Comparison of Chen Medium and Optisol-GS for human corneal preservation at 4 degrees C: results of transplantation.

PURPOSE: To compare results after transplantation of donor corneas stored in Chen Medium (containing beta-hydroxybutyrate without sodium bicarbonate or chondroitin sulfate) to corneas stored in Optisol-GS medium (containing sodium bicarbonate and 2.5% chondroitin sulfate). METHODS: We performed 32 consecutive penetrating keratoplasties with donor corneas stored at 4 degrees C in either Chen Medium or Optisol-GS by random assignment. Corneal thickness measurements were made at 1 day, 1 week, 3 weeks, 2 months, and 1 year postkeratoplasty. Specular microscopic images of the donor endothelium were obtained at the beginning of storage and 2 months and 1 year postkeratoplasty. The percentage of intact epithelium 1 day after keratoplasty and the graft epithelialization time were estimated by the surgeons. Donor rim cultures were performed. RESULTS: No statistically significant differences in corneal thickness or endothelial cell loss between the corneas stored in the two media were found at any time, although differences of less than 12% cell loss or 0.09-mm thickness at 2 months or less than 25% cell loss or 0.10-mm thickness at 1 year could not be excluded with 90% certainty in this small series. The mean percentages of intact graft epithelium on day 1, 64% for Chen Medium and 65% for Optisol-GS, were not significantly different. Endothelial cell density 2 months postkeratoplasty was significantly decreased for corneas stored in both media. Endothelial cell loss at 2 months was directly correlated with storage time in both media. CONCLUSIONS: After keratoplasty, no statistically significant differences in corneal thickness, epithelial survival, and endothelial cell loss were found between corneas stored in Chen Medium and Optisol-GS. Endothelial cell loss at 2 months was significantly correlated with storage time in both media.

Hepatic dysfunction associated with moderate ovarian hyperstimulation syndrome. A case report.

BACKGROUND: Liver dysfunction is a rare complication of severe ovarian hyperstimulation syndrome (OHSS). Based on a MEDLINE search from 1966 to September 2000, we report the second case of liver dysfunction associated with moderate OHSS. In addition, this is the first report of moderate OHSS with serum progesterone levels during the first trimester of pregnancy higher than the upper limit of normal for a third-trimester gestation. CASE: A 33-year-old nulligravida with a history of infertility had previously undergone three failed cycles of assisted reproduction. During her fourth attempt at in vitro fertilization and intracytoplasmic sperm injection, she developed moderate OHSS 11 days after embryo transfer. She was managed on an outpatient basis. Her serum progesterone and liver enzyme levels were significantly elevated, as is unusual for the moderate picture of OHSS in this patient. CONCLUSION: Hepatic dysfunction is not limited to the severe forms of OHSS. Liver function should be analyzed even in moderate cases. Further study is needed to understand the role of elevated liver function tests and serum progesterone in the pathogenesis of OHSS.

Estrogen production and action.

