Supplemental Medium Research Articles

Optimization and purification of L-methioninase enzyme purified from clinical samples of Pseudomonas spp.

Background: This study focused on Pseudomonas aeruginosa, which secrete important enzymes in medical and pharmaceutical applications and are isolated from clinical samples. One of these enzymes is L-methioninase, the most desirable enzyme in the medical and industrial fields today since it converted L-methionine into methanethiol, ammonia, and alpha-ketobutyrate. Objectives: Evaluated the optimum conditions of L-methioninase production and then purified it. Methods: Pseudomonas samples were identified by microscopic and biochemical tests and confirmed with the VITEK2 system. After the L-methioninase production was examined, the optimal conditions for the production of L-methioninase (MGL) were established using clinical samples of P. aeruginosa. Additionally, MGL was purified from the supernatant of P.aeruginosa using ammonium sulfate precipitation, DEAE-cellulose, and Sephadex G150 accordingly. Results: There were 33 isolates of P.aeruginosa according to microscopic characteristics and biochemical tests, followed by the VITEK2 system to ensure identification. The optimization conditions were achieved when supplement media with sucrose 1% w\v, casein 1% w\v, pH 7, temperature 37oC, and incubation for 48hrs within the specific activity of 1.6, 3.3, 3.4, 3.4, and 3.5U\mg protein, respectively, and 4.6 fold the amount of the purified enzyme as compared to the crude enzyme. A single 55 kDa band on the SDS-PAGE indicates the enzyme is purified. Conclusions: P.aeruginosa can be an efficient source of L-methioninase, and this enzyme had better and higher specific activity after applying optimum conditions for it and purifying it.

Elimination of systemic viral disease in shallot (Allium ascolonicum L) var. Bima Brebes through unconventional approach

Shallot (Allium ascolonicum L), belonging to the genus Allium are propagated vegetatively through layered bulbs. This research aimed to improve the quality of shallots by eliminating OYDV, SLDV viruses through meristematic tissue combined with the antiviral Ribavirin. The research was conducted in the IVEGRI Tissue Culture Laboratory from April to August 2018. Research materials included the infected shallot bulbs of Bima Brebes variety which were previously confirmed by DAS ELISA serology test. Meristematic tissues were cultured on MS + vitamin, MS + supplement media (sucrose 30 gL−1 + Myo inositol 100 mgL−1 + CaP 2 mgL−1 + NAA 0.15 mgL−1 + Kinetin 2 mgL−1 + GA3 0.1 mgL−1 + Gel gro 2gL−1, pH 5.7). A Completely Randomized Design with ten replications was applied for the combinations between antiviral Ribavirin concentrations i.e., 0.5,10,15, 20, 30 mgL−1, and explant size including meristematic tissue (0.5 mm) and shoot tip (2.5 mm). The results obtained through visual observation showed that the additional antiviral Ribavirin and explant size affected the percentage of growth and development of plantlets. Statistical analysis showed that there were no significant interactions between treatments. DAS Elisa serology test showed that the higher the concentration of antiviral Ribavirin the lower the percentage of virus infection.

Microbiology diagnostic approach in identifying Streptococcus pneumoniae: Case report of Streptococcal meningitis

Introduction: Bacterial meningitis remains a serious global health problem. Streptococcus pneumoniae as the causative organism, is fastidious. Microbiology diagnostic approach is needed to identify S.pneumoniae and antimicrobial susceptibility tests (AST). This study aims to report S. pneumoniae meningitis has been successfully cured strongly related to diagnostic stewardship. Case: A 1 year old child who fell, suffering from vomiting, fever and seizures was brought to the Emergency Department. Multi-slice Computerized Tomography shows cerebral edema and intracerebral hemorrhage. Laboratory blood tests and cerebrospinal fluid analysis strongly indicate bacterial meningitis. Presumptive Gram microscopy of Cerebrospinal Fluid (CSF) and initial identification of CSF cultures on lysed blood agar under microaerophilic conditions, consistently showed lancet-shaped and encapsulated Gram-positive diplococci. Our Clinical Microbiology Laboratory finally identified the growth of S.pneumoniae from blood and CSF cultures. Patients were treated with cefepime 50 mg/kg body weight/8 hours in the first 10 days as empirical and switched later definitively to ciprofloxacin 10 mg/kg body weight/12 hours according to the AST result on day 11 to day 26. On the 31ˢᵗ day, the patient recovered and was discharged. Conclusion: The case of a 1 year-old child with S. pneumoniae meningitis has been successfully cured strongly related to diagnostic stewardship by the Clinical Microbiology Laboratory. Supplementation media with sodium bicarbonate can improve S. pneumoniae recovery as a basis for identification and AST. Proper inoculation and AST need to be accelerated because S.pneumoniae easily undergoes autolysis.

