Ion size effects on the osmotic pressure and electrocapillarity in a nanoslit: Symmetric and asymmetric ion sizes.

We analyze the effect of asymmetric finite ion size in nanoconfinement in the view of osmotic pressure and electrocapillarity. When the confinement width becomes comparable with the Debye length, the overlapped electric double layer is significantly deformed by the steric effects. We derive the osmotic pressure from the modified Poisson-Boltzmann equation in a nanoslit to examine the deviation from the ideal osmotic pressure and the repulsive force on the wall considering the asymmetry of ion sizes. Then the electrocapillarity due to the steric effect is investigated under constant potential condition with the flat interface assumption. Later, the deformation by the electrocapillarity is also considered in the first order approximation.

The evaluation of combined chemical and physical treatments on the reduction of resident microorganisms and Salmonella Typhimurium attached to chicken skin.

This study was conducted to evaluate the efficacy of sodium hypochlorite (NaOCl, 0-200 mg/kg), thiamine dilauryl sulfate (TDS, 1,000 mg/kg), and ultrasound (37 kHz, 380 W) on reducing Salmonella Typhimurim, mesophilic aerobic bacteria (MAB), and coliforms on chicken skin. Chemical and physical treatments were applied for 5 min either singly or jointly, and Salmonella previously inoculated on chicken skin were quantitatively assessed using brilliant green agar, and the populations of MAB and coliforms in the native flora were enumerated using plate count agar and violet red bile agar, respectively. In the evaluation of bacterial attachment/detachment, chicken skin was quantitatively assessed for loosely, intermediately, and tightly attached bacteria. The treatment effects on bacteria detachment were also visualized using field emission scanning electron microscopy. In addition, color and textural properties of the skin after treatments were evaluated using a color difference meter and texture analyzer. Antimicrobial activity of NaOCl increased as the NaOCl concentration was increased, especially for loosely attached cells. The combination of 200 mg/kg NaOCl and ultrasound (NaOCl/ultrasound) significant reduced loosely, intermediately, and tightly attached bacteria populations by 0.75 to 0.47, 0.43 to 0.41, and 0.83 to 0.54 log cfu/g for MAB, coliforms, and Salmonella Typhimurium, respectively. However, the combination of NaOCl and TDS (NaOCl/TDS) did not sufficiently reduce those cells on chicken skins, except for loosely attached MAB and coliforms. The NaOCl/ultrasound combination produced a higher reduction in numbers of inoculated and native bacteria flora than any single application, with no negative effect on skin color or texture. Generally, the loosely attached bacteria were less resistant to the chemical and physical treatments than the intermediately and tightly attached bacteria in chicken skin, presumably due to their location in deeper skin layer and crevices. Further research is needed to investigate how the intermediately and tightly attached microorganisms can be effectively eliminated from chicken skin.

Electric double layer at the interface of ionic liquid-dielectric liquid under electric field.

The structure of the electric double layer (EDL) is analyzed in order to understand the electromechanical behavior of the interface of ionic liquid-dielectric liquid. The modified Poisson-Boltzmann equation proposed by Bazant et al. is solved to see the crowding and the overscreening effects that are the characteristics of an ionic liquid (Bazant, M. Z.; Storey, B. D.; Kornyshev, A. A. Double layer in ionic liquids: Overscreening versus crowding. Phys. Rev. Lett. 2011, 106, 046102.). From the simple one-dimensional (1-D) analysis, it is found that the changes of the composition and the material properties in the EDL are negligible except under some extreme conditions such as strong electric field over O(10(8)) V/m. From the electromechanical view points, an ionic liquid behaves like a pure conductor at the interface with a dielectric liquid. Based on these findings, three specific application problems are considered. In the first, a new method is suggested for measuring the interfacial tension of an ionic liquid-dielectric liquid system. The deformation of a charged ionic liquid droplet translating between two electrodes is used for this measurement. The second is for the Taylor cone problem, which includes an extreme electric field condition near the tip. The size of the critical region, where the EDL effect should be considered, is estimated by using the 1-D analysis result. Numerical computation is also performed to see the profiles of electric potential and the electric stress along the interface of the Taylor cone. Lastly, the electrowetting problem of the ionic liquid is considered. The discrepancies in the results of previous workers are interpreted by using the results of the present work. It is shown that all the results might be consistent if the leaking of the dielectric layer and/or the adsorption of ions is considered.

