Localized synthesis of specific proteins during oogenesis and early embryogenesis inDrosophila melanogaster

Summary Protein synthesis in egg follicles and blastoderm embryos ofDrosophila melanogaster has been studied by means of two-dimensional gel electrophoresis. Up to 400 polypeptide spots have been resolved on autoradiographs. Stage 10 follicles (for stages see King, 1970) were labelled in vitro for 10 to 60 min with³⁵S-methionine and cut with tungsten needles into an anterior fragment containing the nurse cells and a posterior fragment containing the oocyte and follicle cells. The nurse cells were found to synthesize a complex pattern of proteins. At least two proteins were detected only in nurse cells but not in the oocyte even after a one hour labelling period. Nurse cells isolated from stages 9, 10 and 12 follicles were shown to synthesize stage specific patterns of proteins. Several proteins are synthesized in posterior fragments of stage 10 follicles but not in anterior fragments. These proteins are only found in follicle cells. No oocyte specific proteins have been detected. Striking differences between the protein patterns of anterior and posterior fragments persist until the nurse cells degenerate. In mature stage 14 follicles, labelled in vivo, no significant differences in the protein patterns of isolated anterior and posterior fragments could be detected; this may be due to technical limitations. At the blastoderm stage localized synthesis of specific proteins becomes detectable again. When blastoderm embryos, labelled in vivo, are cut with tungsten needles and the cells are isolated from anterior and posterior halves, differences become apparent. The pole cells located at the posterior pole are highly active in protein synthesis and contribute several specific proteins which are found exclusively in the posterior region of the embryo. In this study synthesis of specific proteins could only be demonstrated at those developmental stages which are characterized by the presence of different cell types within the egg chamber, while no differences were detected when stage 14 follicles were cut and anterior and posterior fragments analyzed separately. The differences in the pattern of protein synthesis by pole cells and blastoderm cells indicate that even the earliest stages of determination are reflected by marked changes at the biochemical level.     

Human vaginal epithelial multilayer tissue culture

Summary Fragments of normal human adult vagina, when explanted onto glass slides gave rise to outgrowing sheets of pure epithelium, which had microscopic morphological features in common with normal vaginal epithelium. Infrequent fibroblast contamination was observed. Proliferating epithelial cells formedmultilayers of stratified squamous epithelium and demonstrated a progressive decrease in proliferative activity after 14 days. Continuous lines of epithelial cells were not obtained. Even in the absence of estrogens, transmission electron microscopy revealed evidence of keratinization of the superficial cells of the multilayer. Scanning electron microscopy of the surface of mature epithelial cells in culture revealed ultrastructural features that closely resembled those present on the surface of exfoliated cells obtained by scraping the vagina in vivo. This in vitro tissue culture model of human vaginal epithelium may provide a simple method of studying factors that influence vaginal epithelium growth, maturation and function.     

Iron Deficiency Alters Discrete Proteins in Rat Caudate Nucleus and Nucleus Accumbens

Young rats (21 days old) made nutritionally iron deficient, by feeding them a semisynthetic diet containing skimmed milk for 5 weeks, had significantly lowered hemoglobin levels (5.2 +/- 4 g/100 ml). The nonheme iron content in caudate nucleus was decreased by 47%. The behavioral response of iron-deficient rats to apomorphine (2 mg/kg) and the density of 3,4-dihydroxyphenylethylamine (dopamine) D2 receptors, as measured by [3H]spiperone binding in caudate nucleus, were significantly reduced by 70 and 53%, respectively. The possibility that nutritional iron deficiency may affect protein content in brain was investigated by measuring the apparent concentration of proteins in caudate nucleus and nucleus accumbens from iron-deficient and control animals using two-dimensional gel electrophoresis. The data indicate that iron deficiency can affect content in these two brain regions. Significant changes in the content of 10 proteins were noted in the caudate nucleus and nucleus accumbens in iron-deficient rats. The albumin level was significantly increased in both regions studied, whereas the neuron-specific enolase level was increased in the nucleus accumbens and the glial fibrillary acidic protein level was reduced in the caudate nucleus. The significance of these protein content changes, as well as a reduction in content of a 94-kilodalton protein (a molecular size similar to that of the D2 dopamine receptor), remains to be established.     