INFLUENCE OF PROCESS PARAMETERS ON CELLULASE PRODUCTION BY Trichoderma viride (MTCC 800) EMPLOYING SOLID STATE FERMENTATION

In this study, the effect of physico-chemical parameters on the production of cellulase from Trichoderma viride (MTCC 800) were examined using five different agricultural residues namely banana peel, groundnut shell, corn cob, sugarcane bagasse, and pigeonpea stalk. The process parameters examined were pretreatment method, supplementation media, initial media pH, incubation period, temperature, and leaching agent. Among the parameters studied, maximum cellulase production was obtained on all the substrates treated with 1 N H2SO4 supplemented with Mandel’s medium of pH 5.5-6 after 6-8 days of incubation at 30-36°C and 0.05 M phosphate buffer (pH 7.0) as a leaching agent.

VP9: Human-Platelet Lysate Serum as a Potential Clinical-Translatable Supplement to Support Human Adipose-Derived Stem Cells Neurotrophic Properties

INTRODUCTION: We investigate a completely xenogeneic-free expansion method for adipose-derived stem cells (hADSC) to maintain their properties and enhancing their neurotrophic potential to develop a reliable and safe strategy to be extended to nerve regeneration in vivo. MATERIALS AND METHODS: We compared isolation methods and supplement media (fetal bovine serum (FBS) or human-platelet lysate (hPL)) on hADSC morphology, proliferation rate, immunophenotype, lineage differentiation potential and neurogenic commitment (nerve growth factors secretion). Functional analysis of hADSC expanded in both medium conditions was performed using an in vitro co-culture model with rat dorsal root ganglia (DRG) explants, following neurite outgrowth. RESULTS: We found no significant difference between the means of cell isolation and the supplements except for a higher proliferation rate and more elongated morphology of hPL-hADSC. Neurotrophic factor secretion by hPL-hADSC showed statistically higher levels in all three growth factors compared with FBS-hADSC. DRG showed significantly longer neurite length and higher axonal area when co-cultured with hPL-hADSC. CONCLUSION: We show that hPL provides a clinically translatable means to support hADSC, strengthening cells proliferation and further promoting their neurotrophic properties in vitro.

Mimicking the Organic and Inorganic Composition of Anabolic Bone Enhances Human Mesenchymal Stem Cell Osteoinduction and Scaffold Mechanical Properties.

Engineered bone graft designs have been largely inspired by adult bone despite functionally significant differences from the composition of anabolic bone in both the mineralized and non-mineralized fractions. Specifically, anabolic bone contains hydroxyapatite with ionic substitutions that facilitate bone turnover and relatively rare collagens type VI and XII that are important for normal bone development. In this work, human mesenchymal stem cells (hMSCs) were cultured in lyophilized collagen type I scaffolds mineralized with hydroxyapatite containing Mg2+ substitutions, then induced to deposit an extracellular matrix (ECM) containing collagens VI and XII by exposure to GW9662, a PPARγ inhibitor. Delivery of GW9662 was accomplished through either Supplemented Media or via composite microspheres embedded in the scaffolds for localized delivery. Furthermore, hMSCs and scaffolds were cultured in both static and perfuse conditions to investigate the interaction between GW9662 treatment and perfusion and their effects on ECM deposition trends. Perfusion culture enhanced cell infiltration into the scaffold, deposition of collagen VI and XII, as well as osteogenic differentiation, as determined by gene expression of osteopontin, BMP2, and ALP. Furthermore, scaffold mineral density and compressive modulus were increased in response to both GW9662 treatment and perfusion after 3 weeks of culture. Local delivery of GW9662 with drug-eluting microspheres had comparable effects to systemic delivery in the perfusate. Together, these results demonstrate a strategy to create a scaffold mimicking both organic and inorganic characteristics of anabolic bone and its potential as a bone graft.