Multiple displacement amplification for preimplantation genetic diagnosis of fragile X syndrome.

Preimplantation genetic diagnosis (PGD) has become an assisted reproductive technique for couples that have genetic risks. Despite the many advantages provided by PGD, there are several problems, including amplification failure, allele drop-out and amplification inefficiency. We evaluated multiple displacement amplification (MDA) for PGD of the fragile X syndrome. Whole genome amplification was performed using MDA. MDA products were subjected to fluorescent PCR of fragile X mental retardation-1 (FMR1) CGG repeats, amelogenin and two polymorphic markers. In the pre-clinical tests, the amplification rates of the FMR1 CGG repeat, DXS1215 and FRAXAC1 were 84.2, 87.5 and 75.0%, respectively, while the allele dropout rates were 31.3, 57.1 and 50.0%, respectively. In two PGD treatment cycles, 20 embryos among 30 embryos were successfully diagnosed as 10 normal embryos, four mutated embryos and six heterozygous carriers. Three healthy embryos were transferred to the uterus; however, no clinical pregnancy was achieved. Our data indicate that MDA and fluorescent PCR with four loci can be successfully applied to PGD for fragile X syndrome. Advanced methods for amplification of minuscule amounts of DNA could improve the sensitivity and reliability of PGD for complicated single gene disorders.

AMPK attenuates bupivacaine-induced neurotoxicity.

Bupivacaine has been widely used as a long-acting local anesthetic. However, evidence strongly suggests that bupivacaine causes apoptosis. AMP-activated protein kinase (AMPK) regulates metabolic homeostasis and mediates cellular protection from stress. We hypothesized that AMPK may be cytoprotective in bupivacaine-treated Schwann cells. To explore this, we applied bupivacaine to the RT4-D6P2T Schwann cell line. The expression of phosphorylated AMPK was compared after bupivacaine treatment. Bupivacaine induced cell death in a time- and dose- [50% lethal dose (LD(50)) = 316 microM] dependent manner, and increased expression of phosphorylated AMPK after bupivacaine treatment. Bupivacaine-induced cytotoxicity was attenuated by AICAR (an AMPK activator), whereas compound C (an AMPK inhibitor) enhanced it. The cytoprotective effect of AICAR was reversed in the presence of iodotubercidin, an AICAR inhibitor. Our results suggest that the AMPK pathway may protect Schwann cells from bupivacaine-induced cytotoxicity.

Steric effects of ions in the charge-related wetting phenomena.

Steric effects of ions on the charge-related wetting phenomena are studied. Along with a general treatment, three specific problems in two-dimensional system are considered: a droplet on an electrode, a droplet on a charged surface, and an electrowetting phenomenon on a dielectric. For computation of wetting tension, the electromechanical approach is adopted with the principle of mechanical force balance for each phase. The modified Poisson-Boltzmann equation, which was originally proposed by Bikerman [Philos. Mag. 33, 384 (1942)], is adopted for the analysis of the steric effects. It is found that the steric hindrance reduces significantly both the osmotic pressure and the electrical stress near the triple contact line. This reduction results in a considerable decrease in the wetting tension when the ratio of the capacitance per unit area of the electrical double layer to that of the dielectric layer is small.

Clinical and genetic analysis of Korean patients with Marfan syndrome: possible ethnic differences in clinical manifestation.

