Use Of Preparations Research Articles

High cell density cultivation of a recombinant Escherichia coli strain expressing a 6-O-sulfotransferase for the production of bioengineered heparin.

One of six heparin biosynthetic enzymes, cloned and expressed in Escherichia coli as a soluble fusion protein, requires large-scale preparation for use in the chemoenzymatic synthesis of heparin, an important anticoagulant drug. The 6-O-sulfotransferase isoform-3 (6-OST-3) can be conveniently prepared at mg/L levels in the laboratory by culturing E. coli on Luria-Bertani medium in shake flasks and inducing with isopropyl β-D-1-thiogalactopyranoside at an optical density of 0·6-0·8. The production of larger amounts of 6-OST-3 required fed-batch cultivation of E. coli in a stirred tank fermenter on medium containing an inexpensive carbon source, such as glucose or glycerol. The cultivation of E. coli on various carbon sources under different feeding schedules and induction strategies was examined. Conditions were established giving yields (5-20 mg g-cell-dry weight(-1)) of active 6-OST-3 with excellent productivity (2-5 mg l(-1) h(-1)). The production of 6-OST-3 in a fed-batch fermentation on an inexpensive carbon source has been demonstrated. The ability to scale-up the production of heparin biosynthetic enzymes, such as 6-OST-3, is critical for scaling-up the chemoenzymatic synthesis of heparin. The success of this project may someday lead to a commercially viable bioengineered heparin to replace the animal-sourced anticoagulant product currently on the market.

Androgen Therapy in Women: A Reappraisal: An Endocrine Society Clinical Practice Guideline

To update practice guidelines for the therapeutic use of androgens in women. A Task Force appointed by the Endocrine Society, American Congress of Obestricians and Gynecologists (ACOG), American Society for Reproductive Medicine (ASRM), European Society of Endocrinology (ESE), and International Menopause Society (IMS) consisting of six experts, a methodologist, and a medical writer. The Task Force commissioned two systematic reviews of published data and considered several other existing meta-analyses and trials. The GRADE methodology was used; the strength of a recommendation is indicated by a number "1" (strong recommendation, we recommend) or "2" (weak recommendation, we suggest). Multiple e-mail communications and conference calls determined consensus. Committees of the Endocrine Society, ASRM, ACOG, ESE, and IMS reviewed and commented on the drafts of the guidelines. We continue to recommend against making a diagnosis of androgen deficiency syndrome in healthy women because there is a lack of a well-defined syndrome, and data correlating androgen levels with specific signs or symptoms are unavailable. We recommend against the general use of T for the following indications: infertility; sexual dysfunction other than hypoactive sexual desire disorder; cognitive, cardiovascular, metabolic, or bone health; or general well-being. We recommend against the routine use of dehydroepiandrosterone due to limited data concerning its effectiveness and safety in normal women or those with adrenal insufficiency. We recommend against the routine prescription of T or dehydroepiandrosterone for the treatment of women with low androgen levels due to hypopituitarism, adrenal insufficiency, surgical menopause, pharmacological glucocorticoid administration, or other conditions associated with low androgen levels because there are limited data supporting improvement in signs and symptoms with therapy and no long-term studies of risk. Evidence supports the short-term efficacy and safety of high physiological doses of T treatment of postmenopausal women with sexual dysfunction due to hypoactive sexual desire disorder. Importantly, endogenous T levels did not predict response to therapy. At present, physiological T preparations for use in women are not available in many countries including the United States, and long-term safety data are lacking. We recommend that any woman receiving T therapy be monitored for signs and symptoms of androgen excess. We outline areas for future research. Ongoing improvement in androgen assays will allow a redefinition of normal ranges across the lifespan; this may help to clarify the impact of varying concentrations of plasma androgens on the biology, physiology, and psychology in women and lead to indications for therapeutic interventions.

