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1.
Clin Biochem ; 121-122: 110678, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37866698

RESUMO

BACKGROUND: LDL-C, a cardiovascular disease risk assessment biomarker, is commonly calculated using the Friedewald equation. The NIH equation overcomes several limitations of the Friedewald equation. Consistent with the Canadian Society of Clinical Chemists (CSCC) lipid reporting recommendations, we assessed the NIH LDL-C equation in Alberta prior to its provincial implementation. METHODS: 1-year (01/01/2021-12/31/2021) of lipid results (n = 1,486,584 after data cleaning) were obtained from five analytical instrument groups used across Alberta. Analyses were performed on all data and after separating by age, analytical instrument group, and fasting status. The correlation between Friedewald- and NIH-calculated LDL-C and between Friedewald- and NIH-calculated LDL-C difference and each lipid parameter, was determined. The frequency of unreportable/inaccurate LDL-C results was compared between the two equations. The concordance between the two equations and with non-HDL-C was determined at LDL-C thresholds. Lastly, LDL-C calculated by Friedewald, NIH, and Martin-Hopkins equations was compared to density-gradient ultracentrifugation. RESULTS: Friedewald- and NIH-calculated LDL-C exhibit the strongest correlation when triglycerides ≤ 4.52 mmol/L. The difference between Friedewald- and NIH-calculated LDL-C increases with decreasing LDL-C concentration. The NIH equation yields fewer inaccurate results (0.35 % vs. 22.0 %). The percent agreement between equations was > 96 % at all LDL-C thresholds, suggesting most patients will not require treatment changes. NIH-calculated LDL-C exhibited better agreement with non-HDL-C when triglycerides ≤ 9.04 mmol/L and better correlated with LDL-C measured by ultracentrifugation (r2 = 0.926 vs. 0.775 (Friedewald) and 0.863 (Martin-Hopkins)). Results were consistent across age, analytical instrument group, and fasting status. CONCLUSIONS: Our findings demonstrate the benefits of implementing the NIH equation across Alberta.


Assuntos
LDL-Colesterol , Humanos , LDL-Colesterol/análise , Alberta , Triglicerídeos , Biomarcadores , Ultracentrifugação
2.
Clin Biochem ; 118: 110589, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37236295

RESUMO

BACKGROUND: Hemoglobinopathies include thalassemia syndromes, where production of one or more globin subunits of hemoglobin (Hb) is reduced, and structural Hb variants. Over 1000 disorders of Hb synthesis and/or structure have been identified and characterized, with phenotypes ranging from having severe clinical manifestations to clinically silent. Various analytical methods are used to phenotypically detect Hb variants. However, molecular genetic analysis is a more definitive method for Hb variant identification. CASE REPORT: Here, we report a case of a 23-month-old male with results from capillary electrophoresis, gel electrophoresis (acid and alkaline), and high-performance liquid chromatography most consistent with HbS trait. Specifically, capillary electrophoresis showed slightly elevated HbF and HbA2, HbA of 39.4% and HbS of 48.5%. The HbS percentage was consistently higher than expected (typically 30-40%) for HbS trait with no concurrent thalassemic indices. The patient has not experienced any clinical complications due to the hemoglobinopathy and he is thriving. CONCLUSION: Molecular genetic analysis revealed the presence of compound heterozygosity for HbS and Hb Olupona. Hb Olupona is an extremely rare beta-chain variant that appears as HbA on all three common methods used for phenotypic Hb analysis. When the fractional concentration of Hb variants is unusual, more definitive methods should be used, such as mass spectrometry or molecular genetic testing. In this case, incorrectly reporting this result as HbS trait is unlikely to have a significant clinical impact, as current evidence suggests Hb Olupona is not a clinically significant variant.


