RESUMO
Various cheese products are involved in outbreaks of listeriosis worldwide due to high consumption and prolonged refrigerated storage. The objective of this study was to determine the efficacy of using lactic acid bacteria and packaging with grapefruit seed extract (GSE) for controlling Listeria monocytogenes growth in soft cheese. Leuconostoc mesenteroides and Lactobacillus curvatus isolated from kimchi were used as a starter culture to make a soft cheese, which was inoculated with a cocktail strain of L. monocytogenes. The soft cheese was packed with low-density polyethylene, biodegradable polybutylene adipate-co-terephthalate (PBAT), low-density polyethylene with GSE, or PBAT with GSE and stored at 10°C and 15°C. Leuconostoc mesenteroides (LcM) better inhibited the growth of L. monocytogenes than Lb. curvatus. The PBAT with GSE film showed the best control for the growth of L. monocytogenes. When both LcM and PBAT with GSE were applied to the soft cheese, the growth of L. monocytogenes was inhibited significantly more than the use of LcM or PBAT with GSE alone. In all test groups, water activity, pH, and moisture on a fat-free basis decreased, and titratable acidity increased compared with the control group. These results suggest that LcM isolated from kimchi and PBAT with GSE packaging film can be used as a hurdle technology to lower the risk of L. monocytogenes in soft cheese at the retail market.
Assuntos
Queijo/microbiologia , Citrus paradisi/química , Lactobacillales/fisiologia , Listeria monocytogenes/efeitos dos fármacos , Extratos Vegetais/farmacologia , Animais , Queijo/análise , Microbiologia de Alimentos , Lactobacillus/fisiologia , Listeria monocytogenes/crescimento & desenvolvimento , PoliésteresRESUMO
Previously, we observed increased transcription levels of specific cytochrome P450 monooxygenase (P450) and adenosine triphosphate binding cassette (ABC) transporter genes in human body lice, Pediculus humanus humanus, following exposure to ivermectin using the non-invasive induction assay, which resulted in tolerance. To confirm the roles of these genes in induction and tolerance, the robust genetic model insect Drosophila melanogaster was chosen. Orthologous genes corresponding to the body louse P450 (Cyp9f2, Cyp6g2 and Cyp9h1) and ABC transporter (Mrp1, GC1824 as an ABCB type and CG3327 as an ABCG type) genes were selected for in vivo bioassay. Following a brief treatment with a sublethal dose of ivermectin, the mortality response was significantly slower, indicating the presence of tolerance. Concurrently, the transcription levels of Cyp9f2 and Mrp1 at 3 h and those of Cyp6g2, Cyp9h1, Mrp1, CG1824 and CG3327 at 6 h post-treatment were upregulated, indicating gene induction. In behavioural bioassay using GAL4/UAS-RNA interference transgenic fly lines, increased susceptibility to ivermectin was observed following heat shock in the Cyp9f2 , Cyp6g2 , Cyp9h1 , Mrp1 or CG3327-knockdown flies. Considering that these five genes are orthologous to those which had the largest over-expression level following ivermectin-induced tolerance in the body louse, the current results suggest that they are also associated with ivermectin detoxification in D. melanogaster and that body lice and D. melanogaster are likely to share, in part, similar mechanisms of tolerance to ivermectin.
