The Lived Experiences of Palliative Care Professionals in Cultivating Mindfulness: A Phenomenological Study.

Purpose: The aim of this study was to explore the essence of the lived experiences of palliative care professionals in cultivating mindfulness, with a focus on the meaning of mindfulness in their lives and how mindfulness is experienced throughout their process of caring for others. Design: This was a qualitative study using a phenomenological approach. Methods: Eleven palliative care professionals (three physicians, four nurses, three psychologists, and one spiritual care provider) partook in in-depth interviews. Data were collected from the in-depth interviews and analyzed according to the method of Giorgi. Findings: Two major themes emerged from this study. First, the palliative care professionals realized the need for self-care amid emotional burden, including recognizing their feelings of guilt and self-doubt, emotional contagion of grief, reflections of others' fragility on themself, and their self-imposed limitations. Second, they noticed the transformative impact of mindfulness on them, including detecting reconnection with their body, changes in their personal values, self-acceptance, and liberation. Conclusion: Palliative care professionals can cultivate self-acceptance and facilitate entirely new life experiences through the practice of mindfulness. For them, mindfulness is not merely a self-regulation technique but an existential epiphany, offering hope for self-care and empowerment.

Effects of a Multiple-Intervention Program on the Depressive Tendencies, Social Support, and Life Satisfaction of Older Adults in Taiwan.

PURPOSE: This study evaluated the effectiveness of a multiple-intervention program (MIP) on the life satisfaction, social support, and depressive tendencies of older adults. DESIGN: A cross-sectional, pretest-posttest design involving a questionnaire survey was adopted. METHODS: Thirty-seven older adults were recruited from 2 daycare centers for an 8-week MIP comprising 60-min sessions conducted once a week. The questionnaire comprised the Short Portable Mental Status Questionnaire, Geriatric Depression Short Form-15 (GDS-SF15), Life Satisfaction Scale (LSS), and Inventory of Social Support Scale (ISSB). FINDINGS: The participants' average GDS-SF15 score decreased considerably (by 35.3%), their average ISSB score increased from 61.9% to 80.9%, and their average LSS score increased from 48% to 64%. CONCLUSIONS: The outcomes for depressive tendencies, social support, and life satisfaction were measured before and after the intervention, and the participants successfully engaged in the MIP throughout its duration and exhibited improvement. The MIP can serve as a basis for planning group activities for older adults. CLINICAL EVIDENCE: The MIP benefits older adults at daycare centers. Incorporating multiple activities into daily care can help increase the diversity, uniqueness, and innovativeness of a daycare center.

Effectiveness of simulation-based education on nursing students' professional knowledge, attitude and self-confidence in handling child abuse cases.

AIM: This study aimed to enhance nursing students' professional knowledge, attitudes and self-confidence in handling child abuse cases through a Child Abuse Simulation-Based Education (CASE). BACKGROUND: Given that nursing students seldom learn about child abuse case management in textbooks, simulation-based education could assist them in appropriately handling child abuse cases when they become nurses. DESIGN: Using a quasi-experimental design. METHODS: 190 nurses enrolled in a night school in-service program at a Taiwanese university were recruited for this study. Ninety-four nurses in the experimental group took a simulation-based education on child abuse while 96 nurses in the contrast group took a general nursing course. The pretest and posttest professional knowledge, attitudes and self-confidence of both groups were analyzed using generalized estimating equations in SPSS V22 software. RESULTS: Following the introduction of the CASE course, the experimental group's posttest professional knowledge, attitudes and self-confidence in handling child abuse cases were higher than those of the contrast group (p < .001). Further analysis showed that professional knowledge increased with tenure, positivity in attitude increased with clinical ladder rating and male nurses were more confident than female nurses when handling child abuse cases. CONCLUSIONS: Based on the results, this study proposes recommendations for implementing child abuse simulation-based education for new nurses and those undergoing continuing education.

Development of an attenuated vaccine against Koi Herpesvirus Disease (KHVD) suitable for oral administration and immersion.