Marfan syndrome (MFS) is an autosomal dominant disorder of the fibrous connective tissue caused by mutations in the fibrillin-1 (FBN1) gene. Although clinical and genetic analyses have been performed in various populations, there have been few studies in Korea. The aim of this study was to investigate the clinical characteristics and genetic background of Korean patients with MFS. In 39 Korean patients with MFS who met the Ghent criteria, the most common clinical finding was aortic dilatation and/or dissection (94.9%), whereas only 35.9% of patients had ectopia lentis. The majority of MFS patients had fewer than four of the skeletal findings required to fulfill the major skeletal Ghent criterion for MFS. Only 21% of Korean patients had major skeletal abnormalities and most cases showed only minor skeletal involvement. FBN1 gene mutations were detected in 35 out of 39 patients (89.7%), which is similar to rates presented in the previous reports. These results suggest that some clinical features in Korean patients with MFS differed from those reported in Western MFS patients.

Deformation and motion of a charged conducting drop in a dielectric liquid under a nonuniform electric field.

As a tool for transporting a drop inside another fluid, a charged conducting drop driven by Coulombic force is considered. Specifically, deformation and motion of a charged conducting drop under nonuniform electric fields are studied using the perturbation method. For simplicity in analysis, the applied electric field is assumed to be expressed as the sum of a uniform field and a linear field and the flow is assumed to be in the Stokes flow range. The deformed drop shape due to electrical stress is computed to the first order of the electrical Weber number (W). Then the electric force and the hydrodynamic drag are computed to derive the formula of the translation velocity, which is valid up to O(W). Several important results have also been obtained for the effect of drop deformation on the electric and hydrodynamic forces exerted on the drop.

MR imaging of congenital heart diseases in adolescents and adults.

Echocardiography and catheterization angiography suffer certain limitations in the evaluation of congenital heart diseases in adults, though these are overcome by MRI, in which a wide field-of view, unlimited multiplanar imaging capability and three-dimensional contrast-enhanced MR angiography techniques are used. In adults, recently introduced fast imaging techniques provide cardiac MR images of sufficient quality and with less artifacts. Ventricular volume, ejection fraction, and vascular flow measurements, including pressure gradients and pulmonary-to-systemic flow ratio, can be calculated or obtained using fast cine MRI, phase-contrast MR flow-velocity mapping, and semiautomatic analysis software. MRI is superior to echocardiography in diagnosing partial anomalous pulmonary venous connection, unroofed coronary sinus, anomalies of the pulmonary arteries, aorta and systemic veins, complex heart diseases, and postsurgical sequelae. Biventricular function is reliably evaluated with cine MRI after repair of tetralogy of Fallot, and Senning's and Mustard's operations. MRI has an important and growing role in the morphologic and functional assessment of congenital heart diseases in adolescents and adults.

Protracted diarrhea: results of the five-year survey in a tertiary hospital in Korea.

The syndrome of protracted diarrhea (PD) includes several diseases with diverse etiologies. This study was conducted to characterize the spectrum of causes, clinical manifestations, and the outcomes of PD. A retrospective analysis of the clinical and pathological findings was performed on 25 patients with diarrhea starting within the first 2 yr of life and a requirement of parenteral nutrition (PN). According to the intestinal histopathology, patients were classified into four groups: immune enteropathy (12 cases), lymphangiectasia (6 cases), epithelial dysplasia (5 cases), and unclassified (2 cases). All patients with epithelial dysplasia had earlier onset of diarrhea and longer duration of PN than those in the other groups. Three patients (12%) had an evidence of a familial condition. Five patients (three with microvillous inclusion disease and two with immune enteropathy) died. Sixteen patients recovered, and three (two with primary lymphangiectasia and one with microvillous inclusion disease) still had diarrhea. One patient underwent intestinal transplantation for tufting enteropathy. In conclusion, infants with PD should be referred to specialized centers where advanced diagnostic and therapeutic facilities are available, because histological analysis is critical for the diagnosis of PD, and PN or intestinal transplantation is the only therapeutic option in a subset of cases.

Expression of the second isoform of gonadotrophin-releasing hormone (GnRH-II) in human endometrium throughout the menstrual cycle.