Estradiol production is most commonly thought of as an endocrine product of the ovary; however, there are many tissues that have the capacity to synthesize estrogens from androgen and to use estrogen in a paracrine or intracrine fashion. In addition, other organs such as the adipose tissue can contribute significantly to the circulating pool of estrogens. There is increasing evidence that in both men and women extraglandular production of C(18) steroids from C(19) precursors is important in normal physiology as well as in pathophysiologic states. The enzyme aromatase is found in a number of human tissues and cells, including ovarian granulosa cells, the placental syncytiotrophoblast, adipose and skin fibroblasts, bone, and the brain, and it locally catalyzes the conversion of C(19) steroids to estrogens. Aromatase expression in adipose tissue and possibly the skin primarily accounts for the extraglandular (peripheral) formation of estrogen and increases as a function of body weight and advancing age. Sufficient circulating levels of the biologically active estrogen estradiol can be produced as a result of extraglandular aromatization of androstenedione to estrone that is subsequently reduced to estradiol in peripheral tissues to cause uterine bleeding and endometrial hyperplasia and cancer in obese anovulatory or postmenopausal women. Extraglandular aromatase expression in adipose tissue and skin (via increasing circulating levels of estradiol) and bone (via increasing local estrogen concentrations) is of paramount importance in slowing the rate of postmenopausal bone loss. Moreover, excessive or inappropriate aromatase expression was demonstrated in adipose fibroblasts surrounding a breast carcinoma, endometriosis-derived stromal cells, and stromal cells in endometrial cancer, giving rise to increased local estrogen concentrations in these tissues. Whether systemically delivered or locally produced, elevated estrogen levels will promote the growth of these steroid-responsive tissues. Finally, local estrogen biosynthesis by aromatase activity in the brain may be important in the regulation of various cognitive and hypothalamic functions. The regulation of aromatase expression in human cells via alternatively used promoters, which can be activated or inhibited by various hormones, increases the complexity of estrogen biosynthesis in the human body. Aromatase expression is under the control of the classically located proximal promoter II in the ovary and a far distal promoter I.1 (40 kilobases upstream of the translation initiation site) in the placenta. In skin, the promoter is I.4. In adipose tissue, 2 other promoters (I.4 and I.3) located between I.1 and II are used in addition to the ovarian-type promoter II. In addition, promoter use in adipose fibroblasts switches between promoters II/I.3 and I.4 upon treatments of these cells with PGE(2) versus glucocorticoids plus cytokines. Moreover, the presence of a carcinoma in breast adipose tissue also causes a switch of promoter use from I.4 to II/I.3. Thus there can be complex mechanisms that regulate the extraglandular production of estrogen in a tissue-specific and state-specific fashion.

Use of third generation gonadotropin-releasing hormone antagonists in in vitro fertilization-embryo transfer: a review.

Before gonadotropin-releasing hormone agonists (GnRHa) became available, approximately 20% of stimulated cycles within an in vitro fertilization (IVF) program were cancelled due to premature LH surges. By using the GnRHa to prevent LH surges via gonadotrope GnRH receptor down-regulation and desensitization, this percentage decreased to about 2%, and concomitantly, the IVF and pregnancy rates per cycle initiated were increased. Several treatment schedules currently are in use, including the so-called "long protocol," in which the GnRHa is begun in the luteal phase and down-regulation occurs before the start of the gonadotropin-stimulation treatment phase. This is generally the most effective regimen and is presently the most frequently used protocol. However, it has some disadvantages, such as hypoestrogenic side effects and an increase in the number of ampules of FSH or hMG required for adequate stimulation. There is a new generation of GnRH antagonists now clinically available, that has been able to minimize the potential side effects and provide reliable antagonism at the GnRH receptor. These agents seem better suited than GnRHa for assisted reproductive technology (ART) cycles inasmuch as they can prevent LH surges without requiring complete gonadotropin suppression. We have reviewed the current literature concerning their use in IVF cycles.

Uncovering the facts: parental behaviors and knowledge regarding sun protection.

PURPOSE: A descriptive study was conducted to examine the knowledge of and behaviors related to sun-protection among parents of youth soccer players. DATA SOURCES: A convenience sample of 56 parents at community soccer events completed an 18-item instrument designed by the researchers. CONCLUSIONS: Results indicated that female respondents were more responsive to skin protection than males. In addition, advice from health care providers was shown to make an impact on the behavior of parents related to skin self-examinations and the use of sunscreen. Family history of skin cancer significantly promoted the use of protective clothing in the sun. IMPLICATIONS FOR PRACTICE: Nurse practitioners can make a difference by educating clients about sun protection and practices that can lower the risk of skin cancer and by teaching parents how to perform skin self-examinations.

POC testing: is it right for everyone?

Point-of-care testing takes select specimen analysis out of the laboratory and to the patient's bedside. To ensure its success, nurse managers must consider the method's appropriateness, staff support, and follow-up.

What's new at the point of care?

Preview the latest in near-patient specimen analysis technology, including blood glucose and occult blood tests, and ways to measure heparin's anti-coagulation.

In vitro comparison of Chen medium and Optisol-GS medium for human corneal storage.

