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BACKGROUND Spoilage of fishery products begins immediately following filleting due to microbial growth that degrades fish tissue quality prior to consumption. Extensive research has been conducted to identify such bacterial populations. A better understanding of the mechanisms involved in fish spoilage is necessary as a novel remedy for microbial spoilage inhibition has yet to be established for fish tissue. The present study identified, for the first time, bacterial populations that produce extracellular DNase and protease from Delaware and local retail distributed channel catfish ( Ictalurus punctatus) fillets. RESULTS A clear trend was identified between bacteria derived from catfish filleted under aseptic conditions where Pseudomonas was the dominant genus. Bacteria isolated from retail catfish contained high quantities of DNase-producing isolates, in contrast to aseptic-filleted catfish tissue which had none. Both types of catfish sample maintained high populations of protease-producing bacterial colonies throughout the duration of the study. Most bacteria isolated from catfish intestines exhibited DNase production with no protease production. CONCLUSION Specific spoilage organism populations were significantly higher on retail-derived catfish in comparison to lab-filleted Delaware cultured catfish tissue. It is suggested that DNase production and protease production contribute to the spoilage of fish tissue as a result of mishandling and septic filleting being the major cause of rapid catfish tissue spoilage. © 2014 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

In this study, lactic acid bacteria ( LAB) were isolated from kimchi prepared in the U.S.A. and from broiler chickens. The resulting isolates were examined for potential probiotic characteristics. A total of 488 LAB strains were isolated, and among them, 16 originating from kimchi and 14 from broilers produced bacteriocin-like substances, which showed antimicrobial effects against S almonella Enteritidis, Heidelberg, Newport and Typhimurium. These strains were investigated with regard to additional probiotic properties such as tolerance to gastric juice and bile, resistance to enzymes, and antibiotics susceptibilities. L actobacillus casei or paracasei Cab-18, L b. saniviri Cuc-1, and L euconostoc mesenteroides Com-54 isolated from kimchi and L b. johnsonii F-6 and L b. crispatus F-59 isolated from broiler chicken demonstrated promising probiotic properties with the ability to produce bacteriocin-like substances with relatively strong antimicrobial activity against S almonella. These strains could potentially be applied as supplements to poultry in order to reduce S almonella contamination. Practical Applications Salmonellosis associated with consumption of contaminated meats, especially poultry products, is a major concern due to the common outbreaks in the U.S.A. Although it has been shown that lactic acid bacteria ( LAB) could reduce S almonella contaminations, LAB isolated from kimchi have not been studied for the potential to inhibit the persistence of S almonella spp. in live poultry. The present study screened LAB, which showed antimicrobial effects against four clinically important serotypes of S almonella. Additionally, we selected LAB that had additional probiotic properties, which would allow for the strains use as potential feed additives. Therefore, the LAB strains isolated in this study could be applied as feed supplements to poultry for the purpose of reducing colonization of S almonella in poultry as well as contamination during processing. [ABSTRACT FROM AUTHOR]

The effects of γ -irradiation on the destruction of Vibrio vulnificus in quahog clam tissue (Mercenaria mercenaria) by real-time PCR (Rti-PCR) were studied. The Rti-PCR methodology used in this study employed four sets of V. vulnificus specific primers, which were utilized to amplify 1000, 700, 300, and 70-bp sequences of the vvhA to determine the percent detection of V. vulnificus genomic targets after γ -irradiation. Tissue homogenates seeded with V. vulnificus were irradiated to gradually reduce the viable CFU by accumulating dosage. The use of these four pairs of primers to amplify DNA target sequences of varying length in seeded tissue exposed to 1.4 KGy failed to discriminate between irradiated and nonirradiated cells. In contrast, with doses of 2.0, 3.0, and 5.0 KGy, the amplification of 700, 300, and 70-bp sequences resulted in higher Ct values, respectively, and lower levels of amplification with each primer pair, reflecting increased DNA fracture with increasing dose. When varying numbers of CFU in tissue homogenates (1.0 × 103 to 1.0 × 106 per gram of tissue) were exposed to 3.0 KGy of γ-irradiation, DNA amplicons of 1000 and 700-bp derived from 1 × 105 CFU/g and below failed to be detected in contrast to 300 and 70-bp amplicons, which were detected with CFU levels of 1.0 × 105/g. This study showed that with increasing dose, the larger amplicons (100 and 700-bp) failed to be detected, which corresponded to zero percent survival of CFU. [ABSTRACT FROM AUTHOR]

