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1.
Meat Sci ; 204: 109289, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37531898

RESUMO

The objectives were to determine the effects of post-thermal processing nitrite-embedded film (NEF) packaging on the quality attributes of alternatively-cured (nitrite from celery juice powder (AC)) and nitrite-free bologna. Attributes evaluated included lipid oxidation, instrumental color, pigment concentration, and sensory properties such as cured meat flavor, aroma, and color. Three bologna formulations, each packaged with two packaging films were produced. A conventionally-cured control formulation (with nitrite from sodium nitrite; CON), a nitrite-free formulation (UCC), and an alternatively cured formulation (nitrite from cultured celery juice powder; AC) were packaged in conventional (CF) or nitrite-embedded (NEF) film. Instrumental a* values (measured during both light and dark storage at intervals of 7 or 14 days over 126 days of storage) and cured pigment concentration (measured at 14-day intervals over 84 days of storage) were significantly greater (P < 0.05) for the UCC-NEF treatment compared to its conventional film counterpart, UCC-CF. No significant differences (P > 0.05) for lipid oxidation (TBARS values) were observed with NEF. Trained sensory panelists, who evaluated samples at 14-day intervals over 70 days of storage, found significantly greater (P < 0.05) cured aroma, cured flavor, pink color and less off-flavor for uncured bologna packaged in NEF compared to conventional film. For the uncured bologna formulation, NEF packaging provided cured meat attributes comparable to the control formulation that included nitrite. This is the first time that cured aroma and flavor have been observed when nitrite from packaging film is added to a cooked meat product under anaerobic conditions.


Assuntos
Produtos da Carne , Carne , Pós , Carne/análise , Produtos da Carne/análise , Nitrito de Sódio , Lipídeos
2.
J Food Prot ; 86(1): 100018, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36916598

RESUMO

Large, renowned outbreaks associated with low-moisture foods (LMFs) bring to light some of the potential, inherent risks that accompany foods with long shelf lives if pathogen contamination occurs. Subsequently, in 2013, Beuchat et al. (2013) noted the increased concern regarding these foods, specifically noting examples of persistence and resistance of pathogens in low-water activity foods (LWAFs), prevalence of pathogens in LWAF processing environments, and sources of and preventive measures for contamination of LWAFs. For the last decade, the body of knowledge related to LMF safety has exponentially expanded. This growing field and interest in LMF safety have led researchers to delve into survival and persistence studies, revealing that some foodborne pathogens can survive in LWAFs for months to years. Research has also uncovered many complications of working with foodborne pathogens in desiccated states, such as inoculation methods and molecular mechanisms that can impact pathogen survival and persistence. Moreover, outbreaks, recalls, and developments in LMF safety research have created a cascading feedback loop of pushing the field forward, which has also led to increased attention on how industry can improve LMF safety and raise safety standards. Scientists across academia, government agencies, and industry have partnered to develop and evaluate innovate thermal and nonthermal technologies to use on LMFs, which are described in the presented review. The objective of this review was to describe aspects of the extensive progress made by researchers and industry members in LMF safety, including lessons-learned about outbreaks and recalls, expansion of knowledge base about pathogens that contaminate LMFs, and mitigation strategies currently employed or in development to reduce food safety risks associated with LMFs.


Assuntos
Surtos de Doenças , Microbiologia de Alimentos , Surtos de Doenças/prevenção & controle , Inocuidade dos Alimentos , Manipulação de Alimentos/métodos , Alimentos , Contaminação de Alimentos/análise
3.
Meat Sci ; 184: 108687, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34656001

RESUMO

Pork bellies were injected with four different alternative curing brines. The bellies were inoculated on the surface and at a depth of 1 cm with multiple strains of Clostridium perfringens, Staphylococcus aureus and Salmonella enterica. The bellies were processed using either a standard process cycle or an interrupted process cycle to simulate a process deviation. Additionally, laboratory simulation of the same cycles was conducted where surface inoculated pork belly samples (22 ± 1 g) were processed in a circulating water bath. Microbiological populations were determined at the beginning, mid-point and end of the cycles, and the change in population was calculated for each bacterium at each time point, by comparing the population to the initial inoculated population. Irrespective of the brine or process cycle, the populations of all of the inoculated bacteria on both the surface and interior samples had decreased by the end of the process. There was no difference in the reductions in bacterial populations for all of the inoculated bacteria by brine type or by sample location (P > 0.30). There were differences in the microbial population reductions for C. perfringens attributable to the processing cycle (P < 0.001), with less population reductions associated with the standard cycle when compared to the interrupted cycle. However, no differences (P > 0.10) were observed in the population reductions between the two processing cycles for either S. aureus or S. enterica.


