Feasibility study of absolute activity measurement with metallic magnetic microcalorimeters

We report on a feasibility study on precise determination of mass-specific activity of low-energy emitting radioisotopes. Conventional methods of activity measurement suffer from source self-absorption and a strong decrease in detection efficiency for low-energy electrons and photons. We propose a new method based on metallic magnetic microcalorimeters with the source embedded in the detector target in a 4π geometry. First results with a ⁵⁵Fe source show that electrons and photons are detected with a detection efficiency close to unity and with little loss of energy for electrons. The aim of this study is to provide standards of activity with very low uncertainties in the framework of radiation metrology.     

A comparative study of thin film insulation techniques for gold electrodes

The increasing use of microelectrodes in small sample volumes and biological environments require the use of thin film insulators. The insulating ability of the thin film insulators electrophoretic paint, polyimide, allylphenol phenol copolymer and an epoxy dip coat have been assessed using gold wires coated in the relevant film. Insulation was assessed by recording the current from the wires in a solution of known redox couple. The effect of prolonged storage in aqueous solutions was also investigated. The most successful insulating approach, with regards to insulative ability, prolonged use in aqueous environments and speed of application, was the combination of cathodic electrophoretic paint followed by polyimide.     

The microbiology of mixed salad containing raw and cooked ingredients without dressing

Growth of aerobic mesophilic bacteria in mixed salad without dressing, containing cooked sweet corn and raw endive, was similar to that of raw endive alone. At 9°C, sweet corn permitted the growth of the lactic acid bacterium Leuconostoc mesenteroides , whenever the bacterium was present on raw endive, whereas no growth of lactic acid bacteria was recorded on raw endive alone. Listeria monocytogenes , artificially inoculated in the samples, grew more in the mixed salad than in the raw endive alone, for products stored at 6°C and 9°C. Acidification of sweet corn to pH 5.0 was proposed to reduce the development of L. monocytogenes in the mixed salad to a level similar to that recorded in the raw endive alone. Acidification with citric acid reduced spoilage of sweet corn, whereas acetic acid caused necrosis on the leaves of raw endive.     

Binding characteristics of novel nonsteroidal antiestrogens to the rat uterine estrogen receptors

Tamoxifen (TAM), the only antiestrogen currently available for the endocrine therapy of breast cancer behaves as a mixed agonist/antagonist of estrogen action, thus limiting its therapeutic potential. We report the binding characteristics of a novel series of nonsteroidal antiestrogens to the rat uterine estrogen receptor. As measured by competition studies, the affinity of EM-652, the active metabolite of the prodrug EM-800, for the estrogen receptor is 7-11 times higher than that of 17beta-estradiol (E2), ICI 182780, and hydroxy-tamoxifen (OH-TAM), the active metabolite of Tamoxifen. EM-652 is 20x more potent than ICI 164384 and Droloxifene while it is 400 times more potent than Toremifene in displacing [3H]E2 from the rat uterine estrogen receptor. On the other hand, the prodrug EM-800 and Tamoxifen have respectively 150-fold and 410-fold less affinity for the estrogen receptor than the pure antiestrogen EM-652. No significant binding of EM-652, EM-800, TAM or OH-TAM was observed to the rat uterine progesterone receptor at concentrations up to 10,000 nM except for TAM that caused a 50% displacement of labeled R5020 at 4000 nM. No significant binding of EM-652 or EM-800 was observed on the rat ventral prostate androgen receptor or the rat uterine progesterone receptor. The present data demonstrate the high affinity and specificity of the new antiestrogen, EM-652, for the rat uterine estrogen receptor. The antiestrogen EM-652 thus becomes the compound having the highest known affinity for the estrogen receptor. Due to its unique potency and its pure antiestrogenic activity already demonstrated in many systems, this antiestrogen could well offer an important advance for the endocrine therapy of breast cancer, uterine cancer, and other estrogen-sensitive diseases in women.     