PURPOSE: To compare paired human corneas after storage at 4 degrees C in Chen medium (CM) and Optisol-GS medium (OM) for 7, 10, 14, and 21 days. METHODS: One cornea of each pair from nine human donors was randomly stored in either CM or OM, with its mate cornea stored in the other medium. Three pairs of corneas were stored for 7 days and two pairs each were stored for 10, 14, and 21 days at 4 degrees C. Baseline corneal thickness measurements and endothelial photographs were obtained with a specular microscope. Corneal thickness measurements were also taken on days 7, 10, 14, and 21 of storage. At the end of storage, the corneas were warmed 2 hours before endothelial photographs were taken and were then placed in fixative. A corneal endothelial analysis system was used to compare changes in endothelial size and shape after storage. After fixation, the corneal endothelium was examined by scanning electron microscopy (SEM), and TdT-dUTP terminal nick-end labeling (TUNEL) assays with 4'6-diamidino-2-phenylindole (DAPI) counterstaining were performed on tissue sections of each cornea. A laser scanning confocal microscope and an automated digital analysis system were used to detect the presence of TUNEL-positive apoptotic cells in each cell layer and to determine keratocyte densities. RESULTS: Mean corneal thickness at 0, 7, 10, 14, 21 days of storage was 0.69 +/- 0.05 mm, 0.69 +/- 0.06 mm, 0.73 +/- 0.08 mm, 0.87 +/- 0.04 mm, and 0.87 +/- 0.03 mm, respectively, for CM and 0.65 +/- 0.06 mm, 0.59 +/- 0.07 mm, 0.63 +/- 0.03 mm, 0.60 +/- 0.03 mm, and 0.69 +/- 0.02 mm, respectively, for OM (p < 0.0001). The mean decrease in endothelial cell density at the end of the 7-, 10-, and 14-day storage periods was 11 +/- 10% for the CM corneas and 5 +/- 5% for the OM corneas (p = 0.18). SEM showed an intact endothelial monolayer in all corneas. The mean percentages of TUNEL-positive cells in epithelium, stroma, and endothelium of CM-stored corneas were 4 +/- 4%, 2 +/- 3%, and 0.1 +/- 0.3%, respectively, and did not differ from the OM-stored corneal values of 4 +/- 3%, 2 +/- 4%, and 0.9 +/- 1.5%. The percentage of TUNEL-positive cells did not increase with storage time. Keratocyte density was 368 +/- 130 cells/mm2 for CM-stored corneas and 447 +/- 96 cells/mm2 for OM-stored corneas (p = 0.13). CONCLUSIONS: Corneas stored in CM were thicker during storage than those stored in OM. The two storage media did not differ with respect to endothelial cell loss during storage or to the percentage of TUNEL-positive cells or keratocyte density at the end of the storage period.

Comparison of three methods for human corneal cryopreservation that utilize dimethyl sulfoxide.

We compared endothelial cell survival in human corneas after cryopreservation by three methods that utilize dimethyl sulfoxide. Twenty-eight human cadaver corneas were cryopreserved by one of three methods, stored briefly over liquid nitrogen, thawed, cultured at 37 degrees C for 3 days, and fixed for scanning electron microscopy. Seventeen control corneas underwent identical cryoprotectant immersion and culture protocols but were not frozen. Endothelial photographs taken after 1 and 3 days of culture were analyzed. Endothelial cell losses in cryopreserved corneas by Methods 1, 2, and 3, respectively, were 36, 22, and 10% after 1 day of culture and 57, 36, and 27% after 3 days of culture. Cryopreservation by Method 3 had less cell loss than Methods 1 or 2 (P<0.02) but greater cell loss than the control corneas for Method 3 (P<0.001). No loss of cells occurred in the control corneas for Methods 1 and 3 but substantial cell loss (26%) occurred in the control corneas for Method 2. Polymegethism and pleomorphism of the endothelial cells were seen in the corneas that lost cells. The endothelial cell loss of 10% seen after 1 day of culture in human corneas cryopreserved by Method 3 is similar to the loss that occurs during organ culture storage as currently used clinically and therefore would be acceptable for clinical use. After 3 days of culture, however, the cell loss had increased significantly to 27%. This additional decrease in cell number that occurs in culture may represent latent cryodamage and must be understood and overcome in vivo before the technique can be used clinically.

Comparison of recording systems and analysis methods in specular microscopy.

