Applicability of the one-step DVS method for the determination of amorphous amounts for further different hydrophilic and hydrophobic drugs

In a former publication the authors showed that low amounts of amorphous content (LOQ of 0.5%) in a hydrophobic model API (Ciclesonide) can be measured with an individually adjusted one-step dynamic organic vapor sorption (DVS). In this investigation the applicability is tested on various APIs which differ in lipophilicity (poor water solubility) and hygroscopicity (absorption of water).The vapor sorption method proved to be applicable in almost all cases. Moisture sorption isotherms were determined for all five investigated crystalline and amorphous APIs. However, it was necessary to select the parameters individually for each API. The used solvents (water, methanol, isopropanol and methylene chloride) and the humidity-levels (0.05 p/p₀ until 0.5 p/p₀) were chosen carefully because otherwise the amorphous amounts switch to their crystalline counterparts and are not detectable. The production of fully amorphous samples (absence of crystalline material measured by DSC, mDSC and XRPD) was optimized over several trials. As successfully methods proved ball-milling, freeze-drying, spray-drying and/or quench cooling. In the next step these fully amorphous amounts were blended with crystalline starting material to calibration curves (Turbula blender, influence of electrostatic charge to homogeneity) for the calculation of amorphous content.In summary, the following presented methods were used to determine and quantify low amorphous amounts (between 1.5% and 17.0%) in jet-milled powders (grinding pressure of 8 bar, 1–3 grinding cycles), respectively.     

Association of <Emphasis Type="Italic">SLC22A4/5</Emphasis> Polymorphisms with Steroid Responsiveness of Inflammatory Bowel Disease in Japan

Purpose We investigated the association between steroid responsiveness and single nucleotide polymorphisms of SLC22A4/A5 located within inflammatory bowel disease 5 locus. Our goal is personalized steroid therapy adjusted to match individual variations in drug responsiveness in each inflammatory bowel disease patient. Methods Unrelated Japanese cohorts of 94 patients with Crohn’s, 94 patients with ulcerative colitis, and 257 healthy control subjects were consecutively enrolled in this study. Genotyping and haplotype analysis focusing on steroid responsiveness was performed by using 15 single nucleotide polymorphisms. Results The G allele of −368T > G in SLC22A5, in which strong linkage disequilibrium was observed and the limited diversity of three haplotypes was estimated, was significantly associated with steroid resistance in Japanese patients with Crohn’s disease (P = 0.016). Haplotype analysis between −446C > T and −368T > G in the SLC22A5 promoter region showed that the CG allele appeared to be a risk haplotype for steroid resistance (CG: odds ratio, 4.13; 95 percent confidence interval, 1.41–12.1; P = 0.016). Conclusions This extensive linkage disequilibrium may form a general risk haplotype for steroid resistance in Crohn’s disease in Japanese. Further analyses of the pharmacogenomics of steroid responsiveness are warranted to achieve the goal of individualized steroid therapy against inflammatory bowel disease. Supported by a grant-in-aid from the Ministry of Health, Labour and Welfare (K.I.), Japan. Address of correspondence: Yoshiaki Arimura, M.D., First Department of Internal Medicine, Sapporo Medical University, S-1, W-16, Chuo-ku, Sapporo, 060-8543, Japan. E-mail: arimura@sapmed.ac.jp     

肾脏损伤时有机阴离子转运子表达变化及其意义

有机阴离子转运子(OAT)在机体清除各种内、外源性有机阴离子的过程中发挥极其重要的作用.在肾脏,OAT参与排泄许多外源性有机阴离子,如药物、环境中的化学物质、生物毒素以及许多内源性有机阴离子包括多种尿毒症毒素等.近年研究发现,各种肾脏疾病条件下OAT表达发生变化,并由此而产生相应病理生理效应.本文讨论了OAT在近端肾小管上皮细胞的生物学特征、肾脏病变状态下的表达变化及其病理生理学意义.     