Assuntos
Clostridium perfringens/crescimento & desenvolvimento , Culinária , Manipulação de Alimentos/métodos , Produtos da Carne/microbiologia , Staphylococcus aureus/crescimento & desenvolvimento , Animais , Microbiologia de Alimentos , Salmonella enterica , Suínos
4.
Epidemiol Infect ; 149: e173, 2021 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-34668464

RESUMO

New Zealand has a strategy of eliminating SARS-CoV-2 that has resulted in a low incidence of reported coronavirus-19 disease (COVID-19). The aim of this study was to describe the spread of SARS-CoV-2 in New Zealand via a nationwide serosurvey of blood donors. Samples (n = 9806) were collected over a month-long period (3 December 2020-6 January 2021) from donors aged 16-88 years. The sample population was geographically spread, covering 16 of 20 district health board regions. A series of Spike-based immunoassays were utilised, and the serological testing algorithm was optimised for specificity given New Zealand is a low prevalence setting. Eighteen samples were seropositive for SARS-CoV-2 antibodies, six of which were retrospectively matched to previously confirmed COVID-19 cases. A further four were from donors that travelled to settings with a high risk of SARS-CoV-2 exposure, suggesting likely infection outside New Zealand. The remaining eight seropositive samples were from seven different district health regions for a true seroprevalence estimate, adjusted for test sensitivity and specificity, of 0.103% (95% confidence interval, 0.09-0.12%). The very low seroprevalence is consistent with limited undetected community transmission and provides robust, serological evidence to support New Zealand's successful elimination strategy for COVID-19.


Assuntos
Doadores de Sangue/estatística & dados numéricos , COVID-19/epidemiologia , COVID-19/prevenção & controle , Erradicação de Doenças/estatística & dados numéricos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Algoritmos , Anticorpos Antivirais/sangue , COVID-19/sangue , COVID-19/transmissão , Teste Sorológico para COVID-19 , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Nova Zelândia/epidemiologia , Prevalência , SARS-CoV-2/imunologia , Estudos Soroepidemiológicos , Adulto Jovem
5.
Clin Transl Immunology ; 10(3): e1261, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33747511

RESUMO

OBJECTIVES: Circulating antibodies are important markers of previous infection and immunity. Questions remain with respect to the durability and functionality of SARS-CoV-2 antibodies. This study explored antibody responses in recovered COVID-19 patients in a setting where the probability of re-exposure is effectively nil, owing to New Zealand's successful elimination strategy. METHODS: A triplex bead-based assay that detects antibody isotype (IgG, IgM and IgA) and subclass (IgG1, IgG2, IgG3 and IgG4) responses against Nucleocapsid (N) protein, the receptor binding domain (RBD) and Spike (S) protein of SARS-CoV-2 was developed. After establishing baseline levels with pre-pandemic control sera (n = 113), samples from PCR-confirmed COVID-19 patients with mild-moderate disease (n = 189) collected up to 8 months post-infection were examined. The relationship between antigen-specific antibodies and neutralising antibodies (NAbs) was explored with a surrogate neutralisation assay that quantifies inhibition of the RBD/hACE-2 interaction. RESULTS: While most individuals had broad isotype and subclass responses to each antigen shortly after infection, only RBD and S protein IgG, as well as NAbs, were relatively stable over the study period, with 99%, 96% and 90% of samples, respectively, having responses over baseline 4-8 months post-infection. Anti-RBD antibodies were strongly correlated with NAbs at all time points (Pearson's r ≥ 0.87), and feasibility of using finger prick sampling to accurately measure anti-RBD IgG was demonstrated. CONCLUSION: Antibodies to SARS-CoV-2 persist for up to 8 months following mild-to-moderate infection. This robust response can be attributed to the initial exposure without immune boosting given the lack of community transmission in our setting.