Ferritin is an iron storage protein found in most living organisms. Human heavy chain (H-) and light chain (L-) ferritin were amplified from human heart cDNA library. Each ferritin gene was inserted down stream of the T7 promoter of bacterial expression, and finally four types of H-, L-, and co-expression vectors were constructed. Recombinant human ferritins were overexpressed and identified with SDS-PAGE and western blotting. The expression levels of recombinant ferritin proteins ranged about 29-36% of whole cell total protein. From atomic absorption spectrometry (AAS) analysis, the rate of iron uptake for H-ferritin was significantly faster than that for the HL-, LH-, and L-ferritin, respectively. From AAS analysis, the levels of iron content in cells progressively increased in transformants with 0-30 mM ferric citrate in the media. Among these ferritin transformants, the highest amount of cellular iron was observed with H-ferritin transformant. The total amounts of iron content in E. coli could be sequentially ranked as H-ferritin > HL-ferritin > LH-ferritin > L-ferritin > negative transformants. Expressed recombinant human ferritins indicated that their assembled subunits were able to store iron in the cells. The results of this study further enhances our understanding of iron uptake properties in vivo and suggest similar strategies for using food-grade ferritins for functional foods or iron-fortified food ingredients. [ABSTRACT FROM AUTHOR]

Objectives: This study was conducted to investigate the current status of handoffs, perception of patient safety culture, and degrees of handoff evaluation in small and medium-sized hospitals and identified factors that make a difference in handoff evaluation., Methods: This is a descriptive study. 425 nurses who work at small and medium-sized hospitals in South Korea were included in our study. They completed a set of self-reporting questionnaires that evaluated demographic data, handoff-related characteristics, perception of patient safety culture, and handoff evaluation., Results: Results showed that the overall score of awareness of a patient safety culture was 3.65 ± 0.45, the level was moderate. The score of handoff evaluation was 5.24 ± 0.85. Most nurses experienced errors in handoff and most nurses had no guidelines and checklist in the ward. Handoff evaluation differed significantly according to the level of education, work patterns, duration of hospital employment, handoff method, degree of satisfaction with the current handoff method, errors occurring at the time of handoff, handoff guidelines, and appropriateness of handoff education time ( P  < 0.05)., Conclusion: For handoff improvement, guidelines and standards should be established. It is necessary to develop a structured handoff education system. And formal handoff education should be implemented to spread knowledge uniformly., Competing Interests: The authors declare no conflict of interest., (© 2020 The authors.)

Background: Whereas most studies have focused on euthanasia and physician-assisted suicide, few have dealt comprehensively with other critical interventions administered at the end of life. We surveyed cancer patients, family caregivers, oncologists and members of the general public to determine their attitudes toward such interventions., Methods: We administered a questionnaire to four groups about their attitudes toward five end-of-life interventions--withdrawal of futile life-sustaining treatment, active pain control, withholding of life-sustaining measures, active euthanasia and physician-assisted suicide. We performed multivariable analyses to compare attitudes and to identify sociodemographic characteristics associated with the attitudes., Results: A total of 3840 individuals--1242 cancer patients, 1289 family caregivers and 303 oncologists from 17 hospitals, as well as 1006 members of the general Korean population--participated in the survey. A large majority in each of the groups supported withdrawal of futile life-sustaining treatment (87.1%-94.0%) and use of active pain control (89.0%-98.4%). A smaller majority (60.8%-76.0%) supported withholding of life-sustaining treatment. About 50% of those in the patient and general population groups supported active euthanasia or physician-assisted suicide, as compared with less than 40% of the family caregivers and less than 10% of the oncologists. Higher income was significantly associated with approval of the withdrawal of futile life-sustaining treatment and the practice of active pain control. Older age, male sex and having no religion were significantly associated with approval of withholding of life-sustaining measures. Older age, male sex, having no religion and lower education level were significantly associated with approval of active euthanasia and physician-assisted suicide., Interpretation: Although the various participant groups shared the same attitude toward futile and ameliorative end-of-life care (the withdrawal of futile life-sustaining treatment and the use of active pain control), oncologists had a more negative attitude than those in the other groups toward the active ending of life (euthanasia and physician-assisted suicide).

