The effect of collection temperature, cooling rate and warming rate on chilling injury and cryopreservation of mouse spermatozoa.

The experiments presented here identify several factors that affect survival (motility) of cryopreserved mouse spermatozoa after freezing and thawing. Among these factors are: (i) the temperature at which spermatozoa are collected, (ii) the cooling rate to 0 degrees C and (iii) the warming rate from -196 degrees C to ambient. When excised epididymides were cooled to near 0 degrees (1-4 degrees C) and spermatozoa collected and mixed with cryoprotectant at that temperature, motilities after subsequent freezing and thawing were 8-10 times higher than when the spermatozoa were collected from the epididymides at 22 degrees C.

Fracture phenomena in an isotonic salt solution during freezing and their elimination using glycerol.

Thermal stress and consequent fracture in frozen organs or cell suspensions have been proposed to be two causes of cell cryoinjury. A specific device was developed to study the thermal stress and the fracture phenomena during a slow cooling process of isotonic NaCl solutions with different concentrations of glycerol (cryoprotectant) in a cylindrical tube. It was shown from the experimental results that glycerol significantly influenced the solidification process of the ternary solutions and reduced the thermal stress.

Growth of mouse pups derived from biopsied blastocysts.

Objective: To test the hypothesis that removal of cells from the pre-embryo affects intrauterine growth, as reflected in birth weight or growth to weaning.

Methods: ICR Swiss albino and C57Bl/6 mice served as embryo donors to provide two strains of distinct coat colors. Alternate strains were assigned to control treatment or blastocyst biopsy of trophectoderm.

Intravenous cholangiography and the management of choledocholithiasis prior to laparoscopic cholecystectomy.

The preoperative diagnosis of choledocholithiasis simplifies the laparoscopic management of biliary tract disease. Slow infusion intravenous cholangiography (SI-IVC) may be an accurate and cost-effective screening test for choledocholithiasis, and it is safer than traditional intravenous cholangiography. Forty-nine patients underwent SI-IVCs for suspected choledocholithiasis.

Molluscum contagiosum presenting as cellulitis in an AIDS patient: cytologic and ultrastructural features.

This report describes the cytologic features of molluscum contagiosum presenting clinically as a left thigh abscess in a 29-yr-old male with AIDS. The fine-needle aspiration findings are correlated with the histologic and ultrastructural findings. Molluscum contagiosum's clinical presentation, natural course, and induction of host immune responses in immunocompetent and immunosuppressed individuals as identified by light microscopy, immunocytochemically and ultrastructurally are reviewed.

Prevention of osmotic injury to human spermatozoa during addition and removal of glycerol.

Use of a cryoprotective agent is indispensable to prevent injury to human spermatozoa during the cryopreservation process. However, addition of cryoprotective agents to spermatozoa before cooling and their removal after warming may create severe osmotic stress for the cells, resulting in injury. The objective of this study was to test the hypothesis that the degree (or magnitude) of human sperm volume excursion can be used as an independent indicator to evaluate and predict possible osmotic injury to spermatozoa during the addition and removal of cryoprotective agents.

Visual disturbance secondary to clomiphene citrate.

Objective: To identify a distinctive constellation of persistent visual abnormalities secondary to treatment with clomiphene citrate.

Design: Description of the clinical findings in three patients with visual disturbance secondary to clomiphene treatment.

Setting: A neuro-ophthalmology referral center.

High water permeability of human spermatozoa is mercury-resistant and not mediated by CHIP28.

A novel integral membrane protein with an apparent molecular mass of 28 kDa (CHIP28) was first isolated from human erythrocytes and is now recognized as a water channel protein. The expression of this protein has been found in several other cell types that all require high water permeability for their functions. Recent studies have shown that the water permeability (Lp) of human spermatozoa is among the highest reported for mammalian cells.

Adie's tonic pupil secondary to migraine.

A 46-year-old woman observed transient unilateral mydriasis during a classic migraine attack. One week later she experienced a similar episode after which anisocoria was persistent. Subsequent examination showed the clinical and pharmacologic features of a postganglionic parasympathetic paresis (Adie's tonic pupil).

Fulminant viral hepatitis.

Fulminant viral hepatitis is the most dramatic and life- threatening complication of acute viral hepatitis. Although our understanding of the diverse agents capable of producing this syndrome has increased, effective, specific therapies are lacking. Liver transplantation as a therapeutic modality for this condition has been widespread application and underscored the need to improve our ability to predict outcome.

Selective T cell depletion with CD8-conjugated magnetic beads in the prevention of graft-versus-host disease after allogeneic bone marrow transplantation.

The effects of a new immunomagnetic method of selectively depleting CD8+ lymphocytes from donor bone marrow were studied in 29 patients undergoing transplantation from HLA-identical sibling (n = 20) or alternative (n = 9) donors. The direct immunomagnetic depletion method consistently removed > 95% of CD8+ cells and the non-specific loss of other cell subsets was only about 15%. Recovery of CFU-GM and BFU-e was on average > 100%.

