Evaluation of senescent cells in intervertebral discs by lipofuscin staining

Intervertebral disc (IVD) degeneration is considered an important contributor of low back pain, a major age related disease. Interestingly, an unprecedented high number of senescent cells has been reported in aged and degenerated IVDs, most probably affecting tissue homeostasis. In previous studies classical markers of cellular senescence have been used, such as SA-beta-gal staining or p16(Ink4a) expression. Aim of the presented study was a reevaluation of the number of senescent IVD cells by using a newly established staining procedure for lipofuscin, based on a Sudan Black-B analogue (GL13), which can be used in fresh, as well as in fixed and embedded tissues. In cultures of senescent rat and human IVD cells both SA-beta-gal and GL13 gave similar percentages of senescent cells. Similarly, in fresh tissues from old rats the ratios of senescent cells were high with both detection procedures. Finally, in formalin-fixed and paraffin-embedded tissues from humans, a significant increased number of GL13-positive cells was found in herniated tissues, as compared to apparently normal ones, while similar numbers of p16(Ink4a)-positive cells were observed. These data confirm the significantly enhanced number of senescent cells in aged and degenerated IVDs, most probably contributing to the degeneration of this tissue

Ambient Intelligence in the Living Room

The emergence of the Ambient Intelligence (AmI) paradigm and the proliferation of Internet of Things (IoT) devices and services unveiled new potentials for the domain of domestic living, where the line between “the computer” and the (intelligent) environment becomes altogether invisible. Particularly, the residents of a house can use the living room not only as a traditional social and individual space where many activities take place, but also as a smart ecosystem that (a) enhances leisure activities by providing a rich suite of entertainment applications, (b) implements a home control middleware, (c) acts as an intervention host that is able to display appropriate content when the users need help or support, (d) behaves as an intelligent agent that communicates with the users in a natural manner and assists them throughout their daily activities, (e) presents a notification hub that provides personalized alerts according to contextual information, and (f) becomes an intermediary communication center for the family. This paper (i) describes how the “Intelligent Living Room” realizes these newly emerged roles, (ii) presents the process that was followed in order to design the living room environment, (iii) introduces the hardware and software facilities that were developed in order to improve quality of life, and (iv) reports the findings of various evaluation experiments conducted to assess the overall User Experience (UX)

Octreotide regulates CC but not CXC LPS-induced chemokine secretion in rat Kupffer cells

1. Kupffer cells (KC) and lipopolysaccharide (LPS) interaction is the initial event leading to hepatic inflammation and fibrosis in many types of liver injury. We studied chemokine secretion by KC activated with LPS and the possible effect of the somatostatin analogue octreotide, in the regulation of this process. 2. KC isolated from Sprague–Dawley rats were cultured in the presence of LPS added alone or with different concentrations of octreotide for 24 and 48 h, and chemokine production was assessed in culture supernatants by ELISA. CC chemokine mRNA expression was assessed by semiquantitative RT–PCR. 3. Vehicle-stimulated KC produced a basal amount of CC and CXC chemokines. LPS-stimulated KC secreted significantly increased amounts of IL-8 (GRO/CINC-1) (P<0.001), MIP-2 (P<0.001), MCP-1 (P<0.001), and RANTES (P<0.01). 4. Octreotide inhibited LPS-induced secretion of the CC chemokines MCP-1 (P<0.05) and RANTES (P<0.05), but not the CXC chemokines IL-8 (GRO/CINC-1) and MIP-2, in a concentration-dependent manner. Downregulation of basal and LPS-induced mRNA expression of the CC chemokines was also observed in the presence of octreotide. 5. Pretreatment with phosphatidylinositol 3 (PI3)-kinase inhibitors reduced chemokine production by LPS-treated KC in both the mRNA and protein level. Furthermore, it prevented the octreotide inhibitory effect on LPS-induced chemokine secretion, indicating a possible involvement of the PI3-kinase pathway. 6. In conclusion, these data demonstrate that chemokine secretion by KC can be differentially regulated by octreotide, and suggest that this somatostatin analogue may have immunoregulatory effects on resident liver macrophages