Since the end of the1990ies, Cyprinid herpesvirus 3 (also known as koi herpesvirus, KHV) has caused mass mortality events of koi and common carp all over the globe. This induced a high economic impact, since the KHV disease cannot be cured up to now, but only prevented by vaccination. Unfortunately, there is only one commercial vaccine available which is not approved in most countries. Therefore, there is an urgent need for new, safe and available vaccines. In this study, a live attenuated vaccine virus was generated by cell culture passages of virulent KHV, and shown to protect carp or koi after immersion or oral application against wild type challenge. An advantage of boost immunization was demonstrated, especially after oral application. Vaccination induced no or mild clinical signs and protecting antibodies have been measured. Additionally, the vaccine virus allowed differentiation of infected from vaccinated animals (DIVA) by PCR. The attenuation of the newly generated vaccine was tracked down to a partial deletion of open reading frame 150. This was confirmed by the generation of engineered ORF150 deletion mutants of wild-type KHV which exhibited a similar attenuation in vivo.

Lived Experiences of Domestic Violence in Women and Their Children: A Phenomenological Study.

This study explores women and their children's lived experience of domestic violence (DV). A qualitative phenomenological research approach was used. Data were collected by semi-structured interviews. Five women and five adult children participated in this study. COREQ reporting guidelines were utilized. Three main themes and six subthemes emerged from the interviews with the women; the main themes were "living with suffering", "insecurity in daily life", and "conformity in coping behaviors". Two main themes and four subthemes were identified from the interviews with the children; the main themes were "barriers to learning and interactions with peers" and "a sense of threat to life". The findings show that the women and their children had to cope with not only the DV itself but also the negative consequences of the violence. Several recommendations are made for the police and first-line healthcare and social work systems and to prevent DV by integrating the care provided to the family. The findings provide prevention and integration care for mothers and their children experiencing DV systematically.

[Patient's Personal Data Protection Awareness: A Survey of Clinical Nurses].

BACKGROUND: The aim of the Personal Data Protection Act (PDPA) is to regulate the collection, processing, and use of personal information; to avoid the infringement of personal rights; and to promote the reasonable use of personal information. Clinical nurses are frontline patient caregivers, and they are the most likely to have access to patients' personal information. If these nurses do not clearly understand the PDPA, they may violate the law and affect patients' rights. PURPOSE: The purpose of this study was to investigate the knowledge level of clinical nurses regarding the PDPA and related factors, with the findings intended to serve as a reference for continuing education. METHODS: A cross-sectional research design was adopted. A purposive sample of nurses working at a regional hospital in southern Taiwan was selected. A self-administered survey incorporating the self-developed Nurses Knowledge Scale for Patient Personal Data Protection Act (NKSPPDPA) was used to collect data from May to June 2017. RESULTS: A total of 269 valid responses were received (return rate: 89.67%). The mean score on the NKSPPDPA was 68.80 out of a total-possible 100 points. Knowledge related to patient privacy and penalties was relatively low. Moreover, working department, job title, and participation in PDPA-related on-the-job education were found to be significant predictors of NKSPPDPA score, while years of experience was found to have a low correlation only. CONCLUSIONS: The results suggest that clinical nurses have knowledge gaps regarding PDPA, especially in terms of privacy and penalties. Nurses should participate in continuing education to address these knowledge gaps.

Serological responses to koi herpesvirus (KHV) in a non-cyprinid reservoir host.

Koi herpesvirus (KHV) is a highly contagious virus that causes KHV disease (KHVD) inducing high mortality in carp and koi (Cyprinus carpio L.). In the late stage, latency occurs with very low, often non-detectable virus concentrations, which represents a challenge for virus detection. After validation according to OIE recommendations, an antibody ELISA was established to recognize antibodies of C. carpio against KHV infection. In this study, the ELISA was modified to detect anti-KHV antibodies from a non-cyprinid fish. Experimentally infected rainbow trout (Oncorhynchus mykiss) were able to transmit KHV to naïve carp at two different temperatures, demonstrating their potential as a reservoir host. At 20°C, KHVD was induced in carp but not at 15°C. Unexpectedly, rainbow trout developed humoral response against KHV at both temperatures. In contrast to carp, at 15°C trout produced neutralizing antibodies but not at 20°C. While antibodies obtained from infected carp sera reacted in a similar way against all KHV, antibodies from rainbow trout sera reacted differently to the same isolates by ELISA. The data show that even when non-cyprinid fish species are infected with KHV, they can produce antibodies that differ from those observed in carp.

Extensive-Stage Small Cell Carcinoma Transformation From EGFR Del19-Mutant Lung Adenocarcinoma on Gefitinib at the Twelfth-Year Follow-Up Case Report.