PURPOSE: To compare corneal endothelial cell images from contact and automated noncontact specular microscopes and to compare endothelial image analysis by the Konan Robo Center Method and the Bio Optics Bambi Corners Method. METHODS: Twenty-six normal corneas of 13 subjects and 41 penetrating keratoplasties (PKs) of 38 patients were photographed with a Keeler-Konan contact specular microscope and a Konan Noncon Robo automated noncontact specular microscope. (i) After measuring and calibrating the magnification of each instrument, we digitized the cellular apices and analyzed the images from both instruments by using the Corners Method modified to accept x and y calibrations. (ii) Using the internal calibration marks of the Konan Noncon Robo specular microscope for calibration of magnification (as required for the Center Method), we evaluated identical cells on images from this microscope by both the Center Method and the Corners Method. (iii) We evaluated the reproducibility of both methods by repeating measurements on the same image. RESULTS: (i) When the images were properly calibrated for magnification by using an external scale, endothelial cell density (ECD) of normal corneas was 2,703 +/- 354 (mean +/- SD) cells/mm2 by contact and 2,685 +/- 357 cells/mm2 by noncontact techniques (p = 0.51). ECD of PK corneas was 1,767 +/- 773 cells/mm2 by contact and 1,807 +/- 775 cells/mm2 by noncontact techniques (p = 0.31). (ii) When images from the Konan Noncon Robo specular microscope were calibrated for magnification on the internal marks, the measured ECD from the same noncontact photographs was 6% less (p < 0.001). ECD was then 2,519 +/- 294 cells/mm2 (means +/- SD) by the Center Method and 2,523 +/- 305 cells/mm2 by the Corners Method (p = 0.55) in normal corneas and 1,715 +/- 748 cells/mm2 by the Center Method and 1,731 +/- 763 cells/mm2 by the Corners Method (p = 0.04) in PK corneas. (iii) The coefficient of variation of repeated measurements on the same normal image was 0.0025 for the Centers Method and 0.0099 for the Corners Method. CONCLUSIONS: (i) Images from the automated noncontact specular microscope may be used interchangeably with those from the contact specular microscope to measure ECD, but only if both are properly calibrated by measuring an external scale. (ii) As a method of analysis, the Center Method is equivalent to the Corners Method in normal corneas, but the proprietary internal calibration of the Center Method, which is required for its use, yields ECDs approximately 6% less than when an external scale is used for distance calibration. (iii) Cell density measurements by both the Center Method and the Corners Method were reproducible within 1%.

Automated quantification of keratocyte density by using confocal microscopy in vivo.

PURPOSE: To compare keratocyte density determined by using confocal microscopy with keratocyte density determined in the same corneas by histology. METHODS: Digital en face images of central corneas were recorded three times by using confocal microscopy in vivo in six New Zealand White rabbits. Bright objects (keratocyte nuclei) in the images were automatically identified by using a custom algorithm to estimate total and regional stromal keratocyte densities. The corneas were then excised, fixed, and sectioned in a sagittal plane for histology. Keratocyte nuclei were manually counted from digitized images of 50 histologic sections per cornea. Total and regional keratocyte densities were estimated from the histologic sections by using stereologic methods based on nuclei per unit area, mean nuclear diameter, and section thickness. Histologic cell densities were corrected for tissue shrinkage. RESULTS: By confocal microscopy, total keratocyte density was 39,000 +/- 1,200 cells/mm3 (mean +/- SE; n = 6); cell density was 47,100 +/- 1,300 cells/mm3 in the anterior stroma and decreased to 27,900 +/- 2,700 cells/mm3 in the posterior stroma (P = 0.004). Analysis of the three separate confocal images of each cornea produced repeatable total cell densities (mean coefficient of variation = 0.035). By histology, total keratocyte density was 37,800 +/- 1,100 cells/mm3, not significantly different from that estimated by confocal microscopy (P = 0.43); anterior cell density was 48,300 +/- 900 cells/mm3 and decreased to 29,400 +/- 900 cells/mm3 posteriorly (P < 0.001). CONCLUSIONS: Rabbit keratocyte density estimated by automated analysis of confocal microscopy images in vivo is repeatable and agrees with keratocyte density estimated from histologic sections.

Ten-year postoperative results of penetrating keratoplasty.