Assuntos
Drosophila melanogaster/genética , Tolerância a Medicamentos/genética , Inativação Metabólica/genética , Inseticidas , Ivermectina , Animais , Feminino , Resistência a Inseticidas , Interferência de RNARESUMO
We previously observed that ivermectin-induced detoxification genes, including ATP binding cassette transporter C4 (PhABCC4) and cytochrome P450 6CJ1 (CYP6CJ1) were identified from body lice following a brief exposure to a sublethal dose of ivermectin using a non-invasive induction assay. In this current study, the functional properties of PhABCC4 and CYP6CJ1 were investigated after expression in either X. laevis oocytes or using a baculovirus expression system, respectively. Efflux of [3 H]-9-(2-phosphonomethoxyethyl) adenine ([3 H]-PMEA), a known ABCC4 substrate in humans, was detected from PhABCC4 cRNA-injected oocytes by liquid scintillation spectrophotometric analysis and PhABCC4 expression in oocytes was confirmed using ABC transporter inhibitors. Efflux was also determined to be ATP-dependent. Using a variety of insecticides in a competition assay, only co-injection of ivermectin and dichlorodiphenyltrichloroethane led to decreased efflux of [3 H]-PMEA. PhABCC4-expressing oocytes also directly effluxed [3 H]-ivermectin, which increased over time. In addition, ivermectin appeared to be oxidatively metabolized and/or sequestered, although at low levels, following functional expression of CYP6CJ1 along with cytochrome P450 reductase in Sf9 cells. Our study suggests that PhABCC4 and perhaps CYP6CJ1 are involved in the Phase III and Phase I xenobiotic metabolism of ivermectin, respectively, and may play an important role in the evolution of ivermectin resistance in lice and other insects as field selection occurs.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Sistema Enzimático do Citocromo P-450/genética , Proteínas de Insetos/genética , Inseticidas/metabolismo , Ivermectina/metabolismo , Pediculus/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Inativação Metabólica , Proteínas de Insetos/metabolismo , Pediculus/metabolismoRESUMO
Human body and head lice are highly related haematophagous ectoparasites but only the body louse has been shown to transmit Bartonella quintana, the causative agent of trench fever. The mechanisms by which body lice became a vector for B. quintana, however, are poorly understood. Following oral challenge, green fluorescent protein-expressing B. quintana proliferated over 9 days postchallenge with the number of bacteria being significantly higher in whole body vs. head lice. The numbers of B. quintana detected in faeces from infected lice, however, were approximately the same in both lice. Nevertheless, the viability of B. quintana was significantly higher in body louse faeces. Comparison of immune responses in alimentary tract tissues revealed that basal transcription levels of peptidoglycan recognition protein and defensins were lower in body lice and the transcription of defensin 1 was up-regulated by oral challenge with wild-type B. quintana in head but not in body lice. In addition, the level of cytotoxic reactive oxygen species generated by epithelial cells was significantly lower in body lice. Although speculative at this time, the reduced immune response is consistent with the higher vector competence seen in body vs. head lice in terms of B. quintana infection.
Assuntos
Bartonella quintana/fisiologia , Insetos Vetores/microbiologia , Pediculus/microbiologia , Febre das Trincheiras/transmissão , Animais , Trato Gastrointestinal/metabolismo , Proteínas de Fluorescência Verde , Humanos , Pediculus/imunologia , Pediculus/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Head and body lice are both blood-feeding parasites of humans although only the body louse is a potent disease vector. In spite of numerous morphological and life history differences, head and body lice have recently been hypothesized to be ecotypes of the same species. We took a comparative genomics approach to measure nucleotide diversity by comparing expressed sequence tag data sets from head and body lice. A total of 10 771 body louse and 10 770 head louse transcripts were predicted from a combined assembly of Roche 454 and Illumina sequenced cDNAs from whole body tissues collected at all life stages and during pesticide exposure and bacterial infection treatments. Illumina reads mapped to the 10 775 draft body louse gene models from the whole genome assembly predicted nine presence/absence differences, but PCR confirmation resulted in a single gene difference. Read per million base pair estimates indicated that 14 genes showed significant differential expression between head and body lice under our treatment conditions. One novel microRNA was predicted in both lice species and 99% of the 544 transcripts from Candidatus riesia indicate that they share the same endosymbiont. Overall, few differences exist, which supports the hypothesis that these two organisms are ecotypes of the same species.
Assuntos
Pediculus/metabolismo , Transcriptoma , Animais , Éxons , Etiquetas de Sequências Expressas , Feminino , Perfilação da Expressão Gênica , Humanos , Pediculus/genética , Reação em Cadeia da Polimerase , Análise de Sequência de DNARESUMO
Transcriptional profiling results, using our non-invasive induction assay {short exposure intervals (2-5 h) to sublethal amounts of insecticides [< lethal concentration 3% (LC(3)) at 24 h] administered by stress-reducing means (contact vs. immersion screen) and with induction assessed in a time frame when tolerance is still present [~lethal concentration 90% (LC(90)) in 2-4 h]}, showed that ivermectin-induced detoxification genes from body lice are identified by quantitative real-time PCR analyses. Of the cytochrome P450 monooxygenase and ATP binding cassette transporter genes induced by ivermectin, CYP6CJ1, CYP9AG1, CYP9AG2 and PhABCC4 were respectively most significantly over-expressed, had high basal expression levels and were most closely related to genes from other organisms that metabolized insecticides, including ivermectin. Injection of double-stranded RNAs (dsRNAs) against either CYP9AG2 or PhABCC4 into non-induced female lice reduced their respective transcript level and resulted in increased sensitivity to ivermectin, indicating that these two genes are involved in the xenobiotic metabolism of ivermectin and in the production of tolerance.