Electromagnetic Image-Guided Orbital Decompression: Technique, Principles, and Preliminary Experience with 6 Consecutive Cases

Purpose: To assess the feasibility of routine use of electromagnetic image guidance systems in orbital decompression.Methods: Six consecutive patients underwent stereotactic-guided three wall orbital decompression using the novel Fusion ENT Navigation System (Medtronic), a portable and expandable electromagnetic guidance system with multi-instrument tracking capabilities. The system consists of the Medtronic LandmarX System software-enabled computer station, signal generator, field-generating magnet, head-mounted marker coil, and surgical tracking instruments.In preparation for use of the LandmarX/Fusion protocol, all patients underwent preoperative non-contrast CT scan from the superior aspect of the frontal sinuses to the inferior aspect of the maxillary sinuses that includes the nasal tip.Results: The Fusion ENT Navigation System (Medtronic™) was used in 6 patients undergoing maximal 3-wall orbital decompression for Graves’ orbitopthy after a minimum of six months of disease inactivity. Preoperative Hertel exophthalmometry measured more than 27 mm in all patients. The navigation system proved to be no more difficult technically than the traditional orbital decompression approach.Conclusion: Electromagnetic image guidance is a stereotactic surgical navigation system that provides additional intraoperative flexibility in orbital surgery. Electromagnetic image-guidance offers the ability to perform more aggressive orbital decompressions with reduced risk.

Anti-inflammatory activities and mechanisms of Artemisia asiatica ethanol extract

Ethnopharmacological relevanceArtemisia asiatica Nakai (Compositae) is a representative herbal plant used to treat infection and inflammatory diseases. Although Artemisia asiatica is reported to have immunopharmacological activities, the mechanisms of these activities and the effectiveness of Artemisia asiatica preparations in use are not known. Materials and methodsTo evaluate the anti-inflammatory activities of Artemisia asiatica ethanol extract (Aa-EE), we assayed nitric oxide (NO), tumor necrosis factor (TNF)-α, and prostaglandin E2 (PGE2) in macrophages and measured the extent of tissue injury in a model of gastric ulcer induced in mice by treatment with HCl in EtOH. Putative enzymatic mediators of Aa-EE activities were identified by nuclear fractionation, reporter gene assay, immuoprecipitation, immunoblotting, and kinase assay. Active compound in Aa-EE was identified using HPLC. ResultsTreatment of RAW264.7 cells and peritoneal macrophages with Aa-EE suppressed the production of NO, PGE2, and TNF-α in response to lipopolysaccharide (LPS) and induced heme oxygenase-1 expression. The Aa-EE also ameliorated symptoms of gastric ulcer in HCl/EtOH-treated mice. These effects were associated with the inhibition of nuclear translocation of nuclear factor (NF)-κB and activator protein (AP)-1, implying that the anti-inflammatory action of the Aa-EE occurred through transcriptional inhibition. The upstream regulatory signals Syk and Src for translocation of NF-κB and TRAF6 for AP-1 were identified as targets of this effect. Analysis of Aa-EE by HPLC revealed the presence of luteolin, known to inhibit NO and PGE2 activity. ConclusionThe anti-inflammatory activities attributed to Artemisia asiatica Nakai in traditional medicine may be mediated by luteolin through inhibition of Src/Syk/NF-κB and TRAF6/JNK/AP-1 signaling pathways.

Determination of the optimal flax fibre preparation for use in unidirectional flax–epoxy composites

The specific stiffness of flax fibres is comparable to that of glass fibres. However, the application of flax fibres in composites still poses two important problems. The orientation of the fibres in the composite is not impeccable and the fibre–matrix adhesion is too weak. This research focuses on the former. To produce a continuous yarn for the textile industry, technical flax fibres are spun. The twisting of the fibres increases the yarn strength but the orientation of the fibres with regard to the fibre axis decreases. Experiments show that the slightly twisted yarns and the spun yarns possess the best processability but have lower mechanical properties than untwisted ribbons. The values fit very well to certain models which predict stiffness with regard to the twist angle.

