Antimicrobial activity of the essential oil from Lippia sidoides, carvacrol and thymol against oral pathogens

Dental caries and periodontal disease are associated with oral pathogens. Several plant derivatives have been evaluated with respect to their antimicrobial effects against such pathogenic microorganisms. Lippia sidoides Cham (Verbenaceae), popularly known as "Alecrim-pimenta" is a typical shrub commonly found in the Northeast of Brazil. Many plant species belonging to the genus Lippia yield very fragrant essential oils of potential economic value which are used by the industry for the commercial production of perfumes, creams, lotions, and deodorants. Since the leaves of L. sidoides are also extensively used in popular medicine for the treatment of skin wounds and cuts, the objective of the present study was to evaluate the composition and antimicrobial activity of L. sidoides essential oil. The essential oil was obtained by hydro-distillation and analyzed by GC-MS. Twelve compounds were characterized, having as major constituents thymol (56.7%) and carvacrol (16.7%). The antimicrobial activity of the oil and the major components was tested against cariogenic bacterial species of the genus Streptococcus as well as Candida albicans using the broth dilution and disk diffusion assays. The essential oil and its major components thymol and carvacrol exhibited potent antimicrobial activity against the organisms tested with minimum inhibitory concentrations ranging from 0.625 to 10.0 mg/mL. The most sensitive microorganisms were C. albicans and Streptococcus mutans. The essential oil of L. sidoides and its major components exert promising antimicrobial effects against oral pathogens and suggest its likely usefulness to combat oral microbial growth.

Griscelli syndrome-type 2 in twin siblings: case report and update on RAB27A human mutations and gene structure

Griscelli syndrome (GS) is a rare autosomal recessive disorder caused by mutation in the MYO5A (GS1, Elejalde), RAB27A (GS2) or MLPH (GS3) genes. Typical features of all three subtypes of this disease include pigmentary dilution of the hair and skin and silvery-gray hair. Whereas the GS3 phenotype is restricted to the pigmentation dysfunction, GS1 patients also show primary neurological impairment and GS2 patients have severe immunological deficiencies that lead to recurrent infections and hemophagocytic syndrome. We report here the diagnosis of GS2 in 3-year-old twin siblings, with silvery-gray hair, immunodeficiency, hepatosplenomegaly and secondary severe neurological symptoms that culminated in multiple organ failure and death. Light microscopy examination of the hair showed large, irregular clumps of pigments characteristic of GS. A homozygous nonsense mutation, C-T transition (c.550C>T), in the coding region of the RAB27A gene, which leads to a premature stop codon and prediction of a truncated protein (R184X), was found. In patient mononuclear cells, RAB27A mRNA levels were the same as in cells from the parents, but no protein was detected. In addition to the case report, we also present an updated summary on the exon/intron organization of the human RAB27A gene, a literature review of GS2 cases, and a complete list of the human mutations currently reported in this gene. Finally, we propose a flow chart to guide the early diagnosis of the GS subtypes and Chédiak-Higashi syndrome.

Increased immunohistochemical expression of YKL-40 in the spleen of patients with portal hypertension

YKL-40 has been identified as a growth factor in connective tissue cells and also a migration factor in vascular smooth muscle cells. To a large extent, the increase of serum YKL-40 is attributed to liver fibrosis and asthma. However, the relationship of the expression and clinical/prognostic significance of YKL-40 to the splenomegaly of patients with portal hypertension is unclear. In the present study, the expression of YKL-40 was studied by immunohistochemistry in 48 splenomegaly tissue samples from patients with portal hypertension and in 14 normal spleen specimens. All specimens were quickly stored at -80°C after resection. Primary antibodies YKL-40 (1:150 dilution, rabbit polyclonal IgG) and MMP-9 (1:200 dilution, rabbit monoclonal IgG) and antirabbit immunoglobulins (HRP K4010) were used in this study. The relationship of clinicopathologic features with YKL-40 is presented. The expression of YKL-40 indicated by increased immunochemical reactivity was significantly up-regulated in splenomegaly tissues compared to normal spleen tissues. Overexpression of YKL-40 was found in 68.8% of splenomegaly tissues and was significantly associated with Child-Pugh classification (P = 0.000), free portal pressure (correlation coefficient = 0.499, P < 0.01) and spleen fibrosis (correlation coefficient = 0.857, P < 0.01). Further study showed a significant correlation between YKL-40 and MMP-9 (correlation coefficient = -0.839, P < 0.01), indicating that YKL-40 might be an accelerator of spleen tissue remodeling by inhibiting the expression of MMP-9. In conclusion, YKL-40 is an important factor involved in the remodeling of spleen tissue of portal hypertension patients and can be used as a therapeutic target for splenomegaly.