INTRODUCTION: The acquired resistance mechanisms in patients with epidermal growth factor receptor (EGFR)-mutant lung cancer, particularly adenocarcinoma (ADC), following treatment with an EGFR tyrosine kinase inhibitor (TKI) have received extensive investigations. The phenotypic transformation to small cell carcinoma (SCCT) has been estimated to occur in approximately 3 to 10% of patients treated with an EGFR-TKI. The prognosis after SCCT is extremely poor. CASE STUDY: We report about SCCT that occurred 45 months after the initial diagnosis of ADC in an East Asian never-smoker woman with advanced-stage EGFR Del-19-mutant lung ADC treated with combined chemoradiotherapy before the era of insurance coverage for EGFR-TKIs in this country and subsequently gefitinib; deletion at codon 746-750 in exon 19 of the EGFR gene was ascertained in the original formalin-fixed paraffin-embedded lung biopsy tissue. Spinal cord compression at thoracic-12 level from SCCT was successfully relieved with neurosurgical treatment, chemotherapy with etoposide and cisplatin, and radiotherapy, while gefitinib treatment was maintained. Eleven months later, SCCT relapsed in the lung parenchyma, which was resected and was found to be sensitive to second-line weekly topotecan. Prophylactic cranial irradiation was subsequently administered. SCCT was confirmed by MALDI-TOF MS analysis of formalin-fixed paraffin-embedded tissues demonstrating the same exon 19 deletion. At the 12th-year follow-up, the patient remains relapse free with very good performance status. The novelty of this case is the successful interdisciplinary team effort to correct the spinal cord compression by maintaining the patient in an ambulatory state, non-stop use of gefitinib justified by the presence of activating EGFR mutation in SCCT tumor cells, and aggressive dose-intensive chemotherapy and radiotherapy for the SCCT that leads to an unprecedented prolonged remission and survival. This case also supports the observation that SCCT is chemotherapy sensitive, and thus, re-biopsy or complete tumor excision is recommended to understand the mutation profiles of the current tumor. Aggressive prudent administration of systemic chemotherapy obtaining optimal dose intensity leads to the successful management of the patient.

The Smoking Paradox: A Twist in the Tale of Vasospastic Angina.

Cigarette smoking is undoubtedly the single most important risk factor and trigger for vasospastic angina, a condition also known as Prinzmetal angina secondary to coronary artery vasospasm. Even decades before vasospastic angina was first described by Dr. Myron Prinzmetal and his colleagues in 1959, there had been suspected connections between smoking and coronary artery vasospasm in what was alluded to then as "tobacco angina." The intimate relationship between smoking and vasospastic angina has since been extensively researched and validated through decades of epidemiological and clinical studies. The fact that smoking would aggravate vasospastic angina comes with very little surprise, as it has been shown to adversely impact many of the disease processes thought to underlie vasospastic angina, including autonomic dysfunction, endothelial dysfunction, smooth muscle hyperactivity, and genetic susceptibility. While avoidance of smoking is the first logical step in managing smokers with vasospastic angina, there have been reported cases of vasospastic angina paradoxically triggered by smoking cessation or relieved with smoking resumption or nicotine replacement therapy. Thus, there appears to be patient-specific factors that could significantly alter the close connection between smoking and vasospastic angina, warranting further mechanistic investigations. In this review, we will examine this complicated relationship between smoking and vasospastic angina from multiple perspectives (historical, mechanistic, and clinical) and call attention to the "smoking paradox," which, with further elucidation, may provide additional insight into the complex mechanisms of VSA and potentially new strategies to treat medically refractory VSA, at least in selected individuals.

Family caregivers' lived experiences of caring for epidermolysis bullosa patients: A phenomenological study.