6.
Microb Drug Resist ; 27(8): 1144-1154, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33539269

RESUMO

Antimicrobial resistance (AMR) developed by Salmonella within animals used for food products is a major global issue. Monitoring AMR in animals destined for slaughter is, therefore, critical. Abattoirs may serve as potential candidate checkpoints for monitoring resistance patterns on farms. A complicating factor, however, is the impact of lairage on Salmonella detected in pigs at slaughter. This study sought to compare AMR patterns in Salmonella spp. in swine collected upon arrival (fecal samples) at the abattoir with those at postslaughter (cecal samples) and evaluate the feasibility of using slaughterhouse samples for surveillance of prevailing AMR Salmonella on farms. Eighty-four Salmonella isolates were recovered from a large, midwestern U.S. abattoir between September and November 2013. Isolates were tested for phenotypic AMR to 12 antimicrobials using the broth microdilution assay. Whole-genome sequencing identified the AMR genes harbored by the strains. Significant differences were observed in the isolate phenotypes and genotypes; however, no significant difference was observed in genotypic resistance patterns. Hence, the AMR profiles of Salmonella spp. postslaughter cannot be predicted from preslaughter samples. Further research considering the genetic diversity of isolates and statistical power of the genotypic analysis is warranted to improve the performance of WGS-inferred antimicrobial susceptibility.


Assuntos
Matadouros , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/fisiologia , Salmonella/efeitos dos fármacos , Animais , Farmacorresistência Bacteriana/genética , Genes Bacterianos , Testes de Sensibilidade Microbiana , Carne Vermelha/microbiologia , Salmonella/genética , Salmonelose Animal/microbiologia , Suínos , Doenças dos Suínos/microbiologia , Sequenciamento Completo do Genoma
7.
PeerJ ; 8: e9863, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32953275

RESUMO

BACKGROUND: Serological assays that detect antibodies to SARS-CoV-2 are critical for determining past infection and investigating immune responses in the COVID-19 pandemic. We established ELISA-based immunoassays using locally produced antigens when New Zealand went into a nationwide lockdown and the supply chain of diagnostic reagents was a widely held domestic concern. The relationship between serum antibody binding measured by ELISA and neutralising capacity was investigated using a surrogate viral neutralisation test (sVNT). METHODS: A pre-pandemic sera panel (n = 113), including respiratory infections with symptom overlap with COVID-19, was used to establish assay specificity. Sera from PCR­confirmed SARS-CoV-2 patients (n = 21), and PCR-negative patients with respiratory symptoms suggestive of COVID-19 (n = 82) that presented to the two largest hospitals in Auckland during the lockdown period were included. A two-step IgG ELISA based on the receptor binding domain (RBD) and spike protein was adapted to determine seropositivity, and neutralising antibodies that block the RBD/hACE­2 interaction were quantified by sVNT. RESULTS: The calculated cut-off (>0.2) in the two-step ELISA maximised specificity by classifying all pre-pandemic samples as negative. Sera from all PCR-confirmed COVID-19 patients were classified as seropositive by ELISA ≥7 days after symptom onset. There was 100% concordance between the two-step ELISA and the sVNT with all 7+ day sera from PCR­confirmed COVID-19 patients also classified as positive with respect to neutralising antibodies. Of the symptomatic PCR-negative cohort, one individual with notable travel history was classified as positive by two-step ELISA and sVNT, demonstrating the value of serology in detecting prior infection. CONCLUSIONS: These serological assays were established and assessed at a time when human activity was severely restricted in New Zealand. This was achieved by generous sharing of reagents and technical expertise by the international scientific community, and highly collaborative efforts of scientists and clinicians across the country. The assays have immediate utility in supporting clinical diagnostics, understanding transmission in high-risk cohorts and underpinning longer­term 'exit' strategies based on effective vaccines and therapeutics.