Assuntos
Hemoglobinopatias , Hemoglobinas Anormais , Talassemia , Masculino , Humanos , Hemoglobinas Anormais/genética , Hemoglobinopatias/diagnóstico , Hemoglobinopatias/genética , Talassemia/genética , Hemoglobina A2 , Eletroforese Capilar/métodos
3.
Clin Biochem ; 116: 105-112, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37100108

RESUMO

OBJECTIVES: Multiple sclerosis is diagnosed based on clinical and laboratory findings, including cerebrospinal fluid (CSF) oligoclonal banding (OCB) analysis. The lack of updated CSF OCB laboratory guidelines in Canada has likely led to variation in processes and reporting across clinical laboratories. As a first step to developing harmonized laboratory recommendations, we examined current CSF OCB processes, reporting, and interpretation across all Canadian clinical laboratories currently performing this test. DESIGN AND METHODS: A survey of 39 questions was sent to clinical chemists at all 13 Canadian clinical laboratories performing CSF OCB analysis. The survey included questions regarding quality control processes, reporting practices for CSF gel electrophoresis pattern interpretation, and associated tests and calculated indices. RESULTS: The survey response rate was 100%. Most (10/13) laboratories use ≥2 CSF-specific bands (2017 McDonald Criteria) as their CSF OCB positivity cut-off and only 2/13 report the number of bands with every report. Most (8/13 and 9/13) laboratories report an inflammatory response pattern and monoclonal gammopathy pattern, respectively. However, the process for reporting and/or confirming a monoclonal gammopathy varies widely. Variation was observed for reference intervals, units, and the panel of reported associated tests and calculated indices. The maximum acceptable time interval between paired CSF and serum collections varied from 24 h to no limit. CONCLUSIONS: Profound variation exists in processes, reporting, and interpretation of CSF OCB and associated tests and indices across Canadian clinical laboratories. Harmonization of CSF OCB analysis is required to ensure continuity and quality of patient care. Our detailed assessment of current practice variation highlights the need for clinical stakeholder engagement and further data analysis to support optimal interpretation and reporting practices, which will aid in developing harmonized laboratory recommendations.


Assuntos
Gamopatia Monoclonal de Significância Indeterminada , Esclerose Múltipla , Paraproteinemias , Humanos , Laboratórios Clínicos , Canadá , Bandas Oligoclonais , Esclerose Múltipla/diagnóstico , Esclerose Múltipla/líquido cefalorraquidiano
4.
Issues Ment Health Nurs ; 41(2): 138-145, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31322982

RESUMO

The expanding practice of multi-disciplinary care to address the complex nature of Autism Spectrum Disorder (ASD) suggests that there is a need for a means of coordinating care that transcends the disciplinary distinctions of relevant ASD treatment providers. As ASD services become more specialized, there is a growing need for effective care coordination with providers across the systems of care. Nursing professionals are ideally qualified to support families affected by ASD, as they provide a necessary holistic lens of health and wellbeing to obtain the appropriate treatments. Solution-focused brief therapy has been applied to a growing number of clinical settings, indicating solution-focused techniques are applicable to the various contexts associated with ASD treatments. We provide a case presentation to demonstrate a solution-focused approach to address ASD-related concerns within the family that are generalizable to coordination of care.


Assuntos
Transtorno do Espectro Autista/psicologia , Transtorno do Espectro Autista/terapia , Equipe de Assistência ao Paciente/organização & administração , Psicoterapia Breve/organização & administração , Adolescente , Criança , Humanos , Masculino
5.
Clin Exp Allergy ; 47(11): 1456-1467, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28703865

