BPA (10 μg, and 5.0 mg/kg/bw) over a period of six days and control group with vehicle. Each male was cohabited with untreated females sequentially over the period of eight weeks. The mated females were sacrificed on 15th day of gestation. The results revealed a significant increase in dominant lethal mutation rate during fourth and sixth week of mating intervals at 5.0 mg/kgbw dose of BPA. These findings demonstrate that mid-spermatids and spermatocytes are more sensitive to BPA exposure. The male rats sacrificed at the end of mating study showed an increase in the sperm DNA damage, and decrease in motility at higher dose. However, significant reductions in sperm production effects were observed at both lower and higher doses of BPA. These preliminary results indicate that BPA may be a weak male germ cell mutagen.     

双酚A对人子宫内膜基质细胞增殖及雌/雄激素受体表达的影响

目的 观察双酚A对人子宫内膜基质细胞(hESCs)增殖和雌/雄激素受体(ERα/AR)表达的影响.方法 体外培养hESCs,在6孔板贴壁达70%～80%后鉴定细胞纯度.采用无血清培养同步化,然后给予不同浓度的双酚A(0、10~(-8)、10~(-6)、10~(-4) mol/L)处理48 h.Trizol裂解细胞提取总RNA,实时荧光定量PCR法测定基质细胞ERα和AR mRNA表达水平;采用流式细胞技术测定细胞周期.结果 本文纯化培养方法得到的hESCs,波形蛋白染色阳性率达95%.10~(-6)、10~(-4)mol/L的双酚A对基质细胞增殖周期有抑制作用.10~(-4) mol/L的双酚A使基质细胞ERαmRNA表达显著升高(P<0.05);10~(-6)、10~(-4)mol/L的双酚A也显著升高AR mRNA的表达水平(P<0.05);但是,10~(-8)mol/L的双酚A则有抑制ERα/AR mRNA表达的趋势(P>0.05).结论 低剂晕双酚A抑制人子宫内膜ERα/AR表达,而高浓度双酚A则升高ERα/AR表达,进而影响子宫内膜的功能.     

双酚A对颗粒细胞激素生成及相关甾体生成酶的影响

目的 探讨双酚A(BPA)对大鼠颗粒细胞雌、孕激素生成及相关甾体生成酶的影响.方法 采用大鼠卵巢颗粒细胞进行体外培养,72 h后给予不同浓度的BPA(0、10-7、10-6、10-5、10-4 M)处理,继续培养48 h后,收集细胞培养液,用超敏感的固相放射免疫方法测定孕酮和雌二醇(E2)的浓度;用荧光实时定量PCR法测定颗粒细胞内芳香化酶(P450arom)、胆固醇侧链裂解酶(P450Scc)和类固醇合成急性调节蛋白(StAR)的mRNA表达水平.结果 在BPA作用下,颗粒细胞孕酮分泌量在10-7M到10-5M组逐渐升高(P＜0.05),但在10-4M剂量组出现意外陡降(P＜0.01);而E2的分泌量则随BPA浓度的增加而下降(P＜0.01).P450arom mRNA表达显著降低;P450Scc表达量亦在10-7M到10-5M组逐渐升高(P＜0.05),而10-4M组出现显著降低(P＜0.01).StAR mRNA表达在10-4M组增高(P＜0.05).结论 BPA能直接作用于卵巢颗粒细胞,通过作用于P450arom,P450Scc等相关酶的表达以及类固醇合成的限速因子-StAR的表达,而影响颗粒细胞雌孕激素的分泌.     

