Antiproteinuric effect of angiotensin-converting enzyme inhibition and C5b-9 urinary excretion in membranous glomerulonephritis

BACKGROUND: Angiotensin-converting enzyme (ACE) inhibitors have an antiproteinuric effect in membranous glomerulonephritis (MGN). However, no studies have investigated whether this antiproteinuric effect is influenced by urinary C5b-9 excretion, a marker of immunological activity in this disease. METHODS: Eleven patients with biopsy-proven MGN were treated with captopril for 8 weeks. The evolution of several clinical and biochemical parameters, including 24-h urinary protein excretion was evaluated every 4 weeks. Urinary C5b-9 excretion was measured at the onset and at the end of captopril treatment. RESULTS: Patients with MGN had significantly higher C5b-9 excretions than a group of 14 healthy controls (89 +/- 23 vs 3.7 +/- 1.4 ng/mg UCr; P < 0.001). A significant correlation was found between urinary C5b-9 and the magnitude of proteinuria, both at the onset and at the end of treatment. After 8 weeks of captopril treatment, proteinuria had decreased from 8 +/- 1.8 to 5.2 +/- 1.3 g/day (P < 0.05). Four weeks after captopril discontinuation, proteinuria rose to 7.3 +/- 1.7 g/day (P < 0.05). A marked variability in the antiproteinuric response was observed, ranging from 0 to 85% with respect to baseline values. No correlation between decrease in proteinuria and baseline urinary C5b-9 levels was observed. Several patients with elevated urinary C5b-9 levels had captopril-induced decrease in proteinuria. CONCLUSIONS: ACE inhibition induces an antiproteinuric effect in patients with MGN. The urinary C5b-9 excretion does not predict the magnitude of this response.     

Trypanosoma cruzi: effect of immunization on the risk of vector-delivered infection in guinea pigs

The protective effect of experimental immunization was studied in guinea pigs exposed to vectorial infection by Trypanosoma cruzi. Immunized animals received an inoculum of live-attenuated T. cruzi epimastigotes into a granuloma previously induced by Freund's complete adjuvant in the hind footpad. Seven days later, a delayed-type hypersensitivity reaction was triggered by reinjection of the parasites in the front footpad. The animals were then placed in Triatoma infestans-colonized corrals and exposed to vectorial T. cruzi transmission of the parasite for up to 200 days. The effectiveness of this immunizing protocol was controlled in terms of the number of bites necessary for infection (NBNI) in immunized as compared with control animals. Periodic entomological census allowed for the determination of vector biting and infection rates and the calculation of NBNI. Although this measurement was quite variable between yards, an overall average of 4,973 bites was enough to infect a control guinea pig in 4 separate experiments. The corresponding figure for the experimental group was 21,307 bites, implying that immunized animals could resist a 4.28-fold increase (range: 1.99-8.32) in the number of vector bites before becoming infected.     

Hybrid Photopatterned Enzymatic Reaction (HyPER) for in Situ Cell Manipulation

The ability to design artificial extracellular matrices as cell‐instructive scaffolds has opened the door to technologies capable of studying the fate of cells in vitro and to guiding tissue repair in vivo. One main component of the design of artificial extracellular matrices is the incorporation of biochemical cues to guide cell phenotype and multicellular organization. The extracellular matrix (ECM) is composed of a heterogeneous mixture of proteins that present a variety of spatially discrete signals to residing cell populations. In contrast, most engineered ECMs do not mimic this heterogeneity. In recent years, photo‐deprotection has been used to spatially immobilize signals. However, this approach has been limited mostly to small peptides. Here we combine photo‐deprotection with enzymatic reaction to achieve spatially controlled immobilization of active bioactive signals that range from small molecules to large proteins. A peptide substrate for transglutaminase factor XIII (FXIIIa) was caged with a photo‐deprotectable group, which was then immobilized to the bulk of a cell‐compatible hydrogel. With focused light, the substrate can be deprotected and used to immobilize patterned bioactive signals. This approach offers an innovative strategy to immobilize delicate bioactive signals, such as growth factors, without loss of activity and enables in situ cell manipulation of encapsulated cells.     