On Fertility

Click to increase image sizeClick to decrease image size AcknowledgmentsI would like to thank Kelly Murphy for her research assistance and many insights into the topic.Additional informationNotes on contributorsJane HuttonJane Hutton is a landscape architect and Assistant Professor at the Harvard Graduate School of Design, where she is faculty director to the Loeb Library Materials Collection. Her work focuses on the extended relationships of material practice in landscape architecture, looking at links between the landscapes of production and consumption of common construction materials. Hutton is a founding editor of the journal Scapegoat: Architecture/Landscape/Political Economy and is coeditor of Issues: 01 Service, 02 Materialism, and 06 Mexico D.F./NAFTA.Notes1 The Committee on Civic Cleanliness and the Economic Disposition of the Refuse of Cities, appointed in 1859, was charged with plans for the disposal of “Offal, Refuse, Street-cleanings, and Nightsoil of cities.” See Egbert Viele, Report of the Committee on Civic Cleanliness and the Economic Disposition of the Refuse of Cities (New York: Edmund Jones, 1860).2 Richard A. Wines, Fertilizer in America: From Waste Recycling to Resource Exploitation (Philadelphia: Temple University Press, 1985).3 Frederick Law Olmsted and Calvert Vaux, “A Review of Recent Changes, and Changes Which Have Been Projected, in the Plans of the Central Park,” in Forty Years of Landscape Architecture: Central Park, edited by Frederick Law Olmsted Jr. and Theodora Kimball (Cambridge, MA: MIT Press, 1973), 250.4 George Waring, Earth Closets: How to Make Them and How to Use Them (New York: Tribune Association, 1869).5 Board of Commissioners of the Central Park, Seventh Annual Report of the Board of Commissioners of the Central Park 1862 (New York: Wm. C. Bryant, 1864), 87.6 Board of Commissioners of the Central Park, Fifth Annual Report of the Board of Commissioners of the Central Park 1862 (New York: Wm. C. Bryant, 1862), 120.7 Board of Commissioners of the Central Park, Seventh Annual Report (note 5).8 Joel Tarr and Clay McShane, “The Centrality of the Horse to the Nineteenth-Century American City,” in The Making of Urban America, edited by Raymond Mohl (New York: SR Publishers, 1997), 121.9 George Waring, “The Disposal of a City's Waste,” North American Review (July 1895), 55 (reprint; copyright 1895, by Lloyd Bryor).10 Roy Rosenzsweig and Elizabeth Blackmar, The Park and the People: A History of Central Park. (Ithaca, NY: Cornell University Press, 1992), 296.11 Ibid., 222.12 Board of Commissioners of the Central Park, Thirteenth Annual Report of the Board of Commissioners of the Central Park, for the Year Ending December 31, 1869 (New York: Evening Post Steam Presses, 1870).13 Board of Commissioners of the Central Park, Fifth Annual Report (note 6), 120.14 “Fertilizing the Park Lawns,” New York Tribune, February 15, 1885, 4.15 Joan H. Geismar, “Where Is Night Soil? Thoughts on an Urban Privy,” Historical Archaeology 27, no. 2 (1993), 61.16 Joanne Abel Goldman, Building New York's Sewers: Developing Mechanisms of Urban Management (West Lafayette, IN: Purdue University Press, 1997), 25.17 Peter Baldwin, In the Watches of the Night: Life in the Nocturnal City, 1820–1930 (Chicago: University of Chicago Press, 2012).18 “Poudrette—Soiling,” The Cultivator, June 1842, 93.19 “Poudrette as a Fertilizer—The N.Y. Poudrette Co. &c.,” The American Farmer, and Spirit of the Agricultural Journals of the Day (1839–1850), June 23, 1841, 39.20 Joel Tarr. “City to Farm: Urban Wastes and the Farmer.” Agricultural History, 49, no. 4 (October 1975): 605.21 “Night-soil—Its Value and Preparation for Use,” The Cultivator, November 1857, 352.22 Richard A. Wines, Fertilizer in America: From Waste Recycling to Resource Exploitation (Philadelphia: Temple University Press, 1985).23 Jimmy M. Skaggs, The Great Guano Rush: Entrepreneurs and American Overseas Expansion. (New York: St. Martin's, 1994), 2.24 Ibid.25 Viele, Report of the Committee on Civic Cleanliness (note 1), 26.26 . Skaggs, The Great Guano Rush (note 23), 15.27 “Guano. Its History, Traffic, Uses, Abuses, and Frauds,” New York Daily Tribune, September 29, 1860, 8.28 Edmund Rodman, “American Guano,” Maine Farmer, June 2, 1859, I.29 “Life on a Guano Island,” New York Times, September 9, 1866, 2.30 “Baker's Island Guano,” New York Daily Tribune, May 11, 1857, 3.31 Board of Commissioners of the Central Park, Seventh Annual Report (note 5), 87.32 Viele, Report of the Committee on Civic Cleanliness (note 1), 26.

