Effects of supplementation of sea buckthorn press cake on mycelium growth and polysaccharides of Inonotus obliquus in submerged cultivation.

AIMS: Investigation of the influence of cultivation time and sea buckthorn press cake (Hippophaë rhamnoides) dosage on mycelium yield of Inonotus obliquus in submerged cultivation and on the yield, monomer composition, and macromolecular properties of the exopolysaccharides (EPS) from culture media and intracellular polysaccharides (IPS) extracted from mycelia. METHODS AND RESULTS: Supplementation at 5 g l-1 combined with cultivation time of 250 h granted highest yield increase in mycelia (by 122%). The supplementation reduced extraction yield and decreased the molecular weight of the main IPS population. The supplementation increased production and molecular weight of EPS. The relative content of arabinose and rhamnose in EPS positively correlated with dosage of the press cake. The press cake supplementation increased the content of galacturonic acid in IPS, but not in EPS. CONCLUSION: Sea buckthorn press cake is a food industry fibrous side stream with high oil content. It increases the cultivation yield of Inonotus obliquus mycelium and influences the produced polysaccharides. SIGNIFICANCE AND IMPACT OF THE STUDY: Mycelium is a resource of bioactive polysaccharides, attracting the interest of nutraceutical companies. Sea buckthorn press cake is a promising supplement for increasing mycelium production. The utilization of this agricultural side stream would therefore favour circular economy.

Oiling of American white pelicans, common loons, and northern gannets in the winter following the Deepwater Horizon (MC252) oil spill.

The Natural Resource Damage Assessment and Restoration Trustees for the Deepwater Horizon oil spill assessed the external oiling of migratory bird species dependent on open water in the Gulf of Mexico following the aforementioned spill. The assessment was designed to evaluate birds that use open water during the winter within 40 km of the Gulf shoreline. We focused on the American white pelican (Pelecanus erythrorhynchos), common loon (Gavia immer), and northern gannet (Morus bassanus). Point counts (pelican, loon) or strip transects (gannet) were used and each target species was assessed for oiling (unoiled, trace, light, moderate, or heavy amounts) and photographed. Due to distance at sighting and/or poor visibility, not all visible birds were assessed. The percentage of birds oiled varied by species, with the common loon being the highest (23.6%), followed by American white pelican (16.9%), and northern gannet (6.9%). Most of the American white pelicans and common loons had trace (83% and 72%, respectively) or light levels (11% and 24%, respectively) of oiling. The northern gannet had just trace levels of oiling. Some pelicans (6%) and loons (4%) had moderate amounts of oiling. Based on expert derived-mortality estimates and our estimates of oil exposure, we used Monte Carlo simulations to predict expected decreases of 2.5%, 4%, and 11% in the observed population for the northern gannet, American white pelican, and common loon, respectively. While these values are underestimates of the true values given the long time lag (10-12 months) between the oil spill and the assessment, these data represent some of the few estimates of exposure for these species and describe minimum risk estimates to these species.

Pelagic seabird density and vulnerability in the Gulf of Mexico to oiling from the Deepwater Horizon/MC-252 spill.

Using ship-based surveys, the Natural Resource Damage Assessment (NRDA) Trustees assessed the external oiling of offshore and pelagic marine birds inhabiting the northern Gulf of Mexico (Gulf) in the year following the Deepwater Horizon oil spill (DWH spill). Study objectives were to (1) collect data on pelagic seabirds that were visibly oiled, (2) collect data to estimate abundance of seabirds in offshore and pelagic waters, and 3) document the location and condition of any bird carcasses encountered. Methods employed included strip line transects and station point counts. Surveys were conducted within a study area bound by the Texas-Mexico border and the Dry Tortugas of Florida to the south, and the nearshore coastal waters of the northern Gulf of Mexico. A total of 5665 strip line transects and 386 station point-counts of variable duration were collected during the study. More than 23,000 individual seabirds comprising 45 estuarine, coastal, offshore, and pelagic species were tallied. Average daily abundance of seabirds detected varied from a low of approximately 7 birds/day in November 2010 along regions of the mid- and outer continental shelf to a high of more than 580 birds/day in June 2011 within the near-shore, coastal waters of the northern Gulf.