Assuntos
Antiparasitários/administração & dosagem , Resistência a Medicamentos/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Ivermectina/administração & dosagem , Pediculus/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Antiparasitários/metabolismo , Antiparasitários/toxicidade , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Dexametasona , Feminino , Perfilação da Expressão Gênica , Genes de Insetos , Humanos , Inativação Metabólica/genética , Ivermectina/metabolismo , Ivermectina/toxicidade , Pediculus/efeitos dos fármacos , Pediculus/genética , Filogenia , Interferência de RNA , VerapamilRESUMO
The majority of reports concern external snapping hips caused by the iliotibial band. Comparatively, little information is available regarding snapping hips caused by a gluteus maximus. Here we show that hip problems caused by a tight gluteus maximus can be treated using a modified Z-plasty technique. Fourteen hips in seven patients were diagnosed as snapping hips caused by a tight gluteus maximus. The main functional impairment is that when the hips were flexed, legs were abducted widely and could not be adducted. All had functional impairments irresponsive to conservative treatments besides snapping, whether painful or not, and all patients underwent surgery using a modified Z-plasty technique on the iliotibial band. All patients were followed up and the mean follow-up was 7 years. All patients had complete resolution of functional impairments, snapping, and pain after surgery. No patient needed revision surgery, and there were no complications, such as, abductor weakness, or irritation over the greater trochanter. We suggest that the intrinsic tendon contracture can cause serious functional impairment in patients with snapping due to a tight gluteus maximus. In this context, a modified Z-plasty technique offers a good surgical approach.
Assuntos
Nádegas/cirurgia , Articulação do Quadril/cirurgia , Músculo Esquelético/cirurgia , Adulto , Feminino , Humanos , Masculino , Amplitude de Movimento Articular/fisiologia , Estudos Retrospectivos , Resultado do TratamentoRESUMO
The molecular mechanisms and genetics of abamectin resistance mediated by target site insensitivity in the two-spotted spider mite, Tetranychus urticae, were investigated by comparing two isogenic abamectin-susceptible (AbaS) and abamectin-resistant (AbaR) strains. Cloning and sequencing of full-length cDNA fragments of gamma-amino butyric acid (GABA)-gated chloride channel genes revealed no polymorphisms between the two strains. However, sequence comparison of the full-length cDNA fragment of a T. urticae glutamate-gated chloride channel gene (TuGluCl) identified a G323D point mutation as being tentatively related with abamectin resistance. In individual F(2) progenies obtained by backcrossing, the G323D genotype was confirmed to correlate with abamectin resistance. Bioassays using progeny from reciprocal crossings revealed that the abamectin resistance trait resulting from TuGluCl insensitivity is incompletely recessive.