Development of two potential diagnostic monoclonal antibodies against human cytomegalovirus glycoprotein B

Human cytomegalovirus glycoprotein B (gB) represents a target for diagnosis and treatment in view of the role it plays in virus entry and spread. Nevertheless, to our knowledge, rare detection of a gB antigen has been reported in transplant patients and limited information is available about diagnostic gB monoclonal antibodies (mAbs). Our aim was to develop gB mAbs with diagnostic potential. Hydrophilic gB peptides (ST: amino acids 27-40, SH: amino acids 81-94) of favorable immunogenicity were synthesized and used to immunize BALB/c mice. Two mAbs, named ZJU-FH6 and ZJU-FE6, were generated by the hybridoma technique and limited serial dilution and then characterized by indirect ELISA, Western blotting, immunoprecipitation, and immunohistochemical staining. The mAbs displayed high titers of specific binding affinities for the ST and SH synthetic peptides at an mAb dilution of 1:60,000 and 1:240,000, respectively. Western blotting and immunoprecipitation indicated that these mAbs recognized both denatured and native gB of the Towne and AD169 strains. The mAbs, when used as the primary antibody, showed positive staining in cells infected with both Towne and AD169 strains. The mAbs were then tested on patients submitted to allogeneic hematopoietic stem cell transplantation. The gB antigen positivity rates of the patients tested using ZJU-FH6 and ZJU-FE6 were 62.0 and 63.0%, respectively. The gB antigen showed a significant correlation with the level of pp65 antigen in peripheral blood leukocytes. In conclusion, two potential diagnostic gB mAbs were developed and were shown to be capable of recognizing gB in peripheral blood leukocytes in a reliable manner.

Expression, production, and renaturation of a functional single-chain variable antibody fragment (scFv) against human ICAM-1

Intercellular adhesion molecule-1 (ICAM-1) is an important factor in the progression of inflammatory responses in vivo. To develop a new anti-inflammatory drug to block the biological activity of ICAM-1, we produced a monoclonal antibody (Ka=4.19×10−8 M) against human ICAM-1. The anti-ICAM-1 single-chain variable antibody fragment (scFv) was expressed at a high level as inclusion bodies in Escherichia coli. We refolded the scFv (Ka=2.35×10−7 M) by ion-exchange chromatography, dialysis, and dilution. The results showed that column chromatography refolding by high-performance Q Sepharose had remarkable advantages over conventional dilution and dialysis methods. Furthermore, the anti-ICAM-1 scFv yield of about 60 mg/L was higher with this method. The purity of the final product was greater than 90%, as shown by denaturing gel electrophoresis. Enzyme-linked immunosorbent assay, cell culture, and animal experiments were used to assess the immunological properties and biological activities of the renatured scFv.

Physical properties of structured lipids from lard and soybean oil produced by enzymatic interesterification

The main goal of the present research was to evaluate the physical properties of blends of lard and soybean oil modified by enzymatic interesterification catalyzed by two different commercial (microbial) lipases, viz. from Candida cylindracea (AY30TM) and from Mucor circinelloides (M10TM). Pure lard exhibited a softening point of ca. 31.8 °C before interesterification, and this value shifted towards 29.1 °C after interesterification by AY30 lipase and towards 28.8 °C after interesterification by M10 lipase The interesterified lard exhibited lower consistency after reaction with both lipases, and this decrease was more pronounced for the reaction catalyzed by M10 lipase. This result was most likely due to the sn-1,3-specificity of M10 lipase. Pure lard displayed a lower SFC after interesterification, and M10 lipase proved to be more effective than AY30 lipase. The non-interesterified lard had a SFC of 31.3% at 10 °C, which was reduced to 23.8 and 19.9% after interesterification with AY30 lipase and M10 lipase, respectively. The lard and soybean oil blends were affected by the enzymatic interesterification and dilution with soybean oil.