RESUMO

BACKGROUND: Atopic dermatitis (AD), psoriasis (PS), and contact dermatitis (CD) are common skin diseases, characterized by barrier disruption and systemic inflammation, with unique epidermal signatures and common inflammatory pathways identified by transcriptomic profiling. This study profiled proteomic signatures in serum from subjects with AD, PS, and CD compared with healthy controls (HC). OBJECTIVE: Identify unique proteomic signatures to distinguish between inflammatory diseases with similar epidermal disruption and overlapping epithelial inflammation. METHODS: Sera from 20 subjects with moderate to severe AD, 10 subjects with CD, 12 subjects with moderate to severe PS, 10 subjects with both AD and CD, and 10 HC with no history of skin disease was analysed using high-throughput proteomic analysis that detects expression of 1129 protein targets. Protein expression was compared between disease and HC, and across diseases for statistical significance (fold change≥1.5 and false discovery rate≤0.05), to identify unique proteomic signatures for each disease. RESULTS: Complement C5A anaphylatoxin (C5A), lipopolysaccharide binding protein (LBP), C-reactive protein (CRP), ILT-4, C-C motif ligand 18 (PARC), and sialic acid-binding Ig-like lectin 14 (SIG14) were significantly modulated in all three diseases compared with HC. We identified unique signatures for AD (Immunoglobulin E (IgE), thymus- and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC)), CD (10 proteins), and PS (kynureninase (KYNU)). Proteomic profiling in subjects with both AD and CD identified additional dysregulated proteins compared with subjects with either condition alone, indicating an exacerbated inflammation reaction. CONCLUSIONS AND CLINICAL RELEVANCE: Unique sera proteomic signatures may distinguish between inflammatory skin diseases despite similar epidermal barrier disruption and epithelial inflammation. This may provide insight into disease pathogenesis, diagnosis, and therapeutic intervention in difficult-to-treat subjects.


Assuntos
Dermatite Atópica/imunologia , Dermatite Atópica/metabolismo , Proteoma , Proteômica , Dermatopatias/metabolismo , Estudos de Casos e Controles , Análise por Conglomerados , Dermatite de Contato/imunologia , Dermatite de Contato/metabolismo , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Mediadores da Inflamação/sangue , Mediadores da Inflamação/metabolismo , Masculino , Proteômica/métodos , Dermatopatias/etiologia
6.
Allergy ; 70(8): 887-96, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25879391

RESUMO

Atopic dermatitis (AD) is a debilitating disease that significantly alters the quality of life for one in four children and one in 10 adults. Current management of AD utilizes combinations of treatments to symptomatically alleviate disease by suppressing the inflammatory response and restoring barrier function in the skin, reducing disease exacerbation and flare, and preventing secondary skin infections. Resolution is temporary and long-term usage of these treatments can be associated with significant side-effects. Antibody therapies previously approved for inflammatory diseases have been opportunistically evaluated in patients with atopic dermatitis; however, they often failed to demonstrate a significant clinical benefit. Monoclonal antibodies currently in development offer hope to those individuals suffering from the disease by specifically targeting immune and molecular pathways important for the pathogenesis of atopic dermatitis. Here, we review the underlying biological pathways and the state of the art in therapeutics in AD.


Assuntos
Terapia Biológica/tendências , Dermatite Atópica/tratamento farmacológico , Dermatite Atópica/imunologia , Qualidade de Vida , Dermatite Atópica/psicologia , Feminino , Previsões , Humanos , Imunoterapia/tendências , Masculino , Índice de Gravidade de Doença , Pele/efeitos dos fármacos , Pele/imunologia , Resultado do Tratamento
7.
Plant Dis ; 98(12): 1695-1701, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30703878

RESUMO

Rhizoctonia solani is a damaging soilborne pathogen, which affects most field crops in the Canadian provinces of Alberta, Manitoba, and Saskatchewan. The objective of this study was to conduct a phylogenetic comparison of isolates of R. solani collected from a previous survey in the major canola- and wheat-growing regions of western Canada. A total of 128 multinucleate isolates from a previous survey were identified by internal transcribed spacer (ITS) sequence and compared to anastomosis group (AG) results. The multinucleate isolates of R. solani were grouped into eight distinct clades. Each clade corresponded to a specific AG with the exception of two distinct clades that were observed for isolates classified as AG 2-1 by anastomosis testing. While most isolates of AG 5 clustered together according to ITS sequences, three isolates classified by anastomosis grouping as AG 5 grouped with AG 2-1, AG 4, and a binucleate Rhizoctonia sp. in the phylogenetic analysis. In most instances, the results from AG tests were consistent with ITS sequence, but there were still several cases where isolates were inconsistently classified or failed to undergo anastomosis with any of the tester strains used in this study. This provides support for the use of the ITS region as a valuable tool for rapid identification of R. solani isolates to their respective AGs.