8.
Front Nutr ; 7: 619023, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33644106

RESUMO

Enteric pathogens such as Salmonella enterica can survive in low pH conditions and pose a food safety threat during marinating of raw poultry meat. A study was conducted to investigate the effectiveness of thyme oil for killing S. enterica on raw chicken during marination in lemon juice containing yucca extract. Samples of raw chicken breast were inoculated with a five-serovar mixture of S. enterica (~108 CFU/mL) and immersed for 2, 4, 6, and 8 h in four lemon-based marinades at 22°C: lemon juice alone (L), L with added 0.5% yucca extract (L + Y), L + Y and 0.5% thyme oil (L + Y + 0.5% TO) and L + Y + 1.0% TO. The L and L + Y served as controls. Survivors were determined by surface plating chicken homogenates on xylose-lysine tergitol-4 (XLT4) agar and XLT4 agar overlaid with non-selective agar (TAL) and counting bacterial colonies after 48 h of incubation (35°C). Marinades containing Y and TO significantly reduced initial viable populations of S. enterica compared to control (L and L + Y) solutions (P < 0.05). Based on S. enterica survivors on TAL medium, the L and L + Y reduced initial populations by 1.12 and 1.42 Log CFU/sample, respectively, after 8 h whereas, Log reductions caused by L + Y + 0.5% TO and L + Y + 1.0% TO, respectively, were 2.62 and 3.91 (P < 0.05). Numbers of survivors were higher on TAL compared to XLT4 agar (P < 0.05); however, the extent of sub-lethal injury caused by the marinades was not statistically significant (P > 0.05). The death rate of S. enterica increased significantly (P < 0.05) in the marinades containing TO (0.5 or 1.0%) compared to control (L + Y). Based on these results, thyme oil has good potential to increase the antimicrobial efficacy of lemon juice marinade against Salmonella on raw chicken breast and enhance the microbial safety of this popular poultry product.

9.
PLoS One ; 14(10): e0223752, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31665165

RESUMO

The Iceman site is unique in the bryology of the Quaternary. Only 21 bryophytes (mosses and liverworts) grow now in the immediate vicinity of the 5,300 year old Iceman discovery site at 3,210m above sea level in the Ötztal Alps, Italy. By contrast 75 or more species including at least ten liverworts were recovered as subfossils frozen in, on and around the Iceman from before, at and after his time. About two thirds of the species grow in the nival zone (above 3,000m above sea level) now while about one third do not. A large part of this third can be explained by the Iceman having both deliberately and inadvertently carried bryophytes during his last, fatal journey. Multivariate analyses (PCA, RDA) provide a variety of explanations for the arrivals of the bryophytes in the rocky hollow where the mummy was discovered. This is well into the nival zone of perennial snow and ice with a very sparse, non-woody flora and very low vegetation cover. Apart from the crucial anthropochory (extra-local plants), both hydrochory (local species) and zoochory (by wild game such as ibex of both local and extra-local species) have been important. Anemochory of mainly local species was of lesser importance and of extra-local species probably of little or no importance. The mosses Neckera complanata and several other ecologically similar species as well as a species of Sphagnum (bogmoss) strongly support the claim that the Iceman, took northwards up Schnalstal, South Tyrol, as the route of the last journey. A different species of bogmoss, taken from his colon is another indication the Iceman's presence at low altitude south of Schnalstal during his last hours when he was first high up, low down and finally at over 3,000m.


Assuntos
Briófitas , Fenômenos Ecológicos e Ambientais , Hepatófitas , Gelo , Arqueologia , Humanos
10.
Risk Anal ; 39(10): 2237-2258, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31039285

RESUMO

The inclusion of deep tissue lymph nodes (DTLNs) or nonvisceral lymph nodes contaminated with Salmonella in wholesale fresh ground pork (WFGP) production may pose risks to public health. To assess the relative contribution of DTLNs to human salmonellosis occurrence associated with ground pork consumption and to investigate potential critical control points in the slaughter-to-table continuum for the control of human salmonellosis in the United States, a quantitative microbial risk assessment (QMRA) model was established. The model predicted an average of 45 cases of salmonellosis (95% CI = [19, 71]) per 100,000 Americans annually due to WFGP consumption. Sensitivity analysis of all stochastic input variables showed that cooking temperature was the most influential parameter for reducing salmonellosis cases associated with WFGP meals, followed by storage temperature and Salmonella concentration on contaminated carcass surface before fabrication. The input variables were grouped to represent three main factors along the slaughter-to-table chain influencing Salmonella doses ingested via WFGP meals: DTLN-related factors, factors at processing other than DTLNs, and consumer-related factors. The evaluation of the impact of each group of factors by second-order Monte Carlo simulation showed that DTLN-related factors had the lowest impact on the risk estimate among the three groups of factors. These findings indicate that interventions to reduce Salmonella contamination in DTLNs or to remove DTLNs from WFGP products may be less critical for reducing human infections attributable to ground pork than improving consumers' cooking habits or interventions of carcass decontamination at processing.