AIMS AND OBJECTIVES: To explore the lived experiences of family caregivers of epidermolysis bullosa (EB) patients. BACKGROUND: Patients with EB need long-term care. Their family experiences physical and mental stress while proving that care. However, very little research has explored the family caregivers' actual lived experiences when caring for EB patients. DESIGN: This is a qualitative research using the phenomenological approach. METHODS: Data were collected with the assistance of the Taiwan Foundation for Rare Disorders (TFRD). Interviews were conducted between 2015 and 2017 with primary family caregivers (n = 10) who had cared for EB patients for at least five years. Data were analysed using Colaizzi's (1978) seven-step phenomenological method. COREQ reporting guidelines were utilised. RESULTS: Four themes and eleven subthemes emerged after data analysis: (a) transformation and helplessness: changing expectations and helplessness associated with the rare, unknown disease; (b) mother's responsibility: mothers loving and protecting their children, surviving through adversity and challenges and self-experiencing a child's pain; (c) physical and mental fatigue experienced by the whole family: physical and mental exhaustion, and unbalanced lives; and (d) adjustment and social support: providing mutual assistance to mitigate adversity, hopeful search for treatments, the importance of patient associations and rare disease foundations, and social assistance requirements. CONCLUSION: Family caregivers experience many challenges when caring for EB patients and experience substantial stress and overwhelming burdens. Consequently, they would benefit greatly from various support systems to reduce the burden of caring for EB patients. RELEVANCE TO CLINICAL PRACTICE: Healthcare professionals can use the findings from this study to understand the circumstances faced by family caregivers and tailor care and education specifically to support each family's financial, social and household needs, thereby helping them reduce the stress of caring for EB patients.

Clinical, Demographic, and Biochemical Characteristics of Patients with Acute ST-Segment Elevation Myocardial Infarction: An Analysis of Acute Coronary Syndrome Registry Data of a Single Medical Center from 2005 to 2016.

BACKGROUND: ST-segment elevation myocardial infarction (STEMI) is one of the leading causes of morbidity and mortality in developed countries. Therefore, understanding the prevalence and trends of major risk factors may facilitate primary and secondary prevention of STEMI. METHODS: In the present study, 2446 consecutive patients with STEMI admitted to Far Eastern Memorial Hospital from 2005 to 2016 were enrolled. A comprehensive analysis of the prevalence, distribution, and trends over time of major risk factors as well as Framingham risk scores of all patients was performed. RESULTS: The most prevalent risk factors were male sex, hypertension (HTN), smoking, age, dyslipidemia, and diabetes mellitus. Furthermore, 95%-97% of the patients had at least one modifiable risk factor, and < 1% of the patients did not have any identifiable risk factors. The prevalence trends of smoking, HTN, dyslipidemia, and metabolic syndrome increased significantly from 2005 to 2016. Seasonal variation analysis revealed a 15% increase in STEMI cases between January and March compared with those between April and December. Isolated low high- density lipoprotein-cholesterol syndrome was the second most common type of dyslipidemia, with a prevalence rate of 16.6%. Moreover, only 56.8% of the male and 32% of the female patients were in the Framingham high-risk group. CONCLUSIONS: A high prevalence rate and an increasing trend of modifiable risk factors resulted in a high number of STEMI cases at our hospital. Controlling modifiable risk factors and improving nontraditional risk factor detection could facilitate primary and secondary preventions for STEMI.

Antenatal sleep quality associated with perinatal outcomes in women of advanced maternal age.

OBJECTIVES: The increasing prevalence of advanced maternal age (AMA) coupled with poor sleep quality among pregnant women makes it important to study their association with perinatal outcomes. However, little is known about the interaction of AMA and maternal antenatal sleep on perinatal outcomes. Here, we examined whether associations between AMA and perinatal outcomes are modified by antenatal sleep quality. PARTICIPANTS: Data were collected from 446 women, with a singleton pregnancy and no pregnancy complications, who participated in the Growing Up in Singapore Towards healthy Outcomes (GUSTO) birth cohort study. MEASUREMENTS: Participants completed the Pittsburgh Sleep Quality Index (PSQI) at 26-28 weeks gestation and had perinatal outcome data collected upon delivery. Interactions between AMA and maternal sleep quality on perinatal outcomes were investigated and where significant, analyses were further stratified by maternal age. All analyses were adjusted for maternal BMI at 26-28 weeks gestation, ethnicity, and maternal education. RESULTS: Neonates of mothers of AMA and poor sleep quality (PSQI score >5) had increased odds of stay in the neonatal intensive care unit (adjusted odds ratio = 3.53, 95% CI: -1.21 to 10.27) and shorter birth length (adjusted mean difference = -1.05 cm, 95% CI: -1.82 to -0.20), as compared with women of AMA and good sleep quality (PSQI score ≤5). In women <35 years, sleep quality did not associate with perinatal outcomes. CONCLUSION: Poor sleep quality in women of AMA was associated with neonatal health outcomes. Improving maternal antenatal sleep may potentially improve perinatal outcomes in offspring of women of AMA.

Cyprinid herpesvirus 3 Evolves In Vitro through an Assemblage of Haplotypes that Alternatively Become Dominant or Under-Represented.