Assuntos
Canais de Cloreto/genética , Resistência a Inseticidas/efeitos dos fármacos , Resistência a Inseticidas/genética , Ivermectina/análogos & derivados , Mutação Puntual/genética , Tetranychidae/efeitos dos fármacos , Tetranychidae/genética , Sequência de Aminoácidos , Substituição de Aminoácidos/genética , Animais , Canais de Cloreto/química , Cruzamentos Genéticos , Esterases/metabolismo , Feminino , Genótipo , Padrões de Herança/efeitos dos fármacos , Padrões de Herança/genética , Ivermectina/farmacologia , Masculino , Oxigenases de Função Mista/metabolismo , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Tetranychidae/enzimologia , Transcrição Gênica/efeitos dos fármacosRESUMO
The human body louse, Pediculus humanus humanus, has one of the smallest insect genomes, containing â¼10 775 annotated genes. Annotation of detoxification [cytochrome P450 monooxygenase (P450), glutathione-S-transferase (GST), esterase (Est) and ATP-binding cassette transporter (ABC transporter)] genes revealed that they are dramatically reduced in P. h. humanus compared to other insects except for Apis mellifera. There are 37 P450, 13 GST and 17 Est genes present in P. h. humanus, approximately half the number found in Drosophila melanogaster and Anopheles gambiae. The number of putatively functional ABC transporter genes in P. h. humanus and Ap. mellifera are the same (36) but both have fewer than An. gambiae (44) or Dr. melanogaster (65). The reduction of detoxification genes in P. h. humanus may be a result of this louse's simple life history, in which it does not encounter a wide variety of xenobiotics. Neuronal component genes are highly conserved across different insect species as expected because of their critical function. Although reduced in number, P. h. humanus still retains at least a minimum repertoire of genes known to confer metabolic or toxicokinetic resistance to xenobiotics (eg Cyp3 clade P450s, Delta GSTs, B clade Ests and B/C subfamily ABC transporters), suggestive of its high potential for resistance development.
Assuntos
Genes de Insetos , Modelos Animais , Pediculus/genética , Pediculus/metabolismo , Xenobióticos/metabolismo , Transportadores de Cassetes de Ligação de ATP/metabolismo , Sequência de Aminoácidos , Animais , Sistema Enzimático do Citocromo P-450/genética , Esterases/química , Esterases/genética , Genes Mitocondriais , Glutationa Transferase/genética , Inativação Metabólica , Insetos/genética , Resistência a Inseticidas , Dados de Sequência Molecular , Família Multigênica , Pediculus/enzimologia , Filogenia , Receptores Nicotínicos/metabolismo , Alinhamento de Sequência , Canais de Sódio/metabolismoRESUMO
Indenone KR-62776 acts as an agonist of PPAR gamma without inducing obesity in animal models and cells. X-ray crystallography reveals that the indenone occupies the binding pocket in a different manner than rosiglitazone. 2-Dimensional gel-electrophoresis showed that the expression of 42 proteins was altered more than 2.0-fold between KR-62776- or rosiglitazone-treated adipocyte cells and control cells. Rosiglitazone down-regulated the expression of ERK1/2 and suppressed the phosphorylation of ERK1/2 in these cells. However, the expression of ERK1/2 was up-regulated in KR-62776-treated cells. Phosphorylated ERK1/2, activated by indenone, affects the localization of PPAR gamma, suggesting a mechanism for indenone-inhibition of adipogenesis in 3T3-L1 preadipocyte cells. The preadipocyte cells are treated with ERK1/2 inhibitor PD98059, a large amount of the cells are converted to adipocyte cells. These results support the conclusion that the localization of PPAR gamma is one of the key factors explaining the biological responses of the ligands.
Assuntos
Adipócitos/citologia , Diferenciação Celular/efeitos dos fármacos , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Indanos/farmacologia , Oximas/farmacologia , PPAR gama/metabolismo , Células 3T3-L1 , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Animais , Cristalografia por Raios X , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , MAP Quinases Reguladas por Sinal Extracelular/genética , Regulação da Expressão Gênica , Indanos/química , Indanos/metabolismo , Camundongos , Modelos Moleculares , Estrutura Molecular , Oximas/química , Oximas/metabolismo , PPAR gama/agonistas , PPAR gama/química , Ligação Proteica , Estrutura Terciária de Proteína , Proteoma/análise , Proteoma/efeitos dos fármacos , Rosiglitazona , Tiazolidinedionas/química , Tiazolidinedionas/metabolismo , Tiazolidinedionas/farmacologiaRESUMO
We developed and validated secondary models that can predict growth parameters of Salmonella Typhimurium and Staphylococcus aureus in cooked-pressed ready-to-eat (RTE) pork as a function of concentrations (0 to 3%) of a commercial potassium lactate and sodium diacetate mixture (PL+SDA) and temperature (10 to 30°C). The primary growth data were fitted to a Gompertz equation to determine the lag time (LT) and growth rate (GR). At 10°C, the growth of Salmonella Typhimurium and S. aureus in cooked-pressed RTE pork containing 2% and 3% PL+SDA was completely inhibited. The effects of temperature and concentration of PL+SDA on the growth kinetics of Salmonella Typhimurium and S. aureus in cooked-pressed RTE pork were modeled by response surface analysis using polynomial models of the natural logarithm transformation of both LT and GR. Model performance was also evaluated by use of the prediction bias (B(f)) and accuracy (A(f)) factors, median relative error, and mean absolute relative error, as well as the acceptable prediction zone method. The results showed that LT and GR models of Salmonella Typhimurium and S. aureus in cooked-pressed RTE pork are acceptable models. Thus, both the LT and GR growth models developed herein can be used for the development of tertiary models for Salmonella Typhimurium and S. aureus in cooked-pressed RTE pork in the matrix of conditions described in the present study.