Oxacillin-susceptible methicillin-resistant Staphylococcus aureus (OS-MRSA), a hidden resistant mechanism among clinically significant isolates in the Wessex region/UK.

PURPOSE: Methicillin-resistant Staphylococcus aureus (MRSA) is defined as S. aureus genetically having the mecA or mecC genes or phenotypically showing minimum inhibitory concentration (MIC) of oxacillin higher than 2 mg/L. However, recently, cefoxitin/oxacillin-susceptible mecA-positive S. aureus (OS-MRSA) has been reported worldwide. Little is known about the prevalence and virulence of these strains among clinically significant isolates in the UK. The aims were to (1) investigate the prevalence of OS-MRSA in seven major hospitals in the Wessex region/UK from a cohort of 500 clinically significant phenotypically identified MSSA isolates, (2) genetically characterise OS-MRSA strains by pulsed-field gel electrophoresis (PFGE) and compare these to common UK epidemic strains; and (3) to determine Panton-Valentine leukocidin (PVL; lukFS) gene carriage rates among these isolates. RESULTS: OS-MRSA was found in six isolates (1.2 %) of phenotypically identified and reported MSSA isolates by conventional methods. PFGE showed OS-MRSA strains to be genetically diverse and distinct from the common UK epidemic strains EMRSA-15 and EMRSA-16. None of these OS-MRSA stains carried the genes encoding PVL; however, overall positivity rate for PVL was 4.4 %, much higher than the nationally reported rates of 2 % in the UK. CONCLUSION: There are still many unknowns regarding phenotypic and/or genetic characterization of the emerging OS-MRSA isolates in the UK and worldwide. Data regarding their epidemiology and optimal therapy for infection are limited and need further investigation not only in the UK, but also worldwide, as it is likely to have an impact on the empirical treatment of S. aureus infections.

Evaluation of water quality threats to the endangered Okaloosa darter (Etheostoma okaloosae) in East Turkey Creek on Eglin Air Force Base.

The threatened Okaloosa darter (Etheostoma okaloosae) is found almost exclusively on the Eglin Air Force Base in the Choctawhatchee Bay watershed of Florida. Portions of this limited habitat are threatened with soil erosion, altered hydrology, and impaired water quality. In the present study, general water quality parameters (i.e., dissolved oxygen, specific conductance, pH, temperature, relative turbidity, and primary productivity) were characterized in East Turkey Creek, which is a body of water potentially impacted by treated wastewater sprayfields, and Long Creek, an adjacent reference stream that does not border the sprayfields. Water quality was assessed during a 30-day exposure using passive samplers for both non-polar and polar effluent parameters. Because the Okaloosa darter was listed as endangered at the time of sampling we chose a closely related species from the same creeks, the sailfin shiner (Pteronotropis hypseleotris) in which to measure metal body burdens. Additionally, fathead minnows (Pimephales promelas) were used for microarray analysis on gonad and liver tissues after 48 h exposures to water collected from the two creeks and brought into the laboratory. Waters from all sites, including reference sites, affected the expression of genes related to various biological processes including transcription and translation, cell cycle control, metabolism, and signaling pathways, suggesting that the sum of anthropogenic compounds in the site waters may cause a generalized stress response in both liver and testis, an effect that could be related to the generally low populations of the Okaloosa darter. Furthermore, effects of site waters on fish gene expression may be related to the impact of human activities other than the wastewater sprayfields, as nearby areas are closed to the public for military testing, training, and administrative activities and due to ordnance contamination.

Intra-operative frozen section analysis for suspected early-stage ovarian cancer: 11 years of Gateshead Cancer Centre experience.