8.
Plant Dis ; 98(1): 32-42, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30708590

RESUMO

Detection and quantification of airborne ascospores as a component of the Sclerotinia rot of carrot (SRC) forecast model is currently accomplished using the blue plate test (BPT), which uses Sclerotinia semiselective medium (SSM). A quantitative polymerase chain reaction (qPCR) assay was developed to reduce the time to specifically quantify ascospores of Sclerotinia sclerotiorum from air samples collected using a Burkard Multi-Vial Cyclone Sampler. The qPCR assay was highly sensitive and detected DNA from 0.5 to 5 × 104 ascospores within a linear range (R2 = 0.99). The qPCR assay was used to quantify ascospores of S. sclerotiorum in air samples collected over three growing seasons. Initial SRC disease was observed 8 and 34 days following detection of 9.5 and 2 ascospores m-3 of air, respectively. Results from air samples collected using an Andersen N6 Sampler and the qPCR assay were compared with the BPT. Ascospore counts from a Burkard Sampler coupled with the qPCR assay and the BPT followed similar trends. In general, fewer ascospores were detected and bioaerosol sampling efficiency was low using an Anderson Sampler. Three days were required to confirm the number of ascospores using SSM in the BPT and with an Andersen Sampler, whereas results from a Burkard Sampler coupled with the qPCR assay can provide results within 5 h of air sampling. The choice of method will depend on the available resources.

9.
J Appl Microbiol ; 105(5): 1239-45, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18399976

RESUMO

AIMS: The aims of the current study were to explore the site of bacterial attachment to vegetable tissues and to investigate the hypothesis that Salmonella must be living in order to attach to this site(s). METHODS AND RESULTS: Scanning electron micrographs of intact potato cells showed that Salm. serotype Typhimurium attached to cell-wall junctions; suggesting a high-level of site selectivity. Inactivation of Salm. Typhimurium using heat, ethanol, formalin or Kanamycin resulted in cells that could be no longer attached to these sites. Attachment of a Gfp(+) strain of Salm. Typhimurium to cell-wall material (CWM) was examined via flow cytometric analysis. Only live Salm. Typhimurium attached to the CWM. CONCLUSIONS: Salmonella serotype Typhimurium must be metabolically active to ensure attachment to vegetable tissues. Attachment preferentially occurs at the plant cell-wall junction and the cell-wall components found here, including pectate, may provide a receptor site for bacterial attachment. SIGNIFICANCE AND IMPACT OF THE STUDY: Further studies into individual plant cell-wall components may yield the specific bacterial receptor site in vegetable tissues. This information could in turn lead to the development of more targeted and effective decontamination protocols that block this site of attachment.


Assuntos
Aderência Bacteriana/fisiologia , Parede Celular/microbiologia , Salmonella typhimurium/fisiologia , Solanum tuberosum/microbiologia , Contagem de Colônia Microbiana , Etanol , Citometria de Fluxo/métodos , Microbiologia de Alimentos , Microscopia Eletrônica de Varredura , Pectinas/análise , Salmonella typhimurium/crescimento & desenvolvimento , Solanum tuberosum/ultraestrutura , Temperatura
10.
Cell Microbiol ; 10(4): 958-84, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18031307

RESUMO

The biogenesis of the Salmonella-containing vacuole within mammalian cells has been intensively studied over recent years. However, the ability of Salmonella to sense and adapt to the intracellular environment of different types of host cells has received much less attention. To address this issue, we report the transcriptome of Salmonella enterica serovar Typhimurium SL1344 within epithelial cells and show comparisons with Salmonella gene expression inside macrophages. We report that S. Typhimurium expresses a characteristic intracellular transcriptomic signature in response to the environments it encounters within different cell types. The signature involves the upregulation of the mgtBC, pstACS and iro genes for magnesium, phosphate and iron uptake, and Salmonella pathogenicity island 2 (SPI2). Surprisingly, in addition to SPI2, the invasion-associated SPI1 pathogenicity island and the genes involved in flagellar biosynthesis were expressed inside epithelial cells at later stages of the infection, while they were constantly downregulated in macrophage-like cells. To our knowledge, this is the first report of the simultaneous transcription of all three Type Three Secretion Systems (T3SS) within an intracellular Salmonella population. We discovered that S. Typhimurium strain SL1344 was strongly cytotoxic to epithelial cells after 6 h of infection and hypothesize that the time-dependent changes in Salmonella gene expression within epithelial cells reflects the bacterial response to host cells that have been injured by the infection process.