Cultivation of microalgae Spirulina platensis (Arthrospira platensis) from biological treatment of swine wastewater

The microalgae biomass production from swine wastewater is a possible solution for the environmental impact generated by wastewater discharge into water sources. The biomass can be added to fish feed, which can be used in the formulation of meat products. This work addresses the adaptation of the microalgae Spirulina platensis (Arthrospira platensis) in swine wastewater and the study of the best dilution of the wastewater for maximum biomass production and for removal of Chemical Oxygen Demand (COD), ammonia and phosphorous to the microalgae. The cultivation of Spirulina platensis, strain Paracas presented maximum cellular concentrations and maximum specific growth rates in the wastewater concentration of 5.0 and 8.5%. The highest COD removals occurred with 26.5 and 30.0% of wastewater in the medium. The maximum removal of total phosphorous (41.6%), was with 8.5% of wastewater, which is related to the microalgae growth. The results of Spirulina culture in the swine wastewater demonstrated the possibility of using these microalgae for the COD and phosphorous removal and for biomass production.

Design of automatic control system for the precipitation of bromelain from the extract of pineapple wastes

In this work, bromelain was recovered from ground pineapple stem and rind by means of precipitation with alcohol at low temperature. Bromelain is the name of a group of powerful protein-digesting, or proteolytic, enzymes that are particularly useful for reducing muscle and tissue inflammation and as a digestive aid. Temperature control is crucial to avoid irreversible protein denaturation and consequently to improve the quality of the enzyme recovered. The process was carried out alternatively in two fed-batch pilot tanks: a glass tank and a stainless steel tank. Aliquots containing 100 mL of pineapple aqueous extract were fed into the tank. Inside the jacketed tank, the protein was exposed to unsteady operating conditions during the addition of the precipitating agent (ethanol 99.5%) because the dilution ratio "aqueous extract to ethanol" and heat transfer area changed. The coolant flow rate was manipulated through a variable speed pump. Fine tuned conventional and adaptive PID controllers were on-line implemented using a fieldbus digital control system. The processing performance efficiency was enhanced and so was the quality (enzyme activity) of the product.

Isotope analysis (δ13C) of pulpy whole apple juice

The objectives of this study were to develop the method of isotope analysis to quantify the carbon of C3 photosynthetic cycle in pulpy whole apple juice and to measure the legal limits based on Brazilian legislation in order to identify the beverages that do not conform to the Ministry of Agriculture, Livestock and Food Supply (MAPA). This beverage was produced in a laboratory according to the Brazilian law. Pulpy juices adulterated by the addition of sugarcane were also produced. The isotope analyses measured the relative isotope enrichment of the juices, their pulpy fractions (internal standard) and purified sugar. From those results, the quantity of C3 source was estimated by means of the isotope dilution equation. To determine the existence of adulteration in commercial juices, it was necessary to create a legal limit according to the Brazilian law. Three brands of commercial juices were analyzed. One was classified as adulterated. The legal limit enabled to clearly identify the juice that was not in conformity with the Brazilian law. The methodology developed proved efficient for quantifying the carbon of C3 origin in commercial pulpy apple juices.