OBJECTIVE: In centres in which intra-operative frozen section (FS) analysis is not performed, 'apparent' early-stage ovarian cancer diagnosed after surgery on paraffin section may require further restaging laparotomy or adjuvant chemotherapy. Previous studies on FS analysis have reported high sensitivity, specificity and overall accuracy. The objective of this article is to present the largest published dataset on the accuracy of FS analysis over an 11-year period from a single institution. DESIGN: Diagnostic test accuracy. SETTING: Northern Gynaecological Oncology Centre and Department of Cellular Pathology, Gateshead, UK. POPULATION: 1439 intra-operative FS analyses performed between January 2000 and December 2010 for suspected ovarian cancer. METHODS: Prospectively collected data on FS analysis were compared with gold standard paraffin section. MAIN OUTCOME MEASURES: Sensitivity, specificity, likelihood ratios and post-test probability. RESULTS: The overall sensitivity and specificity of FS analysis were 91.2% and 98.6%, respectively. Positive and negative likelihood ratios were 64.7% and 0.09%, respectively. The pre-test probability of an ovarian tumour being borderline or malignant was 45.8%. When FS analysis was reported to be positive, the post-test probability increased to 98% (confidence interval, 97-99%). Conversely, when FS analysis was reported to be negative, the post-test probability decreased to 7% (confidence interval, 6-9%). The majority of false test results were either borderline tumours or of mucinous differentiation. CONCLUSIONS: Intra-operative FS analysis has excellent diagnostic test accuracy and assists gynaecological oncologists to perform the appropriate surgery in 95% of cases, thereby preventing the morbidity of surgical staging in benign cases and the morbidity of restaging procedures or chemotherapy in early-stage malignant tumours.

Borderline nuclear change, high-grade dyskaryosis not excluded: current concepts and impact on clinical practice.

OBJECTIVE: Borderline nuclear change, high-grade dyskaryosis not excluded (B/HG) is a subcategory of the borderline category recommended by the British Society for Clinical Cytology as warranting direct referral to colposcopy. This subcategory is equivalent to the Bethesda category of atypical squamous cells, cannot exclude high-grade squamous intraepithelial lesion (ASC-H). The purpose of this study was to determine the validity and accuracy of using B/HG to identify potential cervical intraepithelial neoplasia (CIN) grade 2 or worse (CIN2+). METHODS: Data were collected from the hospital pathology database for borderline, B/HG and high-grade cytology (moderate dyskaryosis and above), and their respective histological and colposcopic outcomes. SPSS was used for data analysis. RESULTS: Of the 84,799 total cytology samples screened between July 2006 and December 2009, 5225 (6.1%) were reported as borderline, 309 (0.4%) as B/HG and 1222 (1.4%) as high-grade cytology. Thus, B/HG comprised 5.9% of the overall borderline category, in keeping with national guidelines (<10%). CIN2+ was confirmed in 86.6% of high-grade, 40.8% of B/HG and 3.0% of borderline cytology. Of 309 women reported with B/HG cytology, 239 had colposcopy. Colposcopic appearances showed a positive predictive value (PPV) of 71.8% for detecting CIN2+ and a negative predictive value of 60.7%. CONCLUSIONS: The B/HG category was associated with a significantly higher incidence of CIN2+ compared with borderline cytology as a whole. This refining performance justifies its existence. Colposcopic appearances had a high PPV for detecting CIN2+. Therefore, colposcopy is recommended in patients with B/HG cytology and treatment should be offered if high-grade colposcopic changes are seen.

The Breakthrough Generations Study: design of a long-term UK cohort study to investigate breast cancer aetiology.

