Effect of waterborne exposure to 4-tert-octylphenol and 17beta-estradiol on smoltification and downstream migration in Atlantic salmon, Salmo salar

Groups of Atlantic salmon parr (November, Exp. 1) or pre-smolts (March, Exp. 2) were exposed to estradiol-17beta (E2 conc.: nominal 500 ngl(-1)/actual 8-16 ngl(-1)) and two doses of tert-octylphenol (OP: nominal 25 microgl(-1)/actual 4.5-6.5 microgl(-1) and OP: nominal 100 microgl(-1)/actual 10-30 microgl(-1)) for 26 days in fresh water, and the effects on physiological and behavioural aspects of parr-smolt transformation were investigated. Vitellogenesis was induced by all treatments, as indicated by elevated levels of plasma vitellogenin (Vtg) and hepatosomatic index. Elevated Vtg levels were still found in OP-100 and E2-treated fish 4-5 months after cessation of treatment, indicating a slow clearance of Vtg from circulation. Smolting was compromised by E2 and OP-100 treatment as judged by reduced gill Na(+), K(+)-ATPase activity and impaired ability to regulate plasma osmolality and muscle water content in 24-h sea water (SW) challenge tests during the period of smolting. Downstream migratory behaviour was monitored from late April to July (Exp. 2) by implanting passive integrated transponder tags into subgroups of treated and control smolts and placing them in a stream raceway. Irrespective of treatment, nocturnal downstream movement was initiated in all groups on April 23, switching to diurnal movement in late May. Average swimming speed was estimated to be higher than current speed, indicating active migration. E2 and OP-100 fish migrated at lower frequency than control fish, suggesting a reduced migratory drive. The data suggests that waterborne exposure of salmon to xenoestrogens reduce both physiological and behavioural components of smoltification, even when exposure occurs several months prior to smolting.     

Expression of Fas ligand (CD95L) in primary malignant melanoma and melanoma metastases is associated with overall survival

BACKGROUND: There is increasing evidence that the Fas/Fas ligand (FasL) system is involved in tumor-mediated immune suppression. The purpose of this study was to investigate the effect of Fas (CD95) as well as FasL (CD95L) expression in primary malignant melanoma and melanoma metastases on overall survival (OS). PATIENTS AND METHODS: 19 patients with metastatic malignant melanoma who were treated with different dacarbazine (DTIC)-based chemotherapy regimens were included in this study. From each patient, primary melanoma biopsies and biopsies from metastases were histologically evaluated. Immunohistology was performed with antibodies to Fas/CD95 and FasL/CD95L. Differences in OS were plotted using the Kaplan-Meier method and compared by the log rank test. RESULTS: Fas/CD95 and FasL/CD95L expression was detected in 73.7 and 63.2% of primary melanomas, respectively. In metastases, expression of both Fas/CD95 (63.2%) and FasL/CD95L (47.4%) was markedly decreased. Presence of FasL/ CD95L expression in primary melanoma resulted in significantly (p = 0.024) prolonged OS compared with FasL/CD95L-negative high-risk primary melanomas. In contrast, loss of FasL/CD95L expression in melanoma metastases resected before chemotherapy was associated with significantly prolonged median survival (p = 0.0139). CONCLUSION: Presence of FasL/CD95L expression in primary malignant melanoma and the loss of FasL/ CD95L expression in metastases seem to be positive prognostic factors.     

Wastewater treatment polymers identified as the toxic component of a diamond mine effluent