Assuntos
Linfonodos/microbiologia , Carne Vermelha/microbiologia , Intoxicação Alimentar por Salmonella/epidemiologia , Infecções por Salmonella/epidemiologia , Salmonella/isolamento & purificação , Animais , Humanos , Medição de Risco , Intoxicação Alimentar por Salmonella/prevenção & controle , Suínos
11.
Ann N Y Acad Sci ; 1441(1): 17-30, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30924539

RESUMO

Antimicrobial resistance (AMR) is a significant threat to both human and animal health. The spread of AMR bacteria and genes across systems can occur through a myriad of pathways, both related and unrelated to agriculture, including via wastewater, soils, manure applications, direct exchange between humans and animals, and food exposure. Tracing origins and drivers of AMR bacteria and genes is challenging due to the array of contexts and the complexity of interactions overlapping health practice, microbiology, genetics, applied science and engineering, as well as social and human factors. Critically assessing the diverse and sometimes contradictory AMR literature is a valuable step in identifying tractable mitigation options to stem AMR spread. In this article we review research on the nonfoodborne spread of AMR, with a focus on domesticated animals and the environment and possible exposures to humans. Attention is especially placed on delineating possible sources and causes of AMR bacterial phenotypes, including underpinning the genetics important to human and animal health.


Assuntos
Animais Domésticos , Resistência Microbiana a Medicamentos , Meio Ambiente , Agricultura , Animais , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Bactérias/genética , Bactérias/isolamento & purificação , Portador Sadio , Resistência Microbiana a Medicamentos/genética , Fezes/microbiologia , Humanos , Higiene , Microbiologia do Solo , Microbiologia da Água
12.
Ann N Y Acad Sci ; 1441(1): 40-49, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30924543

RESUMO

Consumers are increasingly interested in the attributes of the food they consume. This includes what is in the food and how it was raised; and at least some consumers are willing to pay a premium for products with specific attributes. However, the current plethora of labels on the market does not adequately address this issue; rather than providing actionable information, most labels add to the consumer confusion. In addition, there is a tendency toward "absence labels" that can contribute to a negative consumer perception of conventional products that may or may not include the attribute in question. Communication with consumers about the complex and highly technical issue of antimicrobial resistance (AMR) is challenging, and experiences from communication efforts about food safety-related issues demonstrate exactly how challenging this is to communicate clearly. General lessons learned from the science of risk communication can help guide efforts to communicate about the challenging issue of AMR. There are efforts underway to chart out a new approach. A new labeled animal production certification program is under development to provide choice for consumers, while reducing consumer confusion, which mandates antibiotic stewardship practices.


Assuntos
Infecções Bacterianas/transmissão , Comportamento do Consumidor , Farmacorresistência Bacteriana , Microbiologia de Alimentos , Inocuidade dos Alimentos , Animais , Antibacterianos/administração & dosagem , Infecções Bacterianas/microbiologia , Rotulagem de Alimentos , Fatores de Risco
13.
Bioorg Med Chem ; 27(8): 1529-1545, 2019 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-30850264

RESUMO

Replacing one of the morpholine groups of the phosphatidylinositol 3-kinase (PI3K) inhibitor ZSTK474 with a variety of sulfonamide-linked solubilizing substituents produced a new class of active and potent PI3Kα inhibitors, with several derivatives demonstrating high PI3Kα enzyme potency and good cellular potency in two human derived cell lines. The overall results suggest a preference for linear and somewhat flexible solubilizing functions. From this series, compound 16, also known as SN32976, was selected for advanced preclinical evaluation.


Assuntos
Fosfatidilinositol 3-Quinases/química , Inibidores de Fosfoinositídeo-3 Quinase/síntese química , Sulfonamidas/química , Triazinas/química , Animais , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Feminino , Humanos , Concentração Inibidora 50 , Camundongos , Neoplasias/tratamento farmacológico , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores de Fosfoinositídeo-3 Quinase/farmacologia , Inibidores de Fosfoinositídeo-3 Quinase/uso terapêutico , Subunidades Proteicas/antagonistas & inibidores , Subunidades Proteicas/metabolismo , Relação Estrutura-Atividade , Sulfonamidas/farmacologia , Sulfonamidas/uso terapêutico , Transplante Heterólogo
14.
Chem Asian J ; 14(8): 1249-1261, 2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30714356

RESUMO

Using a scaffold-hopping approach, imidazo[1,2-a]pyridine analogues of the ZSTK474 (benzimidazole) class of phosphatidylinositol 3-kinase (PI3K) inhibitors have been synthesized for biological evaluation. Compounds were prepared using a heteroaryl Heck reaction procedure, involving the palladium-catalysed coupling of 2-(difluoromethyl)imidazo[1,2-a]pyridines with chloro, iodo or trifluoromethanesulfonyloxy (trifloxy) substituted 1,3,5-triazines or pyrimidines, with the iodo intermediates being preferred in terms of higher yields and milder reaction conditions. The new compounds maintain the PI3K isoform selectivity of their benzimidazole analogues, but in general show less potency.


