Amyloid-β25–35 induces a permanent phosphorylation of HSF-1, but a transitory and inflammation-independent overexpression of Hsp-70 in C6 astrocytoma cells

Two hallmarks of Alzheimer diseases are the continuous inflammatory process, and the brain deposit of Amyloid b (Aβ), a cytotoxic protein. The intracellular accumulation of Aβ_25–35 fractions, in the absence of Heat Shock proteins (Hs?s), could be responsible for its cytotoxic activity. As, pro-inflammatory mediators and nitric oxide control the expression of Hs?s, our aim was to investigate the effect of Aβ_25–35 on the concentration of IL-1β, TNF-α and nitrite levels, and their relation to pHSF-1, Hsp-60, -70 and -90 expressions, in the rat C6 astrocyte cells. Interleukin-specific ELISA kits, immunohistochemistry with monoclonal anti-Hsp and anti pHSF-1 antibodies, and histochemistry techniques, were used. Our results showed that Aβ_25–35 treatment of C6 cells increased, significantly and consistently the concentration of IL-1β, TNF-α and nitrite 3 days after initiating treatment. The immunoreactivity of C6 cells to Hsp-70 reached its peak after 3 days of treatment followed by an abrupt decrease, as opposed to Hsp-60 and -90 expressions that showed an initial and progressive increase after 3 days of Aβ_25–35 treatment. pHSF-1 was identified throughout the experimental period. Nevertheless, progressive and sustained cell death was observed during all the treatment times and it was not caspase-3 dependent. Our results suggest that Hsp-70 temporary expression serves as a trigger to inhibit casapase-3 pathway and allow the expression of Hsp-60 and -90 in C6 astrocytoma cells stimulated with Aβ_25–35.     

Persistent systemic monocyte and neutrophil activation in neonatal encephalopathy

Aim: Circulating immune cell activation is associated with worse outcome in adult and animal models of brain injury. Our aim was to profile the systemic inflammatory response over the first week of life in infants at risk of neonatal encephalopathy (NE) and correlate early neutrophil and monocyte endotoxin and activation responses with outcome.

Methods: Prospective observational study in a tertiary referral university hospital including 22 infants requiring resuscitation at birth who had serial (five time points) neutrophil and monocyte CD11b (marker of cell adhesion), intracellular reactive oxygen intermediates (ROI; cell activation) and Toll-like receptor (TLR; endotoxin recognition) before and after endotoxin stimulation ex vivo compared to neonatal controls.

Results: All neonates requiring resuscitation at delivery (n?=?122 samples) had higher neutrophil and monocyte CD11b and TLR-4 expression compared with adults and neonatal controls. Neonates with abnormal neuroimaging and/or severe NE had increased CD11b, ROI and TLR-4. Increased polymorphonuclear leukocytes TLR-4 expression was associated with increased mortality in infants with NE.

Conclusion: Innate immune dysregulation in the first week of life is associated with severity of outcome in neonatal brain injury in this cohort and may be amenable to immunomodulation.     

The Effects of Bariatric Surgery-Induced Weight Loss on Adipose Tissue in Morbidly Obese Women Depends on the Initial Metabolic Status

Background

Adipose tissue (AT) dysfunction in obesity is commonly linked to insulin resistance and promotes the development of metabolic disease. Bariatric surgery (BS) represents an effective strategy to reduce weight and to improve metabolic health in morbidly obese subjects. However, the mechanisms and pathways that are modified in AT in response to BS are not fully understood, and few information is still available as to whether these may vary depending on the metabolic status of obese subjects.

Methods

Abdominal subcutaneous adipose tissue (SAT) samples were obtained from morbidly obese women (n?=?18) before and 13.3?±?0.37 months after BS. Obese women were stratified into two groups: normoglycemic (NG; Glu?<?100 mg/dl, HbA1c <5.7 %) or insulin resistant (IR; Glu 100–126 mg/dl, HbA1c 5.7–6.4 %) (n?=?9/group). A multi-comparative proteomic analysis was employed to identify differentially regulated SAT proteins by BS and/or the degree of insulin sensitivity. Serum levels of metabolic, inflammatory, and anti-oxidant markers were also analyzed.

Results

Before surgery, NG and IR subjects exhibited differences in AT proteins related to inflammation, metabolic processes, the cytoskeleton, and mitochondria. BS caused comparable weight reductions and improved glucose homeostasis in both groups. However, BS caused dissimilar changes in metabolic enzymes, inflammatory markers, cytoskeletal components, mitochondrial proteins, and angiogenesis regulators in NG and IR women.

Conclusions

BS evokes significant molecular rearrangements indicative of improved AT function in morbidly obese women at either low or high metabolic risk, though selective adaptive changes in key cellular processes occur depending on the initial individual’s metabolic status.

     

A real-time dynamic 3D model of the human inguinal region for surgical education

A suitable dynamic 3D model that allows the simulation of the inguinal region with real-time performance on a personal computer was developed. A geometric model adjusted to real data was created by means of semiautomatic contour segmentation of anatomic units from the visible human project and data generated from classical anatomic information. A dynamic model included converting muscular units from their continuous geometric representation into a set of voxels and then real-time interaction and performance. The current implementation enables deformation of the realistic model associated with pushing and stretching interaction, allowing immersion in the anatomy of the inguinal structures. The model does not allow simulation of surgical interventions.     

Digital autoradiography using room temperature CCD and CMOS imaging technology

CCD (charged coupled device) and CMOS imaging technologies can be applied to thin tissue autoradiography as potential imaging alternatives to using conventional film. In this work, we compare two particular devices: a CCD operating in slow scan mode and a CMOS-based active pixel sensor, operating at near video rates. Both imaging sensors have been operated at room temperature using direct irradiation with images produced from calibrated microscales and radiolabelled tissue samples. We also compare these digital image sensor technologies with the use of conventional film. We show comparative results obtained with (14)C calibrated microscales and (35)S radiolabelled tissue sections. We also present the first results of (3)H images produced under direct irradiation of a CCD sensor operating at room temperature. Compared to film, silicon-based imaging technologies exhibit enhanced sensitivity, dynamic range and linearity.