Objective: The purpose of this study was to re-evaluate post-remission therapy outcomes after first remission according to years of patient enrollment in patients with core binding factor acute myeloid leukaemia., Methods: We conducted a retrospective study on 138 patients aged less than 60 years diagnosed with core binding factor acute myeloid leukaemia between 1994 and 2006, comparing allogeneic stem cell transplantation and high-dose cytarabine chemotherapy as post-remission treatment options after the first remission., Results: The 5-year probabilities of disease-free survival and overall survival were not different between allogeneic stem cell transplantation and high-dose cytarabine groups. However, 3-year probabilities of disease-free survival (86.7% vs. 67.0%) and overall survival (90.0% vs. 67.3%) showed a trend towards improvement in the allogeneic stem cell transplantation group compared with the high-dose cytarabine group in cohort after 2003 (2003-2006), whereas outcomes were not different in cohort before 2003 (1994-2002). Especially, 3-year probabilities of disease-free survival (95.2% vs. 59.3%, P = 0.008) and overall survival (95.2% vs. 59.6%, P = 0.032) of allogeneic stem cell transplantation group were significantly better than high-dose cytarabine group in cohort after 2003 of acute myeloid leukaemia patients with t(8;21). The relative risk of overall survival with allogeneic stem cell transplantation, compared with high-dose cytarabine chemotherapy, was significantly improved in the cohort after 2003 (0.33; 95% CI, 0.07-1.48) when compared with that before 2003 (1.92; 95% CI, 0.77-4.82). In multivariate analysis in cohort after 2003, allogeneic stem cell transplantation as post-remission therapy was associated with better disease-free survival., Conclusions: Allogeneic stem cell transplantation is currently the more effective post-remission therapy than it was prior to 2003 for core binding factor acute myeloid leukaemia achieving first remission. On the contrary to previous findings, allogeneic stem cell transplantation provides significantly improved outcomes than high-dose cytarabine chemotherapy in acute myeloid leukaemia with t(8;21).

Human heavy chain (H-) and light chain (L-) ferritins were amplified from a human cDNA library. Each ferritin gene was inserted downstream of the T7 promoter of bacterial expression vectors, and two types of coexpression vectors were constructed. The expression levels of recombinant ferritins ranged about 26-36% of whole-cell protein. Hferritin exhibited a lower expression ratio compared with L-ferritin, by a coexpression system. However, the coexpression of HL-ferritins was significantly increased above the expression ratio of H-ferritin by cultivation without IPTG induction overnight. Purified recombinant H-, L-, HL-, and LHferritins were shown to be homo- and heteropolymeric high molecular complexes and it was indicated that their assembled subunits would be able to work functionally in the cell. Thus, these results indicate an improvement in the expression strategy of H-ferritin for heteropolymeric production and studies of ferritin assembly in Escherichia coli.

The effects of gamma-irradiation on the destruction of Vibrio vulnificus by real-time PCR were studied. gamma-irradiation was found to result in extensive reduction in the molecular size of DNA. Irradiation of viable cells (1 x 10(6) CFU/ml) at 1.08 KGy resulted in 100% destruction determined by plate counts, with most of the DNA from the irradiated cells having a bp-length of less than 1000. The use of a pair of primers to amplify a 1000-bp sequence of DNA from cells exposed to 1.08 KGy failed to yield amplification. In contrast, primers designed to amplify sequences of 700, 300, and 70-bp yielded amplification with C(t) values resulting in 13.4, 27.6, and 45.4% detection of genomic targets. When viable cells of V. vulnificus were exposed to 1.08, 3.0, and 5.0 kGy, the average molecular size of genomic DNA visualized in an agarose gel decreased with increasing dose, corresponding to an increased probability of amplification with primers targeting sequences of decreasing size.

Ferritin is an iron storage protein found in most living organisms as a natural assembled macromolecule. For studying the functional ability of the ferritin assembly, human H- and L-ferritins were expressed and purified from Pichia pastoris strain GS115. The recombinant H- and L-ferritins showed a globular form with transmission electron microscopy. The rate of iron uptake for H-ferritin was significantly faster than that for the L-ferritin in vitro. By gel permeation chromatography analysis, recombinant ferritins were confirmed as multimeric subunits with high molecular weight and it was indicated that assembled subunits were able to store iron in vivo.

Human heavy chain ferritin (H-ferritin) was cloned from human heart cDNA library and expressed in Pichia pastoris. The H-ferritin transformant was cultivated by fed-batch and the cell mass reached about 52 g cell dry wt l(-1) after 150 h. In atomic absorption spectrometry analysis, intracellular content of iron in H-ferritin transformant was measured to 3038 +/- 72 microg g(-1) which was 9.6-fold more than that of control strain.