Effects of bisphenol A on gap junctions in HaCaT cells as mediated by the estrogen receptor pathway

Bisphenol A (BPA) is widely used as the raw material for the production of plastics and paper products. People can be exposed to BPA through dermal contact, particularly for cashiers in contact with thermal paper every day. BPA is a known endocrine disruptor that has been shown to be carcinogenic. Many tumors show weak gap junctional intercellular communication (GJIC) function. The aim of this study was to investigate the effects and possible mechanisms of BPA's action on GJIC of human HaCaT skin cells. The results showed that BPA increased cell proliferation rates, prolonged GJIC photobleaching fluorescence recovery times and reduced overall fluorescence recovery rates at 0.01, 0.1 and 1 μm . BPA downregulated connexin (Cx)26 mRNA level at 0.1 μm . Estrogen receptor (ER) antagonist ICI 182 780 at 5 nm partially blocked the above effects of BPA indicating involvement of the ER pathway with BPA exposure. However, BPA did not influence Cx43 mRNA and protein levels. Our immunofluorescence data showed that Cx26 was expressed in the cytoplasm and plasma membrane, and was involved in the formation of gap junctions between adjacent cells, while Cx43 was expressed only in the cytoplasm. Therefore, our data indicate that Cx26 gap junctions may be involved in the GJIC inhibition caused by BPA. In conclusion, our results indicate that BPA can promote human skin cell proliferation, inhibit skin cell GJIC function but not formation and downregulate Cx26 mRNA levels partially through the ER pathway. We hypothesize that BPA can exhibit carcinogenicity by inhibiting GJIC.     

Effects of xenoestrogen bisphenol A on uterine and pituitary weight, serum prolactin levels and immunoreactive prolactin cells in ovariectomized Wistar rats

Considerable attention has currently been focused on bisphenol A (BPA), an environmental endocrine disrupting chemical that has oestrogenic activity. In vitro and in vivo short-term assays have shown that BPA is weakly estrogenic. In addition, the issue of species- and strain-differences in susceptibility to BPA was raised. The treatment of ovariectomized (OVX) Wistar rats with BPA at doses of 11–250 mg/kg per day, s.c., for 7 days, resulted in significant dose-dependent re-growth of uterus in uterotrophic assay. Additionally, the stimulation of anterior pituitary gland growth and induction of hyperprolactinaemia, as determined by wet organ weight and radioimmunoassay (RIA), respectively, were also dose-dependent (at 128 and 250 mg/kg per day, P < 0.05). Prolactin immunostaining of anterior pituitary glands revealed that BPA at a dose of 250 mg/kg per day increased the number of prolactin-immunopositive cells by 63% compared to OVX rats. These results demonstrate that the reproductive tract and neuroendocrine axis of Wistar rats are able to respond to BPA. Furthermore, the pituitary gland hypertrophy and hyperprolactinaemia can be mediated, at least partly, by increase in number of prolactin-immunoreactive cells. The long-term consequences of this proliferation are yet unknown but neoplasm formation is an obvious possibility. Received: 27 October 1999 / Accepted: 30 November 1999     

Chemical change of chlorinated bisphenol A by ultraviolet irradiation and cytotoxicity of their products on Jurkat cells

Chlorinated derivatives of bisphenol A (ClBPAs) have been detected in wastewater from waste paper recycling plants. BPA and ClBPAs are always exposed to ultraviolet (UV) radiation in the environment and consequently various photoproducts might be produced. Acute cytotoxicity of photoproducts of BPA and ClBPAs are not known. In this study, we investigated the cytotoxicity and chemical structure of photoproducts of BPA and ClBPAs (3-chlorobisphenol A (3-ClBPA), 3,3′-dichlorobisphenol A (3,3′-diClBPA) and 3,3′,5-trichlorobisphenol A (3,3′,5-triClBPA)) after UV irradiation (UVA, UVB and UVC). The toxicities of photoproducts on Jurkat cells were determined by Alamar Blue assay, and the chemical structures of the photoproducts were identified using GC/MS. The cytotoxicities of 3-ClBPA and 3,3′-diClBPA were higher than that of BPA and 3,3′,5-triClBPA. In addition, the toxicities of ClBPAs were increased by the irradiation of UVB and UVC at 100 J/cm² and decreased at 1000 J/cm² in comparison with those at 100 J/cm², indicating that their structures had changed. 3-Hydroxybisphenol A (3-OHBPA) was detected in the photoproducts of 3-ClBPA irradiated with UVB and UVC at 100 J/cm², and 3-OHBPA and 3-chloro-3′-hydroxybisphenol A (3-Cl-3′-OHBPA) were detected in those of 3,3′-diClBPA. However, these hydroxylated BPAs were not detected in the photoproducts exposed to 1000 J/cm². The cytotoxicity of 3-OHBPA was the almost same as ClBPAs after UVB and UVC irradiation. These results indicate that the formation of hydroxylated BPAs might contribute to the increase in toxicity caused by UV irradiation.     

