Effect of length of gestation on maternal cellular immunity to human trophoblast antigens

Maternal lymphocyte reactivity to human trophoblast antigens was studied in placentas of gestational ages 8 to 14 weeks and 32 to 34 weeks, respectively. Significant trophoblast lysis became apparent after 24 hours' incubation in the latter case compared with a time lag of 72 hours in the terminated gestations. Maternal cellular immunity, therefore, was not detected during the first 3 1/2 months of pregnancy, but was detectable by the time of parturition. The possible significance is discussed with respect to the antigenic stimulus and survival of the fetal allograft.     

Preliminary characterization of glial-secreted factors responsible for the induction of high electrical resistances across endothelial monolayers in a blood-brain barrier model

Low levels of sex hormone-binding globulin (SHBG) are considered to be an indirect index of hyperinsulinemia, predicting the later onset of diabetes mellitus type 2. In the insulin resistance state and in the presence of an increased pancreatic beta-cell demand (e.g. obesity) both absolute and relative increases in proinsulin secretion occur. In the present study we investigated the correlation between SHBG and pancreatic beta-cell secretion in men with different body compositions. Eighteen young men (30.0 +/- 2.4 years) with normal glucose tolerance and body mass indexes (BMI) ranging from 22.6 to 43.2 kg/m2 were submitted to an oral glucose tolerance test (75 g) and baseline and 120-min blood samples were used to determine insulin, proinsulin and C-peptide by specific immunoassays. Baseline SHBG values were significantly correlated with baseline insulin (r = -0.58, P < 0.05), proinsulin (r = -0.47, P < 0.05), C-peptide (r = -0.55, P < 0.05) and also with proinsulin at 120 min after glucose load (r = -0.58, P < 0.05). Stepwise regression analysis revealed that proinsulin values at 120 min were the strongest predictor of SHBG (r = -0.58, P < 0.05). When subjects were divided into obese (BMI > 28 kg/m2, N = 8) and nonobese (BMI < or = 25 kg/m2, N = 10) groups, significantly lower levels of SHBG were found in the obese subjects. The obese group had significantly higher baseline proinsulin, C-peptide and 120-min proinsulin and insulin levels. For the first time using a specific assay for insulin determination, a strong inverse correlation between insulinemia and SHBG levels was confirmed. The finding of a strong negative correlation between SHBG levels and pancreatic beta-cell secretion, mainly for the 120-min post-glucose load proinsulin levels, reinforces the concept that low SHBG levels are a suitable marker of increased pancreatic beta-cell demand.     

Positron emission tomography for the evaluation of pancreatic disease

Efficient techniques for native-labeling of amino acids have been combined successfully with emission tomography to yield significant improvements in pancreatic imaging. Carbon-11-labeled tryptophan appears to be the best agent available currently for imaging the pancreas. Optimum scanning times begin 30 min after tracer administration. Positron emission tomography with 11C-tryptophan is capable of defining both morphological and functional alterations in the pancreas. Tumors as small as 2 cm in diameter can be detected, but reliable differentiation of pancreatic cancer from pancreatis may not be possible even with this improved imaging technique. Longitudinal multiplane emission tomography in single-photon mode with the Pho/Con provides an efficient and satisfactory approach to pancreatic imaging with the positron-emitting radiopharmaceuticals.     

Porous composites of hydroxyapatite‐filled poly[ethylene‐co‐(vinyl acetate)] for tissue engineering

A novel porous composite of hydroxyapatite/poly[ethylene‐co‐vinyl acetate)] (HAP/EVA) having better osteointegration was fabricated by gas foaming technique using a non toxic gas blowing agent intended for bone replacement applications. Combined techniques of scanning electronic microscopy (SEM) and X‐ray microcomputed tomography (µCT) analysis showed that the pore size and pore volume of the porous composite decrease with the increase of HAP content. The gravimetric analysis evidenced for good pore interconnectivity within the porous composites. Energy dispersive X‐ray analysis (EDX) studies inveterated the even scattering of Ca ions which in turn indicate the uniform dispersion of HAP particles in the composites. The significant gradation in Ca ion concentration seen in EDX studies is well accordance with the amount of HAP loading in the sample. Mechanical properties of the porous composite having different HAP content were measured to have the compressive strength varying from 1.06 to 2.2 MPa. Non‐cytotoxic character of the material was observed by the cytocompatibility studies. The metabolic activity of L929 cells seeded on the material assessed by [3‐(4,5‐dimethylthiazol)‐2‐yl]‐2,5‐diphenyltertrazolium bromide (MTT) assay was found to be 91.8%. The adhesion and migration of the cells inside the pore walls were visualized by confocal microscopy. Copyright © 2010 Society of Chemical Industry     

Modulation of stability properties of bovine trypsin after in vitro structural changes with a variety of chemical modifiers

Controlled chemical modification of enzymes, targeting groups not involvedin the active site, can lead to modified catalysts that are intrinsicallymore efficient and resistant to heat and denaturing agents. Bovinepancreatic trypsin was covalently modified up to 75-85% with monomericglutaraldehyde (MGA), polymeric glutaraldehyde (PGA), oxidized sucrose andoxidized sucrose polymers (OSP 70 and OSP 400). Virtually no loss inactivity occurred upon modification. Temperature optima of trypsin shiftsfrom 45-76 degrees C and T50 from 54-76 degrees C for the best modifiedsample made with OSP. The efficiency of the modifiers in stabilization wasranked in the order: OSP 400-T > OSP 70-T > PGA-T > MGA-T >Sucrose-T. Half-life of modified enzymes also followed the same trend. Bothstabilization factor and t1/2 decreased with increasing temperatures. Thefree energy of activation for inactivation delta(deltaG*) varies from 12-20kJ/mol and the activation enthalpy delta(deltaH*) of the modified trypsinby 80-120 kJ/mol indicating stabilization. Inactivation of modified trypsinby urea is less noticeable. The character of the two-step inactivationprocess of trypsin changes with the degree of stabilization in that theduration of phase I one increased noticeably as stabilization increases.Native trypsin fluoresces less intensely showing a red shift under theinfluence of denaturation. Such a fluorescence change is not so obvious forthe modified enzymes indicating conformational stability acquired bymodification.     

The affinity of an oligodeoxynucleotide-peptide conjugate for an RNA hairpin loop depends on stereochemistry at the DNA-peptide junction

Molecular models of an oligodeoxynucleotide-peptide conjugate complexed to an RNA hairpin loop were constructed to assess the effect of stereoisomerism at the point of attachment of the peptide to the oligodeoxynucleotide on the affinity of the conjugate for an RNA target. The peptide portion of the oligodeoxynucleotide-peptide conjugate, (L-lysine)8, was covalently attached to the N-allyl group of (D)- or (L)-aspartic alcohol that was incorporated into the interior of an antisense oligodeoxynucleotide. The stereocenter in the oligodeoxynucleotide interior originates from either (D)- or (L)-aspartic alcohol. The oligodeoxynucleotide portion of the oligodeoxynucleotide-peptide conjugate forms Watson-Crick base pairs with the single-stranded RNA that flanks the RNA hairpin loop. The positively charged peptide makes specific electrostatic contacts with the negatively charged phosphate backbone of the RNA hairpin loop when attached to the N-allyl of (D)-aspartic alcohol but does not have the proper orientation to make these electrostatic contacts when attached to the N-allyl of (L)-aspartic alcohol. This modelling study emphasizes the importance of stereocontrol at the point of branching in synthesizing oligodeoxynucleotide-peptide conjugates for binding of RNA hairpin loops.