Oral mucosa: an alternative epidermic cell source to develop autologous dermal-epidermal substitutes from diabetic subjects.

Oral mucosa has been highlighted as a suitable source of epidermal cells due to its intrinsic characteristics such as its higher proliferation rate and its obtainability. Diabetic ulcers have a worldwide prevalence that is variable (1%-11%), meanwhile treatment of this has been proven ineffective. Tissue-engineered skin plays an important role in wound care focusing on strategies such autologous dermal-epidermal substitutes. Objective The aim of this study was to obtain autologous dermal-epidermal skin substitutes from oral mucosa from diabetic subjects as a first step towards a possible clinical application for cases of diabetic foot.Material and Methods Oral mucosa was obtained from diabetic and healthy subjects (n=20 per group). Epidermal cells were isolated and cultured using autologous fibrin to develop dermal-epidermal in vitro substitutes by the air-liquid technique with autologous human serum as a supplement media. Substitutes were immunocharacterized with collagen IV and cytokeratin 5-14 as specific markers. A Student´s t- test was performed to assess the differences between both groups.Results It was possible to isolate epidermal cells from the oral mucosa of diabetic and healthy subjects and develop autologous dermal-epidermal skin substitutes using autologous serum as a supplement. Differences in the expression of specific markers were observed and the cytokeratin 5-14 expression was lower in the diabetic substitutes, and the collagen IV expression was higher in the diabetic substitutes when compared with the healthy group, showing a significant difference.Conclusion Cells from oral mucosa could be an alternative and less invasive source for skin substitutes and wound healing. A difference in collagen production of diabetic cells suggests diabetic substitutes could improve diabetic wound healing. More research is needed to determine the crosstalk between components of these skin substitutes and damaged tissues.

Collagen and Fractionated Platelet-Rich Plasma Scaffold for Dermal Regeneration.

Current options for in vivo regeneration of dermal tissue remain limited. The purpose of this study was to engineer a unique scaffold capable of recruiting dermal stem cells from adjacent tissue, thus circumventing the need to seed the scaffolds with stem cells before implantation, leading to skin regeneration. A hydrogel scaffold was created through combination of type I collagen along with fractionated platelet-rich plasma. This was compared to a control hydrogel consisting of type I collagen and fetal bovine serum. Hydrogels were cultured with fresh human skin tissue and incubated with supplemental media. Gels were digested weekly for cellular content as examined by flow cytometry at the 4- and 8-week time points. The fractionated platelet-rich plasma and collagen gels were then implanted onto full-thickness skin defects on the backs of rats and compared to wounds healing by secondary intention. Wound area was evaluated for epithelialization and neovascularization. Platelet-rich plasma fractionation increased platelet-derived growth factors. In contrast to collagen scaffolds, fractionated platelet-rich plasma-supplemented scaffolds recruited more dermal-derived stem cells from fresh skin tissue compared with collagen hydrogels at the 4- and 8-week time points. Furthermore, fractionated platelet-rich plasma-supplemented hydrogels accelerated wound healing, angiogenesis, and hair and sweat gland formation, ultimately regenerating a dermis-like tissue. Generation of hydrogels with fractionated platelet-rich plasma was able to improve cellular recruitment and growth and differentiation of dermal-derived stem cells, leading to hair growth and sweat gland formation. This provides a novel approach to regenerate skin for treating large defects.

Concise Guide to Jazz by Mark C. Gridley (review)