We examined the expression of the protein and mRNA of the newly cloned isoform of human gonadotrophin-releasing hormone (GnRH-II) in the normal human endometrium during the menstrual cycle. Nested RT-PCR and sequence analysis revealed that two spliced variants of GnRH-II mRNA were expressed during the entire menstrual cycle, with the shorter transcript having a 21 nucleotide deletion in the region coding for GnRH-associated peptide. Using immunohistochemistry, we identified immunoreactive GnRH-II in both stromal and glandular epithelial cells during the entire menstrual phase. The immunostaining intensity was stronger during the early and mid-secretory phase compared with the proliferative and late-secretory phase. A large amount of immunoreactive GnRH-II was localized in the apical pole of the glandular lumen. Our results show that the second isoform of GnRH (GnRH-II) is expressed in the human endometrium during the entire menstrual phase. We also suggest that an increased expression of endometrial GnRH-II peptide, noted during the early and mid-secretory phase, may play an important role in human embryo implantation.

Homologous monocuspid valve patch in right ventricular outflow tract reconstruction.

BACKGROUND: In the surgical repair of tetralogy of Fallot or pulmonary atresia, pulmonary regurgitation may be detrimental in the postoperative period. We have used homograft monocuspid valve patch to prevent pulmonary insufficiency. METHODS: From September 1996 to December 1998, twenty-five patients, 4 months to 8 years of age (median 10.1 months) had homograft monocuspid valve in the procedure of right ventricular outflow tract reconstruction. The function of the monocuspid valve was assessed by echocardiogram and graded as trivial to mild, mild to moderate, moderate, and severe. We evaluated the degree of pulmonary insufficiency before discharge, at 3-6 months, and at 12 months after the operation. RESULTS: There was one hospital death due to fulminate adeno viral pneumonia. On echocardiogram, 21 patients (88%, 21/24) had no significant pulmonary insufficiency. Only one patient (4.5%) showed a moderate degree of pulmonary insufficiency. At 3-6 months, seventeen of twenty-one (81%) patients had no significant pulmonary insufficiency. There were fourteen patients who had follow-up over 1 year, and no patients showed newly developed significant pulmonary insufficiency. CONCLUSIONS: We concluded that the homograft monocuspid valve patch for right ventricular outflow tract reconstruction has provided excellent early results for the prevention of pulmonary insufficiency. However these effects are limited in duration and further close follow-up should be needed.

Mutation screening of the FSH receptor gene in infertile men.

Follicle stimulating hormone (FSH) is important for controlling spermatogenesis through binding with its receptor. However, little information is available on mutations of the FSH and its receptor gene in infertile men. To study the genetic defects, which caused problems in spermatogenesis, we screened the point mutations of the FSH receptor gene in infertile men with high serum FSH concentrations. Seventy male infertile patients with high FHS levels (> 12 mIU/ml) were screened for mutations in each of the 10 exons of the FSH receptor gene, using genomic DNA PCR and a single-strand conformation polymorphism (SSCP) analysis. From this study, three shifted bands were detected by SSCP. The first shifted band was found in the PCR product of exon 4, including the exon-intron boundary sequence in only one patient. The sequence analysis revealed a nucleotide A to T substitution in intron 3 (IVS3-4A-->T). The second shifted band was detected in exon 10 with high frequency (33%). A nucleotide A to G substitution was found at the position of the 994th nucleotide, predicting a Thr to Ala substitution at the position of the 307th amino acid (Thr307Ala). The third shifted band in the 3' region of exon 10 was detected frequently in infertile patient and normal groups. It was tightly linked to the Thr307Ala variant. Thus, all of the abnormalities represent neutral polymorphisms, and not pathological mutations of the FSH receptor gene. In conclusion, we did not confirm that the genomic mutation of the FSH receptor is a major genetic cause in Korean infertile patients with high FSH levels.

Arrangement of the systemic and pulmonary venous components of the atrial chambers in hearts with isomeric atrial appendages.