OBJECTIVE: To investigate the changes in central corneal endothelial cells and corneal thickness in transplanted corneas from 5 to 10 years after grafting. This study also aimed to investigate the development of glaucoma, graft rejection, and graft failure during the first 10 postoperative years. DESIGN/PARTICIPANTS: Longitudinal cohort study of 500 consecutive penetrating keratoplasties by 1 surgeon. Patients were asked to return for follow-up examinations at 2 months and at 1, 3, 5, and 10 years after grafting. The authors excluded eyes regrafted during the study and the fellow eyes of bilateral cases, leaving 394 grafts in 394 patients for analysis. INTERVENTION: Penetrating keratoplasty was performed. MAIN OUTCOME MEASURES: Using specular microscopy, the authors measured endothelial cell density, coefficient of variation of cell area, percentage of hexagonal cells, and corneal thickness. The authors performed clinical examinations to determine graft rejection or failure and the development of glaucoma. RESULTS: By 10 years postkeratoplasty, 80 of the 394 patients had died and 68 grafts had failed. Of the remaining 246 patients, 119 (48%) returned for their 10-year examinations. For the 72 patients who returned for all of the scheduled postoperative visits and had no rejection episodes, reoperations, or failure, endothelial cell loss from preoperative donor levels at 10 years was 67 +/- 18% (mean +/- standard deviation), endothelial cell density was 958 +/- 471 cells/mm2, coefficient of variation was 0.32 +/- 0.11, hexagonal cells were 56 +/- 12%, and corneal thickness was 0.58 +/- 0.05 mm. The 5- to 10-year changes for all these values were significant (P < or = 0.004). The mean rate of late endothelial cell loss from 5 to 10 years postkeratoplasty was 4.2% per year. Eyes that were aphakic after grafting had the lowest endothelial cell loss (57 +/- 24%) and the lowest interval cell loss from 5 to 10 years postkeratoplasty (4 +/- 19%). Eyes that were phakic had the highest endothelial cell loss (73 +/- 8%) and 5- to 10-year-interval cell loss (17 +/- 31%). Eyes with posterior chamber lenses had a greater endothelial cell loss (71 +/- 9%) than did eyes with anterior chamber lenses (51 +/- 25%, P = 0.03). The 10-year cumulative risk of glaucoma, rejection, or failure was 21%, 21%, and 22%, respectively. Late endothelial failure became the major cause for graft failure, accounting for 9 of the 11 failures after 5 postoperative years. CONCLUSIONS: From 5 to 10 years after penetrating keratoplasty, the annual rate of endothelial cell loss was seven times the normal rate. The endothelial cell loss, pleomorphism, polymegethism, and corneal thickness increased significantly during this time, indicating continued endothelial instability and dysfunction, resulting in an increasing rate of late endothelial failure.

Central corneal endothelial cell changes over a ten-year period.

PURPOSE: To obtain longitudinal data to estimate long-term morphometric changes in normal human corneal endothelia. METHODS: Ten years after an initial study, the authors rephotographed the central corneal endothelium of 52 normal subjects with the same contact specular microscope. The findings for the 10 subjects younger than 18 years of age at the initial examination were considered separately. For the remaining 42 adult subjects, the time between examinations averaged 10.6 +/- 0.2 years (range, 10.1 to 11 years). At the recent examination, these subjects' ages averaged 59.5 +/- 16.8 years (range, 30 to 84 years). Outlines of 100 cells for each cornea were digitized. RESULTS: For the 42 adult subjects, the mean endothelial cell density decreased during the 10.6-year interval from 2715 +/- 301 cells/mm2 to 2539 +/- 284 cells/mm2 (P < 0.001). The calculated exponential cell loss rate over this interval was 0.6% +/- 0.5% per year. There was no statistically significant correlation between cell loss rate and age. During the 10.6-year interval, the coefficient of variation of cell area increased from 0.26 +/- 0.05 to 0.29 +/- 0.06 (P < 0.001), and the percentage of hexagonal cells decreased from 67% +/- 8% to 64% +/- 6% (P = 0.003). For the 10 subjects 5 to 15 years of age at the initial examination, the exponential cell loss rate was 1.1% +/- 0.8% per year. CONCLUSIONS: Human central endothelial cell density decreases at an average rate of approximately 0.6% per year in normal corneas throughout adult life, with gradual increases in polymegethism and pleomorphism.

Dose-related suppression of serum luteinizing hormone in women by a potent new gonadotropin-releasing hormone antagonist (Ganirelix) administered by intranasal spray.