Assuntos
Manipulação de Alimentos/métodos , Conservantes de Alimentos/farmacologia , Produtos da Carne/microbiologia , Modelos Biológicos , Salmonella typhimurium/crescimento & desenvolvimento , Staphylococcus aureus/crescimento & desenvolvimento , Acetatos/farmacologia , Animais , Contagem de Colônia Microbiana , Qualidade de Produtos para o Consumidor , Microbiologia de Alimentos , Conservação de Alimentos/métodos , Humanos , Cinética , Potássio/farmacologia , Suínos , TemperaturaRESUMO
We investigated the growth kinetics and morphological changes in acid-stressed Salmonella Typhimurium as well as the antimicrobial effects of epsilon-polylysine (SAVE-ORY GL610) and combined potassium lactate (PL) and sodium diacetate (SDA) (PURASAL Opti.Form PD Plus) on acid-stressed S. Typhimurium. Exposure to 0.5% acetic or lactic acid injured over 90% of the S. Typhimurium population. Although the lag time of the injured S. Typhimurium was extended, the injured cells were recovered at 10 degrees C and 24 degrees C, indicating a risk of using 10 degrees C as a storage temperature. Additionally, 4.5% PL/SDA mixture or 2% epsilon-polylysine completely inhibited the growth of acid-stressed S. Typhimurium in broth at 10, 24, or 35 degrees C. Although 3% PL/SDA mixture inhibited the growth of lactic acid-stressed S. Typhimurium at 10 degrees C, it did not inhibit the growth of unstressed S. Typhimurium at the same temperature. This finding indicates a different antimicrobial effect due to the physiological status of the pathogen. Furthermore, acid-stressed S. Typhimurium was not resistant to epsilon-polylysine or the PL/SDA mixture, although the antimicrobial effect of these compounds was enhanced at a lower storage temperature. TEM analysis revealed that most of the stressed cells lost their cellular integrity and membranes partially. Both dead and doubling cells were observed after recovery at 30 degrees C for 12 h. The addition of 2% epsilon-polylysine or 4.5% PL/SDA mixture resulted in the collapse of the structure of S. Typhimurium cells and cytoplasmic materials being released. These results provide valuable information regarding the morphological and physiological responses of acid-stressed S. Typhimurium cells in broth and chicken patties followed by antimicrobial stress with epsilon-polylysine or PL/SDA mixture.
Assuntos
Manipulação de Alimentos/métodos , Conservantes de Alimentos/farmacologia , Lactatos/farmacologia , Produtos Avícolas/microbiologia , Salmonella typhimurium/crescimento & desenvolvimento , Acetato de Sódio/farmacologia , Animais , Galinhas , Contagem de Colônia Microbiana , Relação Dose-Resposta a Droga , Microbiologia de Alimentos , Conservação de Alimentos/métodos , Concentração de Íons de Hidrogênio , Microscopia Eletrônica de Transmissão , Polilisina , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/ultraestrutura , Estresse Fisiológico , Temperatura , Fatores de TempoRESUMO
We describe here a general technique for the graded inhibition of cellular excitability in vivo. Inhibition is accomplished by expressing a genetically modified Shaker K(+) channel (termed the EKO channel) in targeted cells. Unlike native K(+) channels, the EKO channel strongly shunts depolarizing current: activating at potentials near E(K) and not inactivating. Selective targeting of the channel to neurons, muscles, and photoreceptors in Drosophila using the Gal4-UAS system results in physiological and behavioral effects consistent with attenuated excitability in the targeted cells, often with loss of neuronal function at higher transgene dosages. By permitting the incremental reduction of electrical activity, the EKO technique can be used to address a wide range of questions regarding neuronal function.