Assuntos
Células Epiteliais/microbiologia , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/genética , Transcrição Gênica , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Células CACO-2 , Linhagem Celular , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos/genética , Células HeLa , Humanos , Imuno-Histoquímica , Macrófagos/citologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Salmonella typhimurium/ultraestrutura
11.
Planta ; 225(6): 1393-403, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17151887

RESUMO

One of the major factors contributing to the failure of new wheat varieties is seasonal variability in end-use quality. Consequently, it is important to produce varieties which are robust and stable over a range of environmental conditions. Recently developed sample preparation methods have allowed the application of FT-IR spectroscopic imaging methods to the analysis of wheat endosperm cell wall composition, allowing the spatial distribution of structural components to be determined without the limitations of conventional chemical analysis. The advantages of the methods, described in this paper, are that they determine the composition of endosperm cell walls in situ and with minimal modification during preparation. Two bread-making wheat cultivars, Spark and Rialto, were selected to determine the impact of environmental conditions on the cell-wall composition of the starchy endosperm of the developing and mature grain, focusing on the period of grain filling (starting at about 14 days after anthesis). Studies carried out over two successive seasons show that the structure of the arabinoxylans in the endosperm cell walls changes from a highly branched form to a less branched form. Furthermore, during development the rate of restructuring was faster when the plants were grown at higher temperature with restricted water availability from 14 days after anthesis with differences in the rate of restructuring occurring between the two cultivars.


Assuntos
Parede Celular/metabolismo , Meio Ambiente , Sementes/citologia , Triticum/citologia , Espectrofotometria Infravermelho , Fatores de Tempo
12.
Planta ; 220(5): 667-77, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15580525

RESUMO

Endosperm cell walls of cultivars of wheat (Triticum aestivum L.) selected for their endosperm texture (two soft and two hard) were analysed in situ by Fourier transform infrared (FTIR) microspectroscopy. FTIR imaging coupled with statistical analysis was used to map the compositional and structural heterogeneity within transverse sections from which cell contents had been removed by sonication. In the majority of grains analysed, two distinct populations of endosperm cells could be identified by spectral features that were related to cell morphology and age, regardless of cultivar. The main cell-wall component responsible for these differences was the polysaccharide arabinoxylan. In a few samples, this heterogeneity was absent, for reasons that are not understood, but this was not correlated to endosperm texture or growth conditions. Within the same population of endosperm cells, cell walls of hard endosperm could be distinguished from those of soft endosperm by their spectral features. Compared to hard cultivars, the peripheral endosperm of soft cultivars was characterised by a higher amount of polymer, whose spectral feature was similar to water-extractable arabinoxylan. In contrast, no specific compound has been identified in the central endosperm: structural differences within the polysaccharides probably contribute to the distinction between hard and soft cultivars. In developing grain, a clear difference in the composition of the endosperm cell walls of hard and soft wheat cultivars was observed as early as 15 days after anthesis.


Assuntos
Sementes/ultraestrutura , Triticum/ultraestrutura , Parede Celular/química , Parede Celular/ultraestrutura , Fenótipo , Sementes/fisiologia , Espectroscopia de Infravermelho com Transformada de Fourier , Triticum/fisiologia
13.
Carcinogenesis ; 25(10): 1945-52, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15155531