BACKGROUND: The rationale, design, recruitment and follow-up methods are described for the Breakthrough Generations Study, a UK cohort study started in 2003, targeted at investigation of breast cancer aetiology. METHODS: Cohort members have been recruited by a participant referral method intended to assemble economically a large general population cohort from whom detailed questionnaire information and blood samples can be obtained repeatedly over decades, with high completeness of follow-up and inclusion of large numbers of related individuals. 'First-generation' recruits were women contacted directly, or who volunteered directly, to join the study. They nominated female friends and family, whom we contacted, and those who joined ('second generation') nominated others, reiterated for up to 28 generations. RESULTS: The method has successfully been used during 2003-2011 to recruit 112,049 motivated participants with a broad geographic and socioeconomic distribution, aged 16-102 years, who have completed detailed questionnaires; 92% of the participants gave blood samples at recruitment. When eligible, 2½ years after recruitment, >98% completed the first follow-up questionnaire. Thirty percent are first-degree relatives of other study members. CONCLUSION: The 'generational' recruitment method has enabled recruitment of a large cohort who appear to have the commitment to enable long-term continuing data and sample collection, to investigate the effects of changing endogenous and exogenous factors on cancer risk.

A new era in supraglottitis? An isolated UK case of supraglottitis secondary to Neisseria meningitidis.

OBJECTIVE: We report the first UK case of supraglottitis secondary to Neisseria meningitidis. METHOD: Case report with review of the current literature on supraglottitis and its aetiology. RESULTS: An 89-year-old woman was referred with worsening symptoms of dysphagia, hoarseness and neck discomfort. After nasopharyngoscopy and neck X-ray, supraglottitis was diagnosed. Prompt treatment comprised nebulised adrenaline, oxygen therapy and intravenous antibiotics. Microbiology samples grew N meningitidis, a notifiable disease in the UK. Public health officials were informed, and full precautions and prophylactic treatment initiated for those at risk. The patient made excellent progress and was discharged several days later. DISCUSSION AND CONCLUSION: Supraglottitis occurs in <4 per 100 000 population. Following a successful UK childhood immunisation programme, most cases occur in adults. Supraglottitis secondary to N meningitidis is exceptionally rare, with only seven other reported cases worldwide. Morbidity is exceptionally high; over 60 per cent of patients require airway intervention. To our knowledge, this is the first reported UK case of supraglottitis secondary to N meningitidis. This case highlights the important clinical, diagnostic and therapeutic interventions required to prevent complications associated with this potentially fatal condition.

Racial and socioeconomic disparities in parkinsonism.

OBJECTIVE: To assess potential racial and socioeconomic disparities in patients with parkinsonism treated at a tertiary Movement Disorders Center. METHODS: Patients with parkinsonism were evaluated for demographics (age, race, annual income, and educational level), medical comorbidities, medication regimen, disability (Older Americans Resources and Services subscale), presence of Parkinson disease, and disease severity (Unified Parkinson Disease Rating Scale). Disability and disease severity measures were compared by race, income, and educational level using analysis of variance for continuous variables and χ(2) tests for dichotomous variables. RESULTS: The sample included 1159 patients with parkinsonism (93.4% white, 6.1% African American, 61.2% who earned more than $50,000 annually, 62.7% who completed college, and 79.2% with a diagnosis of Parkinson disease). Cross-sectional analyses by race, income, and educational level showed greater disability and disease severity in African American compared with white patients (African American vs white Older Americans Resources and Services subscale total score, 29.8 vs 25.3, P = .005; Unified Parkinson's Disease Rating Scale total score, 53.0 vs 42.8; P < .001). African Americans were less likely to be prescribed dopaminergic medications, particularly newer agents (African Americans 20.6% vs whites: 41.1%; P = .01). Lower income and lower educational level were independently associated with greater disease severity and disability (P < .003). CONCLUSION: Racial and socioeconomic disparities exist among patients with parkinsonism being treated at a tertiary Movement Disorders Center. African Americans and those with lower socioeconomic status have greater disease severity and disability than whites. These disparities may be because of problems in diagnosis, access to care, physician referrals, and patient attitudes regarding the appropriate threshold for seeking treatment at a specialized center. Understanding and correction of these disparities may improve outcomes.

Attachment of bovine parvovirus to O-linked alpha 2,3 neuraminic acid on glycophorin A.

