Fine structure of early human embryos frozen with 1,2 propanediol

Previous studies by a French group (Fertil Steril 44:645–651, 1985) have shown that two-to eight-cell human embryos can survive slow freeze-thawing with propanediol in a biological freezer. These embryos were assessed for morphological appearance by phase-contrast microscopy. We assessed the structure of 25 frozen-thawed one- to 12-cell embryos, obtained from our in vitro fertilization (IVF) and GIFT programmes, by phase-contrast and electron microscopy, using the same method of cryopreservation. One-fourth of the embryos examined had all cells intact, and more than one-half the embryos had over 50% of their cells well preserved. Some of these embryos had unequal blastomeres and cytoplasmic fragments. Ultrastructural assessment revealed good preservation of fine structure in the intact blastomeres of all embryos and maintenance of cell-to-cell contacts. Most cytoplasmic organelles, cell membranes, and nuclei were well preserved compared to nonfrozen controls. The cells that were cryoinjured showed varying degrees of disorganization of the cell membrane, cytosol, and cellular membranes, including swelling and disruption of the nuclear envelope. Disruption of the zona was somewhat rare. Small cytoplasmic fragments were less prone to cryoinjury than blastomeres. The use of propanediol for embryo cryopreservation seems to be feasible; frozen embryos with more than 50% cells intact have produced 10 pregnancies after embryo transfer (Fertil Steril 46:268–272, 1986). Replacement of 17 frozen embryos in seven patients has resulted in a twin pregnancy in Singapore. However, the effects of freezing on the mitotic spindles of embryonic cells need to be investigated further.     

The determination of purine levels in human and mouse plasma

Variable levels of acetic anhydride have been recommended for addition to one of two reagents used in the glyoxylic acid method for the determination of tryptophan. For use of this reagent immediately after preparation it was shown that a minimum of 16% (v/v) of acetic anhydride should be included in the formulation to obtain near-maximum sensitivity. It was further demonstrated that reagent formulations with and without acetic anhydride changed with exposure to light. The observed changes are manifest as changes in the relative sensitivities of the assay. Several modifications are recommended to improve the sensitivity and stability of the acetic anhydride-containing reagent in this assay.     

Intensification of alcoholic fermentation upon dehydration-rehydration of the yeast Saccharomyces cerevisiae

Summary In comparison with intact yeast, dehydrated-rehydrated cells of Saccharomyces cerevisiae show significantly higher ethanol production from exogenous substrate under both anaerobic and aerobic conditions, particularly when low concentration (0.1%) of glucose are used. For populations with a higher percentage of viable rehydrated cells (above 70%) a more notable decrease in the Pasteur effect (the difference between the quantity of ethanol formed under anaerobic and aerobic conditions) is observed.     

Tissue factor expression in newt iris coincides with thrombin activation and lens regeneration

Lens regeneration in adult salamanders occurs at the pupillary margin of the mid-dorsal iris where pigmented epithelial cells (PEC) re-enter the cell cycle and transdifferentiate into lens. It is not understood how the injury caused by removal of the lens (lentectomy) in one location is linked to initiating the response in a different spatial location (dorsal iris) and to this particular sector. We propose that the blood provides a link between the localised coagulation and signal transduction pathways that lead to regeneration. A transmembrane protein (tissue factor) is expressed in a striking patch-like domain in the dorsal iris of the newt that localises coagulation specifically to this location, but is not expressed in the axolotl, a related species that does not show thrombin activation after lentectomy and cannot regenerate its lens. Our hypothesis is that tissue factor expression localises the initiation of regeneration through the activation of thrombin and the recruitment of blood cells, leading to local growth factor release. This is the first example of gene expression in a patch of cells that prefigures the location of a regenerative response, and links the immune system with the initiation of a regenerative program.     

Isolation and Pathogenicity of Rhizosphere Fungi of Cocoyam in Relation to Cocoyam Root Rot Disease

Cocoyam is the second most important staple crop of Cameroon and root rot is a destructive disease of this plant. Pythium myriotylum (Pm), Fusarium solani (Fs), and Rhizoctonia solani (Rs) were isolated from the rhizosphere of root rot affected cocoyams and from the soil of a cocoyam experimental field plot temporarily devoid of same in Mamu, Cameroon. Pm was isolated from the above soil by the cocoyam leaf disc baits. Fs and Rs were also isolated from the same soils by the water dilution method and from the roots of diseased cocoyams but were always associated with mycelial growth of Pm. Pathogenicity of Pm and in combinations with Fs or Rs or Fs + Rs all developed cocoyam root rot disease (CRRD) symptoms on 3– and 7–month old cocoyam plantlets 2–7 days after inoculation. Symptoms included rotted roots and wilting with general chlorosis of inoculated plantlets. No symptoms of CRRD were noted on cocoyam plantlets inoculated with Fs, Rs, Fs + Rs, and distilled water. Results indicated that CRRD is not caused by several pathogens but only by Pm. Pm isolates from the soils and roots of diseased cocoyams and those maintained in the ROTREP laboratory have significantly bigger diameter of mycelial colony growth in 24 h–period at 31 °C on lima bean sucrose agar, V–8 juice sucrose agar, and potato sucrose agar than on potato dextrose agar and 2 % water agar. The cocoyam plantlets were raised axenically from tissue culture of explants in the laboratory.