Assuntos
Drosophila melanogaster/genética , Marcação de Genes , Potenciais da Membrana/genética , Sistema Nervoso/metabolismo , Neurônios/metabolismo , Canais de Potássio/genética , Adaptação Fisiológica/genética , Animais , Comportamento Animal/fisiologia , Células Cultivadas , Proteínas de Drosophila , Drosophila melanogaster/citologia , Drosophila melanogaster/metabolismo , Feminino , Dosagem de Genes , Regulação da Expressão Gênica no Desenvolvimento/genética , Genes Letais/fisiologia , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Músculos/embriologia , Músculos/metabolismo , Músculos/fisiopatologia , Mutação/fisiologia , Sistema Nervoso/citologia , Sistema Nervoso/embriologia , Inibição Neural/genética , Neurônios/citologia , Neurônios Aferentes/citologia , Neurônios Aferentes/metabolismo , Fenótipo , Células Fotorreceptoras de Invertebrados/citologia , Células Fotorreceptoras de Invertebrados/metabolismo , Canais de Potássio/metabolismo , Superfamília Shaker de Canais de Potássio , Transmissão Sináptica/genética , Transgenes/fisiologiaRESUMO
Vibrio parahaemolyticus is recognized as the leading cause of human gastroenteritis associated with the consumption of seafood. The objective of this study was to model the growth kinetics of pathogenic and nonpathogenic V. parahaemolyticus in broth and oyster slurry. Primary growth models of V. parahaemolyticus in broth and oyster slurry fit well to a modified Gomperz equation (broth R(2)=0.99; oyster slurry R(2)=0.96). The lag time (LT), specific growth rate (SGR), and maximum population density (MPD) of each primary model were compared. The growth of nonpathogenic V. parahaemolyticus was found to be more rapid than that of pathogenic V. parahaemolyticus, regardless of the model medium. In addition, significant (P<0.05) differences in the growth kinetics between pathogenic and nonpathogenic V. parahaemolyticus in broth were observed at 10 degrees C. When compared to growth in broth, the growth of V. parahaemolyticus was delayed in oyster slurry, and growth was not observed at 10 or 15 degrees C. The Davey and square root models were identified as appropriate secondary models for predicting the LT and SGR, respectively. For the broth model, the average B(f) and A(f) values for LT were found to be 0.97 and 1.3, respectively, whereas the average B(f) and A(f) values for SGR were 1.05 and 1.11, respectively. The model generated in this study predicted an LT that was shorter and an SGR that was similar to those that were actually observed, which indicates that these models provide a reliable and safe prediction of V. parahaemolyticus growth.
Assuntos
Modelos Biológicos , Ostreidae/microbiologia , Frutos do Mar/microbiologia , Temperatura , Vibrio parahaemolyticus/crescimento & desenvolvimento , Animais , Contagem de Colônia Microbiana , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Humanos , Cinética , Coreia (Geográfico) , Vibrio parahaemolyticus/patogenicidadeRESUMO
We have examined the results obtained with 72 NexGen legacy posterior stabilised-flex fixed total knee replacements in 47 patients implanted by a single surgeon between March 2003 and September 2004. Aseptic loosening of the femoral component was found in 27 (38%) of the replacements at a mean follow-up of 32 months (30 to 48) and 15 knees (21%) required revision at a mean of 23 months (11 to 45). We compared the radiologically-loose and revised knees with those which had remained well-fixed to identify the factors which had contributed to this high rate of aseptic loosening. Post-operatively, the mean maximum flexion was 136 degrees (110 degrees to 140 degrees) in the loosened group and 125 degrees (95 degrees to 140 degrees) in the well-fixed group (independent t-test, p = 0.022). Squatting, kneeling, or sitting cross-legged could be achieved by 23 (85%) of the loosened knees, but only 22 (49%) of the well-fixed knees (chi-squared test, p = 0.001). The loosened femoral components were found to migrate into a more flexed position, but no migration was detected in the well-fixed group. These implants allowed a high degree of flexion, but showed a marked rate of early loosening of the femoral component, which was associated with weight-bearing in maximum flexion.