The bovine parvovirus (BPV) hemagglutinates human erythrocytes by binding to glycophorin A (GPA). The purpose of this study was to determine which carbohydrate on GPA binds BPV. Treatment of GPA with alpha2,3,-6,-8 neuraminidase eliminated binding of BPV to GPA. Beta-elimination of O-linked sialic acids on GPA eliminated binding, while removal of N-linked carbohydrates using the N-glycosidase PNGase F failed to eliminate binding. Treatment of GPA with a neuraminidase which specifically cleaved alpha2,3 glycosidic bonds eliminated BPV binding and, following this treatment, virus binding to GPA was restored by reconstitution of alpha2,3-linked neuraminic acids. These results indicated the O-linked alpha2,3 neuraminic acids of GPA bind BPV.

Evolutionary implications of permanent odd polyploidy in the stable sexual, pentaploid of Rosa canina L.

In Rosa canina (2n = 5x = 35), the pollen and ovular parents contribute, respectively, seven and 28 chromosomes to the zygote. At meiosis I, 14 chromosomes form seven bivalents and 21 chromosomes remain as univalents. Fluorescent in situ hybridization to mitotic and pollen mother cells (PMC) of R. canina showed that 10 chromosomes (two per genome) carry ribosomal DNA (rDNA) loci. Five chromosomes carry terminal 18S-5.8S-26S rDNA loci; three of these also carry paracentric 5S rDNA loci and were designated as marker chromosomes 1. Five chromosomes carry only 5S rDNA loci and three of these were designated as marker chromosomes 2. The remaining four of the 10 chromosomes with rDNA loci were individually identifiable by the type and relative sizes of their rDNA loci and were numbered separately. At PMC meiosis, two marker chromosomes 1 and two marker chromosomes 2 formed bivalents, whereas the others were unpaired. In a gynogenetic haploid of R. canina (n = 4x = 28), obtained after pollination with gamma-irradiated pollen, chromosomes at meiosis I in PMC remained predominantly unpaired. The data indicate only one pair of truly homologous genomes in R. canina. The 21 unpaired chromosomes probably remain as univalents through multiple generations and do not recombine. The long-term evolutionary consequence for the univalents is likely to be genetic degradation through accumulated mutational change as in the mammalian Y chromosome and chromosomes of asexual species. But there is no indication that univalents carry degenerate 5S rDNA families. This may point to a recent evolution of the R. canina meiotic system.

Assessment of toxicity reduction in wastewater effluent flowing through a treatment wetland using Pimephales promelas, Ceriodaphnia dubia, and Vibrio fischeri.

Effluent toxicity is regularly assessed with Ceriodaphnia dubia short-term chronic and Vibrio fischeri toxicity tests. Condition factor and hemotocrit of fish have recently been used to assess fish health following exposure to xenoestrogens in complex municipal effluents. To assess the ability of a treatment wetland to reduce or remove toxicity of a municipal effluent, we compared C. dubia and V. fischeri bioassays to Pimephales promelas responses in situ. Final whole effluent was diverted to a constructed wetland and effluent samples were taken daily from four sites, at incremental distances from the inflow, for a 3-week study. Overlapping 7-day C. dubia tests and V. fischeri assays were conducted with samples from each wetland site concurrent with a 3-week fish exposure. C. dubia survival and fecundity were significantly (< 0.0001) reduced at the inflow, but steadily improved with distance from the inflow. Fish condition and hematocrit were lower (alpha < 0.05) at wetland sites closer to the inflow than other wetland sites and laboratory controls. However, effluent toxicity was not detected by V. fischeri bioassays. Our findings indicate that 7-day C. dubia bioassays were most sensitive to effluent toxicity and suggest that other bioassays should be used concurrent with V. fischeri assays for municipal effluent toxicity testing.

Assessment of the estrogenicity and toxicity of a domestic wastewater effluent flowing through a constructed wetland system using biomarkers in male fathead minnows (Pimephales promelas rafinesque, 1820).