Abstracts

Dental enamel is the most mineralized tissue of vertebrate organisms. Enamel biosynthesis is initiated by the secretion, processing, and self-assembly of a complex mixture of proteins. The formation of an ordered enamel organic extracellular matrix (ECM) seems be a crucial step for the proper formation of mineral phase. Polarizing microscopy demonstrates that the ordered supramolecular structure of the secretory-stage enamel organic ECM is strongly birefringent. In the present work we analyzed the birefringence of secretory-stage enamel organic ECM in amelogenin (Amelx)- and enamelysin (Mmp20)-deficient mice. Female Amelx^+/? animals showed significant reduction in optical retardation values when compared with the Amelx^+/+ subgroup (p = 0.0029). The secretory-stage enamel organic ECM of the Amelx^?/? subgroup did not exhibit birefringence. The secretory-stage enamel organic ECM of Mmp20^?/? mice showed a significant decrease in optical retardation as compared with Mmp20^+/+ and Mmp20^+/? mice (p = 0.0000). Mmp20^+/? and Mmp20^+/+ mice exhibited similar birefringence (p = 1.0000). The results presented here support growing evidence for the idea that the birefringence of secretory-stage enamel organic ECM is influenced by the ordered supramolecular organization of its components.     

Metabolic profiling in human SH-SY5Y neuronal cells exposed to perfluorooctanoic acid (PFOA)

Perfluorooctanoic acid (PFOA) is an abundant per- and polyfluoroalkyl substance (PFAS) detected in both indoor and outdoor environments. While studies suggest exposure concerns for humans, studies investigating PFOA-induced neurotoxicity are lacking. To address this gap, we exposed differentiated human SH-SY5Y cells to PFOA (0.1 μM up to 500 μM) at different time points (4, 24, 48, and 72 h) and measured cell viability, Casp3/7 activity, ATP levels, ATP synthase enzyme activity, mitochondrial membrane potential, reactive oxygen species (ROS), oxygen consumption rates for mitochondrial stress test (XFe24 Flux analyzer), glucose utilization, and global metabolome profiles to assess the potential for PFOA-induced neurotoxicity. Treatment with 10 or 100 μM PFOA did not compromise cell viability nor induce cytotoxicity to SH-SY5Y cells over a 48-hour exposure period. However, >250 μM PFOA compromised cell viability, induced cytotoxicity, and induced caspase 3/7 activity at 48 h. ATP levels were reduced in cells treated with 400 μM PFOA for 24 and 48 h, and with 100 μM PFOA and higher at 72 h. ATP synthase activity was inhibited by 250 μM PFOA but was unchanged by PFOA treatment at 200 μM or less. Conversely, mitochondrial membrane potential was reduced by >10 μM PFOA after 24 h. Total ROS was increased with 100 μM PFOA and higher after 4 h of exposure. Several mitochondria-related endpoints (basal respiration, ATP production, maximum respiration) were negatively affected at 250 μM PFOA at both 24- and 48-hour exposure, but were unaltered at concentrations of 100 μM PFOA or less. One exception was mitochondrial spare capacity, which was reduced by 100 μM PFOA after 24-hour exposure. Similarly, glycolysis, glycolytic capacity, and glycolytic reserve of SH-SY5Y cells were not altered by 10 nor 100 μM PFOA. Nontargeted metabolomics was conducted in cells treated with either 10 or 100 μM PFOA for 48 h, as these two concentrations were not cytotoxic and 28 metabolites differed among treatments. Notable was that 10 μM PFOA had little effect on the SH-SY5Y metabolome, and the metabolic profile was not statistically different from media nor solvent controls. On the other hand, 100 μM PFOA shifted the metabolic signature of the neuronal cells, leading to reduced abundance of ATP-related metabolites (adenine, nicotinamide), neurotransmitter precursors (DL-tryptophan, l-tyrosine), and metabolites that protect mitochondria during oxidative stress (betaine, orotic acid, and l-acetyl carnitine). We hypothesize that this metabolic signature may be associated with the reduced mitochondrial membrane potential observed at lower PFOA concentrations. Metabolic shifts appear to precede compromised cell viability, cytotoxicity, and apoptosis. This study generates mechanistic knowledge regarding PFOA-induced neurotoxicity, focusing on mitochondrial oxidative respiration and the neuronal metabolome.     