One-Pot Construction of Functional Mesoporous Silica Nanoparticles for the Tumor-Acidity-Activated Synergistic Chemotherapy of Glioblastoma

Mesoporous silica nanoparticles (MSNs) have proved to be an effective carrier for controlled drug release and can be functionalized easily for use as stimuli-responsive vehicles. Here, a novel intelligent drug-delivery system (DDS), camptothecin (CPT)-loaded and doxorubicin (DOX)-conjugated MSN (CPT@MSN-hyd-DOX), is reported via a facile one-pot preparation for use in synergistic chemotherapy of glioblastoma. DOX was conjugated to MSNs via acid-labile hydrazone bonds, and CPT was loaded in the pores of the MSNs. At pH 6.5 (analogous to the pH in tumor tissues), a fast DOX release was observed that was attributed to the hydrolysis of the hydrazone bonds. In addition, a further burst release of DOX was found at pH 5.0 (analogous to the pH in lyso/endosomes of tumor cells), leading to a strong synergistic effect. In all, CPT and DOX could be delivered simultaneously into tumor cells, and this intelligent DDS has great potential for tumor-trigged drug release for use in the synergistic chemotherapy of tumors.

Antibiotics in chronic suppurative otitis media: A bacteriologic study

ObjectiveThis study sought to determine causative microorganisms of chronic suppurative otitis media (CSOM) and to estimate their susceptibility to antibiotic agents. Materials and methodsOne hundred sixty-four (164) patients with unilateral or bilateral active chronic suppurative otitis media were prospectively studied. They had chronic ear discharge and had not received any antibiotics for the previous 5days. Swabs were taken from all patients, and sent to the Microbiology Department for processing. The specimens were cultured and the isolates were identified using standard microbiological techniques. The antibiotic susceptibilities of the isolates were determined with standard antibiotic discs using the Kirby–bauer disc diffusion method. ResultsThis study analyzes the causal organisms and their sensitivity to various antibiotics. The major organisms isolated were Methicillin sensitive Staphylococcus aureus [MSSA] (45.1%) followed by Pseudomonas aeruginosa (19.5%). The sensitivity of S. aureus (MSSA) was 79.7% to ciprofloxacin, 69% to cotrimoxazole, and 82.5% to gentamicin whereas the sensitivity of P. aeruginosa was 100% to ceftazidime, 84.4% to ciprofloxacin, 90.6% to gentamicin, and 78.1% to Piperacillin. Majority of the isolates of P. aeruginosa were completely resistant to polymixin B (71.9%) whereas 50% of S. aureus (MSSA) showed resistance to Ampicillin. All Methicillin resistant S. aureus (MRSA) isolates showed 100% resistance to Ampicillin, Augmentin and cephalothin. Among the available topical antibiotic preparations for use in the ear, we found gentamicin and ciprofloxacin as the best choices. ConclusionThe study of microbial pattern and their antibiotic sensitivity determines the prevalent bacterial organisms causing CSOM in the local area to start empirical treatment of otitis media and its complications for a successful outcome, and thus to prevent the emergence of resistant strains.

Scientific Opinion on the evaluation of the safety in use of Yohimbe (Pausinystalia yohimbe (K. Schum.) Pierre ex Beille)

The Panel on Food Additives and Nutrient Sources added to Food provides a scientific opinion evaluating the safety in use of yohimbe bark and its preparations originating from Yohimbe (Pausinystalia yohimbe (K. Schum.) Pierre ex Beille when used in food, e.g. in food supplements. The bark of the plant contains a number of indole alkaloids of biological relevance and preparations of yohimbe bark have been traditionally used as general tonic, performance enhancer and as an aphrodisiac. Food supplements containing yohimbe bark preparations are available nowadays, especially via internet retail. Yohimbine, the major alkaloid of yohimbe bark and raubasine, another alkaloid occurring in lower concentrations in the bark, are used as active ingredients in a number of medicinal products for which adverse effects are described. The Panel reviewed the available scientific data on a possible association between the intake of yohimbe bark and its preparations and potential harmful effects on health. When those data were not available, priority was given to yohimbine, as the only alkaloid for which occurrence had been shown and quantified in food supplements containing yohimbe bark. The Panel concluded that the chemical and toxicological characterisation of yohimbe bark and its preparations for use in food are not adequate to conclude on their safety as ingredients of food, e.g. in food supplements. Thus the Panel could not provide advice on a daily intake of yohimbe bark and its preparations that do not give rise to concerns about harmful effects to health. An estimation of exposure to yohimbine from food supplements was performed showing that theoretical maximum daily intake may exceed the maximum approved daily dose of yohimbine from use as a medicinal product.