Assuntos
Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/farmacologia , Inibidores de Fosfoinositídeo-3 Quinase , Piridinas/síntese química , Piridinas/farmacologia , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/química , Humanos , Estrutura Molecular , Fosfatidilinositol 3-Quinases/metabolismo , Piridinas/química , Relação Estrutura-Atividade
15.
Acta Pharmacol Sin ; 39(11): 1816-1822, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29991713

RESUMO

The phosphatidylinositol 3-kinase (PI3K) pathway is involved in many cellular functions including cell growth, metabolism, and transformation. Hyperactivation of this pathway contributes to tumorigenesis, therefore, PI3K is a major target for anticancer drug discovery. Since the PI3Kα isoform is implicated mostly in cancer, we conducted a high-throughput screening (HTS) campaign using a 3-step PI3K homogenous time-resolved fluorescence assay against this isoform bearing the H1047R mutation. A total of 288,000 synthetic and natural product-derived compounds were screened and of which, we identified 124 initial hits that were further selected by considering the predicted binding mode, relationship to known pan-assay interference compounds and previous descriptions as a lipid kinase inhibitor. A total of 24 compounds were then tested for concentration-dependent responses. These hit compounds provide novel scaffolds that can potentially be optimized to create novel PI3K inhibitors.


Assuntos
Isoenzimas/antagonistas & inibidores , Inibidores de Fosfoinositídeo-3 Quinase , Inibidores de Proteínas Quinases/química , Ensaios de Triagem em Larga Escala , Humanos , Ligação de Hidrogênio , Isoenzimas/genética , Isoenzimas/metabolismo , Simulação de Acoplamento Molecular , Mutação , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Ligação Proteica , Inibidores de Proteínas Quinases/metabolismo
16.
Foodborne Pathog Dis ; 15(9): 538-543, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29851358

RESUMO

Nonendospore-forming pathogenic bacteria in the long-term survival (LTS) phase can remain viable for months or years and may show reduced susceptibility to various antimicrobial interventions. In the present study, we investigated the response of LTS phase Salmonella enterica serovar Typhimurium (ATCC 14028) to ultraviolet (UV) radiation in 0.85% (w/v) saline and apple juice and the extent of sublethal injury in LTS phase survivors. The LTS-phase Salmonella Typhimurium cells were cultured at 35°C for 14 days in tryptic soy broth with 0.6% (w/v) yeast extract (TSBYE). Exponential- and stationary-phase cells, cultured in TSBYE (35°C) for 2.5 and 18 h, respectively, served as control samples. Cells (107 CFU [colony-forming unit]/mL) from each physiological state were exposed to UV light in saline (80 µW/cm2) and apple juice (1500 µW/cm2). The Salmonella Typhimurium survivors were plated for enumeration on either tryptic soy agar with 0.6% yeast extract or xylose-lysine-tergitol 4 (XLT4) agar and colonies counted after incubation (35°C, 24 h). Of all the growth phases tested, LTS phase cells were consistently impacted the least by UV treatment (p < 0.05). In saline, D-values of exponential, stationary, and LTS Salmonella Typhimurium were 0.35, 0.38, and 0.49 min, respectively. D-values in apple juice at pH 3.63 and pH 5.65 were 2.52, 3.19, and 3.57 min and 3.24, 3.50, and 4.18 min, respectively. UV radiation (80 µW/cm2) of Salmonella Typhimurium in saline for 2.5 min reduced the number of exponential- and stationary-phase cells by ∼7.19 and 6.30 log10 CFU/mL, respectively. In contrast, LTS cells were only reduced by 5.08 log10 CFU/mL. Among the three physiological states, LTS phase cells had the least sublethal injury in the surviving population (p < 0.05). These results indicate that the LTS state cross-protects Salmonella Typhimurium against UV radiation and should be considered in determination of the UV radiation D-value for this pathogen.


