Ernst Schering Foundation

To evaluate the kinetics of acrosome reaction, sperm samples from four fertile donors were prepared by swim-up and incubated with solutions of human zonae containing 0.1, 0.15, 0.3, 0.5 and 1.0 zonae microliter-1. After 20, 40 and 60 min of incubation at 37 degrees C, aliquots were taken for evaluation of the acrosomal status. The results showed a distinct time- and dose dependence of the acrosome reaction induced by solubilized zona proteins. After 60 min of incubation in 1.0 zonae microliter-1, about 80% of the spermatozoa showed signs of acrosomal loss; about 40% were completely acrosome-reacted. In addition, zona-bound sperm showed the same ratios of acrosome-reacted spermatozoa in control experiments. The velocity of acrosome reaction was calculated by means of a double-reciprocal plot being 2.0-2.5% min-1 for completely reacted spermatozoa and those showing signs of acrosome reaction. However, both subgroups differed considerably in their constants of equilibrium (K = 2.0 ZP microliter-1 and K = 0.2 ZP microliter-1, respectively). In nonreacted and partly reacted spermatozoa results might indicate a disturbed course of acrosome reaction or possibly the existence of different subpopulations in respect of sperm competition.

Acute respiratory distress syndrome/acute lung injury is a serious complication of burn patients with concomitant smoke inhalation injury. Nitric oxide has been shown to play a major role in pulmonary dysfunction from thermal damage. In this study, we have tested the hypothesis that inhibition of neuronal nitric oxide synthase could ameliorate the severity of acute lung injury using our well-established ovine model of cutaneous burn and smoke inhalation. Prospective, randomized, controlled, experimental animals study. Investigational intensive care unit at university hospital. Adult female sheep. Female sheep (n = 16) were surgically prepared for the study. Seven days after surgery, all sheep were randomly allocated into three study groups: sham (noninjured, nontreated, n = 6); control (injured, treated with saline, n = 6); and neuronal nitric oxide synthase (injured, treated with specific neuronal nitric oxide synthase inhibitor, ZK 234238 (n = 4). Control and neuronal nitric oxide synthase groups were given a cutaneous burn (40% of total body surface, third degree) and insufflated with cotton smoke (48 breaths, <40 degrees C) under halothane anesthesia. Animals in sham group received fake injury also under halothane anesthesia. After injury or fake injury procedure, all sheep were placed on ventilators and resuscitated with lactated Ringer's solution. Neuronal nitric oxide synthase group was administered with continuous infusion of ZK 234238 started 1 hr postinjury with a dose of 100 microg/kg/hr. Sham and control groups received same amount of saline. Cardiopulmonary hemodynamics monitored during the 24-hr experimental time period was stable in the sham group. Control sheep developed multiple signs of acute lung injury. This pathophysiology included decreased pulmonary gas exchange and lung compliance, increased pulmonary edema, and inflammatory indices, such as interleukin-8. Treatment of injured sheep with neuronal nitric oxide synthase inhibitor attenuated all the observed pulmonary pathophysiology. The results provide definitive evidence that inhibition of neuronal nitric oxide synthase-derived excessive nitric oxide may be a novel and beneficial treatment strategy for pulmonary pathology in burn victims with smoke inhalation injury.

PURPOSE: Gadofluorine M with a fluorescent dye (GdFMCy3) is a lipophilic paramegnetic contrast agent that is readily absorbed by cultured cells. We hypothesized that this agent would be superior to iron oxide based techniques for cell tracking post cell transplantation. METHODS: Embryonic Stem Cell derived cardiac progenitor cells (ES-CPCs) were generated using previously established methods and incubated for 12 hours with 5 mM GdFMCy3, or transfected with iron oxide using published protocols. Cell survival was &gt;95% for cells incubated with both GdFMCy3 and iron. 500,000 cells labelled with GdFMCy3, iron oxide or control (no contrast agent) were directly injected into the myocardium of mice (n=5/group). Mice were scanned over a two week interval post injection at 9.4T using gated T1-weighted sequences (GdFMCy3), T2* weighted GRE sequences (iron oxide) or the positive contrast sequence GRASP (iron oxide). Mice were sacrificed and the hearts sectioned for microscopy. Perl staining and fluorescence microscopy were used to identify iron oxide and GdFMCy3 within the myocardium, respectively. RESULTS: GdFMCy3 labelled cells were successfully identified in vivo at 9.4T (figure panel B). Good correlation between MRI and histology was observed for both cell labels (figure ). Contrast to noise ratios were significantly higher in the GdFMCy3 group relative to the iron oxide group (figure ). CONCLUSIONS: GdFM-Cy3 is readily taken up by stem cells and easily identified by both MRI and fluorescence microscopy. Given its superior contrast to noise ratio GdFM-Cy3 may be an excellent alternative to iron oxide for in vivo detection and tracking of transplanted cells.