The toxicity and estrogenicity of a final treated municipal effluent was examined while flowing through a constructed wetland in north-central Texas, USA. Fish data were collected, and a baseline wetland characterization was performed to assess wetland treatment potential for these effluent properties. Vitellogenin (VTG), gonadosomatic index (GSI), hepatosomatic index (HSI), and secondary sexual characteristics were biomarkers used in fish models to assess aqueous estrogenicity. Biological indicators used to assess overall fish health included hematocrit and condition factor. Estrogenic nature of final sewage treatment works effluent was screened, concurrent with a three-week fish exposure, via gas chromatography/mass spectrometry for target estrogenic compounds, including 17beta-estradiol, ethynylestradiol, bisphenol A, nonylphenolic compounds, phthalates, and DDT. The VTG in Pimephales promelas was measured after exposure at four sites in a treatment wetland and was significantly elevated (p < 0.0001) in fish exposed at the inflow site. The GSIs were significantly less (alpha = 0.001) at the inflow site. At wetland sites closest to the inflow, secondary sexual characteristics, tubercle numbers, and fatpad thickness were less (alpha = 0.0001) than in laboratory controls. The HSIs and density of male breeding stripes were not significantly different from those of laboratory controls. However, elevated HSIs were found at the inflow site. Hematocrit and condition factors were both less (alpha = 0.001) in effluent-exposed fish at wetland sites closer to the inflow than in control fish or fish further downstream.

Use of the monoclonal antibody DAKO-ERbeta (8D5-1) to measure oestrogen receptor beta in breast cancer cells.

BACKGROUND: Oestrogen receptor beta (ERbeta) is highly homologous with the classical ER (known now as ERalpha). The exact role of ERbeta in breast cancer and its contribution in influencing patient response to endocrine therapy remains unclear. The aim of this study was to develop and evaluate a flow cytometric method for the detection of ERbeta in breast cancer cells using the DAKO monoclonal anti-ERbeta 8D5-1 antibody. METHODS: MCF7 cells were used as a positive control and U937 as a negative control for titration of the antibody. Cell lines and tumour samples were fixed with 1% paraformaldehyde and permeabilised with 0.5% saponin prior to flow cytometric analysis. RESULTS: A ten fold difference in expression of ERbeta within the different breast cell lines studied was found. Confirmation of antibody specificity against ERbeta protein by Western blot analysis detected a single band at approximately 65kDa. ERbeta immunopositive nuclei were identified in MCF7 cells by immunohistochemistry. CONCLUSIONS: DAKO ERbeta 8D5-1 antibody is specific for ERbeta protein and does not cross react with ERalpha protein. Using this antibody, ERbeta can be detected and accurately quantified in cell lines and solid breast tumours by flow cytometry.

The feasibility of a one-stop colposcopy clinic in the management of women with low grade smear abnormalities: a prospective study.

OBJECTIVES: To assess the feasibility of a one-stop colposcopy clinic for the management of women with low-grade smear abnormalities. Secondly, to determine whether the approach of immediate information of biopsy results combined with treatment if indicated helps to reduce patient anxiety and improve overall patient satisfaction with the colposcopy process. DESIGN: Prospective study following the introduction of a "one-stop" process for the management of women with low grade smear abnormalities. SUBJECTS: First 118 women managed in a "one-stop" clinic during an 8-month period. METHODS: Assessment of patient anxiety via self-completed questionnaires and comparison of anxiety scores with a control group managed via a standard clinic. RESULTS: The median waiting time for results in the one-stop clinic was 120 min (range: 100-165). All women in both groups felt anxious at the time of the clinic visit. However, after 1 week the majority of patients managed via the one-stop process felt slight anxiety only (P=0.0001) as opposed to those patients in the control group who remained anxious (P=NS). In addition, all women said they would prefer the one-stop approach for further smear abnormalities if a further colposcopic examination was warranted. CONCLUSION: A one-stop colposcopy clinic is feasible for the management of women with low-grade smear abnormalities. In addition, it delivers a quality service, optimises patient management, reduces anxiety and is the patient's choice.