Live imaging and analysis of postnatal mouse retinal development

BackgroundThe explanted, developing rodent retina provides an efficient and accessible preparation for use in gene transfer and pharmacological experimentation. Many of the features of normal development are retained in the explanted retina, including retinal progenitor cell proliferation, heterochronic cell production, interkinetic nuclear migration, and connectivity. To date, live imaging in the developing retina has been reported in non-mammalian and mammalian whole-mount samples. An integrated approach to rodent retinal culture/transfection, live imaging, cell tracking, and analysis in structurally intact explants greatly improves our ability to assess the kinetics of cell production.ResultsIn this report, we describe the assembly and maintenance of an in vitro, CO2-independent, live mouse retinal preparation that is accessible by both upright and inverted, 2-photon or confocal microscopes. The optics of this preparation permit high-quality and multi-channel imaging of retinal cells expressing fluorescent reporters for up to 48h. Tracking of interkinetic nuclear migration within individual cells, and changes in retinal progenitor cell morphology are described. Follow-up, hierarchical cluster screening revealed that several different dependent variable measures can be used to identify and group movement kinetics in experimental and control samples.ConclusionsCollectively, these methods provide a robust approach to assay multiple features of rodent retinal development using live imaging.

Bone Degradation and Environment: Understanding, Assessing and Conducting Archaeological Experiments Using Modern Animal Bones

AbstractArchaeological experiments that use modern bones to replicate past animal bone assemblages have often failed to consider the effects of environment, storage and preparation on modern bones. Often, these experiments make little mention of the conditions to which bones were subject during their storage and preparation for use in experiments. In other instances, these variables are reported but not considered as factors that contribute to the nature of the results obtained. This study considers previously reported data concerning the degradation of frozen bones (−20°C), and bones exposed to hot, dry conditions (40°C), and presents new data for bones exposed to room temperature environments (22°C) and refrigerated environments (2°C), and bones that are frozen (−20°C) and then thawed (22°C). These conditions are all relevant to understanding the nature of bone degradation and the use of bones in modern archaeological experimentation. This article also surveys a range of previously reported experiments that utilise modern bones to create analogies to the past and considers different methodological approaches and their relationship to the condition of bones at the time of their fracture and fragmentation. The longitudinal data presented in this study demonstrate differential rates of bone degradation over time in various environmental conditions. This degradation results in dramatic changes in bone fracture morphology, bone strength and utility for bone tool production. These observations have significant implications for experiments that utilise modern bones, especially when experimental data are used to create analogies to the archaeological past. Copyright © 2012 John Wiley & Sons, Ltd.

Utilization of Metal Halide Species Ambiguity to Develop Amorphous, Stabilized Pharmaceutical Agents As Ionic Liquids

A new strategy employs amorphous melts of active pharmaceutical agents, which are formed with metal chlorides, including ZnCl2, through speciation ambiguity. The Lewis acidic, bioinorganic ionic liquid formulations improve shelf life and water solubility and eliminate issues related to crystal polymorphism. This broad-ranging approach produces thermally robust chlorometallate preparations for use as lotions, antibiofilm agents, and in situations without refrigeration.