Assuntos
Irradiação de Alimentos/métodos , Sucos de Frutas e Vegetais/microbiologia , Tolerância a Radiação , Salmonella typhimurium/efeitos da radiação , Contagem de Colônia Microbiana , Microbiologia de Alimentos , Malus , Solução Salina , Salmonella typhimurium/crescimento & desenvolvimento , Temperatura , Fatores de Tempo
17.
J Food Prot ; 81(7): 1068-1072, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29893583

RESUMO

Validated surrogates are a useful tool for studying the response of pathogens to food safety interventions, but better surrogates are needed for studies using high pressure processing. Ground beef (85% lean, 15% fat) was inoculated separately with mixed cultures of Escherichia coli O157, non-O157 Shiga toxin-producing E. coli, nontyphoidal Salmonella, and nonpathogenic E. coli surrogate bacteria. The inoculated ground beef was subjected to high hydrostatic pressures of 200, 400, and 600 MPa for 4, 6, and 8 min at each pressure. High pressure processing at 200 MPa reduced the inoculated populations of the pathogenic bacteria by 0.9 to 1.8 log CFU/g, 400 MPa reduced the inoculated populations by 2.5 to 3.6 log CFU/g, and 600 MPa reduced the inoculated populations by 4.5 to 5.6 log CFU/g. The nonpathogenic E. coli surrogates were more resistant to the effects of high pressure processing than were the inoculated pathogen populations. This finding suggests that the nonpathogenic E. coli surrogates could be used as process control indicators for high pressure processing of ground beef to predict a specific level of pathogen reduction. The surviving populations of the potential surrogate bacteria were proportional to the surviving populations of the pathogenic bacteria. The models allow for an estimation of the potential surviving populations of the pathogenic bacteria based on quantitative results of the populations of the surrogate bacteria.


Assuntos
Escherichia coli O157 , Pressão Hidrostática , Carne/microbiologia , Escherichia coli Shiga Toxigênica , Animais , Bovinos , Contagem de Colônia Microbiana , Escherichia coli O157/crescimento & desenvolvimento , Manipulação de Alimentos , Microbiologia de Alimentos , Conservação de Alimentos , Salmonella/crescimento & desenvolvimento , Escherichia coli Shiga Toxigênica/crescimento & desenvolvimento
18.
Biochem J ; 474(13): 2261-2276, 2017 06 26.
Artigo em Inglês | MEDLINE | ID: mdl-28526744

RESUMO

Phosphoinositide 3-kinases (PI3Ks) are major regulators of many cellular functions, and hyperactivation of PI3K cell signalling pathways is a major target for anticancer drug discovery. PI3Kα is the isoform most implicated in cancer, and our aim is to selectively inhibit this isoform, which may be more beneficial than concurrent inhibition of all Class I PI3Ks. We have used structure-guided design to merge high-selectivity and high-affinity characteristics found in existing compounds. Molecular docking, including the prediction of water-mediated interactions, was used to model interactions between the ligands and the PI3Kα affinity pocket. Inhibition was tested using lipid kinase assays, and active compounds were tested for effects on PI3K cell signalling. The first-generation compounds synthesized had IC50 (half maximal inhibitory concentration) values >4 µM for PI3Kα yet were selective for PI3Kα over the other Class I isoforms (ß, δ and γ). The second-generation compounds explored were predicted to better engage the affinity pocket through direct and water-mediated interactions with the enzyme, and the IC50 values decreased by ∼30-fold. Cell signalling analysis showed that some of the new PI3Kα inhibitors were more active in the H1047R mutant bearing cell lines SK-OV-3 and T47D, compared with the E545K mutant harbouring MCF-7 cell line. In conclusion, we have used a structure-based design approach to combine features from two different compound classes to create new PI3Kα-selective inhibitors. This provides new insights into the contribution of different chemical units and interactions with different parts of the active site to the selectivity and potency of PI3Kα inhibitors.