BACKGROUND: The morphological definition of atrial chambers, and the determination of atrial laterality, are based on analysis of the structure of the atrial appendages. The systemic and pulmonary venous connections to the heart, nonetheless, are important in the management of patients having isomeric appendages. In this study, therefore, we analysed the morphology of the postero-superior walls of the atrial chambers so as to provide evidence concerning the morphogenetic background of those hearts, and to improve operative management. METHODS: We reviewed 15 autopsied specimens with isomeric right appendages, and 10 with isomeric left appendages, paying particular attention to the morphology of the systemic and pulmonary venous connections. The postero-superior walls of the atrial chambers can be made up of the atrial body, the systemic venous components, or the pulmonary venous component. We analysed the contributions made by each of these components. RESULTS: The postero-superior walls of the atrial chambers were markedly variable, but could be grouped into five patterns. Bilaterally well-developed systemic venous components and absence of the pulmonary venous component within the hypoplastic atrial body were present in 9 hearts with extracardiac pulmonary venous connections in the setting of right isomerism. Bilaterally well-developed systemic venous components, and a hypoplastic pulmonary venous component within the hypoplastic atrial body, were present in 5 hearts with intracardiac pulmonary venous connections in right isomerism. Bilaterally well-developed systemic venous components, and a hypoplastic pulmonary venous component within the sizable atrial body, were present in 1 heart with an intracardiac pulmonary venous connection in right isomerism. A well-developed pulmonary venous component within the atrial body, and hypoplasia of one systemic venous component, were present in 7 hearts with left isomerism. A well-developed pulmonary venous component within the atrial body, and hypoplasia of bilateral systemic venous components, were present in 3 hearts with left isomerism. CONCLUSIONS: The postero-superior walls of the atrial chambers in hearts with isomeric atrial appendages can be analysed on the basis of a compound structure made of bilateral systemic venous components, a central pulmonary venous component, and the body of the atrium. Hearts with isomeric right appendages have absence or hypoplasia of the pulmonary venous component, while hearts with isomeric left appendages have hypoplastic systemic venous components.

Expression pattern of germ cell-specific genes in the testis of patients with nonobstructive azoospermia: usefulness as a molecular marker to predict the presence of testicular sperm.

OBJECTIVE: To analyze the expression pattern of testis-specific genes of patients with various spermatogenic defects and their usefulness as a molecular marker to predict the presence of testicular spermatozoa in patients with nonobstructive azoospermia undergoing IVF. DESIGN: Prospective, controlled study. SETTING: Hospital-based infertility research laboratory. PATIENT(S): Fifty-eight men with azoospermia or severe oligozoospermia. INTERVENTION(S): Testicular biopsy was done in the patients with obstructive or nonobstructive azoospermia, including Sertoli cell-only syndrome, maturation arrest, severe hypospermatogenesis, and normal spermatogenesis. MAIN OUTCOME MEASURE(S): Reverse transcriptase polymerase chain reaction was performed using 1 microgram of total RNA extracted from testicular tissues. Three pairs of primers were used for amplification of male germ cell-specific genes (DAZ, transcribed in male germ cells; PGK2, in late spermatocytes and spermatids; protamine-2, in spermatids) as molecular markers. Testicular sperm was obtained by multiple testicular sperm extraction. RESULT(S): The DAZ, PGK2, and protamine-2 genes were expressed in 38, 30, and 21 of the 43 patients with nonobstructive azoospermia, respectively. Testicular spermatozoa were successfully extracted in 4 of 43 patients with nonobstructive azoospermia with the use of multiple testicular sperm extraction. Detection of protamine-2 transcripts predicted the presence or absence of spermatozoa in the testicular tissue in 39 of 43 patients (91%). CONCLUSION(S): Expression of the protamine-2 gene may be a useful molecular marker to predict the presence of testicular sperm in patients with nonobstructive azoospermia.

Antiperistaltic ileostomy using the long terminal ileal segment.

