Aspartic Proteinase Members Secreted by the Ruminant Placenta: Specificity of Three Radioimmunoassay Systems for the Measurement of Pregnancy-associated Glycoproteins

Pregnancy‐associated glycoproteins (PAGs) isolated from the placenta of various ruminant species are enzymatically inactive members of the aspartic proteinase family. The measurement of these proteins in the maternal blood can be a good indicator of the presence of a live embryo. As certain aspartic proteinases are present in biological fluids in physiological and pathological conditions at various concentrations, it was necessary to determine the specificity of three radioimmunoassay (RIA) systems currently used for the detection of PAG molecules. Commercially available members of the aspartic proteinase family like pepsinogen, pepsin, chymosin, rennet, cathepsin D and renin were tested in a wide concentration range (10 ng/ml – 1 mg/ml). Pepsinogen cross‐reacted in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 50 μg/ml and 500 μg/ml concentrations, respectively. In the presence of pepsin, cross‐reaction was observed in RIA 1, RIA 2 and RIA 3 over 1 mg/ml, 500 μg/ml and 1 mg/ml concentrations, respectively. Chymosin and rennet could cross‐react in RIA 2 and RIA 3, while renin and cathepsin D did not decrease the binding of the tracer to antisera more, than that of the minimal detection limit. As the plasma/serum concentrations of the examined aspartic proteinases reported in the literature were outside the concentration range where cross‐reaction was observed, it can be concluded that these RIA systems were specific for the detection of PAGs in biological fluids.     

Sulfur diagenesis in everglades peat and origin of pyrite in coal

The pattern of sulfur transformation in peat across the Everglades basin indicates that pyrite formation in organic-rich swamps depends on the use of organic oxysulfur compounds in dissimilatory respiration by sulfur-reducing bacteria. This paragenesis explains the primary distribution of sulfur compounds in low-sulfur coals and possibly in most coals and many organic-rich soils and sediments. It also accounts for the occurrence of framboidal pyrite bound in fossil tissue in coal and sediments.     

Synaptonemal complex investigations on lamas (Lama glama) with differing fertility recordings

The present study involved three male lamas ( Lama glama ). Mitotic preparations from fibroblast culture and an electron microscopic observation on the synaptonemal complexes (Scs) were reported. Special attention was given to the morphology and behaviour of the chromosomes at the zygotene and pachytene substages of prophase I in primary spermatocytes from lamas. Analysis of mitotic preparations show diploid and triploid cells, with a relatively high frequency of polyploidy. Analyses of synaptonemal complexes in primary spermatocytes were carried out on 89 cells. Pairing abnormalities were only recorded in an average of 63% of the cells of the animal Tabasco. The other two animals were normal. The photographed cells give an upper limit estimate of the existing abnormalities, as there was a deliberate tendency towards selecting abnormal cells for photography. The presence of degenerating primary spermatocytes in SC preparations as well as in testicular sections, and the absence of spermatozoa in ejaculates confirm the chromosomally derived male sterility of one animal.     

Transformation of Montmorillonite to Kaolinite during Weathering

Extensive deposits of kaolinite in Florida are formed by transformation of montmorillonite during low-temperature supergene weathering. The transformation occurs by intracrystalline leaching of interlayer cations and tetrahedral silica layers. Interposition of stripped layers within montmorillonite creates a regular 1:1 mixedlayered montmorillonite-kaolinite, a new clay structure. Kaolin-like layers are nourished by lateral epitaxy, as the iron-rich montmorillonite decomposes. Hexagonal outgrowths of new kaolinite develop at the edges of montmorillonite flakes and nucleate new vertical growth. Kaolinitic sands impregnated with goethite are ultimately formed, and the released silica enriches groundwater and forms secondary chert.     

Multidimensional drug profiling by automated microscopy

We present a method for high-throughput cytological profiling by microscopy. Our system provides quantitative multidimensional measures of individual cell states over wide ranges of perturbations. We profile dose-dependent phenotypic effects of drugs in human cell culture with a titration-invariant similarity score (TISS). This method successfully categorized blinded drugs and suggested targets for drugs of uncertain mechanism. Multivariate single-cell analysis is a starting point for identifying relationships among drug effects at a systems level and a step toward phenotypic profiling at the single-cell level. Our methods will be useful for discovering the mechanism and predicting the toxicity of new drugs.     

Spontaneous cell polarization through actomyosin-based delivery of the Cdc42 GTPase

Cell polarization can occur in the absence of any spatial cues. To investigate the mechanism of spontaneous cell polarization, we used an assay in yeast where expression of an activated form of Cdc42, a Rho-type guanosine triphosphatase (GTPase) required for cell polarization, could generate cell polarity without any recourse to a preestablished physical cue. The polar distribution of Cdc42 in this assay required targeted secretion directed by the actin cytoskeleton. A mathematical simulation showed that a stable polarity axis could be generated through a positive feedback loop in which a stochastic increase in the local concentration of activated Cdc42 on the plasma membrane enhanced the probability of actin polymerization and increased the probability of further Cdc42 accumulation to that site.     

Comparison of the Ability of Three Radioimmunoassay to Detect Pregnancy-associated Glycoproteins in Bovine Plasma

Pregnancy‐associated glycoproteins (PAGs) constitute a large family of glycoproteins that are synthesized in the superficial layer of the ruminant placenta according to a spatial and temporal expression pattern. When PAGs are released in the maternal blood they can be used for pregnancy diagnosis, pregnancy follow‐up and for the monitoring of the trophoblastic function. Three different radioimmunoassay systems (RIA 1, RIA 2 and RIA 3) using antisera produced against PAG I₆₇ (RIA 1), PAG₅₅₊₆₂ (RIA 2) and PAG₅₅₊₅₉ (RIA 3) were used in this investigation in order to measure the PAG concentration in plasma samples withdrawn from pregnant cows and heifers during different periods following artificial insemination (AI). These systems were able to detect PAG molecules in the maternal blood as early as 21 days after AI in different concentrations (RIA 1: 0.43 ± 0.24 ng/ml, mean ± SD; RIA 2: 0.48 ± 0.24 ng/ml; RIA 3: 0.64 ± 0.37 ng/ml). On days 32 and 42 RIA 2 (4.30 ± 1.32 ng/ml and 5.56 ± 1.95 ng/ml) and RIA 3 (4.17 ± 1.15 ng/ml and 5.60 ± 1.89 ng/ml) presented significantly (p < 0.0001) higher PAG concentrations than those of RIA 1 (2.43 ± 0.81 ng/ml and 4.01 ± 1.48 ng/ml), respectively. After day 21, significant correlations (p < 0.0001; r ≥ 0.929) were determined between the three systems. Additionally the three individual PAG profiles presented in this study showed that PAG molecules secreted in the maternal blood between 21 and 50 days after AI were better recognized by the RIA 2 and RIA 3 systems. This study clearly indicated that the ability of a RIA test to recognize PAG molecules in the maternal blood can be improved by carefully selecting the antiserum.