A primer on evidence‐based plasma therapy

Background and Objectives Although traditionally fresh‐frozen plasma (FFP) has been the product of choice for reversing a significant coagulopathy, the modern blood bank will have several different plasma preparations for use in reversing a significant coagulopathy or arresting coagulopathic bleeding. These products include the aforementioned FFP which is frozen within 8 h of collection, plasma frozen within 24 h after phlebotomy (FP24), and thawed plasma (TP) prepared from either FFP or FP24. Based on their factor levels, these products should all be equally efficacious in reversing a significant coagulopathy although a head‐to‐head comparison between these products has not been performed. Evidence from two systematic literature reviews and numerous observational studies suggest that for a stable patient with a mild coagulopathy, transfusing plasma for an INR ≤ 1·5 does not confer a hemostatic benefit. This lack of benefit is due to the exponential relationship that exists between factor levels and a mildly elevated INR, that is, a large quantity of plasma would be required to produce a reduction in the INR when its pre‐transfusion value is <1·6. By contrast, the relationship becomes more linear when the INR is more significantly prolonged. Thus, even if a reduction in a mildly elevated INR could be achieved with plasma transfusion, the clinical significance of the decrease is dubious and the transfusion itself exposes the recipient to a variety of adverse events such as allergic reactions and transfusion‐related acute lung injury (TRALI). Furthermore, emerging evidence indicates that transfusion‐associated circulatory overload (TACO) following plasma transfusion is an important and under reported adverse event. This review will discuss the various plasma products that are available, describe why the commonly used markers of hemostasis (such as the PT/INR) are not effective markers of perioperative hemostasis, and present some of the current literature on the clinical uses of plasma. Strategies to reduce non‐evidence‐based plasma transfusion, ranging from automated interventions at the time plasma is ordered to more labour‐intensive manual attempts to dissuade providers from ordering plasma unnecessarily, will also be addressed.

Development of a Cryogenic Temperature Confocal Laser Scanning Microscope for Investigating Molecular Electric Fields

Molecular electric fields and potentials govern the chemical and physical behavior of molecules. In many biological systems, processes involving charge separation and transport such as enzymatic activity [1, 2], photosynthesis [3] and the electrostatic steering of ligands towards active sites, have been attributed to these fields. Gaining a detailed, quantitative description of the effects molecular electric fields play, will provide for a more precise mechanistic understanding of such chemical phenomena. Until now, obtaining information concerning these fields and potentials from experiments has been a problematic task. It is the principle objective of this investigation to quantitatively determine and describe the molecular electric fields at the oxygen binding site in the heme proteins myoglobin and hemoglobin. By utilizing single molecule and hole-burning spectroscopies, Stark effect measurements will be employed; to study the impact internal electric field distributions play. To this end, we have developed and constructed a cryogenic temperature version of a confocal laser scanning microscope; to be used for Stark spectroscopy investigations on these heme proteins using high resolution techniques such as single molecule or hole burning spectroscopy. At this stage, room and low temperature imaging and calibration of the microscope have been performed using 1μm fluorescent beads and AFM grids. The next step entails imaging single molecules of Protoporphyrin IX and Stark investigations of various metal substituted porphyrins in glasses. The primary focus of work thus far has been on assembly and design of the microscope and on sample preparation for use in molecular electric field examinations.1) E.D. Getzoff, et.al, Nature 358, (1992) 347.2) J. P. Hosler, et.al,Biochemistry 35, (1996) 10776.3) A. P. de Silva, et.al.,Chem. Commun., (1999) 163.

The Impact of Thromboemboli Histology on the Performance of a Mechanical Thrombectomy Device

Recently we published a novel method of thrombus preparation for use in a swine model for evaluation of thrombectomy designs. The clot (fibrin rich clot) is characterized by its similarity in histologic characteristics to the thromboemboli recovered from stroke patients. The purpose of this latest study was to evaluate if the performance of a mechanical thrombectomy device was affected by the histologic characteristics of thromboembolus. Erythrocyte rich clot, which was created using exogenous thrombin, and the novel experimental clot with abundance of fibrin/cellular component were used for comparison. The Merci clot retriever was used for the evaluation and the angiographic outcomes were analyzed. Two histologically different types of experimental clot, a conventionally used thrombin-induced clot (erythrocyte-rich clot) and a novel experimental clot that is similar in histologic characteristics to the thromboemboli recovered from patients with stroke (fibrin-rich clot), were prepared. Eight extracranial arteries in swine were occluded with erythrocyte-rich clot (group A), and 8 were occluded with fibrin-rich clot (group B), and MT by using the Merci clot retriever device was performed. Angiographic results in each group were evaluated. A total of 48 attempts at MT were made. The average number of attempts to achieve TIMI grade II or III recanalization was 2.75 times in group A and 4.5 times in group B (P < .001), respectively. The mean time to achieve recanalization was 15.5 minutes in group A and 81.5 minutes in group B (P < .01). Every vessel in group A showed recanalization (100%), whereas only 3 of 8 samples (37.5%) achieved recanalization in group B. In this model, arteries occluded by fibrin-rich clot demonstrated a significantly lower recanalization rate, lower final TIMI score, and a longer mean recanalization time than did arteries occluded by erythrocyte-rich clot. The angiographic outcome of MT by using the Merci clot retriever system was influenced by the histologic characteristics of the occluding thromboembolus.