OBJECTIVE: To determine if the GnRH antagonist (GnRH-a) Ganirelix (Syntex Research, Palo Alto, CA), administered by intranasal (IN) spray to normal women, is absorbed into the systemic circulation and suppresses LH secretion. DESIGN: A single center, open label, nonrandomized, dose-escalation study. SETTING: Academic research environment. PATIENT(S): Normal female volunteers ages 23 to 43 years. INTERVENTION(S): Ganirelix was administered as a single dose by IN spray. The administered doses and the number of women receiving each of them were 0.1 mg (n = 1), 0.3 mg (n = 1), 1 mg (n = 2), 3 mg (n = 5), and 6 mg (n = 5). Blood samples were collected from -15 minutes to 24 hours after dosing. MAIN OUTCOME MEASURE(S): Serum concentrations of Ganirelix and LH. RESULT(S): Ganirelix was absorbed rapidly. The mean time to maximal serum levels in the 3- and 6-mg groups was 0.67 and 0.53 hour, respectively. Mean serum LH levels were suppressed by > or = 35% relative to baseline from 2 to 12 hours after dosing in both groups. The mean maximal percent decrease in serum LH was -62% (at 8 hours after dosing) and -74% (at 6 hours after dosing) in the 3- and 6-mg groups, respectively. CONCLUSION(S): Single dose IN administration of 3 or 6 mg of Ganirelix suppressed serum LH levels in women, further enhancing the potential clinical utility of this potent GnRH-a. This is the first clinical report of a GnRH-a reducing the secretion of a pituitary gonadotropin when administered by an IN delivery system. Based on the duration and extent of LH suppression observed in this study, Ganirelix, administered by twice daily IN spray, may be effective for the treatment of gonadal hormone-dependent disorders in women.

Lysosomal enzymes in corneal storage media and corneal graft outcome.

PURPOSE: The purpose of this study was to relate lysosomal enzyme activities in corneal storage media to the outcome of the transplanted corneas. METHODS: Corneal storage media from 358 transplanted corneas were frozen at -70 degrees C and kept for enzyme analysis. Corneas were stored in K-Sol (28), CSM (35), Dexsol (80), Index medium (five), Optisol (158), and Optisol GS (52). Activities of alpha-D-mannosidase, beta-glucuronidase, alpha-glucosidase, and N-acetyl-beta-glucosaminidase were assayed fluorometrically. Mayo Clinic records were examined for donor information, including cause of death and 2-month graft follow-up data. RESULTS: For all corneas, there was a low but significant correlation between activities of each enzyme and storage time (rs = 0.13-0.35; p = 0.02-0.0001), and donor age (rs = -0.14 to -0.23; p = 0.009-0.0001). There was no significant correlation of enzyme activity with 2-month endothelial cell density, structure, cell loss, or corneal thickness. Enzyme activities for four primary donor failures and six grafts with > 65% 2-month endothelial cell loss were not significantly different from those for the rest of the transplanted corneas. Enzyme activities were higher for corneas from donors with renal failure but not from those with diabetes mellitus. There was no significant difference in graft outcome for different cause-of-death groups. CONCLUSIONS: The activities of lysosomal enzymes released into corneal storage media are not useful as predictors of graft outcome.

The contralateral corneal endothelium in the iridocorneal endothelial syndrome.

OBJECTIVE: To evaluate the corneal endothelial morphometric measures of the contralateral, clinically uninvolved eye of patients with the iridocorneal endothelial (ICE) syndrome. DESIGN: A retrospective review of the specular microscopic photographs of the contralateral corneal endothelium of all patients with ICE syndrome seen at Mayo Clinic, Rochester, Minn. SETTING: Ophthalmology department, Mayo Clinic. PARTICIPANTS: Twenty-eight patients with unilateral ICE syndrome who had bilateral endothelial photographs (ICE group) and 28 normal, age-matched control subjects (control group). MAIN OUTCOME MEASURES: Percentage of hexagonal cells, coefficient of variation of cell area, and endothelial cell density. METHODS: For each patient and control, 100 endothelial cells were digitized from projected endothelial photomicrographs of the central corneas in the uninvolved eyes. RESULTS: A statistically significant decrease was noted in the mean percentage of hexagonal cells (ICE, 62%; control, 69%; P = .002), and an increase was noted in the mean coefficient of variation of cell area (ICE, 0.28; control, 0.25; P = .02) in the patients with ICE syndrome compared with normal, age-matched controls. The mean endothelial cell density did not differ significantly between the 2 groups (ICE, 2588; control, 2759; P = .10). CONCLUSION: Our data suggest that the clinically uninvolved, contralateral eyes in patients with ICE syndrome have subclinical endothelial abnormalities as evidenced by a relatively low percentage of hexagonal cells and a relatively high coefficient of variation of cell area.