Concise Guide to By Mark C. Gridley. 7th ed. Upper Saddle River, NJ: Pearson, 2014. [viii, 312 p. ISBN 0205937004. $98.80.] Music examples, illustrations, photographs, bibliogra- phies, discographies, videographies, index. Supplemental media: Jazz Classics 2-CD Set, Prentice Hall Jazz CD Collection, Listening to Jazz Demonstration CD. Further media at companion Web site: http://www .pearsonhighered.com/mediaproducts /mysearchlab/.This book attempts to give a general overview of jazz history, from the late- nineteenth century to today. The author, Mark Gridley, designed it for a college-level music course for non-majors, as either the text for a semester-long class on the sub- ject, or as a supplement to a general music course. Gridley is a jazz musician and edu- cator, and this book, in its previous edi- tions, has established itself as an important resource in jazz pedagogy.The book's supplemental media are many and varied. The primary audio sup- plement is the Jazz Classics two-compact-disc anthology of performances which are dis- cussed and analyzed in the text. The Prentice Hall Jazz Collection compact disc contains more recorded performances, some of which are cited in the book (but not ana- lyzed). In addition, there is the Listening to Jazz Demonstration compact disc. Narrated by Gridley, it contains examples cited throughout the book of instruments em- ployed in jazz, as well as performance styles. These include the components of the drum set; trumpets, trombones, and the different types of mutes employed on them; the clar- inet and the saxophone family; walking bass; and more complex concepts, such as chord progressions and types of piano ac- companiment.New to this edition of the book is MySearchLab®, a Web site available from Pearson. (Buyers of the book receive access to the Web site, while the compact discs must be purchased separately.) It brings to- gether all the music on the Jazz Classics set of discs, as well as analyses of that music from the book. Both elements are pre- sented in a multimedia setting in which each section of the tune is highlighted as the music streams, along with analytical comments from the text. Students may pre- fer this method of studying the material, es- pecially given that many of them (perhaps most) may not own compact disc players, or prefer that resources be available online rather than on discs that require either purchase or a visit to the music library. (The content of the Prentice Hall disc, however, is not on the Web site.)MySearchLab® also contains animated flashcards for self-testing; a quiz for each chapter, results of which can be emailed to both student and instructor; and a large video component composed of perfor- mances of significant musicians and bands, brief historical clips about key musicians, and short clips duplicating some (but not all) of the material from the Listening to Jazz Demonstration compact disc, offering the ad- vantage of watching musicians perform it.Gridley divides the book and online con- tent into eleven chapters, of which the first two are preparatory but vital: What is Jazz? and How to Listen to Jazz. The first of these sets up themes that are repeated throughout the book and addresses some of the vexing challenges of writing a brief survey of jazz.The idea of a short textbook covering all of the music, even one for non-music- majors, invariably leads an aficionado to suspect that something important will be left out. This is, in part, because so many jazz people have trouble agreeing on what the music is: Is it always supposed to swing? Must the blues always be present? Can through-composed music be jazz (a re- quirement that would disqualify chunks of Duke Ellington's oeuvre)? Did Miles Davis quit making jazz in 1968? Was Keith Jarrett's American quartet jazzier than his European quartet? And so on.Perhaps the single most refreshing aspect of the book is the extent to which Gridley not only tackles these issues, but does so immediately, in the first few pages. …

Role of magnesium ions on Improvement of Fermentative activity of local yeast isolates Saccharomyces cerevisiae for ethanol production

The interest in bio-ethanol increase as biofuel, it seems to be a good alternative for fossil fuels. The growth of yeast Saccharomyces cerevisiae is influence in micronutrient like ions. This research aims to evaluate the influence different amount of magnesium as co-enzyme in growth and fermentation of yeast S. cerevisiae which isolated from different local sources. The batch fermentation process was followed at different magnesium concentrations as magnesium chloride at incubation temperature 30C○ and pH 4.5, best supplement concentration was 1.25 gm/l by measuring the mass of the released carbon dioxide. Supplement media with magnesium prolonged exponential growth, reduced the decline in fermentation activity resulting in reduced the time required for the conversion glucose into ethanol.

Callus Induction and Plantlet Regeneration Ability of Selected Sri Lankan Traditional Rice Cultivars (Oryza Sativa L.) in Caryopses Culture