Interferon beta-1b (IFNβ-1b; Betaferon®/Betaseron®) is an effective treatment for relapsing forms of multiple sclerosis (MS). It was the first immunomodulatory therapy to receive approval for this indication in 1993 in the USA and in 1995 in Europe. It remains the only high-dose, high-frequency treatment with proven efficacy in patients with a clinical demyelinating event, RRMS and SPMS. Several large controlled studies have been performed, including 16 years follow-up of patients, providing a unique data set to evaluate long-term safety and efficacy. Results obtained from several studies have shown an inconsistent impact of neutralizing antibodies (NAb) to IFNβ-1b, unlike NAb to other IFNβ therapies. Although patient heterogeneity and the use of different NAb assays are involved, this reflects intrinsic differences between the respective IFNβ-1b versus IFNβ-1a molecules as well as their sensitivity to NAb effects. When evaluating the occurrence and the impact of NAb during different IFNβ therapies, standardization is a necessity. Three phase III trials of IFNβ-1b, in which the same assay methodology was applied, suggest that NAb to IFNβ-1b are transient, mostly of low to moderate titre and have no effect on disability progression with only limited effect on relapse rate. Two controlled studies showed better efficacy of high dose, high frequency beta-interferon treatment even in NAb-positive patients over low dose, low frequency beta-interferon treatment in NAb-negative individuals. Given the overall clinical efficacy of IFNβ-1b, the inconsistent data regarding the effect of NAb to IFNβ-1b on treatment outcome, and the unpredictable course of disease in a given patient, treatment decisions should be based on patient response to therapy, not on NAb status. This position is endorsed by current regulatory and neurology association guidelines and is reflected in the package insert information. Multiple Sclerosis 2007; 13: S21—S27 http://msj.sagepub.com

Clinicians routinely prescribe progestins along with estrogens during menopausal hormone therapy (HT) to block estrogen-dependent endometrial proliferation. Breakthrough bleeding (BTB) can negate the utility of this treatment. Because progestin antagonists also inhibit estrogen-dependent endometrial proliferation in women and macaques, we used a menopausal macaque model to determine whether a potent progestin antagonist (ZK 230 211, Schering AG; ZK) combined with estrogen would provide a novel mode of HT. Ovariectomized rhesus macaques were treated for 5 months with either estradiol (E(2)) alone, E(2) + progesterone (two doses) or E(2) + ZK (0.01, 0.05 or 0.25 mg/kg). In the E(2) + progesterone groups, progesterone suppressed endometrial proliferation and induced a thick decidualized endometrium. In the E(2) + ZK 230 211 groups, all doses of ZK blocked endometrial proliferation and induced endometrial atrophy. In all ZK-treated groups, the atrophied endometrium contained some dilated glands lined by an inactive, flattened, non-mitotic epithelium. BTB was much lower in the E(2) + ZK groups (17 days of spotting, all groups) than in the E(2) and E(2) + progesterone groups (155 bleeding days, all groups). ZK suppressed E(2) effects in the cervix, but not in the vagina, oviduct or mammary glands. All serum chemistry and lipid profiles were normal. The ability of ZK to block estrogen-dependent endometrial proliferation, induce endometrial atrophy and suppress BTB in a menopausal macaque model indicates that progestin antagonists may provide a novel mode of HT.

We investigated the expression pattern of the breast cancer associated gene LIV-1 on mRNA and protein level in 111 human breast cancer patients by in situ hybridization as well as immunohistochemistry and focused on the unknown potential of LIV-1 expression levels as a prognostic marker. To our knowledge, this is the first study on endogenous LIV-1 protein expression. Results of our study indicate that LIV-1 mRNA and protein expression levels are only weakly correlated, suggesting posttranscriptional regulatory mechanisms. Furthermore, LIV-1 mRNA quantity in combination with a positive ER status seem to represent a better marker than the progesterone receptor status according to the prognostic significance for relapse free survival (RFS). A negative correlation of LIV-1 protein levels with tumor size, grade and stage reflects an association of LIV-1 protein expression with less aggressive tumors. High LIV-1 protein expression seems to be associated with a longer relapse free and overall survival in breast cancer patients with invasive ductal carcinoma. This association, however, seems to be dependent from other prognostic markers. Our data suggest that LIV-1 is a promising candidate for a novel marker for breast cancer patients with better outcome. Furthermore, our study presents a revised cDNA sequence of LIV-1 and demonstrates the localization of endogenous LIV-1 in the endoplasmic reticulum.