Potential application of commercial enzyme preparations for industrial production of short-chain fructooligosaccharides

Abstract Twenty-five commercial enzyme preparations for use in the food industry were assayed for transfructosylation activity. Three preparations showed high transfructosylation activity from sucrose as well as the ratio of transferase and hydrolase activities. Short-chain fructooligosaccharides (sc-FOS) were not hydrolyzed by the three enzyme preparations after a 12 h reaction time. At a 6 h reaction time, yield and volumetric productivity were in the range from 58.8 to 62.6% (g sc-FOS/100 g initial sucrose) and 52.5 to 55.9 g sc-FOS/L h, respectively. One enzyme preparation was then evaluated for sc-FOS synthesis. Thus, environmental factors influencing the reaction were studied on products. Total sc-FOS concentration was not affected by temperature, pH and enzyme concentration at the studied levels, but high concentrations of sucrose affected the sc-FOS formation. The results suggest that these enzyme preparations can be exploited as a source of food-grade fructosyltransferase, in addition to Pectinex Ultra SP-L.

Home-Based Computer-Assisted Cognitive Training

Initial evidence suggests that computer-assisted cognitive rehabilitation may improve cognitive performance among people with multiple sclerosis (MS). Most studies of computer-assisted cognitive training have incorporated an individualized in-office/clinic approach for training. The purpose of this study was to explore the feasibility of home-based computer-assisted training and systematically examine the perceptions of people with MS regarding home use of the program. Qualitative data (written and verbal) obtained as part of a larger randomized clinical trial of a cognitive rehabilitation intervention were analyzed. The computer training component of the intervention included prescribed tracks and exercises on the Internet-based Neuropsychonline program. The majority of the participants used the program the recommended number of times per week and for the minimum number of minutes over the 8-week intervention. Although participants had notable negative perceptions and complaints about the program during the training experience, most of them acknowledged during training that use of the program helped them recognize cognitive limitations, create and practice strategies to increase cognitive function, and improve the quality of their daily life. Data from focus groups 3 months after the training included similar criticisms regarding lack of feedback and difficulties with the program as well as acknowledgment of the positive effects and a desire for continued access to the program. The findings suggest that while home training is feasible, careful preparation for use of the program is required.

Antimicrobial Activity of Grapefruit Seeds Extracts (In vitro Study)

Aims: The purpose of this study is to determine the antimicrobial activity of aqueous and alcoholic extract of grapefruit (Citrus Paradisi Rutaceae) seeds. Materials and Methods: Aqueous and alcoholic (ethanol) extracts of grapefruit seeds "GSE" in 20% (w/v) concentration were investigated for activity against Staphylococcus aureus, Proteus vulgaris, Klebsiella pneumonia, Candida albicans and a mixed oral flora. The level of antimicrobial effects was established using in vitro disc diffusion method. Their antibacterial and antifungal activity was compared to the activity of Chlorhexidine (CHX) solution in two concentrations (0.12 and 0.2%) as a control. Results: The aqueous GSE (20%) solution used in this study gave positive results with lethal effect on the tested organisms with zones of inhibition ranging from 10mm-18mm in diameter, which is comparable to that of chlorhexidine (7mm - 22mm) and (13mm - 20mm) for 0.12% and 0.2% solutions respectively. Ethanolic GSE did not show any antimi-crobial activity. Conclusions: Aqueous GSE has a good antimicrobial effect, which makes it a good natural preparation for use as antiseptic or disinfectant.