The Ekati Diamond Mine, located approximately 300 km northeast of Yellowknife in Canada's Northwest Territories, uses mechanical crushing and washing processes to extract diamonds from kimberlite ore. The processing plant's effluent contains kimberlite ore particles (< or =0.5 mm), wastewater, and two wastewater treatment polymers, a cationic polydiallydimethylammonium chloride (DADMAC) polymer and an anionic sodium acrylate polyacrylamide (PAM) polymer. A series of acute (48-h) and chronic (7-d) toxicity tests determined the processed kimberlite effluent (PKE) was chronically, but not acutely, toxic to Ceriodaphnia dubia. Reproduction of C. dubia was inhibited significantly at concentrations as low as 12.5% PKE. Toxicity identification evaluations (TIE) were initiated to identify the toxic component of PKE. Ethylenediaminetetraacetic acid (EDTA), sodium thiosulfate, aeration, and solid phase extraction with C-18 manipulations failed to reduce PKE toxicity. Toxicity was reduced significantly by pH adjustments to pH 3 or 11 followed by filtration. Toxicity testing with C. dubia determined that the cationic DADMAC polymer had a 48-h median lethal concentration (LC50) of 0.32 mg/L and 7-d median effective concentration (EC50) of 0.014 mg/L. The anionic PAM polymer had a 48-h LC50 of 218 mg/L. A weight-of-evidence approach, using the data obtained from the TIE, the polymer toxicity experiments, the estimated concentration of the cationic polymer in the kimberlite effluent, and the behavior of kimberlite minerals in pH-adjusted solutions provided sufficient evidence to identify the cationic DADMAC polymer as the toxic component of the diamond mine PKE.     

County level responses to the introduction of DRG rates for "extended choice" hospital patients in Denmark

Choice of hospitals is being discussed in a number of European health systems. The Danish case provides interesting lessons because patients' free choice has been in effect since 1993. This paper explores the responses at the supply side after the introduction of DRG rates for extended choice patients in the Danish hospital system in 2000. The main question is whether the introduction of DRG rates and the resulting changes in incentives have affected county management of health care. How has the county-based governance system, which traditionally has emphasised budget control, co-operation and equity, reacted to the introduction of DRG rates and stronger incentives for "extended choice patients"?     

Plasticity of the acoustic startle reflex in currently abstinent ecstasy (MDMA) users

Rationale 3,4-Methylenedioxymethamphetamine (MDMA, ecstasy) is neurotoxic upon central serotonin systems in experimental animals and probably also in humans. Serotonin is involved in the habituation, sensitization and prepulse inhibition (PPI) of the startle reflex.Objectives To study the plasticity of startle reflex in currently abstinent MDMA users.Methods Electromyographic responses to acoustic startle stimuli (pulse alone and prepulse-pulse trials) were recorded in 23 currently abstinent ecstasy users and 20 matched control subjects. Depending on the extent of their previous drug use ecstasy users were divided into two groups [life-time dose <90 (n=11) and 90 pills (n=12), respectively].Results There were no significant differences in habituation, sensitization or PPI of the startle reflex between the entire group of ecstasy users and controls. However, sensitization of the startle reflex was stronger in the 90 compared with either the <90 MDMA pills or the control group. Correlations between patterns of drug use and startle parameters did not reach the level of significance, although users with a younger age at the onset of MDMA (and other drug) use tended to present with higher sensitization of the startle reflex.Conclusions Heavy users of MDMA (and other recreational drugs) present with strong sensitization of the startle reflex. Nevertheless, it is unclear whether this finding is secondary to the use of MDMA and its well-recognized neurotoxic potential. Alternatively, strong sensitization might reflect a pre-existing trait predisposing to drug use. A clearer picture of the impact of ecstasy on startle plasticity may be obtained from longitudinal investigations.     

Prevalence of familial pancreatic cancer in Germany

Based on several case-control studies, it has been estimated that familial aggregation and genetic susceptibility play a role in up to 10% of patients with pancreatic cancer, although conclusive epidemiologic data are still lacking. Therefore, we evaluated the prevalence of familial pancreatic cancer and differences to its sporadic form in a prospective multicenter trial. A total of 479 consecutive patients with newly diagnosed, histologically confirmed adenocarcinoma of the pancreas were prospectively evaluated regarding medical and family history, treatment and pathology of the tumour. A family history for pancreatic cancer was confirmed whenever possible by reviewing the tumour specimens and medical reports. Statistical analysis was performed by calculating odds ratios, regression analysis with a logit-model and the Kaplan-Meier method. Twenty-three of 479 (prevalence 4.8%, 95% CI 3.1-7.1) patients reported at least 1 first-degree relative with pancreatic cancer. The familial aggregation could be confirmed by histology in 5 of 23 patients (1.1%, 95% CI 0.3-2.4), by medical records in 9 of 23 patients (1.9%, 95% CI 0.9-3.5) and by standardized interviews of first-degree relatives in 17 of 23 patients (3.5%, 95% CI 2.1-5.6), respectively. There were no statistical significant differences between familial and sporadic pancreatic cancer cases regarding sex ratio, age of onset, presence of diabetes mellitus and pancreatitis, tumour histology and stage, prognosis after palliative or curative treatment as well as associated tumours in index patients and families, respectively. The prevalence of familial pancreatic cancer in Germany is at most 3.5% (range 1.1-3.5%) depending on the mode of confirmation of the pancreatic carcinoma in relatives. This prevalence is lower than so far postulated in the literature. There were no significant clinical differences between the familial and sporadic form of pancreatic cancer.     

