Distinctive Capillary Action by Micro-channels in Bone-like Templates can Enhance Recruitment of Cells for Restoration of Large Bony Defect

Without an active, thriving cell population that is well-distributed and stably anchored to the inserted template, exceptional bone regeneration does not occur. With conventional templates, the absence of internal micro-channels results in the lack of cell infiltration, distribution, and inhabitance deep inside the templates. Hence, a highly porous and uniformly interconnected trabecular-bone-like template with micro-channels (biogenic microenvironment template; BMT) has been developed to address these obstacles. The novel BMT was created by innovative concepts (capillary action) and fabricated with a sponge-template coating technique. The BMT consists of several structural components: inter-connected primary-pores (300-400 µm) that mimic pores in trabecular bone, micro-channels (25-70 µm) within each trabecula, and nanopores (100-400 nm) on the surface to allow cells to anchor. Moreover, the BMT has been documented by mechanical test study to have similar mechanical strength properties to those of human trabecular bone (~3.8 MPa)¹².The BMT exhibited high absorption, retention, and habitation of cells throughout the bridge-shaped (Π) templates (3 cm height and 4 cm length). The cells that were initially seeded into one end of the templates immediately mobilized to the other end (10 cm distance) by capillary action of the BMT on the cell media. After 4 hr, the cells homogenously occupied the entire BMT and exhibited normal cellular behavior. The capillary action accounted for the infiltration of the cells suspended in the media and the distribution (active migration) throughout the BMT. Having observed these capabilities of the BMT, we project that BMTs will absorb bone marrow cells, growth factors, and nutrients from the periphery under physiological conditions.The BMT may resolve current limitations via rapid infiltration, homogenous distribution and inhabitance of cells in large, volumetric templates to repair massive skeletal defects.     

IgG Western Blot for Confirmatory Diagnosis of Equivocal Cases of Toxoplasmosis by EIA-IgG and Fluorescent Antibody Test

The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique.     

Anti-epithelial cell adhesion molecule RNA aptamer-conjugated liposomal doxorubicin as an efficient targeted therapy in mice bearing colon carcinoma tumor model

To overcome the lack of selectivity and nonspecific biodistribution of drugs in the body, targeted delivery of chemotherapeutic agents with aptamers is a very effective method. In this strategy, aptamers could be specifically identified and attach to targeted molecules on the cancerous cells and deliver the chemotherapeutic agents to desired tissue with minimal or no damage to the normal cells. In this study, we designed anti-epithelial cell adhesion molecule (EpCAM) RNA aptamer conjugated PEGylated liposomal doxorubicin (ER-lip) to investigate its in vitro and in vivo anticancer abilities. Data showed that EpCAM aptamer was able to enhance cell uptake and cytotoxic effects of Dox in C26 cell line. The biodistribution study indicated that ER-lip enhanced the tumor accumulation of Dox compared to Caelyx. Also, double staining of isolated tumor cells with anti-CD44-PE-cy5 and anti-EpCAM Cy-7 antibodies indicated that tumor cells expressed a high level of EpCAM⁺ CD44⁺ cells (p ≤ .001) compared to cultured C26 cell line. in vivo results showed that ER-lip promoted survival and reduced tumor growth rate in animal model compared to Caelyx. In conclusion, these results suggested that the ER-lip could be served as promising formulation for the treatment of cancers with the high expression of EpCAM.     

Restoration of a normal phenotype, microtubular pattern and DNA synthesis in embryonic fibroblasts concanavalin A pretreated

The present research investigated the time required for reconstructing normal microtubular pattern and the phenotype of cells after short-term (30 min.) and long-term (24 hrs.) pretreatment with the lectin concanavalin A (Con A). Short-term pretreatment led to the formation of incomplete tubules within 2 and 24 hrs. in cells cultured in 199 medium alone. The addition of serum to the medium reversed the globular phenotype and allowed the formation of normal microtubules, even after prolonged pretreatment with Con A. Whereas long-term Con A treatment provoked a reduction in DNA synthesis in 199 alone, in serum-added 199 the percentage of 3H-thymidine incorporation of treated cells tended to reach that of controls over time.     

Fractional reabsorption of calcium,magnesium and phosphate in the kidneys of freshwater North American eels (Anguilla rostrata LeSueur) following removal of the corpuscles of Stannius

Renal function was observed in freshwater North American eels (Anguilla rostrata LeSueur) 2 weeks after the removal of the corpuscles of Stannius. There was a positive linear correlation between glomerular filtration rates and urine flow rates in both sham-operated and stanniectomized eels but there was no difference in slope or elevation between the two groups nor did urine flow rates ever exceed glomerular filtration rates. Osmolar clearance and free-water clearance were unchanged following stanniectomy. Plasma Ca²⁺ and K⁺ concentrations increased and plasma Mg²⁺, phosphate, Na⁺ and Cl^- concentrations decreased following stanniectomy. Plasma ultrafilterable Ca increased and ultrafilterable Mg decreased after stanniectomy but neither changed relative to its total plasma concentration. Stanniectomy was followed by a decreased renal tubular reabsorption of Mg²⁺ relative to the amount filtered (C _Mg/C _In); the same applies to C _Na/C _In. Even though the filtered load of Ca increased in conjuction with the predictable hypercalcemia, there was no change in the fraction of filtered Ca reabsorbed. Net tubular secretion of phosphate was observed in both sham-operated and stanniectomized cels together with a slight increase in C _phos/C _In following stanniectomy. Some or all of these changes in plasma electrolytes and/or the modified renal transport of Na⁺, Mg²⁺ and possibly phosphate may be caused by the changes in cardiovascular function that were recently shown to follow stanniectomy.Abbreviations CSX Stanniectomized/stanniectomy - ER endoplasmatic reticulum - FW fresh water - GFR glomerular filtration rate - Pi inorganic phosphate - RAS renin-angiotensin system - SHM sham-operated - SW sea water - UFR urine flow rate - U/P ratio urine/plasma ratio     

Revised transthyretin Ile 122 allele frequency in African-Americans

The transthyretin (TTR) Ile 122 variant is associated with cardiac amyloidosis in individuals of African descent. To determine the prevalence of the allele encoding TTR Ile 122 in African-Americans, we have used PCR and restriction analysis to test DNA from African-Americans from various geographic areas, and found an allele frequency of 66/3376 (0.020), which is higher than the value we previously reported in a much smaller pilot study. Our data indicate that this TTR variant is present at equal frequency in African-Americans throughout the U.S., and suggest that this mutation may be a common, often unrecognized cause of cardiac disease in African-Americans. Received: 23 January 1996