Use of real-time PCR to evaluate two DNA extraction methods from food

The DNA extraction is a critical step in Genetically Modified Organisms analysis based on real-time PCR. In this study, the CTAB and DNeasy methods provided good quality and quantity of DNA from the texturized soy protein, infant formula, and soy milk samples. Concerning the Certified Reference Material consisting of 5% Roundup Ready® soybean, neither method yielded DNA of good quality. However, the dilution test applied in the CTAB extracts showed no interference of inhibitory substances. The PCR efficiencies of lectin target amplification were not statistically different, and the coefficients of correlation (R²) demonstrated high degree of correlation between the copy numbers and the threshold cycle (Ct) values. ANOVA showed suitable adjustment of the regression and absence of significant linear deviations. The efficiencies of the p35S amplification were not statistically different, and all R² values using DNeasy extracts were above 0.98 with no significant linear deviations. Two out of three R² values using CTAB extracts were lower than 0.98, corresponding to lower degree of correlation, and the lack-of-fit test showed significant linear deviation in one run. The comparative analysis of the Ct values for the p35S and lectin targets demonstrated no statistical significant differences between the analytical curves of each target.

A novel procedure to measure the antioxidant capacity of Yerba maté extracts

Yerba maté extracts have in vitro antioxidant capacity attributed to the presence of polyphenolic compounds, mainly chlorogenic acids and dicaffeoylquinic acid derivatives. DPPH is one of the most used assays to measure the antioxidant capacity of pure compounds and plant extracts. It is difficult to compare the results between studies because this assay is applied in too many different conditions by the different research groups. Thus, in order to assess the antioxidant capacity of yerba maté extracts, the following procedure is proposed: 100 µL of an aqueous dilution of the extracts is mixed in duplicate with 3.0 mL of a DPPH 'work solution in absolute methanol (100 µM.L-1), with an incubation time of 120 minutes in darkness at 37 ± 1 °C, and then absorbance is read at 517 nm against absolute methanol. The results should be expressed as ascorbic acid equivalents or Trolox equivalents in mass percentage (g% dm, dry matter) in order to facilitate comparisons. The AOC of the ethanolic extracts ranged between 12.8 and 23.1 g TE % dm and from 9.1 to 16.4 g AAE % dm. The AOC determined by the DPPH assay proposed in the present study can be related to the total polyphenolic content determined by the Folin-Ciocalteu assay.

In vitro and in vivo antifungal activity of natural inhibitors against Penicillium expansum

Penicillium expansum is the causative agent of apple blue mold. The inhibitory effects of the capsaicin derived from Capsicum spp. fruits and yeast Hansenula wingei against P. expansum were evaluated in an in vitro and in in vivo assay using Fuji apples. The minimum inhibitory concentration of capsaicin determined using the broth micro-dilution method was 122.16 µg mL-1. Capsaicin did not reduce blue mold incidence in apples. However, it was able to delay fungal growth in the first 14 days of the in vivo assay. The in vivo effect of the yeast Hansenula wingei AM2(-2), alone and combined with thiabendazole at low dosage (40 µg mL-1), on the incidence of apple diseases caused by P. expansum was also described. H. wingei AM2(-2) combined with a low fungicide dosage (10% of the dosage recommended by the manufacturer) showed the best efficacy (100%) up to 7 days of storage at 21 ºC, later showing a non-statistically different decrease (p > 0.05) after 14 (80.45%) and 21 days (72.13%), respectively. These results contribute providing new options for using antifungal agents against Penicillium expansum.