Viruses are able to evolve in vitro by mutations after serial passages in cell cultures, which can lead to either a loss, or an increase, of virulence. Cyprinid herpesvirus 3 (CyHV-3), a 295-kb double-stranded DNA virus, is the etiological agent of the koi herpesvirus disease (KHVD). To assess the influence of serial passages, an isolate of CyHV-3 (KHV-T) was passaged 99 times onto common carp brain (CCB) cells, and virus virulence was evaluated during passages through the experimental infections of common carp. After 78 CCB passages, the isolate was much less virulent than the original form. A comparative genomic analysis of these three forms of KHV-T (P0, P78 and P99) revealed a limited number of variations. The largest one was a deletion of 1363 bp in the predicted ORF150, which was detected in P78, but not in P99. This unexpected finding was confirmed by conventional PCR and digital PCR. The results presented here primarily suggest that, CyHV-3 evolves, at least in vitro, through an assemblage of haplotypes that alternatively become dominant or under-represented.

A fully automated sample-to-answer PCR system for easy and sensitive detection of dengue virus in human serum and mosquitos.

BACKGROUND: The insulated isothermal PCR (iiPCR) technology enables consistent PCR amplification and detection in a simple heating device. A pan-dengue virus (DENV) RT-iiPCR, targeting the 5' untranslated region, was validated previously on the semi-automated POCKIT combo system (involving separate devices for nucleic acid extraction and PCR amplification/detection) to offer performance comparable to a laboratory real-time PCR. Working on the same technologies, a compact automated sample-in-answer-out system (POCKIT Central Nucleic Acid Analyser) has been available commercially for iiPCR, minimizing human error risks and allowing easy molecular bio-detection near points of need. Here, we evaluated the analytical and clinical performance of the pan-DENV RT-iiPCR on the fully automated system by comparison to those on the semi-automated system. METHODOLOGY/PRINCIPAL FINDINGS: Testing sera containing serial diluted DENV-1, -2, -3, or -4 cell culture stock, the pan-DENV RT-iiPCR system had similar 100% detection endpoints on the two systems; i.e. at 1, 10, 1 and 10 PFU/ml, respectively, on the fully automated system, and at 10, 1, 10 and 10 PFU/ml, respectively, on the semi-automated system. Furthermore, both fully automated and semi-automated PCR system can detect all four DENV serotypes in mosquitos. Clinical performance of the reagent on the two systems was evaluated by testing 60 human serum samples. Both systems detected the same 40 samples (ten DENV-1, -2, -3, and -4 positive each) and did not detect the other 20; 100% agreement (κ = 1) was found between the two systems. CONCLUSIONS/SIGNIFICANCE: With performance comparable to a previously validated system, the fully-automated PCR system allows applications of the pan-DENV reagent as a useful tool near points of need to facilitate easy, fast and effective detection of dengue virus and help mitigate versatile public health challenges in the control and management of dengue disease.

Development and evaluation of a real-time RT-PCR and a field-deployable RT-insulated isothermal PCR for the detection of Seneca Valley virus.

BACKGROUND: Seneca Valley virus (SVV) has emerged in multiple countries in recent years. SVV infection can cause vesicular lesions clinically indistinguishable from those caused by other vesicular disease viruses, such as foot-and-mouth disease virus (FMDV), swine vesicular disease virus (SVDV), vesicular stomatitis virus (VSV), and vesicular exanthema of swine virus (VESV). Sensitive and specific RT-PCR assays for the SVV detection is necessary for differential diagnosis. Real-time RT-PCR (rRT-PCR) has been used for the detection of many RNA viruses. The insulated isothermal PCR (iiPCR) on a portable POCKIT™ device is user friendly for on-site pathogen detection. In the present study, SVV rRT-PCR and RT-iiPCR were developed and validated. RESULTS: Neither the SVV rRT-PCR nor the RT-iiPCR cross-reacted with any of the vesicular disease viruses (20 FMDV, two SVDV, six VSV, and two VESV strains), classical swine fever virus (four strains), and 15 other common swine viruses. Analytical sensitivities of the SVV rRT-PCR and RT-iiPCR were determined using serial dilutions of in vitro transcribed RNA as well as viral RNA extracted from a historical SVV isolate and a contemporary SVV isolate. Diagnostic performances were further evaluated using 125 swine samples by two approaches. First, nucleic acids were extracted from the 125 samples using the MagMAX™ kit and then tested by both RT-PCR methods. One sample was negative by the rRT-PCR but positive by the RT-iiPCR, resulting in a 99.20% agreement (124/125; 95% CI: 96.59-100%, κ = 0.98). Second, the 125 samples were tested by the taco™ mini extraction/RT-iiPCR and by the MagMAX™ extraction/rRT-PCR system in parallel. Two samples were positive by the MagMAX™/rRT-PCR system but negative by the taco™ mini/RT-iiPCR system, resulting in a 98.40% agreement (123/125; 95% CI: 95.39-100%, κ = 0.97). The two samples with discrepant results had relatively high CT values. CONCLUSIONS: The SVV rRT-PCR and RT-iiPCR developed in this study are very sensitive and specific and have comparable diagnostic performances for SVV RNA detection. The SVV rRT-PCR can be adopted for SVV detection in laboratories. The SVV RT-iiPCR in a simple field-deployable system could serve as a tool to help diagnose vesicular diseases in swine at points of need.