A microdialysis study of glycinamide, glycine and other amino acid neurotransmitters in rat frontal cortex and hippocampus after the administration of milacemide, a glycine pro-drug

Milacemide is a glycinamide derivative which readily enters the brain and is metabolised to glycine. As its mechanism of action as an anticonvulsant drug is unknown we used the technique of microdialysis to study the temporal inter-relationship of glycinamide, glycine and other amino acid neurotransmitters in the extracellular fluid of rat hippocampus and frontal cortex. After milacemide administration (400 or 800 mg/kg i.p.), glycinamide concentrations rose linearly and dose-dependently in both hippocampus and frontal cortex. In contrast, whilst glycine concentrations rose in the hippocampus, glycine was unaffected in the frontal cortex. Concomitant increases in taurine hippocampal concentrations were observed. An increase in serine and a decrease in alanine concentrations was only observed at the highest milacemide dose (800 mg/kg). Other amino acids were unaffected. Thus, while glycinamide appears to be universally distributed throughout the brain, its metabolism to glycine and its effects on brain amino acids appear to be region specific.     

Effects of excitatory and inhibitory amino acid agonists and antagonists on ventral horn cells in slices of spinal cord isolated from adult rats

Excitatory amino acid agonists increased the probability of discharge of ventral horn cells in slices of spinal cord isolated from adult rats. The order of potency for the facilitatory action was kainate > quisqualate > (NMDA) > -glutamate > -aspartate. (±)-2-Amino-5-phosphonovalerate (2-APV) markedly reduced the facilitatory effects produced by -aspartate and NMDA, and slightly decreased the effect produced by -glutamate but did not alter the effect of kainate. Inhibitory amino acid agonists decreased the probability of cell discharge. The order of potency for the inhibitory action was (−)-baclofen > muscimol > GLYCINE = GABA. The effects produced by glycine, muscimol or GABA and (−)-baclofen were selectively reduced by strychnine, bicuculline and phaclofen, respectively. This type of preparation may be useful for in vitro pharmacological studies on the action of transmitters and drugs on ventral horn cells in the adult spinal cord.     

双酚A对小鼠睾丸间质细胞毒性及miR-203-3p和PI3K/AKT/FOXO1信号通路的影响

目的研究双酚A对小鼠睾丸间质细胞的毒性作用,及对miR-203-3p和PI3K/AKT/FOXO1信号通路的影响。方法不同浓度BPA(0、2、10、50、250μmol/L)处理小鼠睾丸间质细胞24 h,CCK8法检测细胞活力,Real time PCR检测miR-203-3p和FOXO1、AKT、PI3K的相对表达水平。结果不同浓度BPA处理细胞后,细胞活力随BPA剂量的增大而减少,50、250μmol/L组与对照组比较,差异有统计学意义(P<0.001)。各处理组miR-203-3p表达量均较对照组升高,10、250μmol/L组与对照组比较,差异有统计学意义(P<0.05)。2、10μmol/L组FOXO1表达量较对照组升高,50、250μmol/L组表达量较对照组降低,2、50、250μmol/L组FOXO1相对表达量与对照组比较差异有统计学意义(P<0.001)。各处理组AKT水平均出现下降趋势,10、50、250μmol/L组与对照组比较,差异有统计学意义(P<0.001)。各处理组PI3K水平均出现下降趋势,50、250μmol/L组与对照组比较,差异有统计学意义(P<0.001)。结论双酚A致睾丸间质细胞损伤,影响miR-203-3p和FOXO信号通路相关基因的改变。     