Chromosomal alterations during lymphatic and liver metastasis formation of colorectal cancer

Comparative genomic hybridization (CGH) was used to screen colorectal carcinomas for chromosomal aberrations that are associated with metastatic phenotype. In total, 63 tumor specimens from 40 patients were investigated, comprising 30 primary tumors, 22 systemic metastases (12 liver, 6 brain, and 4 abdominal wall metastases) and 11 lymph node tumors. Using statistical analysis and histograms to evaluate the chromosomal imbalances, overrepresentations were detected most frequently at 20q11.2-20q13.2, 7q11.1-7q12, 13q11.2-13q14, 16p12, 19p13, 9q34, and 19q13.1-19q13.2. Deletions were prominent at 18q12-18q23, 4q27-4q28, 4p14, 5q21, 1p21-1p22, 21q21, 6q16-6q21, 3p12, 8p22-8p23, 9p21, 11q22, and 14q13-14q21. Hematogenous metastases showed more alterations than lymph node tumors, particularly more deletions at 1p, 3, 4, 5q, 10q, 14, and 21q21 and gains at 1q, 7p, 12qter, 13, 16, and 22q. Comparing liver metastases with their corresponding primary tumors, particularly deletions at 2q, 5q, 8p, 9p, 10q, and 21q21 and gains at 1q, 11, 12qter, 17q12-q21, 19, and 22q were more often observed. The analysis suggested that the different pathways of tumor dissemination are reflected by a nonrandom accumulation of chromosomal alterations with specific changes being responsible for the different characteristics of the metastatic phenotype.     

Solid Lipid Nanoparticles (SLN) and Oil-Loaded SLN Studied by Spectrofluorometry and Raman Spectroscopy

Purpose Recently, colloidal dispersions made of mixtures from solid and liquid lipids have been described to overcome the poor drug loading capacity of solid lipid nanoparticles (SLN). It has been proposed that these nanostructured lipid carriers (NLC) are composed of oily droplets, which are embedded in a solid lipid matrix. High loading capacities and controlled release characteristics have been claimed. It is the objective of the present paper to investigate these new NLC particles in more detail to obtain insights into their structure. Methods Colloidal lipid dispersions were produced by high-pressure homogenization. Particle sizes were estimated by laser diffraction and photon correlation spectroscopy. The hydrophobic fluorescent marker nile red (NR) was used as model drug, and by fluorometric spectroscopy, the molecular environment (polarity) was elucidated because of solvatochromism of NR. The packaging of the lipid nanoparticles was investigated by Raman spectroscopy and by densimetry. The light propagation in lipid nanodispersions was examined by refractometry to obtain further insights into the nanostructural compositions of the carriers. Results Fluorometric spectroscopy clearly demonstrates that NLC nanoparticles offer two nanocompartments of different polarity to accommodate NR. Nevertheless, in both compartments, NR experiences less protection from the outer water phase than in a nanoemulsion. In conventional SLN, lipid crystallization leads to the expulsion of the lipophilic NR from the solid lipid. Measurements performed by densimetry and Raman spectroscopy confirm the idea of intact glyceryl behenate lattices in spite of oil loading. The lipid crystals are not disturbed in their structure as it could be suggested in case of oil incorporation. Refractometric data reveal the idea of light protection because of incorporation of sensitive drug molecules in NLC. Conclusion Neither SLN nor NLC lipid nanoparticles did show any advantage with respect to incorporation rate compared to conventional nanoemulsions. The experimental data let us conclude that NLC lipid nanoparticles are not spherical solid lipid particles with embedded liquid droplets, but they are rather solid platelets with oil present between the solid platelet and the surfactant layer.     

