Cytoplasmic Halo Research Articles

A Graduated Embryo Scoring (GES) System Predicts Blastocyst Development From Cleavage-Stage Embryos

Objective: To evaluate a graduated embryo scoring system for predicting blastocyst formation from cleavage-stage embryos. Design: Ongoing, prospective analysis. Materials and Methods: Two hundred and sixty-eight fertilized embryos were derived from 22 patients following IVF/ICSI and were cultured individually in P1 until day 3 and then in Blastocyst Medium until day 5. Weighted scores out of 50 were awarded at each of four evaluations based on the presence of critical developmental milestones. The first evaluation at 16–18 hours post ICSI rated cytoplasmic halo and vacuoles, pronuclear size and juxtaposition, nucleolar alignment, and polar body apposition and fragmentation. At 25–27 hours the embryos were evaluated for dissolution of the pronuclear membrane, blastomere cleavage, symmetry and the degree of fragmentation. At 40–43 hours, the embryos were evaluated for blastomere number, symmetry and percent fragmentation. A final evaluation of blastomere number and morphology at 64–67 hours generated a total GES score out of a possible 200 points. This GES score was correlated with blastocyst development. Results: A total GES score of 150–200 on day 3 was achieved by 35% (93/268) of embryos, 100–149 by 45% (121/268), and <100 by 20% (54/268). Seventy-five percent (70/93) of embryos scoring 150–200 developed to blastocyst, compared to 45% (55/121) of embryos scoring 100–149 (p<0.0001), and to 13% (7/54) scoring <100 (p<0.00001). The difference in blastocyst development between embryos scoring 100–149 and those scoring <100 was also significant (p<0.0001). Fifty-seven embryos were transferred into the 22 patients on day 5 and an additional 20 were frozen, yielding 77 (29%) “usable” embryos. Based on traditional morphology characteristics 184 would have been deemed transferable/freezable on day 3. The mean GES value for “usable” embryos was 146, compared to 129 for those not transferred or frozen. Conclusions: Graduated embryo scoring based on early developmental milestones is an accurate predictor of blastocyst development from cleavage-stage embryos. The ability to identify those embryos on day 3 which are likely to develop to blastocysts may decrease laboratory culture expense, allow earlier cryopreservation of supernumeray embryos, and could be useful in situations where the ability to culture more embryos than are intended for transfer is limited. Our early experience also indicates that there was no effect on the rate blastocyst development from individual embryo culture or from the repeated removal of the embryos from the incubator. We hope to better define those characteristics that are critical to blastocyst development, so that “usable” embryos could be identified even earlier. Data correlating GES with IVF outcome will also be presented.

Histologic effects of ruby laser hair removal in Japanese patients.

Hair removal by lasers has recently become a popular method to remove unwanted hair. However, histologic changes in human skin before and after exposure to lasers have not been thoroughly investigated. The aim of this study was to clarify the differences that occur immediately after laser exposure and 1 month after laser exposure. Eight adult Japanese volunteers were recruited for this study. They were treated with a long pulsed ruby laser at 20 J/cm(2). A single 3-mm punch biopsy of the laser-treated sites was obtained immediately after laser irradiation and at the 1-month follow-up visit, and they were analyzed using hematoxylin and eosin, PAM, and immunohistological staining. Immediately after laser exposure, hair follicles were very damaged and had extensive eosinophilic degeneration. One month after laser therapy, one type of hair follicle showed cystlike formations with negative proliferating cell nuclear antigen reactions (PCNA). Another type of hair follicle showed follicular mitotic figures with cytoplasmic halos. Early anagen hair follicles were apparently not treated effectively by ruby laser. Ruby laser leads to extensive follicular damage, and hair follicles considered to be at early anagen phase were not effectively treated. This may be the reason several courses of laser therapy are required to obtain satisfactory results.

Original Article: Relaxin Reduces the Number of Circulating Platelets and Depresses Platelet Release from Megakaryocytes: Studies in Rats

In previous studies we have shown that relaxin (RLX), a peptide hormone produced predominantly during pregnancy, is a powerful inhibitor of platelet aggregation. The current study shows that RLX, given systemically to rats, also affects the number of circulating platelets and their release by spleen megakaryocytes. Male albino rats were treated acutely or chronically with RLX, and the effects of the peptide evaluated by platelet count and morphological analysis of spleen megakaryocytes. The results obtained show that RLX causes a decrease of circulating platelets, which is already appreciable 1 h after a single administration and becomes significant upon a May treatment with the peptide (790000 ± 18000/ml in the RLX-treated rats versus 865 000 ± 23 000/ml in the controls; P<0.01). Coincidentally, spleen megakaryocytes show clearcut changes consistent with a depression of platelet release, namely the appearance of a thick peripheral cytoplasmic halo, filled with actin microfilaments and without platelet territories, and a virtual absence of images of thrombocytopoiesis. Based on the properties of RLX in inhibiting hemostasis, recognized in previous investigations and further proved by the present study, a role of the peptide is proposed in counteracting the hypercoagulable state which currently takes place during normal pregnancy.

Temperature sensitivity on proliferation and morphologic alteration of human esophageal carcinoma cells in culture

As basic studies of hyperthermia and hypothermia on malignant tumor, the kinetics of proliferative activity, the morphologic changes in the two cell lines, SGF-3 and SGF-5, established in our department after the change of culture temperature were examined. The results obtained were: a) A significant difference was found in the sensitivity to temperatures between the two cell lines originated from human esophageal squamous cell carcinoma. The temperature range allowing cultured cell to proliferate were from 31 degrees to 39 degrees C in SGF-3 and from 29 degrees to 41 degrees C in SGF-5. b) Minor difference occurred in the results between the two cell lines examined during the recovery of proliferative activity, but no proliferative activity was discovered after the cells were exposed to 42 degrees C for 72 h. Two cell lines resumed their proliferation after having been exposed to 27 degrees or 28 degrees C for 72 h. c) Morphologic changes of the cell lines cultured at high temperature were cytoplasmic vacuolation and cell aggregation by phase contrast microscope and the increase of heterochromatin, the decrease of granular formation in nucleoli, and nucleolar vacuolation by transmission electron microscopy (TEM). At low temperatures the changes observed included cytoplasmic ballooning and circumnuclear halo formation by phase contrast microscope, and the increase of heterochromatin, nucleolar segregation, swelling of mitochondria, and dilatation of rough endoplasmic reticulum (rER) by TEM.