职业性有机溶剂中毒15例临床表现和影像学特征分析

目的 探讨职业性有机溶剂中毒的临床表现、神经影像学特征、治疗及预后。方法 回顾性分析15例职业性有机溶剂中毒患者的临床资料。结果 15例患者均急性或亚急性起病,有明确的有机溶剂职业接触史,主要临床表现包括头晕(73.3%)、认知功能下降(60%)、锥体束损害(53.3%)、颅内高压综合征(46.7%)、意识障碍(33.3%); 其中14例患者头颅磁共振成像(Magnetic resonance imaging,MRI)显示为弥漫性脑组织肿胀,以双侧大脑半球白质为主,呈“火焰状”,10例累及双侧基底节(苍白球),8例累及双侧小脑齿状核,呈“肾形”改变,5例累及双侧丘脑; 所有患者使用脱水剂、糖皮质激素及高压氧等综合治疗后,其中12例患者好转出院,无或仅遗留轻微后遗症,3例患者无明显改善,预后差。结论 职业性有机溶剂中毒以头晕、认知功能下降、锥体束损害、颅内高压综合征及意识障碍等中毒性脑病表现为主,典型头颅MRI为双侧大脑半球白质、苍白球及小脑齿状核对称性病变,结合患者职业接触史需考虑本病; 早期、长程、足量给予脱水剂、糖皮质激素及高压氧等综合治疗,大多数患者预后良好。     

不同直径半夏药材的质量差异比较研究

目的:考察不同直径半夏药材的质量差异,为其资源的合理利用提供参考。方法:以来自国内四大药材市场、2家公司的65批商品半夏为样品,采用游标卡尺测定其直径后,用分药筛将其筛分为不同直径范围(0.5 cm≤d≤0.8 cm、0.8 cm0.05)。73批半夏对口药材的直径主要分布在0.6~1.8 cm之间,平均重量占比为95.54%。若以2015年版《中国药典》(一部)规定的直径标准(1~1.5 cm),73批半夏对口药材中仅8批符合要求,合格率为10.96%;若适当扩大直径范围为0.7~1.5 cm,有54批符合要求,合格率为73.97%;当直径范围扩大为0.7~1.6 cm时,有68批符合要求,合格率达93.15%。结论:不同直径的半夏药材之间质量无明显差异。结合各产地半夏的直径分布情况,可将相关标准中规定的直径大小由"1~1.5 cm"扩大为"0.7~1.6 cm",避免资源浪费。     

Nutrient recovery from pigeon dropping by using exotic earthworm Eisenia fetida

Pigeon dropping is a kind of organic waste which can be transformed easily into organic manure by vermicomposting with exotic earthworm Eisenia fetida. In the present study, a pigeon excreta was collected from roof tops. The pigeon excreta was mixed with cattle dung with the ratios as 10%, 20%, 30%, 40% and 50% respectively, in plastic containers; meanwhile, those proposal were name as C₁₀, C₂₀, C₃₀, C₄₀ and C₅₀ respectively. Feed mixtures were allowed to decompose (pre-vermicomposting without earthworm) for 20–25 days. After decomposition, 20 hatchlings of E. fetida were put in each concentration after weighing. Cattle dung (without pigeon excreta) was kept as the control. The physico-chemical analysis of initial feed mixture and final product after vermicomposting was also done to measure the changes in the feed mixtures from the initial to the final states. There were increases in the content of N (52.35%), P (115.79%), K (39.97%) and EC (56.16%) but decrease in those of OC (-15.49%), OM (-15.49%), pH (-7.92%) and Ca (-39.62%) from the initial to the final product. The highest population size of earthworms and minimum mortality rate was observed in C₃₀ feed mixture. Thus, the present study indicated that cattle dung mixed with 30% (w/w) pigeon excreta waste may be a good choice for vermicomposting.