RESUMO

Epidemiological studies suggest that the use of NSAIDs and/or a high intake of fruit and vegetables reduce the risk of oesophageal adenocarcinoma. Since COX-2 is up-regulated in Barrett's oesophageal carcinogenesis, the protective effect of NSAIDs and natural food components might reflect COX-2 inhibition. We explored the effects of quercetin, a natural flavonoid with a potent COX-2 inhibitory activity, and two commercially available selective COX-2 inhibitors (NS-398 and nimesulide) on cell proliferation, apoptosis, PGE2 production and COX-2 mRNA expression in a human oesophageal adenocarcinoma cell line (OE33). Changes in the relative numbers of adherent and floating cells were quantified and apoptotic cells were identified using ethidium bromide and acridine orange staining under fluorescence microscopy. Flow cytometric analysis of adherent and floating cells was used to quantify apoptosis and to examine the effects of the agents on the cell cycle. After 48 h exposure at concentrations of > or =1 microM both COX-2 inhibitors and quercetin suppressed cell proliferation (P < 0.01) and increased the fraction of floating apoptotic cells. At higher concentrations (50 microM) and longer exposure (48 h) the effects of quercetin were significantly greater than those of the selective COX-2 inhibitors (P < 0.01). Cell cycle analyses showed that quercetin blocked cells in S phase, while the selective COX-2 inhibitors blocked cells in G1/S interphase. COX-2 mRNA expression was suppressed by quercetin and the synthetic COX-2 inhibitors in a time- and dose-dependent manner. Quercetin and the synthetic COX-2 inhibitors (10 microM) suppressed PGE2 production by approximately 70% after 24 h exposure (P < 0.001). We conclude that OE33 is a useful model for the study of COX-2 expression and associated phenomena in human adenocarcinoma cells. Synthetic COX-2 inhibitors and the food-borne flavonoid quercetin suppress proliferation, induce apoptosis and cell cycle block in human oesophageal adenocarcinoma cells in vitro, and future studies should assess their effects in vivo.


Assuntos
Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase/farmacologia , Neoplasias Esofágicas/patologia , Isoenzimas/antagonistas & inibidores , Adenocarcinoma/enzimologia , Adenocarcinoma/patologia , Ciclo-Oxigenase 2 , Inibidores de Ciclo-Oxigenase 2 , Neoplasias Esofágicas/enzimologia , Humanos , Proteínas de Membrana , Prostaglandina-Endoperóxido Sintases/metabolismo , Células Tumorais Cultivadas
14.
Compr Rev Food Sci Food Saf ; 2(4): 128-146, 2003 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33451229

RESUMO

Plant cell walls constitute the key structural components of plants and many plant-based foods. They are well known for contributing to a range of "quality" characteristics, from organoleptic texture to the properties of dietary fiber. Much of the research on cell walls has focused on the physiological aspects of plant growth and development with the belief that this route holds the key to controlling quality characteristics. In addition, consideration of quality has often been determined by what is easily measurable. This review assesses critically the role of plant cell walls in relation to the ultimate determinant of quality - the consumer, but within a whole food-chain context. We conclude that effective exploitation of cell-wall research in relation to optimizing quality requires an integrated approach taking into account the multi-functional roles of plant cell walls, and the diversity of consumer-related quality dimensions.

15.
Lett Appl Microbiol ; 34(3): 173-7, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-11874537

RESUMO

AIMS: To establish the site of microbial growth on naturally black fermented table olives, and to monitor the population dynamics of yeasts and selected micro-organisms together with the changes in organic acid profile and pH in the cover brine during fermentation. METHODS AND RESULTS: During fermentation, the numbers of Enterobacteriaceae and Pseudomonas spp. in the brine decreased whilst lactic acid bacteria and yeast populations increased. Scanning electron microscopy showed that a yeast-rich biofilm developed on the epicuticular wax of the olive skin during fermentation. Yeasts also predominated in the stomatal openings, but bacteria were more numerous in intercellular spaces in the sub-stomatal flesh. Citric, malic and tartaric acids were the major organic acids accumulating in the brine during fermentation. CONCLUSIONS: Micro-organisms associated with the skin, stomata and flesh in fermenting black olives may experience different local conditions to those prevailing in the cover brine. SIGNIFICANCE AND IMPACT OF THE STUDY: These are the first observations of the micro-organisms associated with the fruit of naturally fermented black olives and of the accumulation of specific organic acids during fermentation.