An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system.

BACKGROUND: Dengue fever, a mosquito-borne disease, is caused by dengue virus (DENV) which includes four major serotypes (DENV-1, -2, -3, and -4). Some serotypes cause more severe diseases than the other; severe dengue is associated with secondary infections by a different serotype. Timely serotyping can provide early warning of dengue epidemics to improve management of patients and outbreaks. A mobile insulated isothermal PCR (iiPCR) system is available to allow molecular detection of pathogens near points of need. METHODOLOGY/PRINCIPLE FINDINGS: In this study, side-by-side comparison with the CDC DENV-1-4 Real Time RT-PCR (qRT-PCR) was performed to evaluate the performance of four singleplex DENV-1-4 serotyping reverse transcription-iiPCR (RT-iiPCR) reagents for DENV subtyping on the mobile PCR system. The four RT-iiPCRs did not react with Zika virus and chikungunya virus; tests with serial dilutions of the four DENV serotypes made in human serum showed they had detection endpoints comparable to those of the reference method, indicating great analytical sensitivity and specificity. Clinical performance of the RT-iiPCR reagents was evaluated by testing 40 serum samples each (around 20 target serotype-positive and 20 DENV-negative); all four reagents had high agreement (97.5-100%) with the reference qRT-PCR. Moreover, testing of mosquitoes separately infected experimentally with each serotype showed that the four reagents detected specifically their target DENV serotypes in mosquito. CONCLUSIONS/SIGNIFICANCE: With analytical and clinical performance comparable to the reference qRT-PCR assay, the four index RT-iiPCR reagents on the field-deployable PCR system can serve as a useful tool for DENV detection near points of needs.

Generation of a potential koi herpesvirus live vaccine by simultaneous deletion of the viral thymidine kinase and dUTPase genes.

Koi herpesvirus (KHV, Cyprinidherpesvirus 3) causes a fatal disease of koi and common carp. To obtain safe and efficacious live vaccines, we generated deletion mutants of KHV lacking the nonessential genes encoding two enzymes of nucleotide metabolism, thymidine kinase (TK, ORF55) and deoxyuridine-triphosphatase (DUT, ORF123). Since single-deletion mutants based on a KHV isolate from Israel (KHV-I) only exhibited partial attenuation (Fuchs W, Fichtner D, Bergmann SM, Mettenleiter TC. Arch Virol 2011;156 : 1059-1063), a corresponding double mutant was generated and tested in vivo, and shown to be almost avirulent but still protective. To overcome the low in vitro virus titres of KHV-I (≤105 p.f.u. ml-1), single and double TK and DUT deletions were also introduced into a cell culture-adapted KHV strain from Taiwan (KHV-T). The deletions did not affect in vitro virus replication, and all KHV-T mutants exhibited wild-type-like plaque sizes and titres exceeding 107 p.f.u. ml-1, as a prerequisite for economic vaccine production. Compared to wild-type and revertant viruses, the single-deletion mutants of KHV-T were significantly attenuated in vivo, and immersion of juvenile carp in water containing high doses of the double mutant caused almost no fatalities. Nevertheless, the deletion mutants induced similar levels of KHV-specific serum antibodies to the parental wild-type virus, and conferred solid protection against disease after challenge with wild-type KHV. For the convenient differentiation of DNA samples prepared from gill swabs of carp infected with wild-type and TK-deleted KHV we developed a triplex real-time PCR. Thus, KHV-TΔDUT/TK might be suitable as a genetic DIVA vaccine in the field.