A total of forty Sri Lankan traditional rice varieties ( Oryza sativa L) were examined for their callus induction, growth and regeneration ability at Laboratory of Agricultural Biology, Faculty of Agriculture, University of Ruhuna. Surface sterilized rice seeds (caryopses) were cultured on four different callus induction media supplemented with 1.5, 2.5, 3mg/l 2,4 – dichlorophenoxy acetic acid (2,4-D) and 0.1 mg/l 6 – benzyladenine (BA) on Murashige and Skoog’s (MS) basal medium. Proliferated calli were regenerated on 4 different regeneration media supplemented with 1.5, 2, 2.5, 3mg/l BA with 0.1 mg/l naphthaleneacetic acid (NAA). Eighty seven point five % of rice cultivars were able to produce callus on callus induction medium and among them 12.5 % of rice cultivars regenerated shoots. 100% callus induction was recorded by 75% of tested rice cultivars on 2.5mg/l 2,4-D supplemented to MS basal medium while the callus induction percentages were 30% and 32.5% in 1.5mg/l 2,4-D and 3mg/l 2,4-D supple-mented media respectively. Callus growth of rice cultivars showed significant difference with the cultivar and medium. By the 4th week of callus induction maximum diameter (> 2cm) of callus recorded by rice cultivar Herath on 2.5 mg/l 2,4-D + 0.1mg/l BA + MS basal medium. The regeneration ability of rice cultivars varied from 0 % to 100% while the highest regeneration was recorded in rice cultivar Herath on regeneration medium supplemented with 2.5mg/l BA and 0.1mg/l NAA. Callus induction varied from 0% - 100% and signifi-cantly high callus formation was offered in Dik wee, Herath, Pachchai Perumal and Rathu wee . Sudu bala wee, Kahata wee (long grain), Sudu heenati, Kahata samba and Muthumala failed to initiate callus on any of the medium. Significantly highest shoot regeneration percentage was found in Herath (100%) while Mas samba, Seedevi, Dostara heenati and Rathel were also categorized in to the same DMRT (Duncan's Multiple Range Test) group with Herath . The highest number of shoots (8 shoots/callus) was recorded in Herath compared to other varieties (0.1- 8 per explant). There was a significant interaction between callus induction medium and rice cultivar as well as regeneration medium and rice cultivar. Tropical Agricultural Research and Extension 15(1): 2012: page 15-18 DOI: http://dx.doi.org/10.4038/tare.v15i2.5244

Your Hero Is a Ghost, 2010

Stainless steel, wood, conduit, black paint, speakers, sound. Photo by Jason Mandella, courtesy Sculpture Center, New York, and the artists Lovett/Codagnone, an artist team based in New York, have worked together since 1995 using photography, performance, video, sound, and installation. Their ongoing exploration of relations of power, as manifested in explicit cultural signifiers as well as clandestine or unconscious practices, investigates the way power comes to play within social structures (relationships, family) to focus on intimacy and the construction of desire. Recent solo exhibitions: Museo Marino Marini, Florence, 2012; LA><ART, Los Angeles, 2012; September Galerie, Berlin, 2011; Sculpture Center, New York, 2010; Sara Meltzer Gallery, New York, 2009; MoMA PS1, New York, 2007–2008. Their performances have been presented at the ICA Philadelphia (2010); Judson Memorial Church, New York (2010); and the ICA Boston (2007). Their work has been shown in galleries and museums in France, Germany, Italy, and The Netherlands. Supplemental media related to this piece can be found at www.mitpressjournals.org/doi/suppl/10.1162/DRAM_a_00205

Organizational Use of a Media Campaign Booklet to Encourage Parent–Child Communication about Waiting to Have Sex

Promoting parent—child communication through the use of print materials may be an important health communication approach for preventing teen sexual activity in the United States. Although prior work has suggested successful methods of disseminating print materials, research has not examined dissemination approaches across organization types. Understanding the use and dissemination of print materials among different organizations is crucial to guiding materials development in a way that maximizes uptake and effectiveness among consumers. Accordingly, this study examined the use of a booklet for parents that encouraged parent–child communication about waiting to have sex as collateral material for a national media campaign. We interviewed staff at 9 community organizations, 9 health care facilities, 5 school-based organizations, and 3 campaign outreach centers. Results suggested variability within and across organization types regarding use of the booklet. Community and outreach center staff tended to walk through the booklet content with parents; health care facilities and education-oriented organizations usually reported both direct and indirect distribution approaches. Staff identified useful elements of the booklet and made suggestions for dissemination in line with prior research. A better understanding of how print materials are utilized to supplement media campaigns can improve their usefulness and potential influence on health behaviors.

Sulphate reduction and biomass growth rates for Desulfobacterium autotrophicum in yeast extract – Supplemented media at 38°C

Water pollution by heavy metals and sulphates is common in areas with mineral deposits. Sulphate-reducing bacteria (SRB) are microorganisms widely distributed, and can be used as heavy metals removal agents. Furthermore, the use of organic substrates or wastes in SRB systems could reduce the cost and increase the treatment efficiency. The aim of the study was to assess the sulphate reduction capacity and growth rates of the bacterium Desulfobacterium autotrophicumin batch culture media supplemented with different concentrations of yeast extract. At 38°C, in the growth phase, there was a direct relationship between the concentration of yeast extract employed and the biomass growth and sulphate reduction rates. In the presence of low sulphide concentrations, supplementary 0.5 g/L of yeast extract maximized sulphate reduction on a per-cell basis, but at high concentrations of H 2S it would be advisable to use 2 g/L to reduce the product inhibition.