Specular microscopy before and after enucleation of live donor eyes.

PURPOSE: To compare the results of specular microscopic examination of corneal endothelium before and after enucleation of eyes from live donors. METHODS: Endothelial cell density (ECD), coefficient of variation of cell area (CV), and percent hexagonal cells were compared for 34 cornea donors before enucleation of their eyes and after excision of the corneoscleral rims and placement in preservative media. RESULTS: There was no statistically significant difference in ECD, CV, or percent six-sided cells after enucleation. The pre-enucleation and post-enucleation ECD measurements were significantly correlated (rs = .85, P < .0001). Mean percentage change in ECD was -0.7% +/- 6.0%. CONCLUSIONS: There was no significant difference in ECD, CV, or percent six-sided cells between measurements taken from the epithelial side in vivo and those taken of the same corneas from the endothelial side in vitro after enucleation and corneoscleral rim excision. These findings suggest that it is reasonable to compare postkeratoplasty clinical measurements with those of the donor corneas taken in the eye bank.

Localization and regulation of corticotropin receptor expression in the midgestation human fetal adrenal cortex: implications for in utero homeostasis.

Developmental changes in the responsiveness of the fetal adrenals to corticotropin (ACTH) play an important role in the regulation of the fetal hypothalamic-pituitary-adrenal axis. Responsiveness of adrenal cortical cells to ACTH is dependent on the extent of ACTH receptor expression. Therefore, we examined the localization and regulation of ACTH receptor expression in the midgestation (16-24 weeks) human fetal adrenal cortex. In situ hybridization analysis was used to localize messenger RNA (mRNA) encoding the ACTH receptor in sections of human fetal adrenal glands. Messenger RNA encoding the ACTH receptor was localized in cells from all cortical zones; abundance was higher in definitive zone than in fetal zone cells and was least abundant in the more central portions of the cortex. Regulation of ACTH receptor expression was studied using Northern blot analysis of total RNA extracted from primary cultures of fetal and definitive zone cells. Two major (1.5 and 3.5 kilobases) and, upon stimulation with ACTH, 3 minor (4.0, 6.0 and 10.0 kb) ACTH receptor mRNA transcripts were detected in RNA from fetal and definitive zone cells. In both cell types, ACTH-(1-24) increased the abundance of mRNA encoding the ACTH receptor 10- to 20-fold compared with untreated cells. The effects of ACTH-(1-24) on ACTH receptor expression in fetal zone cells were time- and dose-dependent. The ED50 for the stimulation of ACTH receptor expression by ACTH-(1-24) was 1-10 pM, and maximal response to 0.1 nm ACTH-(1-24) was detected after 12-16 h. Eight-bromoadenosine cAMP and forskolin also stimulated ACTH receptor expression in fetal zone cells and closely mimicked the effects of ACTH-(1-24). In contrast, stimulation of protein kinase C with 12-O-tetradecanoyl phorbol 13-acetate had no effect on ACTH receptor expression. Changes in ACTH receptor expression in response to ACTH-(1-24), cAMP and forskolin were paralleled by changes in expression of the P450 cholesterol side chain cleavage (P450scc) enzyme. These data demonstrate that expression of the ACTH receptor by the human fetal adrenal cortex is up-regulated by its own ligand and that this effect is mediated by a cAMP-dependent mechanism. In addition, the coordinate stimulation of ACTH receptor and P450scc expression by ACTH indicates that the gene for the ACTH receptor is one of a specific cohort of genes regulated by ACTH that are required to facilitate fetal adrenal cortical response to ACTH. ACTH regulation of its own receptor may represent a mechanism by which fetal adrenal responsiveness to ACTH is maintained and possibly enhanced during fetal development.