Mouse blastocysts hatch in vitro by using a trypsin-like proteinase associated with cells of mural trophectoderm

The mammalian blastocyst must hatch from its extracellular coat, or zona pellucida, to implant in the uterus and continue development normally. Results of experiments described here strongly suggest that a proteinase (74K Mr), called "strypsin," is directly involved in hatching of isolated mouse blastocysts in vitro. Strypsin is a trypsin-like proteinase, based on its substrate specificity and sensitivity to inhibitors, that is present in mouse blastocysts and exhibits certain properties characteristic of membrane-associated enzymes. Histochemical and autoradiographic evidence suggests that, prior to hatching of blastocysts, strypsin is found with cells of mural trophectoderm; not with polar trophectoderm or inner cell mass. Following hatching, strypsin is also found associated with empty zonae pellucidae, specifically at the opening through which the embryo emerged. These and other observations suggest that hatching of mouse blastocysts in vitro is initiated by limited proteolysis of the region of zona pellucida overlying mural trophectoderm.     

Laser Doppler blood flow measurements of common cutaneous donor sites for reconstructive surgery

The purpose of this study was to evaluate cutaneous blood flow in regions commonly used as donor sites in reconstructive surgery in order to better establish normal flow ranges. Flow was measured with the TSI Laserflo BPM 403 in 27 healthy volunteers and compared to the flow in uncomplicated postoperative autologous tissue transplants. The forehead produced the highest flow, with an average value of 6.50 +/- 0.31 (mean +/- SE), and the dorsalis pedis had the lowest flow, with an average value of 0.60 +/- 0.04. Gender differences were noted in the latissimus dorsi, pectoralis major, and rectus abdominis areas. There were no significant differences between smokers and nonsmokers, hand dominance, musculocutaneous and fasciocutaneous tissues, or supine and sitting body positions. Flow levels in volunteers were similar to those in postoperative surviving autologous tissue transplants. The site-specific flow and flow changes over long time periods (hours) have helped clinical monitoring of 77 patients in the last 24 months. In every case identified by the flowmeter as decreased perfusion, a definite etiology for low reduction was documented. Complications occurred in 12 patients, and the rate of salvaging compromised tissue has increased from 50 percent using temperature monitoring and clinical observation to 83 percent with the computerized laser Doppler flowmeter.     

Structural basis of substrate specificity in the serine proteases.

Structure-based mutational analysis of serine protease specificity has produced a large database of information useful in addressing biological function and in establishing a basis for targeted design efforts. Critical issues examined include the function of water molecules in providing strength and specificity of binding, the extent to which binding subsites are interdependent, and the roles of polypeptide chain flexibility and distal structural elements in contributing to specificity profiles. The studies also provide a foundation for exploring why specificity modification can be either straightforward or complex, depending on the particular system.