PURPOSE: This study was undertaken to determine whether reversed terminal ileal segments can be used to decrease ileostomy output in patients who have undergone total proctocolectomy and ileostomy for ulcerative colitis or familial adenomatous polyposis. METHODS: An approximately 25-cm length of terminal ileum was reversed in an antiperistaltic manner, and the new terminal ileal end was used for the ileostomy constructed in the usual manner. Six patients underwent this procedure and were compared with six patients who had conventional total proctocolectomy and ileostomy. Variables studied included weight of ileostomy output and the weight of the filtered fluid component. Data were obtained on seven different occasions during a two-month period beginning three months after the operation. Analysis was done using Student's t-test. RESULTS: There was a statistically significant decrease in the weight of the average 24-hour ileostomy effluent in those patients undergoing reversed antiperistaltic loop procedures. There was also a statistically significant decrease in the filterable liquid proportions. CONCLUSIONS: The antiperistaltic ileostomy is effective in reducing the daily amount of ileostomy effluent and facilitates stoma care, owing to its diminished liquid component.

A comparison of texture and quality of breast fillets from broilers stunned by electricity and carbon dioxide on a shackle line or killed with carbon dioxide.

To compare the broiler breast muscle quality resulting from three different slaughter methods, 36 broilers in each of two replicates were randomly divided into three groups receiving CO2 stunning, electrical stunning (ES), or CO2 killing. Carbon dioxide stunning was accomplished in a tunnel with a gradient from 40 to 60% CO2 by allowing the broilers on shackles to pass through the tunnel for 25 s. Electrical stunning was done by passing the bird's head through a charged 1% brine solution (35 mA, 7 s). For CO2 killing, the birds were killed by asphyxiation in an atmosphere of less than 2% oxygen (air displaced by CO2) for 2.5 min. Following slaughter, all breast fillets were harvested at 1.25 h postmortem and analyzed for pH, R value, shear value (SV), expressible moisture, and color (lightness and redness at 1.25 and 24 h postmortem). There were no differences (P<0.05) between treatments in pH, R value, SV, 1.25-h color values, or expressible moisture. There was an increase (P<0.05) in lightness between 1.25 and 24 h postmortem in all treatments, with the CO2 stun exhibiting the greatest increase and resulting in a significantly greater L* value at 24 h postmortem than the CO2 killing treatment. These results suggest that the postmortem metabolism or characteristics of the meat from animals processed with these stunning or killing methods does not differ to a large extent.

Concentration-dependent effects of muscimol to enhance pulsatile GnRH release from GT1-7 neurons in vitro.

Immortalized GT1-7 neurons were used to characterize the effect of muscimol, a GABAA receptor agonist, to enhance pulsatile gonadotropin-releasing hormone (GnRH) release. GT1-7 neurons were grown on Cytodex-3 beads and placed in special superfusion microchambers. The cells were superfused at a rate of 6.2 ml x h-1 with Media 199 (pH 7.35) using a commercially available perfusion system. After a pre-muscimol period of 120 min, the cells were exposed for 5 min to 0.35, 1, 5 or 10 microM muscimol or 5 microM muscimol+20 microM of the GABAA receptor antagonist, bicuculline. Following removal of the muscimol (and bicuculline, in the case of the latter experiment), the superfusion was continued for another 115 min. Sample fractions were collected at 5 min intervals throughout the perfusion. Basal GnRH release from the GT1-7 neurons was pulsatile with an average interpulse interval of 45.4+/-0.5 min and an average pulse amplitude of 191.5+/-22.6 pg x min x ml-1. Our results also demonstrated that the GABAA receptor agonist, muscimol, enhances pulsatile GnRH release from GT1-7 neurons in culture. The response to muscimol was saturable and concentration-dependent with an EC50 of 0.47 microM. The effects of 5 microM muscimol to increase GnRH pulsatility were blocked by co-exposure to the GABAA receptor antagonist, bicuculline. The average GnRH interpulse intervals were 41.7+/-1.8 min, 32.5+/-2.9 min, 30.6+/-0.7 min and 25.5+/-0.4 min in the period following exposure to 0.35, 1, 5 and 10 microM of muscimol, respectively (post-muscimol period). GnRH pulse amplitude (mean-area for each pulse) was increased during exposure to muscimol but not during the pre- or post-muscimol periods. The GABAA receptor antagonist, bicuculline, itself had no effect on pulsatile GnRH release. These results are consistent with previously published reports suggesting that activation of the GABAA receptor stimulates hypothalamic GnRH release in embryonic and neonatal animals.