Internal contamination and spoilage of harvested apples by patulin-producing and other toxigenic fungi

A total of 424 apple samples comprised of six varieties (Gala, Red Delicious, Golden Delicious, Fuji, Granny Smith, and Braeburn) were analyzed for internal fungal contamination. Two hundred sixteen apples were incubated intact for 2–4 weeks at room temperature. The cores of the remaining 208 apples were aseptically removed and incubated without supplemental media at room temperature for 3 weeks. After the incubation period was over, the mycological profiles of the analyzed samples were determined. Twelve per cent of the intact apples showed visible growth after 2–4 weeks of incubation at room temperature. Penicillia (including the patulin producer, Penicillium expansum) were the most frequent, found in 8% of the samples followed by Fusarium and Alternaria spp. (each found in 3% of the samples tested). The highest mould incidence was observed in the Red Delicious and Fuji and the lowest in the Granny Smith variety. A variety of microfungi including members of the toxigenic genera Alternaria, Penicillium and Fusarium were isolated from the apple cores. The predominant moulds were Alternaria, Cladosporium, Penicillium and Fusarium spp. recovered from 50, 22, 33 and 23% of the analyzed samples, respectively. Less common were Ulocladium spp., Botrytis cinerea and Aureobasidium pullulans found in less than 4% of the samples. Yeasts were found only in 2% of the samples. Apple cores from all varieties tested showed a high degree of mould contamination.

Evaluation of itunes U Podcasts on Undergraduate Biology Education

Recent changes in undergraduate science education include online or hybrid courses and electronic material such as textbooks and supplemental media. Electronically enhanced courses are attractive to educators as a way to bolster course material and provide students with more and varied learning options, approaches fitting to the era of internet information access. This study was undertaken to evaluate one such electronic supplement to undergraduate‐level science courses. Audio podcasts of biology class lectures were posted on the College of San Mateo's iTunes University website, provided by the Apple Corporation. Surveys were administered to students of three consecutive semesters to evaluate the use, effectiveness and outcome, as determined by course grade, of the podcasts. The results showed that the majority of students accessed the recorded lectures and those that used them the most felt that doing so improved their performance in the class. Data indicated that students most at risk of poor academic performance used the podcasts most frequently. This study indicates that electronic media supplementation enhanced science student performance. Further studies are underway in order to more effectively implement and use electronic supports and supplements to contribute to student success.

Solid phase microextraction (SPME) analysis of volatile compounds produced by in vitro shoots of Lantana camara L. under the influence of auxins and cytokinins

The effects of the auxin indole-3-acetic acid at 0.44 µmol L-1 and the cytokinins thidiazuron at 0.44 µmol L-1 and benzyladenine at 0.44 and 4.4 µmol L-1 on growth, development and volatile compounds production of Lantana camara L. in vitro plants were investigated. The volatiles were extracted by solid phase microextraction (SPME) and the quantification was performed using GC-MS. A higher content of myrcene, alpha-phellandrene, alpha-copaene, trans-caryophyllene and beta-gurjunene, and the lower levels of alpha-pinene and beta-pinene were observed in in vitro plants cultivated on different supplementation media than in donor plants.