二甲双胍对人肝癌Hep-G2细胞表皮生长因子及其受体表达的影响

目的 观察二甲双胍对人肝癌Hep-G2细胞表皮生长因子（EGF）及其受体（EGFR）的影响.方法 体外培养人肝癌Hep-G2细胞,用不同浓度二甲双胍进行干预,3-（4,5-二甲基噻唑-2）-2,5-二苯基四氨唑溴盐法检测二甲双胍对Hep-G2细胞生长的影响;Hoechst 33342染色荧光显微镜观察细胞的形态学变化,流式细胞术观察细胞凋亡;蛋白质印迹法（Western blot）检测EGF、EGFR的表达.结果 二甲双胍对肝癌细胞的活性具有明显的抑制作用,且呈剂量依赖性;以10 mmol/L作用终浓度抑制效果最明显,其48 h的细胞吸光度为（0.477 ±0.025）,与对照组（0.602±0.026）比较差异有统计学意义（P＜0.01）;逐渐增加二甲双胍的作用浓度,细胞凋亡率随之逐渐增加,药物组细胞凋亡率分别为（10.76±0.96）％、（20.77±1.16）％,与对照组（5.21±0.13）％相比差异均有统计学意义（P＜0.05）;Hoechst33342染色可见明显的细胞皱缩,核染色质浓缩,核碎裂等凋亡形态学变化;Western blot结果显示,EGF及其受体蛋白的表达随着二甲双胍作用浓度的增加而逐渐下调.结论 在体外,二甲双胍能够抑制人肝癌Hep-G2细胞增殖及诱导细胞凋亡,其抗肿瘤机制可能与细胞内EGF及EGFR表达下调有关.     

Effects of various LED light spectra on antioxidant and immune response in juvenile rock bream,Oplegnathus fasciatus exposed to bisphenol A

Bisphenol A (BPA) is a monomer used in plastics and plasticizers. As an environmental toxin included in industrial wastewater, it contaminates the aquatic environment and is known to cause endocrine disruption in fish. Particular wavelengths of light-emitting diodes (LEDs) are known to affect the endocrine regulation of fish. The present study aimed to investigate the effects of green and red LED light on the antioxidant and immune systems in juvenile rock bream (Oplegnathus fasciatus) exposed to BPA. We used green and red LED exposure at two intensities (0.3 and 0.5 W/m²) for 1, 3, and 5 days. We measured liver mRNA expression and plasma levels of antioxidant enzyme superoxide dismutase (SOD) and caspase-3. Furthermore, we measured plasma levels of hydrogen peroxide (H₂O₂), lipid peroxidation (LPO), melatonin, and immunoglobulin M (IgM). DNA damage and apoptotic activity were measured using comet and terminal transferase dUTP nick end labeling (TUNEL) assays, respectively. We found that SOD, H₂O₂, and LPO increased significantly, whereas melatonin and IgM decreased significantly, suggesting that BPA induces oxidative stress and reduces immune function. Likewise, both DNA damage and apoptotic activity increased following BPA exposure. However, we found that exposure to green LED light effectively reduced the detrimental effects induced by BPA, including decreasing DNA damage, apoptotic activity, SOD mRNA expression, and plasma levels of SOD, H₂O₂, and LPO. Likewise, the plasma levels of melatonin and IgM increased. Thus, our results indicate that green light conditions effectively reduces oxidative stress and promotes the immune function in juvenile rock bream.     

Human exposure to bisphenol A by biomonitoring: methods, results and assessment of environmental exposures

Human exposure to bisphenol A is controversially discussed. This review critically assesses methods for biomonitoring of bisphenol A exposures and reported concentrations of bisphenol A in blood and urine of non-occupationally ("environmentally") exposed humans. From the many methods published to assess bisphenol A concentrations in biological media, mass spectrometry-based methods are considered most appropriate due to high sensitivity, selectivity and precision. In human blood, based on the known toxicokinetics of bisphenol A in humans, the expected very low concentrations of bisphenol A due to rapid biotransformation and the very rapid excretion result in severe limitations in the use of reported blood levels of bisphenol A for exposure assessment. Due to the rapid and complete excretion of orally administered bisphenol A, urine samples are considered as the appropriate body fluid for bisphenol A exposure assessment. In urine samples from several cohorts, bisphenol A (as glucuronide) was present in average concentrations in the range of 1-3 microg/L suggesting that daily human exposure to bisphenol A is below 6 microg per person (<0.1 microg/kg bw/day) for the majority of the population.