Evaluation of total contents of Al,As, Ca,Cd, Fe,K, Mg,Ni, Pb,Zn and their fractions leached to the infusions of different tea samples. A multivariate study

ABSTRACT

Total aluminum, arsenic, calcium, cadmium, iron, lead, potassium, magnesium, nickel and zinc were determined in black tea by electro thermal atomic absorption spectrometry after ultrasonic assisted pseudo-digestion with mixture of acid and oxidant. All the metals were also determined in the tea infusions in order to know the percentage of each element leached into the liquor. A conventional acid digestion on electric hot plate was used to obtain total metals under study for comparative purpose. Analytical results for Al, As, Ca, Cd, Fe, K, Mg, Ni, Pb and Zn obtained by ultrasound assisted pseudodigestion, and conventional wet digestion methods showed a good agreement, thus indicating the possibility of using ultrasonic assisted digestion sample preparation instead of intensive treatments inherent with the acid digestion methods on electric hot plate. The validity and accuracy of both procedures were checked by using certified sample of NIES No. 7 (Tea Leaves). Non significant differences were observed for P>0.05 when comparing the values obtained by both methods (paired t-test).

Principal component analysis (PCA) was applied in order to determine the different amount of metals as main sources of variability in the data sets and to establish the relation between samples (branded and nonbranded tea samples) and micronutrient, trace and toxic metals (variables). Hierarchical cluster analysis (HCA) was used to explore the different branded and nonbranded tea samples grouping according to the essential and toxic metals as additional information to the out put obtained by PCA.     

Ultrasound combined with SDF‐1α chemotactic microbubbles promotes stem cell homing in an osteoarthritis model

Osteoarthritis (OA) is a common joint disease in the middle and old age group with obvious cartilage damage, and the regeneration of cartilage is the key to alleviating or treating OA. In stem cell therapy, bone marrow stem cell (BMSC) has been confirmed to have cartilage regeneration ability. However, the role of stem cells in promoting articular cartilage regeneration is severely limited by their low homing rate. Stromal cell‐derived factor‐1α (SDF‐1α) plays a vital role in MSC migration and involves activation, mobilization, homing and retention. So, we aim to develop SDF‐1α‐loaded microbubbles MB(SDF‐1α), and to verify the migration of BMSCs with the effect of ultrasound combined with MB(SDF‐1α) in vitro and in vivo. The characteristics of microbubbles and the content of SDF‐1α were examined in vitro. To evaluate the effect of ultrasound combined with chemotactic microbubbles on stem cell migration, BMSCs were injected locally and intravenously into the knee joint of the OA model, and the markers of BMSCs in the cartilage were detected. We successfully prepared MB(SDF‐1α) through covalent bonding with impressive SDF‐1α loading efficacy loading content. In vitro study, ultrasound combined with MB(SDF‐1α) group can promote more stem cell migration with highest migrating cell counts, good cell viability and highest CXCR4 expression. In vivo experiment, more BMSCs surface markers presented in the ultrasound combined with MB(SDF‐1α) group with or without exogenous BMSCs administration. Hence, ultrasound combined with MB(SDF‐1α) could promote the homing of BMSCs to cartilage and provide a novel promising therapeutic approach for OA.     

甲状腺结节良恶性的彩色多普勒超声特征及其诊断价值分析

目的:研究甲状腺结节良恶性的彩色多普勒超声特征及其诊断价值。方法:选取从2016年3月~2019年2月于我院接受手术治疗的甲状腺疾病患者300例作为研究对象,均予以彩超检查,比较甲状腺良恶性结节的超声特征(主要包括直径、钙化情况、边界、回声、血流状况)。以病理活检结果为金标准,分析彩超诊断甲状腺结节良恶性的敏感性、特异性以及准确度。对比甲状腺良恶性结节的血流分型情况以及各项血流动力学参数。结果:恶性结节超声特征直径≥2 cm、有钙化、边界模糊、无回声/低回声、血流丰富人数占比均高于良性结节(均P<0.05)。以手术病理组织活检结果作为金标准,彩色多普勒超声诊断甲状腺结节的敏感性、特异性以及准确度分别为97.73%、86.11%、96.33%。甲状腺良性结节血流分型为Ⅰ型、Ⅱ型人数占比高于恶性结节,而Ⅲ型、Ⅳ型人数占比低于恶性结节(均P<0.05)。甲状腺良性结节的收缩期峰值血流速度(PSV)、阻力指数(RI)均低于恶性结节,而舒张末期血流速度(EDV)高于恶性结节(均P<0.05)。结论:彩色多普勒超声对甲状腺结节良恶性的鉴别价值较高,且具有较高的敏感性、特异性以及准确度,可为甲状腺良恶性结节的早期诊断、临床治疗提供重要的参考依据。     