Mould and yeast flora in fresh berries, grapes and citrus fruits

Fresh fruits are prone to fungal contamination in the field, during harvest, transport, marketing, and with the consumer. It is important to identify fungal contaminants in fresh fruits because some moulds can grow and produce mycotoxins on these commodities while certain yeasts and moulds can cause infections or allergies. In this study, 251 fresh fruit samples including several varieties of grapes, strawberries, blueberries, raspberries, blackberries, and various citrus fruits were surface-disinfected, incubated at room temperature for up to 14 days without supplemental media, and subsequently examined for mould and yeast growth. The level of contamination (percent of contaminated items/sample) varied depending on the type of fruit. All raspberry and blackberry samples were contaminated at levels ranging from 33% to 100%, whereas 95% of the blueberry samples supported mould growth at levels between 10% and 100% of the tested berries, and 97% of strawberry samples showed fungal growth on 33–100% of tested berries. The most common moulds isolated from these commodities were Botrytis cinerea, Rhizopus (in strawberries), Alternaria, Penicillium, Cladosporium and Fusarium followed by yeasts, Trichoderma and Aureobasidium. Thirty-five percent of the grape samples tested were contaminated and supported fungal growth; the levels of contamination ranged from 9% to 80%. The most common fungi spoiling grapes were Alternaria, B. cinerea and Cladosporium. Eighty-three percent of the citrus fruit samples showed fungal growth at levels ranging from 25% to 100% of tested fruits. The most common fungi in citrus fruits were Alternaria, Cladosporium, Penicillium, Fusarium and yeasts. Less common were Trichoderma, Geotrichum and Rhizopus.

Immortalization of Bone Marrow Mesenchymal Stem Cells from Inbred Pig for Regenerative Medicine

The aim of this experiment is to investigate whether MSCs from Banna Minipig Inbred Line (BMI) could be immortalized by introducing SV40 large T antigen gene. MSCs were isolated from BMI and transfected with pSV3neo plasmid. Normal BMI-MSCs would apoptosis and senescence during proliferation while its population doubling (PD) number exceeded 20. However, SV40-transfected cells were immortal. As same as that of normal BMI-MSCs, transfected MSCs were positive for stem cell markers and negative for differentiated osteoblast specific marker. After cultured in osteogenesis supplement media, cbfa1 and calcium deposit on stimulated cells were enhanced obviously. There were no data to prove the tumorigenicity potential of the immortalized cells. Furthermore, histological analysis demonstrated that bone formation was initiated in the pores of HA/TCP implants loaded immortalized BMI-MSCs 7 weeks postimplantation. BMI-MSCs were immortalized by introducing SV40 large T antigen into the cells and still kept the stem cell characters and might be used as seeding cells for tissue engineering as well as stable test cells for biocompatibility of bone biomaterials.

EFFECT OF LIMING MATERIALS ON MEDIA pH AND OVERALL GROWTH AND NUTRITION OF DWARF NANDINA

ABSTRACT Ground dolomitic limestone (CaCO3/MgCO3) is used throughout the southeastern United States as a fertilizer amendment to soilless media. In addition to neutralizing media acidity, carbonate-based limestone also provides a source of calcium (Ca) and magnesium (Mg) for growth of many woody ornamental species, among these dwarf nandina. This study was conducted to compare the effects of CaO and CaO/MgO/CaCO3 blends to ground dolomitic limestone on the growth, marketable quality, and foliar nutrition of Nandina domestica ‘Nana Purpurea’ grown in soilless media. Materials were preplant incorporated based on the calcium carbonate equivalence (CCE, or acid-neutralizing equivalence) of ground dolomitic limestone (CCE=63) and plants were arranged in a completely randomized design (CRD). Growth indices (GI) were determined and plants harvested for tissue dry matter weights and foliar nutrient content. Plants amended with CaO, agricultural gypsum (CaSO4ċ2H2O), or pelletized dolomitic limestone, were of similar or better quality than plants amended with ground dolomitic limestone. Greatest nutrient uptake [nitrogen (N), Ca, potassium (K), and phosphorus (P)] was likewise observed for these treatments. All liming materials resulted in higher nutrient uptake compared to the unlimed control and highest foliar Mg was reported for plants amended with ground and pelletized dolomitic limestones. In year 1, plants were of similar quality, with the exception of the Ca(OH)2 treatment, which gave the lowest foliar quality ratings. In year 2, all plants were of marketable quality. Throughout the study, and especially during year 2, many foliar nutrient levels fell below established survey averages for dwarf nandina, illustrating the need to supplement media mixes with additional fertilizers during the growing season. All liming materials raised media pH to within the target range for nursery crops (pH=5.5–6.5) within one month from initial application, but lost their most positive neutralizing effect by the end of the third month. Research conducted indicates that CaO, CaO/MgO/ CaCO3 blends, CaSO4ċ2H2O, or pelletized limestone may serve as suitable alternatives to ground dolomitic limestone for use in soilless media.