Assuntos
Artroplastia do Joelho/reabilitação , Falha de Prótese , Amplitude de Movimento Articular/fisiologia , Atividades Cotidianas , Idoso , Artroplastia do Joelho/métodos , Fenômenos Biomecânicos , Feminino , Humanos , Prótese do Joelho , Coreia (Geográfico) , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/diagnóstico por imagem , Complicações Pós-Operatórias/fisiopatologia , Desenho de Prótese , Radiografia , Estudos Retrospectivos , Resultado do TratamentoRESUMO
The objective of this study was to determine the antimicrobial effect of a combination of potassium lactate and sodium diacetate (0, 1.8, 3, and 4.5%; PURASAL P Opti. Form 4, 60% solution) on the survival and growth of Listeria monocytogenes Scott A in pH-adjusted broth (5.5, 6.0, 6.5, and 7.0) stored at 4, 10, 17, 24, 30, and 37 degrees C. Appropriate dilutions of broth were enumerated by spiral plating on tryptose agar and counted with an automated colony counter. Growth data were iteratively fit, using nonlinear regression analysis to a three-phase linear model, using GraphPad PRISM. At pH 5.5, the combination of lactate-diacetate fully inhibited (P < 0.001) the growth of L. monocytogenes at all four levels and six temperatures. At pH 6.0, addition of 1.8% lactate-diacetate reduced (P < 0.001) the specific growth rate of L. monocytogenes and increased lag time; however, 3 and 4.5% completely inhibited the growth at the six temperatures studied. Efficacy of the lactate-diacetate mixture was decreased as pH increased and incubation temperature increased. Thus, at pH 6.5, at least 3% was required to retard (P < 0.001) the growth of L. monocytogenes in broth. There was a limited effect of the lactate-diacetate level on the specific growth rate of the pathogen at pH 7.0. However, 1.8 and 3% significantly lengthened the lag time at 4 and 10 degrees C. These results suggest that 1.8% of lactate-diacetate mixture can be used as a substantial hurdle to the growth of L. monocytogenes when refrigerated temperatures are maintained for products with pH less than 6.5.
Assuntos
Manipulação de Alimentos/métodos , Conservação de Alimentos/métodos , Lactatos/farmacologia , Listeria monocytogenes/crescimento & desenvolvimento , Acetato de Sódio/farmacologia , Contagem de Colônia Microbiana , Meios de Cultura/química , Relação Dose-Resposta a Droga , Microbiologia de Alimentos , Conservantes de Alimentos/farmacologia , Concentração de Íons de Hidrogênio , Listeria monocytogenes/efeitos dos fármacos , Temperatura , Fatores de TempoRESUMO
NAD+ (oxidized form of NAD:nicotinamide adenine dinucleotide)-reducing soluble [NiFe]-hydrogenase (SH) is phylogenetically related to NADH (reduced form of NAD+):quinone oxidoreductase (complex I), but the geometrical arrangements of the subunits and Fe-S clusters are unclear. Here, we describe the crystal structures of SH in the oxidized and reduced states. The cluster arrangement is similar to that of complex I, but the subunits orientation is not, which supports the hypothesis that subunits evolved as prebuilt modules. The oxidized active site includes a six-coordinate Ni, which is unprecedented for hydrogenases, whose coordination geometry would prevent O2 from approaching. In the reduced state showing the normal active site structure without a physiological electron acceptor, the flavin mononucleotide cofactor is dissociated, which may be caused by the oxidation state change of nearby Fe-S clusters and may suppress production of reactive oxygen species.