Bleedout efficiency, carcass damage, and rigor mortis development following electrical stunning or carbon dioxide stunning on a shackle line.

In Experiment 1, 400 male broilers were stunned using a gradient of 40 to 60% CO2 over a period of 25 s or a 1% brine solution that was electrically charged (35 mA) for 7 s. Blood loss during bleeding was measured in 30-s intervals for a total of 120 s. After conventional processing and chilling, carcass damage was subjectively evaluated. Results indicated that the birds stunned with electricity bled faster than the CO2-stunned birds until 60 s. However, the cumulative blood loss was not different after 90 s. Carcass damage evaluation indicated that birds stunned with CO2 had a significantly lower percentage of broken clavicles, and had fewer hemorrhages on the surface of the Pectoralis. However, there was no difference between the two stunning methods in the frequency of damage at the shoulder. In Experiment 2, 256 broilers were stunned using the same conditions as in Experiment 1. Measurements of pH, R-value, sarcomere length (SL), and fragmentation index (FI) were evaluated from the left breast fillets harvested at 0, 1, 2, and 6 h postmortem. Shear values (SV) were determined using the right fillets harvested at the same four postmortem times and aged on ice until 24 h. No significant difference in breast muscle pH value was observed at 0, 2, and 6 h postmortem. However, CO2-stunned fillets had significantly higher pH values than the ES fillets at 1 h postmortem. Carbon dioxide produced greater R values than electricity at 2 and 6 h. Sarcomere length, FI, and SV were not significantly different at any time tested. These data suggest that CO2 stunning reduced carcass damage but did not reduce the need for aging before deboning when compared to the electrical stunning method used.

A second look at the embryotoxicity of hydrosalpingeal fluid: an in-vitro assessment in a murine model.

The purpose of this study was to evaluate whether hydrosalpingeal fluid (HSF) is toxic to the mouse embryo as assessed by the blastocyst development rate (BDR) and by cell counting in vitro. HSF was collected from nine patients undergoing salpingoneostomy to correct hydrosalpinx. Two-cell embryos were obtained from superovulated ICR mice. T6 medium and T6 + 0.4% bovine serum albumin (BSA) were used as control media. T6 medium containing 10% or 50% HSF and 100% HSF from each patient were used as test media. Nine to 15 embryos were cultured in microdrops prepared from each of these media. The BDR was examined after 72 h of culture in these media. To assess the total cell number within each blastocyst, the blastocysts were fixed and stained with Hoechst 33342 to facilitate cell counting. The BDR was affected adversely only by 100% HSF and not in media containing 10% or 50% HSF. The mean BDR using T6 medium and T6 + BSA were 88.7% and 85.3%, respectively. The mean BDR using media containing 10% HSF or 50% HSF were 90.0% and 89.4%, respectively. Mean BDR using 100% HSF was 75.2% (P < 0.05). The overall mean cell counts (+/- SEM) using T6 medium and T6 + BSA were 86.9+/-3.2 and 91.0+/-3.8 respectively. Mean cells counts were decreased significantly only in blastocysts cultured in 100% HSF (63.3+/-4.6; P < 0.01) but not in blastocysts cultured in 10% or 50% HSF (90.8+/-4.2 and 81.9+/-6.1 respectively). Thus, it is concluded that HSF has no embryotoxic effect but has a mildly negative effect on embryonic growth and development.