Assuntos
Microbiologia de Alimentos , Frutas/microbiologia , Sais/química , Sais/metabolismo , Leveduras/química , Leveduras/crescimento & desenvolvimento , Biofilmes , Fermentação , Frutas/crescimento & desenvolvimento , Concentração de Íons de Hidrogênio , Ácido Láctico/metabolismo , Leveduras/isolamento & purificação
16.
J Agric Food Chem ; 49(9): 4364-71, 2001 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-11559139

RESUMO

The crispness of fruits and vegetables is dependent, predominantly, on the maintenance of cell adhesion. There is a growing body of evidence to suggest that cell adhesion in plants is controlled at the edge of cell faces rather than across the entire cell surface. The aim of the current study has been to exploit antibody-labeling techniques in conjunction with methods that induce cell separation to explore the distribution of highly esterified and weakly esterified pectic polysaccharides on the cell surface. Potato parenchyma tissue was subjected to cooking and chemical treatments, which induced softening through cell separation. Scanning electron microscopy (SEM) revealed characteristic patterns on the surface of these separated cells, which outlined the imprint of neighboring cells. Monoclonal antibodies, JIM5 and JIM7, were used to locate weakly esterified and highly esterified pectin by silver-enhanced immunogold SEM. The edge-of-face structures labeled strongly with JIM5 but not JIM7, indicating that they contained polygalacturonic acid of low ester content. In addition, adhesion of the middle lamella to the face of the primary wall was found to differ from adhesion at the edge of each cell face. This, in conjunction with the antibody-labeling observations, complements previous transmission electron microscopy studies and is consistent with the edge-of-face regions having a specialist role in cell adhesion.


Assuntos
Adesão Celular/fisiologia , Parede Celular/ultraestrutura , Pectinas/análise , Solanum tuberosum/citologia , Anticorpos Monoclonais , Culinária , Microscopia Eletrônica de Varredura
17.
Planta ; 212(4): 606-11, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11525518

RESUMO

Previous mechanical studies using algae have concentrated on cell extension and growth using creep-type experiments, but there appears to be no published study of their failure properties. The mechanical strength of single large internode cell walls (up to 2 mm diameter and 100 mm in length) of the charophyte (giant alga) Chara corallina was determined by dissecting cells to give sheets of cell wall, which were then notched and fractured under tension. Tensile tests, using a range of notch sizes, were conducted on cell walls of varying age and maturity to establish their notch sensitivity and to investigate the propagation of cracks in plant cell walls. The thickness and stiffness of the walls increased with age whereas their strength was little affected. The strength of unnotched walls was estimated as 47+/-13 MPa, comparable to that of some grasses but an order of magnitude higher than that published for model bacterial cellulose composite walls. The strength was notch-sensitive and the critical stress intensity factor K1c was estimated to be 0.63+/-0.19 MNm(-3/2), comparable to published values for grasses.


Assuntos
Parede Celular , Clorófitas , Clorófitas/citologia , Modelos Biológicos , Estresse Mecânico , Resistência à Tração , Ferimentos e Lesões
18.
J Agric Food Chem ; 49(7): 3414-9, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11453784

RESUMO

It has been reported that caroubin, a protein mixture obtained from carob seeds, has rheological properties similar to those of gluten. Comparative studies of the effects of hydration and temperature on caroubin and gluten were carried out with the aid of NMR, FTIR, scanning electron microscopy, and differential scanning calorimetry techniques. The results show that caroubin has a more ordered structure than gluten and that hydration has little effect on its secondary structure when compared to gluten. Caroubin is more easily accessible to water than gluten, suggesting that caroubin is more hydrophilic in nature. On hydration, caroubin, like gluten, forms fibrillar structures and sheets.


Assuntos
Glutens/química , Polissacarídeos/química , Sementes/química , Varredura Diferencial de Calorimetria , Galactanos , Glutens/ultraestrutura , Espectroscopia de Ressonância Magnética , Mananas , Microscopia Eletrônica de Varredura , Gomas Vegetais , Polissacarídeos/ultraestrutura , Sementes/ultraestrutura , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura , Água
19.
Plant Cell ; 13(7): 1669-82, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11449058