Assuntos
Desenho de Fármacos , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Inibidores de Fosfoinositídeo-3 Quinase , Antineoplásicos/química , Antineoplásicos/farmacologia , Domínio Catalítico , Proliferação de Células/efeitos dos fármacos , Classe I de Fosfatidilinositol 3-Quinases , Humanos , Ligantes , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Neoplasias/tratamento farmacológico , Neoplasias/enzimologia , Neoplasias/patologia , Ligação Proteica , Conformação Proteica
19.
J Food Sci ; 82(5): 1208-1215, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28407236

RESUMO

Selected processing methods, demonstrated to be effective at reducing Salmonella, were assessed to determine if spice and herb quality was affected. Black peppercorn, cumin seed, oregano, and onion powder were irradiated to a target dose of 8 kGy. Two additional processes were examined for whole black peppercorns and cumin seeds: ethylene oxide (EtO) fumigation and vacuum assisted-steam (82.22 °C, 7.5 psia). Treated and untreated spices/herbs were compared (visual, odor) using sensory similarity testing protocols (α = 0.20; ß = 0.05; proportion of discriminators: 20%) to determine if processing altered sensory quality. Analytical assessment of quality (color, water activity, and volatile chemistry) was completed. Irradiation did not alter visual or odor sensory quality of black peppercorn, cumin seed, or oregano but created differences in onion powder, which was lighter (higher L* ) and more red (higher a* ) in color, and resulted in nearly complete loss of measured volatile compounds. EtO processing did not create detectable odor or appearance differences in black peppercorn; however visual and odor sensory quality differences, supported by changes in color (higher b* ; lower L* ) and increased concentrations of most volatiles, were detected for cumin seeds. Steam processing of black peppercorn resulted in perceptible odor differences, supported by increased concentration of monoterpene volatiles and loss of all sesquiterpenes; only visual differences were noted for cumin seed. An important step in process validation is the verification that no effect is detectable from a sensory perspective.


Assuntos
Cuminum/química , Óxido de Etileno/farmacologia , Piper nigrum/química , Especiarias/análise , Vapor , Monoterpenos/análise , Sementes/química , Sesquiterpenos/análise
20.
Foodborne Pathog Dis ; 14(5): 302-307, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-28398867

RESUMO

The effectiveness of cinnamaldehyde for inactivating Salmonella enterica and Escherichia coli O157:H7 in carrot juice (CRJ) and mixed berry juice (MBJ) was investigated. Brain heart infusion broth (BHI), CRJ, and MBJ, with concentrations of added cinnamaldehyde ranging from 0.15 to 1.5 µL/mL, 0.25 to 2.0 µL/mL, and 0.25 to 1.5 µL/mL, respectively, were each inoculated with a 5-strain mixture of Salmonella enterica or Escherichia coli O157:H7 to give an initial viable count of 5.07 log10 colony-forming units/mL. Inoculated BHI or juices without cinnamaldehyde served as controls. Growth of the pathogens in BHI (35°C) was monitored by taking absorbance readings (optical density [OD] 600 nm) for 24 h. The inoculated juices were held at 4°C or 12°C for 24 h, and numbers of viable pathogens were determined at 0, 2, 4, 8, and 24 h by plating samples on selective agar followed by incubation (35°C) and counting bacterial colonies at 48 h. The minimum inhibitory concentration of cinnamaldehyde for both pathogens in BHI was 0.25 µL/mL. The pathogens were more sensitive to cinnamaldehyde in MBJ compared with CRJ, irrespective of storage temperature (p < 0.05). At 4°C, cinnamaldehyde (1.5 µL/mL) completely inactivated S. enterica and E. coli in MBJ (negative by enrichment) within 2 h and 8 h, respectively; whereas both pathogens were detected in CRJ (4°C; with 2.0 µL/mL cinnamaldehyde) at 8 and 24 h. At 12°C, S. enterica and E. coli were undetected in MBJ (1.5 µL/mL cinnamaldehyde) within 2 and 4 h, respectively; however, in CRJ (12°C; 2.0 µL/mL cinnamaldehyde), complete inactivation of S. enterica and E. coli occurred within 4 and 24 h, respectively. Cinnamaldehyde is an effective antimicrobial from natural sources that can be used for inactivating bacterial pathogens in fruit and vegetable juices to enhance microbial safety of these nutritious food products.


Assuntos
Acroleína/análogos & derivados , Anti-Infecciosos/farmacologia , Escherichia coli O157/efeitos dos fármacos , Salmonella enterica/efeitos dos fármacos , Acroleína/farmacologia , Temperatura Baixa , Contagem de Colônia Microbiana , Daucus carota/microbiologia , Contaminação de Alimentos , Manipulação de Alimentos , Microbiologia de Alimentos , Conservação de Alimentos , Frutas/microbiologia , Sucos de Frutas e Vegetais/microbiologia , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana
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