High endocytotic and lysosomal activities in segments of rat myotubes differentiated in vitro

Summary The entire microvascular architecture in rat foot-pads including that of eccrine sweat glands was studied by scanning electron microscopy using a vascular corrosion-cast replication technique. In the central roofs of the pads, particularly elaborate capillary networks were arranged in rows perpendicular to the long axis of the foot. In the marginal regions of the pads, simple networks of capillaries were arranged in lamellar sheets parallel to the surface of the sole of the foot. Complex spongy networks of vascular trees were observed in the subcutaneous layer of the pads. These vessels were supplied by the pad artery, and then, after forming capillary networks in the roofs of the pads, they drained into the metatarsal vein. Rod-shaped cages of capillaries were observed around the eccrine sweat glands. One descending arteriole, arising from a connecting arteriole, and a few venules were connected with these capillary cages at their upper and lateral sides. Occasional arterio-venous and veno-venous anastomoses were also observed around the eccrine sweat glands. This microvascular architecture may adjust well to the mechanical and physiological conditions encountered in the foot-pads. The relation of the microvascular architecture around the eccrine sweat glands with their development is also discussed.     

Development of functional glow-in-the-dark photoluminescence linen fabrics with ultraviolet sensing and shielding

Linen fibres were coated with a glow-in-the-dark photoluminescence, flame-retarding, and hydrophobic smart nanocomposite using the pad-dry-curing process. Ecologically friendly ammonium polyphosphate and lanthanide-activated strontium aluminium oxide (LSAO) nanoparticles were immobilized into linen fabric using eco-friendly room-temperature-vulcanizing silicone rubber. Different analytical techniques were used to examine the morphological characteristics and elemental compositions of LSAO nanoparticles and treated linen textiles. The self-extinguishing properties of the treated linen textiles were tested for their fire resistance. After 24 washing cycles, the coated linen samples retained their flame-retarding properties. The treated linen's superhydrophobicity rose in direct proportion to the LSAO concentration. After being excited at 365 nm, the colourless luminescent film that was coated on linen surface gave out an emission wavelength of 519 nm. The photoluminescent linen was monitored to create a range of different colours, including off-white in daytime light and green under ultraviolet (UV) light radiation, according to the Commission Internationale de l'éclairage laboratory colorimetric coordinates and photoluminescence spectra. Emission, excitation, and lifetime spectral analysis of the treated linen revealed persistent phosphorescence. For mechanical and comfort evaluation, the coated linen textiles' bending length and air permeability were assessed. Good UV light shielding and enhanced antibacterial activity were detected in the treated linens.     

Gene expression,protein profiling,and chemotactic activity of infrapatellar fat pad mesenchymal stem cells in pathologies of the knee joint

The infrapatellar fat pad (IPFP) is a periarticular adipose knee tissue. This tissue contains a large number of mesenchymal stem cells (MSCs). In the present work, we wanted to study the IPFP MSCs and their relationship and differences in two groups, anterior cruciate ligament (ACL) ruptures knees and ostheoarthrosis (OA). The IPFP of 42 patients with OA or ACL rupture were analyzed. Isolation, primary culture, and a genetic and proteomic study of MSCs from IPFP were performed. Gene expression of IL-6, tumor necrosis factor (TNF), IL-8, HSPA1A (Hsp70), CXCL10, RANTES, MMP1, MMP3, TIMP1, and BMP7 was analyzed by real-time quantitative polymerase chain reaction (RT-qPCR). We analyzed MSCs from from 12 diferents patients in two cellular pools (6 from AO disease and 6 from ALC rupture to form two cell pool), for the iTRAQ Proteomic Assay. The conditional media were used in quantitative analysis of MSC soluble factors by Luminex and for de migration assay. A higher gene expression of IL-6, TNF, CXCL10, RANTES, and MMP1 and OPG in MSCs from OA versus ACL (p < 0.05) was observed. Conversely HSPA1A, TIMP1, and RANKL showed a significant lower expression in OA-MSCs (p < 0.05). In the secretome analysis, adipsin and visfantin levels in the supernatants from OA-MSCs were lower (p < 0.05) respect to ACL-MSCs. Also, the monocytic cells migrated two-folds in the presence of conditioned media from OA-MSCs patients versus patients with ACL-MSC. The infrapatellar pad should be considered as an adipose tissue capable of producing and excreting inflammatory mediators directly in the knee joint, influencing the development and progression of knee joint pathologies.     

Fine structural analysis of the fibrillar adhesion apparatus in ladybird beetle

The attachment system on the ladybird beetle Harmonia axyridis is composed of a pair of pretarsal claws and adhesive pads at the tarsal segments. The claws, which are connected to the pretarsal segment, are mainly used to hold the rough substrates by their apical diverged hooks. In contrast, the adhesive pads have an adhesive function when landing on smooth surfaces. They are interspersed at the ventral adhesive pad of each tarsomere, and are composed of two kinds of hairy setae. The discoid tip seta (DtS) is located at the central region of each adhesive pad. The DtS has a spoon‐shaped endplate with a long and narrow shaft. In contrast, the pointed tip seta (PtS) is interspersed along the marginal regions of each adhesive pad, and has a hook‐shaped spine near the tip. In the present study, we found numerous fine cuticular pores beneath the setae, which seem to be related to the secretion of some adhesive fluids. It may be deduced that ladybird beetles can attach to smooth surfaces more effectively by employing adhesive fluids filling in surface crevices to overcome problems cause by their larger size endplates.     