Organizational creativity – Hegemonic and alternative discourses

SUMMARY Organizational creativity – hegemonic and alternative discourses Over the course of recent developments in the societal and business environment, the concept of creativity has been brought into new arenas. The rise of ‘creative industries’ and the idea of creativity as a form of capital have attracted the interests of business and management professionals – as well as academics. As the notion of creativity has been adopted in the organization studies literature, the concept of organizational creativity has been introduced to refer to creativity that takes place in an organizational context. This doctoral thesis focuses on organizational creativity, and its purpose is to explore and problematize the hegemonic organizational creativity discourse and to provide alternative viewpoints for theorizing about creativity in organizations. Taking a discourse theory approach, this thesis, first, provides an outline of the currently predominant, i.e. hegemonic, discourse on organizational creativity, which is explored regarding themes, perspectives, methods and paradigms. Second, this thesis consists of five studies that act as illustrations of certain alternative viewpoints. Through these exemplary studies, this thesis sheds light on the limitations and taken-for-granted aspects of the hegemonic discourse and discusses what these alternative viewpoints could offer for the understanding of and theorizing for organizational creativity. This study leans on an assumption that the development of organizational creativity knowledge and the related discourse is not inevitable or progressive but rather contingent. The organizational creativity discourse has developed in a certain direction, meaning that some themes, perspectives, and methods, as well as assumptions, values, and objectives, have gained a hegemonic position over others, and are therefore often taken for granted and considered valid and relevant. The hegemonization of certain aspects, however, contributes to the marginalization of others. The thesis concludes that the hegemonic discourse on organizational creativity is based on an extensive coverage of certain themes and perspectives, such as those focusing on individual cognitive processes, motivation, or organizational climate and their relation to creativity, to name a few. The limited focus on some themes and the confinement to certain prevalent perspectives, however, results in the marginalization of other themes and perspectives. The negative, often unintended, consequences, implications, and side effects of creativity, the factors that might hinder or prevent creativity, and a deeper inquiry into the ontology and epistemology of creativity have attracted relatively marginal interest. The material embeddedness of organizational creativity, in other words, the physical organizational environment as well as the human body and its non-cognitive resources, has largely been overlooked in the hegemonic discourse, although thereare studies in this area that give reason to believe that they might prove relevant for the understanding of creativity. The hegemonic discourse is based on an individual-centered understanding of creativity which overattributes creativity to an individual and his/her cognitive capabilities, while simultaneously neglecting how, for instance, the physical environment, artifacts, social dynamics and interactions condition organizational creativity. Due to historical reasons, quantitative as well as qualitative yet functionally- oriented studies have predominated the organizational creativity discourse, although studies falling into the interpretationist paradigm have gradually become more popular. The two radical paradigms, as well as methodological and analytical approaches typical of radical research, can be considered to hold a marginal position in the field of organizational creativity. The hegemonic organizational creativity discourse has provided extensive findings related to many aspects of organizational creativity, although the con- ceptualizations and understandings of organizational creativity in the hegemonic discourse are also in many respects limited and one-sided. The hegemonic discourse is based on an assumption that creativity is desirable, good, necessary, or even obligatory, and should be encouraged and nourished. The conceptualiza- tions of creativity favor the kind of creativity which is useful, valuable and can be harnessed for productivity. The current conceptualization is limited to the type of creativity that is acceptable and fits the managerial ideology, and washes out any risky, seemingly useless, or negative aspects of creativity. It also limits the possible meanings and representations that ‘creativity’ has in the respective discourse, excluding many meanings of creativity encountered in other discourses. The excessive focus on creativity that is good, positive, productive and fits the managerial agenda while ignoring other forms and aspects of creativity, however, contributes to the dilution of the notion. Practices aimed at encouraging the kind of creativity may actually entail a risk of fostering moderate alterations rather than more radical novelty, as well as management and organizational practices which limit creative endeavors, rather than increase their likelihood. The thesis concludes that although not often given the space and attention they deserve, there are alternative conceptualizations and understandings of organizational creativity which embrace a broader notion of creativity. The inability to accommodate the ‘other’ understandings and viewpoints within the organizational creativity discourse runs a risk of misrepresenting the complex and many-sided phenomenon of creativity in organizational context. Keywords: Organizational creativity, creativity, organization studies, discourse theory, hegemony

