Fine mapping and analysis of a candidate gene controlling Phytophthora blight resistance in cucumber.

Cucumber blight is a destructive disease. The best way to control this disease is resistance breeding. This study focuses on disease resistance gene mapping and molecular marker development. We used the resistant cucumber, JSH, and susceptible cucumber, B80, as parents to construct F2 populations. Bulked segregant analysis (BSA) combined with specific length amplified fragment sequencing (SLAF-seq) were used, from which we developed cleaved amplified polymorphic sequence (CAPs) markers to map the resistance gene. Resistance in F2 individuals showed a segregation ratio of resistance:susceptibility close to 3:1. The gene in JSH resistant cucumber was mapped to an interval of 9.25 kb, and sequencing results for the three genes in the mapped region revealed three mutations at base sites 225, 302, and 591 in the coding region of Csa5G139130 between JSH and B80, but no mutations in coding regions of Csa5G139140 and Csa5G139150. The mutations caused changes in amino acids 75 and 101 of the protein encoded by Csa5G139130, suggesting that Csa5G139130 is the most likely resistance candidate gene. We developed a molecular marker, CAPs-4, as a closely linked marker for the cucumber blight resistance gene. This is the first report on mapping of a cucumber blight resistance gene and will provideg a useful marker for molecular breeding of cucumber resistance to Phytophthora blight.

Influence of Nutritional Status on a Family-Centered Care Intervention for Older Adults with Cognitive Impairment following Hip-Fracture Surgery: Secondary Data Analysis of a Randomized Controlled Trial.

OBJECTIVES: In Taiwan, older adults with cognitive impairment who undergo hip-fracture surgery are routinely cared for by family members. This study aimed to determine if nutritional status influenced the effects of a family-centered intervention for older adults with cognitive impairment recovering from hip-fracture surgery. DESIGN: Secondary data analysis of data from a randomized controlled trial was conducted to examine the influences of nutritional status 1 month after hospital discharge on the effects of a family-centered care intervention model, which was designed for older adults with hip fracture and cognitive impairment. Outcomes were compared among participants according to nutrition status (well-nourished/poorly-nourished) and treatment approach (control/intervention). SETTING: The original study was conducted at a 3000-bed medical center from July 2015 to October 2019. PARTICIPANTS: Participants were older adults with cognitive impairment who had undergone hip-fracture surgery. Participants were assessed as poorly-nourished or well-nourished with the Mini-Nutritional Assessment (MNA) 1-month post-discharge and were then randomly assigned to either the intervention group or control group. INTERVENTION: A family-centered intervention model for family caregivers of older adults with cognitive impairment recovering from hip-fracture surgery was implemented. The intervention was delivered by geriatric nurses, which included instructions for family caregivers in overseeing exercises for strengthening the hip, understanding dietary requirements, and managing behavioral problems associated with cognitive impairment. MEASUREMENTS: Outcome measures included activities of daily living (ADLs), instrumental ADLs, hip range of motion, hip muscle strength, depression, measured with the Geriatric Depressive Scale, and physical and mental health related quality of life, measured with the Short Form Survey (SF-36), Taiwanese version. Participants were assessed at 1-, 3-, 6-, and 12-months post-discharge. RESULTS: Most of the 134 participants were assessed as poorly nourished (n = 122); 57 were the control group and 65 received the intervention. For the well-nourished participants (n = 12), four were in the intervention group and eight were controls. There were no significant differences in any outcome variables for poorly nourished participants who received the intervention compared with controls. For the sample of well-nourished participants, those who received the intervention performed significantly better in outcomes of IADLs (b = 1.74, p < .05), hip muscle strength (b = 9.64, p < .01), and physical health related quality of life (b = 10.47, p < .01). CONCLUSION: The family-centered care intervention was only effective for older adults with cognitive impairment recovering from hip-fracture surgery who were well-nourished at 1 month following hospital discharge, but not for those at risk of malnutrition. Interventions should focus on enhancing nutritional status following hip surgery which could allow the family-centered in-home intervention to be beneficial for more older adults with cognitive impairment recovering from hip-fracture surgery.

[Chronic hypoxia increases intracellular Ca(2+) concentration and augments proliferation by enhancing store-operated Ca(2+) entry in pulmonary arterial smooth muscle cells].