Assuntos
Proteínas de Bactérias/química , Hidrogenase/química , NAD/química , Sítios de Ligação , Oxirredução , Conformação Proteica , Subunidades Proteicas/química , SolubilidadeRESUMO
Memory impairments, which occur regularly across species as a result of aging, disease and psychological insults (for example, stress), constitute a useful area for investigation into the neurobiological basis of learning and memory. Memory researchers have identified the hippocampus as a crucial brain structure involved in key aspects of memory formation. The most widely accepted putative mechanisms of memory storage in this structure are LTP and LTD. The hippocampus is enriched with receptors for corticosterone (a glucocorticoid hormone released in response to stress) and it plays a role in glucocorticoid negative feedback and, therefore, some hippocampal functioning might be particularly susceptible to stress. In support of this view, stress-induced modifications in learning, synaptic plasticity and endangerment of neurons have been seen in the hippocampus. Stress and glucocorticoids appear to exert a metaplastic effect through the modulation of Ca2+ levels. We propose a synaptic model that provides a conceptual scaffold to structure our understanding of the manifold actions of stress on the hippocampus. Accordingly, we suggest that stress-induced metaplasticity could disrupt Ca2+ homeostasis and thus endanger hippocampal neurons.
Assuntos
Hipocampo/patologia , Metaplasia/etiologia , Estresse Fisiológico/patologia , Animais , Cálcio/fisiologia , Glucocorticoides/fisiologia , Humanos , Aprendizagem/fisiologia , Potenciação de Longa Duração/fisiologia , Memória/fisiologia , Modelos Biológicos , Plasticidade Neuronal/fisiologia , Estresse Fisiológico/complicaçõesRESUMO
A case-control study was performed to assess the potential influence of catechol O-methyl transferase (COMT) genotype on the risk of breast cancer in Korean women. One hundred and sixty-three histologically confirmed incident breast cancer cases and 163 age- and menopausal status-matched control individuals with no present or previous history of cancer were selected as study subjects. COMT genetic polymorphism was determined by gel electrophoresis after NlaIII enzyme digestion of amplified DNA. Odds ratios and 95% confidence intervals were estimated by unconditional logistic regression after adjustment for known or suspected risk factors of breast cancer. Women with at least one COMT lower enzyme activity associated allele (COMT-L) were at elevated risk for breast cancer (OR = 1.7, 95% CI = 1.04-2.78) compared with those homozygous for high enzyme activity associated COMT-H alleles. Among women with low (> or = 23.1) body mass index the COMT-L allele containing genotypes posed a marginally significant increased risk of breast cancer compared to the COMT-HH genotype (OR = 1.8, 95% CI = 0.95-3.48). Women with at least one COMT-L allele who had experienced a full-term pregnancy when aged over 30 years or were nulliparous had 2.7-fold increased risk; however, this increase did not reach statistical significance (OR = 2.7, 95% CI = 0.64-11.35). Furthermore, never-drinking and never-smoking women with at least one COMT-L allele were at increased risk of breast cancer compared to those with COMT-HH genotype with ORs of 2.0 (95% CI = 1.23-3.38) and 1.7 (95% CI = 1.04-2.62), respectively. These results are consistent with studies showing that COMT genotype of lower enzyme activity might be related to increase in risk of breast cancer, and extend this finding to Korean women.
Assuntos
Neoplasias da Mama/enzimologia , Neoplasias da Mama/genética , Catecol O-Metiltransferase/genética , Polimorfismo Genético , Adulto , Idoso , Alelos , Estudos de Casos e Controles , Estrogênios/metabolismo , Feminino , Genótipo , Humanos , Coreia (Geográfico) , Menopausa , Pessoa de Meia-Idade , Neoplasias Hormônio-Dependentes/enzimologia , Neoplasias Hormônio-Dependentes/genética , Fatores de RiscoRESUMO
Morphometric study was performed on transverse sections of microdissected muscle spindles from rabbit tenuissimus muscles. It showed a statistically significant difference in size and structure between the intrafusal capillaries and those of extrafusal muscle. The former are larger in diameter, circumference, and area due to a proportionate increase in the number of endothelial cells. Vesicles within spindle endothelial cells are fewer, mitochondrial counts are greater, but in proportion to the increased number of endothelial cells, intercellular junctions are tight and pericyte coverage is greater. The basement membrane around endothelial cells and pericytes is thicker and more often multilayered. The endoneurial capillaries of tenuissimus intramuscular nerve are morphologically identical to intrafusal capillaries. The similarity of the capillaries of spindles to those in brain suggests that a blood-nervous system barrier extends from brain into the peripheral nerves and muscle spindles.