Characterization of gene deletion mutants of Cyprinid herpesvirus 3 (koi herpesvirus) lacking the immunogenic envelope glycoproteins pORF25, pORF65, pORF148 and pORF149.

Cyprinid herpesvirus 3 (CyHV-3) or koi herpesvirus is a global pathogen causing mass mortality in koi and common carp, against which improved vaccines are urgently needed. In this study we investigated the role of four nonessential, but immunogenic envelope glycoproteins encoded by members of the ORF25 gene family (ORF25, ORF65, ORF148 and ORF149) during CyHV-3 replication. Single deletion of ORF65 did not affect in vitro replication, and deletion of ORF148 even slightly enhanced virus growth on common carp brain (CCB) cells. Deletions of ORF25 or ORF149 led to reduced plaque sizes and virus titers, which was due to delayed entry into host cells. An ORF148/ORF149 double deletion mutant exhibited wild-type like growth indicating opposing functions of the two proteins. Electron microscopy of CCB cells infected with either mutant did not indicate any effects on virion formation and maturation in nucleus or cytoplasm, nor on release of enveloped particles. The ORF148, ORF149 and double deletion mutants were also tested in animal experiments using juvenile carp, and proved to be insufficiently attenuated for use as live virus vaccines. However, surviving fish were protected against challenge with wild-type CyHV-3, demonstrating that these antibody inducing proteins are dispensable for an efficient immune response in vivo.

Elimination of bromate from water using aluminum beverage cans via catalytic reduction and adsorption.

While zero valent aluminum (ZVAl) is a promising reductant for eliminating bromate from water, ZVAl is typically obtained from reagent grade aluminum. As used aluminum beverage can is the most common aluminum waste, it can be conveniently used to prepare ZVAl. Thus, in this study aluminum beverage cans are employed for the first time as a plentiful and easily accessible aluminum source to afford ZVAl for eliminating bromate from water. As aluminum is easily oxidized to form aluminum oxide, aluminum can pieces (ACPs) are pre-treated with HCl for removing the oxide layer to afford ZVAl. While non-acid-treated ACP is ineffective to remove bromate, the acid-treated ACP successfully eliminates bromate from water completely. Bromate elimination by ACP is attributed to reduction of bromate to bromide by the reactive sites of ACP and adsorption of bromate to the surface of ACP. Bromate elimination by ACP also proceeds much faster at higher temperatures and low pH values, while the alkaline condition causes serious negative effects on bromate elimination. Besides, oxalic acid is found to facilitate bromate elimination not only on the kinetics but also reduction to bromide because the passivation layer is suppressed in the presence of oxalic acid. ACP could also be reused and the acid-washing regeneration could enable used ACP to restore its reactive sites for bromate elimination. This study successfully demonstrates the valorization of aluminum beverage cans for mitigating the toxic bromate and the findings here provide useful information and insights to develop aluminum beverage cans for controlling pollutants in water.

On-site detection of bovine leukemia virus by a field-deployable automatic nucleic extraction plus insulated isothermal polymerase chain reaction system.

Bovine leukemia virus (BLV) is a contagious, oncogenic deltaretrovirus of cattle with a worldwide distribution. In the US, over 40% of dairy cows are infected with the virus, and evidence of its economic impact is growing. This study evaluated the performance of a field-deployable automatic nucleic acid-extraction/insulated isothermal PCR (iiPCR) system for on-site BLV-proviral DNA detection in dairy cows compared with a conventional laboratory real-time PCR (rt-PCR). Assay performance was verified in parallel tests of 36 archived blood samples with 100% agreement (κ = 1.0; n = 36) between the iiPCR and conventional rt-PCR systems, and the limit of detection of the iiPCR assay was estimated to be 4 copies (genome equivalent) per reaction. The field-deployable iiPCR system was subsequently used on-farm to test freshly collected blood samples, and showed 100% agreement (κ = 1.0; n = 32) with the laboratory rt-PCR system. Fresh blood samples were collected on a second farm and tested on both systems, also with 100% agreement (κ = 1.0; n = 34). The field-deployable iiPCR/POCKIT™ combo system performs as well as a conventional laboratory-based rt-PCR system for detection of BLV proviral DNA in whole blood and may be a useful tool for on-farm evaluation of BLV-infection status in dairy cattle.