RESUMO

The gene for a bacterial enoyl-CoA hydratase (crotonase) homolog (HCHL) previously shown to convert 4-coumaroyl-CoA, caffeoyl-CoA, and feruloyl-CoA to the corresponding hydroxybenzaldehydes in vitro provided an opportunity to subvert the plant phenylpropanoid pathway and channel carbon flux through 4-hydroxybenzaldehyde and the important flavor compound 4-hydroxy-3-methoxybenzaldehyde (vanillin). Expression of the Pseudomonas fluorescens AN103 HCHL gene in two generations of tobacco plants caused the development of phenotypic abnormalities, including stunting, interveinal chlorosis and senescence, curled leaf margins, low pollen production, and male sterility. In second generation progeny, the phenotype segregated with the transgene and transgenic siblings exhibited orange/red coloration of the vascular ring, distorted cells in the xylem and phloem bundles, and lignin modification/reduction. There was depletion of the principal phenolics concomitant with massive accumulation of novel metabolites, including the glucosides and glucose esters of 4-hydroxybenzoic acid and vanillic acid and the glucosides of 4-hydroxybenzyl alcohol and vanillyl alcohol. HCHL plants exhibited increased accumulation of transcripts for phenylalanine ammonia-lyase, cinnamate-4-hydroxylase, and 4-coumarate:CoA ligase, whereas beta-1,3-glucanase was suppressed. This study, exploiting the ability of a bacterial gene to divert plant secondary metabolism, provides insight into how plants modify inappropriately accumulated metabolites and reveals the consequences of depleting the major phenolic pools.


Assuntos
Proteínas de Bactérias/genética , Benzaldeídos/metabolismo , Enoil-CoA Hidratase/genética , Hidroliases/genética , Nicotiana/genética , Fenóis/metabolismo , Plantas Tóxicas , Pseudomonas fluorescens/genética , Acil Coenzima A/química , Acil Coenzima A/metabolismo , Antocianinas/biossíntese , Antioxidantes/química , Antioxidantes/metabolismo , Benzaldeídos/análise , Benzaldeídos/química , Enoil-CoA Hidratase/biossíntese , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Hidroliases/biossíntese , Fenóis/análise , Fenótipo , Estruturas Vegetais/química , Estruturas Vegetais/citologia , Plantas Geneticamente Modificadas , RNA Mensageiro , RNA de Plantas , Nicotiana/citologia , Nicotiana/metabolismo , Ácido Vanílico/metabolismo
20.
J Agric Food Chem ; 48(11): 5612-7, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11087527

RESUMO

The structure and mechanical properties of onions are important factors affecting their textural quality. The onion bulb consists of several layers of pigmented, papery scales surrounding fleshy storage scales that comprise an upper epidermis, an intermediate parenchyma tissue, and a lower epidermis. The purpose of this study was to examine the chemical composition of cell walls from the papery scales and outer fleshy scales of onion (Allium cepa L. cv. Sturon) in relation to their mechanical properties. Cell-wall material (CWM) was prepared from the component tissues and analyzed for its carbohydrate and phenolic composition. The CWMs were rich in uronic acid and glucose, with smaller quantities of arabinose, galactose, and xylose. In the fleshy scales, the lower epidermis contained relatively more galactose-rich pectic polysaccharides, whereas the upper epidermis and the papery scales contained virtually no galactose. Analysis of mechanical properties showed that the order of strength of the tissues was papery scales > fleshy scales, which were in the order lower epidermis > upper epidermis > intermediate parenchyma. The upper epidermis of fleshy scales was stronger in the vertical than the horizontal direction, and both orientations showed negligible notch sensitivity. Cyclohexane-trans-1,2-diaminetetraacetate-induced vortex-induced cell separation of the intermediate layer of fleshy scales indicated that calcium cross-linking may play an important role in cell-cell adhesion. A small but significant amount of ferulic acid was found in the walls, predominantly in the thick cuticle of the lower epidermis of fleshy scales. Alkali-labile wall-bound flavonoids were also detected.


Assuntos
Cebolas/química , Cebolas/citologia , Polissacarídeos/análise , Ácidos Carboxílicos/análise , Parede Celular/química , Monossacarídeos/análise , Fenóis/análise , Ácidos Urônicos/análise
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