Tear film function and corneal sensation in the early postoperative period after LASEK for the correction of myopia

Purpose Subepithelial nerves and stromal nerves of the cornea are damaged during LASEK surgery for the correction of myopia. This leads to a reduction in corneal sensation and to alterations of the tear film function in the early postoperative period. The aim of this study was to evaluate tear film function, corneal sensation and subjective symptoms of dry eye in the early postoperative period after LASEK for the correction of myopia.Methods LASEK surgery was performed in 20 eyes of 10 consecutive patients for the correction of myopia (–1.0 to –9.0 D, mean –4.86 D). Subjective symptoms of a dry eye were evaluated with a standardised questionnaire. Schirmer test with and without local anaesthesia, tear film break-up time, fluorescein staining of the cornea and corneal esthesiometry (Cochet–Bonnet) were performed before surgery and 1 week, 1 month, 2 months and 3 months after LASEK. Additionally corneal staining and subjective symptom severity scores were assessed 3 days after surgery.Results Corneal sensation was reduced 1 week after LASEK and recovered during the first month after LASEK. The tear film break-up time was reduced 1 week and 1 month after LASEK and reached preoperative values 2 months after surgery. Fluorescein staining was increased 3 days and 1 week after LASEK. Subjective symptom severity scores were increased 3 days, 1 week, 1 month and 2 months after LASEK. Schirmer tests values with local anaesthesia were significantly reduced at 3 months after surgery, but not at 3 days, 1 month, or 2 months. Schirmer test values without local anaesthesia were significantly decreased 2 and 3 months after LASEK, but not after 3 days and 1 month after LASEK.Conclusions LASEK alters ocular surface haemostasis and reduces corneal sensation in the early postoperative period. Subjective symptoms of dry eye were described during the first 2 months after surgery.     

Spectrum and frequencies of mutations in MSH2 and MLH1 identified in 1,721 German families suspected of hereditary nonpolyposis colorectal cancer

Mutations in DNA MMR genes, mainly MSH2 and MLH1, account for the majority of HNPCC, an autosomal dominant predisposition to colorectal cancer and other malignancies. The evaluation of many questions regarding HNPCC requires clinically and genetically well-characterized HNPCC patient cohorts of reasonable size. One main focus of this multicenter study is the evaluation of the mutation spectrum and mutation frequencies in a large HNPCC cohort in Germany; 1,721 unrelated patients, mainly of German descent, who met the Bethesda criteria were included in the study. In tumor samples of 1,377 patients, microsatellite analysis was successfully performed and the results were applied to select patients eligible for mutation analysis. In the patients meeting the strict Amsterdam criteria (AC) for HNPCC, 72% of the tumors exhibited high microsatellite instability (MSI-H) while only 37% of the tumors from patients fulfilling the less stringent criteria showed MSI-H; 454 index patients (406 MSI-H and 48 meeting the AC of whom no tumor samples were available) were screened for small mutations. In 134 index patients, a pathogenic MSH2 mutation, and in 118 patients, a pathogenic MLH1 mutation was identified (overall detection rate for pathogenic mutations 56%). One hundred sixty distinct mutations were detected, of which 86 are novel mutations. Noteworthy is that 2 mutations were over-represented in our patient series: MSH2,c.942+3A>T and MLH1,c.1489_1490insC, which account for 11% and 18% of the MSH2 and MLH1 mutations, respectively. A subset of 238 patients was screened for large genomic deletions. In 24 (10%) patients, a deletion was found. In 72 patients, only unspecified variants were found. Our findings demonstrate that preselection by microsatellite analysis substantially raises mutation detection rates in patients not meeting the AC. As a mutation detection strategy for German HNPCC patients, we recommend to start with screening for large genomic deletions and to continue by screening for common mutations in exon 5 of MSH2 and exon 13 of MLH1 before searching for small mutations in the remaining exons.