Patient informatics: using a computerized system to monitor patient compliance in the treatment of hypertension.

A group of investigators at a major teaching hospital have prepared a study designed to test the hypothesis that patient outcomes related to hypertension will be improved by increased compliance, as defined by consistent daily compliance with prescribed medical regimen. The study will test the effectiveness of the following interventions in improving compliance: 1) improved access to medication, 2) computerized phone follow-up, and 3) one-on-one follow-up conversations concerning compliance issues. This paper is intended to describe the pending study in detail while focusing on the computerized phone system that will be used in the treatment group of the controlled study for compliance.

Developmental potential of microbiopsied murine blastocysts.

Objective: To evaluate the frequency of mouse pups born following blastocyst biopsy and embryo transfer compared to nonbiopsied controls.

Methods: ICR Swiss albino and C57B1/6 mice served as embryo donors. Pregnant mare serum gonadotropin treatment was followed 46-52 hours later by hCG, when donors were paired with fertile males.

New techniques for liver transplantation: reduced-size, split-liver, living-related and auxiliary liver transplantation.

Mortality among patients on a waiting list for orthotopic liver transplantation continues to be 10-15%; this is of particular concern in the pediatric population and may become more problematic in adult patients as longer waiting lists for cadaveric transplantation accrue. The longer cold ischemia times afforded by use of University of Wisconsin (UW) solution and improved hepatic surgery techniques have allowed the development of reduced-size liver transplantation (RSLT), split-liver transplantation (SLT), and living-related liver transplantation (LRLT). These new surgical techniques have been predominantly employed in children, up to 40% of whom may be candidates for one of these modified procedures.

Maternal cells in chorionic villi from placentae of normal and abnormal human pregnancies.

Problem: We asked if activated macrophages and CD4 positive T lymphocytes in placental chorionic villi with villitis were of maternal or fetal origin.

Method: We employed a double antibody immunocytochemical technique on placental sections from three normal and four abnormal pregnancies with small-for-gestational-age infants. All studied placentae were mismatched for the maternal-fetal HLA-DRw 52 antigen.

Intercellular adhesion molecule-1 (ICAM-1) and HLA-DR antigens are expressed on endovascular cytotrophoblasts in abnormal pregnancies.

Problem: We asked if the lack of normal trophoblastic invasion of spiral arteries in the basal plate of abnormal pregnancies was associated with the expression of HLA-DR antigens and intercellular adhesion molecules (ICAM-1) on endovascular cytotrophoblasts.

Method: The basal plates of placentae from 15 normal and 55 abnormal pregnancies, including preeclampsia, small-for-gestational age infants, and mothers with history of secondary recurrent spontaneous abortion, were studied immunocytochemically by using monoclonal antibodies to HLA-DR and ICAM-1. Spiral and uteroplacental arteries were identified by using a triple antibody technique with antibodies to cytokeratin, alpha-smooth muscle actin, and von Willebrand factor to detect cytotrophoblasts, arterial smooth muscle cells, and endothelium, respectively.

Predictive value of hCG level 14 days after embryo transfer.

Objective: It has been reported that the quantitative serum hCG level 14 days after embryo transfer (ET) correlated with pregnancy outcome as well as a likelihood of a multiple gestation pregnancy. This prospective study was designed to assess the predictive value of a 14-day post-ET hCG level with pregnancy outcome and multiple gestation pregnancies.

Methods: Patients undergoing in vitro fertilization (IVF) and ET were monitored by serum quantitative hCG levels 14 days after ET.

Hemifacial spasm. An unusual manifestation of idiopathic intracranial hypertension.

Increased intracranial pressure may produce a variety of clinical manifestations, some common and others rare. We present a patient with idiopathic intracranial hypertension whose initial symptom was hemifacial spasm. All signs and symptoms of intracranial hypertension resolved with acetazolamide.

Single injection of methotrexate for treatment of ectopic pregnancies.

Objective: Our purpose was to evaluate the efficacy of single-dose methotrexate as a treatment option for ectopic pregnancies.

Study Design: Patients were recruited from a tertiary teaching hospital setting. They had (1) a gestational sac < 3.

Impact of vancomycin therapeutic drug monitoring on patient care.

Objective: To document differences in the outcome of vancomycin therapy in patients managed through a therapeutic drug monitoring (TDM) service and patients managed empirically, without the participation of a TDM service.

Design: Prospective, cohort study.

Setting: An 1100-bed, tertiary-care, teaching hospital.

Membrane transport properties of mammalian oocytes: a micropipette perfusion technique.

A perfusion technique using micropipette methodology was developed to determine quantitatively the membrane transport properties of mammalian oocytes. This method eliminates modelling ambiguities inherent in microdiffusion, a closely related technology, and should prove to be especially valuable for study of the coupled transport of water and cryoprotectant through mammalian oocytes and embryos. The method is described and evidence given for validity of the method for the simple case of uncoupled flow of water through the mouse oocyte membrane.