Hereditary neurocutaneous angiomatosis. Report of four cases

The authors report the coexistence of vascular nevi (hemangiomas and arteriovenous malformations (AVMs) of the skin) with AVMs and venous malformations of the brain in male siblings from two related but nonconsanguineous families of three generations. The proband, his siblings, parents, aunts, uncles, and cousins were examined, underwent magnetic resonance (MR) imaging and MR angiography, and when appropriate, cerebral angiography. A father had vascular nevi and a mother, his sister, had an azygos anterior cerebral artery. No other cutaneous or cerebrovascular malformations were present in the parents. Each of the two families had two boys and one girl, 9 to 18 years of age. All the children had vascular nevi and all of the boys had coexisting cerebrovascular malformations: AVMs in three, and a venous malformation in another. One boy had three cerebral AVMs. Two boys had a cerebral hemorrhage, and one also had focal motor seizures. The skin lesions were not those of the Sturge-Weber-Dimitri, Rendu-Osler-Weber, or Wybum-Mason syndromes. The association of cutaneous and cerebrovascular malformations was seen only in males in these families. but females have also been reported in the literature. The results obtained in these families and three other families reported from Western and Central Europe indicate that the association of cerebral and cutaneous vascular hamartomas constitutes a distinct, hereditary clinicopathological entity with autosomal dominant inheritance and variable penetrance. The clinical manifestations of this syndrome are visible, painful vascular nevi, epilepsy, cerebral hemorrhage, and focal neurological deficits. The preponderance of male patients with the full expression of the syndrome suggests a possible hormonal influence on the expression of the gene.     

Combining teacher-Led and technology-assisted instruction to improve phoneme blending to read words for elementary students with disabilities

Technology-assisted instruction has potential for helping students improve their reading skills. In the current studies, PowerPoint software was used to supplement teacher-led reading instruction for elementary-aged students with disabilities who struggled with phoneme blending to read words. The effectiveness of the intervention was assessed using two multiple probe design studies across sets of words based on an onset-rime (word family) strategy. In Study 1, teacher-led instruction was paired with technology-assisted instruction for all intervention sessions for three students in first and second grade. In Study 2, teacher-led instruction occurred between baseline and technology-assisted intervention sessions for three students in third and fourth grade. During the initial instruction sessions, participants recorded their own voices onto PowerPoint slides so that their technology-assisted instruction included self-modelling. Results of both studies indicated that technology-assisted instruction can be effective for helping students learn how to blend phonemes to read words; however, performance varied and there were limitations in each study that should be addressed in future research. Relevant implications for classroom instruction are emphasized.     

hTERT-Driven Immortalization of RDEB Fibroblast and Keratinocyte Cell Lines Followed by Cre-Mediated Transgene Elimination

The recessive form of dystrophic epidermolysis bullosa (RDEB) is a crippling disease caused by impairments in the junctions of the dermis and the basement membrane of the epidermis. Using ectopic expression of hTERT/hTERT + BMI-1 in primary cells, we developed expansible cultures of RDEB fibroblasts and keratinocytes. We showed that they display the properties of their founders, including morphology, contraction ability and expression of the respective specific markers including reduced secretion of type VII collagen (C7). The immortalized keratinocytes retained normal stratification in 3D skin equivalents. The comparison of secreted protein patterns from immortalized RDEB and healthy keratinocytes revealed the differences in the contents of the extracellular matrix that were earlier observed specifically for RDEB. We demonstrated the possibility to reverse the genotype of immortalized cells to the state closer to the progenitors by the Cre-dependent hTERT switch off. Increased β-galactosidase activity and reduced proliferation of fibroblasts were shown after splitting out of transgenes. We anticipate our cell lines to be tractable models for studying RDEB from the level of single-cell changes to the evaluation of 3D skin equivalents. Our approach permits the creation of standardized and expandable models of RDEB that can be compared with the models based on primary cell cultures.     

Embryonic Trophectoderm Secretomics Reveals Chemotactic Migration and Intercellular Communication of Endometrial and Circulating MSCs in Embryonic Implantation

Embryonic implantation is a key step in the establishment of pregnancy. In the present work, we have carried out an in-depth proteomic analysis of the secretome (extracellular vesicles and soluble proteins) of two bovine blastocysts embryonic trophectoderm primary cultures (BBT), confirming different epithelial–mesenchymal transition stages in these cells. BBT-secretomes contain early pregnancy-related proteins and angiogenic proteins both as cargo in EVs and the soluble fraction. We have demonstrated the functional transfer of protein-containing secretome between embryonic trophectoderm and maternal MSC in vitro using two BBT primary cultures eight endometrial MSC (eMSC) and five peripheral blood MSC (pbMSC) lines. We observed that eMSC and pbMSC chemotax to both the soluble fraction and EVs of the BBT secretome. In addition, in a complementary direction, we found that the pattern of expression of implantation proteins in BBT-EVs changes depending on: (i) their epithelial–mesenchymal phenotype; (ii) as a result of the uptake of eMSC- or pbMSC-EV previously stimulated or not with embryonic signals (IFN-τ); (iii) because of the stimulation with the endometrial cytokines present in the uterine fluid in the peri-implantation period.