Classical dot–blot format implemented as an amperometric hybridisation genosensor

A new electrochemical hybridisation genosensor has been designed. This genosensor is based on a concept adapted from classical dot–blot DNA analysis, but implemented in an electrochemical biosensor configuration. The use of amperometric transduction and the enzyme label method—that increases the genosensor sensitivity—are the main features of this new approach. The analytical procedure consists of five steps: DNA target immobilisation by adsorption onto a nylon membrane, hybridisation between DNA target and biotin–DNA probe, complexation reaction between biotin-DNA probe and an enzyme (horseradish peroxidase) streptavidin conjugate; integration of the modified membrane onto an electrochemical transducer; and finally, amperometric detection using a suitable substrate for the enzyme labelled duplex. Besides the adapted dot–blot format, a competitive assay in which the target is in solution is reported as well. This procedure, based on amperometric transduction, represents certain advantages with respect to dot–blot analysis: labelled hybrid detection is far simpler, quicker and requires more ordinary or simple reactives; the response obtained is a direct analytical signal via low-cost instrumentation, a nonisotopic labelling is used, and the membranes can be reused. These characteristics are ideal in implementing the procedure developed in kit form.     

Constitutive formulation and numerical analysis of the heel pad region

The aim of this work is to provide a numerical approach for the investigation of the mechanical behaviour of the heel pad region. A visco-hyperelastic model is formulated with regard to fat pad tissue, while a fibre-reinforced hyperelastic model is considered for the heel skin tissue. Bone components are defined by means of an orthotropic linear elastic model. Particular attention is paid to the evaluation of constitutive parameters within different models adopted in consideration of experimental tests data. Preliminarily, indentation tests on a skinless cadaveric foot are considered with regard to fat pad tissue. Indentation tests on an intact heel pad of a cadaveric foot are subsequently adopted for the final identification of constitutive parameters of fat pad and skin tissues. A numerical model of the rear foot is defined and different loading conditions are assumed according to experimental data. A comparison between experimental and numerical data leads to the evaluation of the real capability of the procedure to interpret the actual response of the rear foot.     

高分子膜载体固定化酵母催化2-辛酮不对称还原

以戊二醛交联尼龙6膜载体固定化面包酵母DX213,采用固定化酵母细胞催化2-辛酮不对称还原得到(R)-2-辛醇。系统考察了有机溶剂、反应时间、pH、底物、辅助底物和热处理等因素对反应的产率和光学选择性的影响。结果表明,上述因素对酵母细胞催化不对称合成(R)-2-辛醇反应均有显著影响。二氯甲烷为该反应最适有机溶剂,在固定化细胞57 g/L(50℃预热50 min),水相与有机溶剂相体积比4/1,pH 7.0,初始2-辛酮浓度为60 mmoL/L(分别在反应0,10,17 h等分添加),蔗糖5.7 g/L和28℃条件下反应48 h,(R)-2-辛醇的产率和e.e.值分别达到89.3%和96.8%。     

Bisulfite-modified target DNA array for aberrant methylation analysis

Aberrant DNA methylation of CpG islands is among the earliest and most frequent alterations in cancer. It is of great importance to develop simple and high-throughput methods of methylation analysis for earlier cancer diagnosis or the detection of recurrence. In this study, bisulfite-modified target DNA arrays were prepared on positively charged nylon membrane with two different procedures: fixing PCR products and fixing genomic DNA. First, a bisulfite PCR product array was prepared through fixing PCR products amplified in bisulfite sequencing primers from the bisulfite-modified genomic DNA of different clinical samples on membrane. Furthermore, bisulfite-modified genomic DNA of the different samples was directly fixed on membrane to fabricate bisulfite genomic DNA arrays. The two kinds of arrays were hybridized by probes labeled with digoxigenin, and the hybridization signals were obtained through chemiluminescent detection. The methylation statuses of the IGFBP7 gene for breast tumor and normal tissue samples and for normal human blood cell samples were detected successfully by the two procedures. It was shown that the methods are reliable and sensitive and that they have high potential in screening molecular methylation markers from a large number of clinical samples.     

Plasmodium chabaudi: Adoptive transfer of immunity with different spleen cell populations and development of protective activity in the serum of lethally irradiated recipient mice

Groups of lethally X-irradiated NIH mice were injected with either glass wool-filtered (g.w.) immune spleen cells or nylon wool enriched immune T cells from syngeneic mice immune to Plasmodium chabaudi, or g.w. normal spleen cells. After cell recipients were infected with P. chabaudi the three groups reached similar mean peak parasitaemias on Day 11. In passive transfer tests serum obtained from mice sacrificed at this time gave little protection compared to normal serum. On Day 14 g.w. immune spleen cell recipients had subpatent infections and enriched immune T-cell recipients had a lower mean parasitaemia than g.w. normal spleen cell recipients. Serum obtained on Day 14 from g.w. immune spleen cell recipients gave better protection after passive transfer than sera from enriched immune T-cell or g.w. normal spleen cell recipients. Day 14 serum from enriched immune T-cell recipients, but not from g.w. normal spleen cell recipients, produced some initial protection after passive transfer. These results suggest that the transferred immune spleen cells contributed to the observed humoral immunity in lethally irradiated recipient mice.