经会阴二维超声对自然分娩女性产后盆底三腔室运动和早期盆底功能的评估价值

目的:探讨经会阴二维超声对自然分娩女性产后盆底三腔室运动和早期盆底功能的评估价值。方法:选取2019年2月～2019年7月于我院进行自然分娩的产妇284例作为研究组,另选取同期于我院接受诊治的未育女性200例作为对照组。两组均进行经会阴二维超声检查,比较两组静息期、张力期、缩肛期的肛管直肠连接部(ARJ)、宫颈外口(CV)、尿道膀胱连接部(UVJ)的位置及运动变化情况,对比两组静息状态下以及最大Valsaval动作下盆底超声参数。结果:研究组静息期、张力期、缩肛期的CV距离水平参照线的垂直长度(CV-VD)、UVJ距离水平参照线的垂直长度(UVJ-VD)均低于对照组(均P0.05)。研究组张力期对应静息期垂直长度△r-s CV-VD、△r-s UVJ-VD均高于对照组,而缩肛期对应静息期垂直长度△r-s CV-VD、△r-s UVJ-VD均低于对照组(均P0.05)。研究组静息状态下膀胱颈位置(BNP)低于对照组,而膀胱尿道后角(PUA)高于对照组(均P0.05);研究组最大Valsaval动作下膀胱颈移动度(BND)、PUA、尿道旋转角(URA)均高于对照组(均P0.05)。结论:经会阴二维超声可对自然分娩女性产后盆底三腔室运动和早期盆底功能进行有效观察,有助于评估女性盆底结构、功能变化,具有一定的临床推广应用价值。     

经会阴实时三维盆底超声评估不同分娩方式对产后女性前腔室结构和盆膈裂孔的影响

摘要 目的：探讨经会阴实时三维盆底超声评估不同分娩方式对产后女性前腔室结构和盆膈裂孔的影响。方法：回顾性分析2015年1月至2019年10月在我院接受检查的100例产妇的诊治资料。根据分娩方式的不同,将患者分为经阴道分娩组（n=55）和剖宫产分娩组（n=45）。比较两组产妇在静息状态和Valsalva状态下的前腔室和盆膈裂孔超声参数。结果：在静息状态下,两组的膀胱颈位置、逼尿肌厚度、膀胱后角和尿道倾斜角相比无差异（P>0.05）。经阴道分娩组在Valsalva状态下的膀胱颈移动度和尿道旋转角均大于剖宫产分娩组,尿道内口漏斗形成率和膀胱膨出率均高于剖宫产分娩组（P<0.05）。在Valsalva状态下,经阴道分娩组的盆膈裂孔前后径、左右径、面积和周长均大于剖宫产分娩组（P<0.05）;在静息状态下,两组的上述指标相比,差异均无统计学意义（P>0.05）。结论：应用经会阴实时三维盆底超声技术观察产妇前腔室结构和盆膈裂孔的参数变化,可评估产妇盆底功能受损的程度,经阴道分娩对其影响较大。     

Synergized subcritical-ultrasound-assisted aqueous two-phase extraction,purification, and characterization of Lentinus edodes polysaccharides

The effect of a synergized two-step subcritical water extraction (SWE) and simultaneous multi-frequency ultrasound-assisted alcohol/salt aqueous two-phase extraction (MFu-AATPE) on the yield, physicochemical, structural characteristics, and antioxidant activities of Lentinus edodes polysaccharides (LEPs) was investigated. Crude LEP extracted under subcritical extraction (31.14%) was purified using dual and triple frequency ultrasound-assisted aqueous two-phase systems giving a yield of 94.57% and 97% respectively. Compared with the dual-frequency sonication (20/40 kHz), the triple frequency sonication (20/40/60 kHz) gave maximum yield and improved the desalination rate by 33.6%. Congo red analysis revealed triple helix structures. Gas Chromatography analysis showed the existence of mannose, glucose, arabinose, xylose, and galactose whilst spectral similarities were observed through Fourier Transform Infrared Spectroscopy. The average molecular weight (M_w) was 83.6170 Da, 2384 Da and 2387 Da for control, dual and triple frequency samples respectively. Ultrasound treated polysaccharides displayed stronger antioxidant activities against the 2.2-diphenyl;-1 picrylhydrazyl (DPPH), 2, 2ʹ-azino-bis 3-ethylbenzothiazoline-6 sulfonic acid (ABTS) and hydroxyl radicals, thus demonstrating their potency in reducing oxidation. However, the microstructure of the triple frequency treated sample was altered compared to the dual-frequency sample. Therefore, this study demonstrated the efficacy of SWE in crude LEPs extraction and illustrated MFu-AATPE as an efficient technique that selectively isolates polysaccharides and enhances extraction efficiencies.