Field measurement of albedo for limited extent test surfaces

A new method is introduced for field measurement of surface albedo. This method consists of the use of a cylindrical shade ring made of opaque fabric with a known (low) albedo placed over a test surface. The albedo measurement is accomplished using two small pyranometers situated so that the downward-facing pyranometer receives radiation only from the test surface and the shade ring. The upward-facing pyranometer simultaneously records the incoming solar radiation. The radiation received by the downward-facing pyramometer is a combination of reflected radiation from shaded and unshaded portions of these two surfaces, requiring detailed accounting of the resulting view factor geometries. The method presented here improves upon past approaches by allowing for smaller sample sizes, minimizing errors associated with reflective properties of the surroundings, and allowing for accurate measurements even under partially cloudy skies. In addition to these methodological improvements we introduce an approach for estimating the uncertainty in the resulting albedo measurements. Results from field measurements are presented to validate the measurement protocol, and to compare its accuracy with the accuracy of a published standard.     

Delivery of iPS‐NPCs to the Stroke Cavity within a Hyaluronic Acid Matrix Promotes the Differentiation of Transplanted Cells

Stroke is the leading cause of adult disability with ≈80% being ischemic. Stem cell transplantation has been shown to improve functional recovery. However, the overall survival and differentiation of these cells is still low. The infarct cavity is an ideal location for transplantation as it is directly adjacent to the highly plastic peri‐infarct region. Direct transplantation of cells near the infarct cavity has resulted in low cell viability. Here, neural progenitor cells derived from induce pluripotent stem cells (iPS‐NPC) are delivered to the infarct cavity of stroked mice encapsulated in a hyaluronic acid hydrogel matrix to protect the cells. To improve the overall viability of transplanted cells, each step of the transplantation process is optimized. Hydrogel mechanics and cell injection parameters are investigated to determine their effects on the inflammatory response of the brain and cell viability, respectively. Using parameters that balanced the desire to keep surgery invasiveness minimal and cell viability high, iPS‐NPCs are transplanted to the stroke cavity of mice encapsulated in buffer or the hydrogel. While the hydrogel does not promote stem cell survival one week post‐transplantation, it does promote differentiation of the neural progenitor cells to neuroblasts.     

Sialomucins are characteristically O-acylated in poorly differentiated and colloid prostatic adenocarcinomas

Mucinous glycoproteins are secreted by prostatic adenocarcinomas and might play important roles in tumor invasion and metastasis. Their histochemical properties on routine biopsy specimens have not been fully characterized. We present a histochemical study of mucin in 21 prostatic adenocarcinomas, with particular focus on the demonstration of different types of sialomucins. We applied the following histochemical techniques to routinely processed, formalin-fixed, paraffin-embedded tissue sections: Alcian blue (pH 2.5) and periodic acid-Schiff to reveal both acidic and neutral mucins; high iron diamine and Alcian blue (pH 2.5) to show sulfated and acidic nonsulfated mucosubstances simultaneously; periodic acid borohydride, potassium hydroxide, and periodic acid-Schiff to demonstrate O-acylated sialic acids; periodic acid thionine-Schiff, potassium hydroxide, and periodic acid-Schiff to differentiate pre-existing glycols from those revealed after saponification procedures; and periodic acid borohydride and periodic acid-Schiff to show C9-O-acylated sialic acid. These techniques are useful tools for demonstrating neutral and acidic (sialo- and sulfo-) mucins and di(C8,C9- or C7,C9-)-O-acylated, tri(C7,C8,C9-)-O-acylated and mono(C9)-O-acylated sialomucins. Most prostatic adenocarcinomas showed acidic mucins, with sialomucins predominating over sulfomucins. Well-differentiated and moderately differentiated noncolloid tumors had non-O-acylated sialomucins. Poorly differentiated tumors contained mono-O-acylated (C9) sialomucins, and colloid-type tumors secreted mono-, di-, and tri-O-acylated sialoglycoproteins. Acidic mucins, mainly sialomucins, constitute the major secretory component in prostatic adenocarcinomas, and our results show that the O-acylation of these sialoglycoproteins inversely correlates with tumor differentiation. Well-differentiated and moderately differentiated tumors are not O-acylated, whereas the poorly differentiated ones characteristically have O-acylated sialomucins in C9. Adenocarcinomas of the colloid type, thought to bear a poor prognosis, are the most heavily O-acylated.