The impact of temperature on the production and fitness of microsclerotia of the fungal bioherbicide Mycoleptodiscus terrestris

The impact of growth temperature was evaluated for the fungal plant pathogen Mycoleptodiscus terrestris over a range of temperatures (20–36°C). The effect of temperature on biomass accumulation, colony forming units (cfu), and microsclerotia production was determined. Culture temperatures of 24–30°C produced significantly higher biomass accumulations and 20–24°C resulted in a significantly higher cfu. The growth of M. terrestris was greatly reduced at temperatures above 30°C and was absent at 36°C. The highest microsclerotia concentrations were produced over a wide range of temperatures (20–30°C). These data suggest that a growth temperature of 24°C would optimize the parameters evaluated in this study. In addition to growth parameters, we also evaluated the desiccation tolerance and storage stability of air-dried microsclerotial preparations from these cultures during storage at 4°C. During 5 months storage, there was no significant difference in viability for air-dried microsclerotial preparations from cultures grown at 20–30°C (>72% hyphal germination) or in conidia production (sporogenic germination) for air-dried preparations from cultures grown at 20–32°C. When the effect of temperature on germination by air-dried microsclerotial preparations was evaluated, data showed that temperatures of 22–30°C were optimal for hyphal and sporogenic germination. Air-dried microsclerotial preparations did not germinate hyphally at 36°C or sporogenically at 20, 32, 34, or 36°C. These data show that temperature does impact the growth and germination of M. terrestris and suggest that water temperature may be a critical environmental consideration for the application of air-dried M. terrestris preparations for use in controlling hydrilla.

Editorial Commentary: Is it time to abandon progesterone supplementation of early pregnancy after IVF?

It is estimated that a million couples receive IVF treatment globally every year (ICMART, 2009). The majority of women who become pregnant after IVF receive progesterone during the first trimester. A study conducted by Kyrou et al. published in the present issue of Human Reproduction challenges this concept for GnRH-antagonist treatment cycles and thereby adds to the growing concern that progesterone supplementation of early pregnancy after IVF might overall be unnecessary. The scientific evidence on which progesterone supplementation of early pregnancy was introduced to the field is unfortunately weak and stems from only a single randomized study (Prietl et al., 1992) on 120 subjects. IVF patients who received a combination of intra-muscular estrogen and progesterone supplementation until 12 weeks of pregnancy had an ongoing pregnancy rate of 89% compared with only 59% in the group with no supplementation. Although these data may not pass rigorous re-assessment, they have formed the basis for a treatment standard, namely continuation of progesterone administration in early pregnancy after IVF. Accordingly, Crinone, a vaginal progesterone formulation licensed for luteal phase support by the European Medicine Agency (EMEA), has on its label to continue administration ‘for 30 days if there is laboratory evidence of pregnancy’. The US Food and Drug Administration (FDA) label for the same product reads: ‘If pregnancy occurs, treatment may be continued until placental autonomy is achieved, up to 10–12 weeks’. The same is in the posology for Utrogestan, which is supposed to be administered until 12 weeks of gestation. Another vaginal progesterone preparation for use in IVF, Endometrin, is supposed to start the ‘day after oocyte retrieval and continuing for up to 10 weeks total duration’ according to FDA labeling. In this setting, clinicians now need to have a high level of confidence of not doing harm when routinely withdrawing early pregnancy progesterone supplementation from their patients. In other words, a conclusive ‘non-inferiority trial’ is required for that purpose (EMEA, 2006). In contrast to a superiority trial, the null hypothesis of the noninferiority trial is that a difference of at least Dx exists between two treatments. The trial is targeted at disproving this in favor of the alternative that no difference exists. Thus, the task now is to prove that stopping progesterone with a positive pregnancy test is non-inferior to continuing progesterone in terms of live birth achievement. But what does ‘proving of noninferiority’ mean in clinical and statistical terms? In order to demonstrate non-inferiority, the recommended approach is to pre-specify a margin of non-inferiority in the protocol (which was 27% difference in live-birth rate in the trial by Kyrou et al., 2011). After study completion, a one-sided 97.5% interval for the true difference between the two treatments is constructed. This interval should lie entirely on the positive side of the non-inferiority margin with special note on the lower limit of the confidence interval, which represents the degree of inferiority to the reference that can be excluded (with an acceptable low chance of error) based on the data presented.