Lignoselluloosapohjaisten väkevähappohydrolysaattien kromatografinen fraktiointi

Väkevän hapon katalysoiman hydrolyysin avulla lignoselluloosasta on mahdollista valmistaa arvokkaita sokereita. Katalyyttinä toimiva happo voidaan käyttää uudelleen hydrolyysissä, jos se saadaan erotettua sokereista ilman neutralointia. Tämän kandidaatintyön tavoitteena oli selvittää, soveltuuko happoretardaatiotekniikka väkevähappohydrolysaatin fraktiointiin. Työssä verrattiin happoretardaatiotekniikkaa elektrolyyttiekskluusiotekniikkaan. Työn kirjallisuusosassa käsiteltiin happoretardaation ja elektrolyyttiekskluusion teoriaa. Lisäksi esiteltiin elektrolyyttiekskluusioon ja happoretardaatioon liittyviä tutkimuksia. Työn kokeellisessa osassa suoritettiin panoskromatografiakokeita käyttäen syöttöliuoksena rikkihappoa, etikkahappoa, glukoosia ja ksyloosia sisältävää synteettistä liuosta. Erotusmateriaaleina käytettiin neljää eri anionin- ja yhtä kationinvaihtohartsia. Kokeiden perusteella tutkittiin anioninvaihtohartsin tyypin ja kolonnin latauksen vaikutusta happoretardaatiotekniikalla saavutettavaan erotustulokseen sekä verrattiin elektrolyyttiekskluusiota happoretardaatioon. Työn tulosten perusteella rikkihappo laimeni happoretardaatiotekniikalla jopa 20-kertaisesti kromatografiakolonniin syötettyyn liuokseen verrattuna, riippumatta kolonnin latauksesta ja anioninvaihtohartsista. Rikkihapon laimenemisen vuoksi happoretardaatio ei soveltunut lignoselluloosapohjaisten väkevähappohydrolysaattien fraktiointiin. Elektrolyyttiekskluusiotekniikalla rikkihapon laimeneminen oli merkittävästi vähäisempää, minkä vuoksi elektrolyyttiekskluusion todettiin soveltuvan happoretardaatiota paremmin lignoselluloosapohjaisten väkevähappohydrolysaattien fraktiointiin.

Geochemistry and mineralogical association of heavy metal contaminants in fine grained sediment, Welland River, Southern Ontario /

This investigation of geochemistry and mineralogy of heavy metals in fine grained (<63^m) sediment of the Welland River was imdertaken to: 1) describe metal dispersion patterns relative to a source, identify minerals forming and existing at the outfall region and relate sediment particle size to chemistry; 2) to delineate sample handling, preparation and evaluate, modify and develop analytical methods for heavy metal analysis of complex environmental samples. Ajoint project between Brock University and Geoscience Laboratories was initiated to test a contaminated site of the Welland River at the base of Atlas Speciality Steels Co. Methods were developed and utilized for particle size separation and two acid extraction techniques: 1) Partial extraction; 2) Total extraction. The mineralogical assessment identified calcite, dolomite, quartz and clays. These minerals are typical of the carbonate-shale rock basement of the Niagara Peninsula. Minerals such as, mullite and ferrocolumbite were found at the outfall region. These are not typical of the local geology and are generally associated with industrial pollutants. Partial and total extraction techniques were used to characterize the sediments based on chemical distribution, elemental behaviour and analytical differences. The majority of elements were lower in concentration in the partial extraction technique; suggesting these elements are bound in an acid extractable phase (exchangeable, organic and carbonate phases). The total extraction technique yielded higher elemental concentrations taking difficult oxides and silicates into solution. Geochemical analyses of grain size separates revealed that heavy metal (Co, Ni, V, Mn, Fe, Ba) concentrations did not increase with decreasing grain size. This is a function of the anthropogenic mill scale input into the river. The background elements (Sc, Y, Sr, Mg, Al and Ti) showed an increase in concentration to the finest grain size suggesting that it is directly related to the local mineralogy and geology. Dispersion patterns ofmetals fall into two distinct categories: 1) the heavy metals (Co, Cu, Ni, Zn, V and Cr), and 2) the background elements (Be, Sc, Y, Sr, Al and Ti). The heavy metals show a marked increase in the outfall region, while the background elements show a significant decrease at the outfall. This pattern is attributed to a "dilution effect" ofthe natural sediments by the anthropogenic mill scale sediments. Multivariant statistical analysis and correlation coefficient matrix results clearly support these results and conclusions. These results indicate the outfall region ofthe Welland River is highly contaminated with to heavy metals from the industrialized area of Welland. A short distance downstream, the metal concentrations return to baseline geochemical levels. It appears, contaminants rapidly come out of suspension and are deposited in close proximity to the source. Therefore, it is likely that dredging the sediment from the river may cause resuspension of contaminated sediments, but may not distribute the sediment as far as initially anticipated.