OBJECTIVE: To determine whether store-operated Ca(2+) entry (SOCE) is involved in chronic hypoxia-induced alteration of intracellular Ca(2+) concentration ([Ca(2+) ]i) and proliferation in pulmonary arterial smooth muscle cells (PASMC). METHODS: Rat PASMCs were cultured and treated in normoxia (21%O2) or hypoxia (4%O2) condition. The proliferation of PASMC was detected by cell counting kit-8 (CCK-8) assay. [Ca(2+) ]i, SOCE and the effects of store-operated Ca(2+) channel (SOCC) inhibitors, SKF96365 and NiCl2, on SOCE in hypoxic PASMCs were tested by InCyte [Ca(2+) ]i measurement system. RESULTS: Hypoxia for 24-60 h augmented PASMC proliferation (1.12±0.09 vs 0.71±0.05, P<0.05) and [Ca(2+) ]i [(214.8 ± 20.4) nmol/L vs (115.2±13.2) nmol/L, P<0.05] in a time-dependent manner with the maximum effect at 60 h. Perfusion of Ca(2+) -free Krebs solution containing nifedipine (5 µmol/L), cyclopiazonic acid (CPA, 10 µmol/L) in PASMCs caused a small transient increase of [Ca(2+) ]i with peak [Ca(2+) ]i (113.3±49.3) nmol/L.Chronic hypoxia (4% O2, 60 h) enhanced [Ca(2+) ]i level with peak value of (193.2±22.7) nmol/L (P<0.05) in PASMC.After restoration of extracellular Ca(2+) , CPA caused marked increase of [Ca(2+) ]i with peak value of (328.0 ±56.7) nmol/L.Chronic hypoxia strengthened CPA-induced increase of [Ca(2+) ]i with peak value of (526.0±33.7) nmol/L (P<0.05) in PASMCs.Either SKF96365 50 µmol/L or NiCl2 500 µmol/L distinctly attenuated CPA-induced enhancement of [Ca(2+) ]i, the peak value of which dropped from (526.0±33.7) nmol/L to (170.4±26.4) nmol/L (P<0.05) or (177.4±45.9) nmol/L (P<0.05) respectively. CONCLUSION: Chronic hypoxia boosts the release of Ca(2+) from sarcoplasmic reticulum and promotes the activity of SOCC and SOCE, leading to [Ca(2+) ]i elevation and proliferation of rat PASMCs.

14-3-3γ regulates cell viability and milk fat synthesis in lipopolysaccharide-induced dairy cow mammary epithelial cells.

Our previous study demonstrated that 14-3-3γ overexpression was able to inhibit the production of lipopolysaccharide (LPS)-induced cytokines in dairy cow mammary epithelial cells (DCMECs) by inhibiting the activation of nuclear factor-κB (NF-κB) signaling pathways. However, the association between 14-3-3γ overexpression and milk fat synthesis in LPS-induced DCMECs remains unclear. Therefore, the present study investigated the effect of 14-3-3γ on cell viability and milk fat synthesis in LPS-induced DCMECs. The results of the MTT assay and lactate dehydrogenase activity assay demonstrated that 14-3-3γ overexpression was able to attenuate LPS-induced cytotoxicity in DCMECs, and increase the viability of the cells. In addition, the results of reverse transcription-quantitative polymerase chain reaction suggested that mRNA expression levels of genes associated with milk fat synthesis, including sterol regulatory element binding protein (SREBP1), peroxisome proliferator-activated receptor-γ (PPARG), cluster of differentiation 36, acetyl-coA carboxylase (ACC), fatty acid synthase (FAS) and fatty acid binding protein-3, were significantly upregulated in cells overexpressing the 14-3-3γ protein. In addition, as compared with the LPS-treated group, the activities of FAS and ACC were significantly increased. Furthermore, western blotting demonstrated that 14-3-3γ overexpression enhanced the protein expression levels of phosphorylated SREBP1 and PPARG. These results suggested that high levels of 14-3-3γ protein were able to attenuate LPS-induced cell damage and promote milk fat synthesis in LPS-induced DCMECs by increasing the cell viability and upregulating the expression levels of transcription factors associated with milk fat synthesis.

Cost-effective identification of Mycobacterium tuberculosis complex using microscopic morphology and rapid tests.

BACKGROUND: The rapid diagnosis of tuberculosis (TB) by detecting and identifying Mycobacterium tuberculosis complex (MTC) in clinical culture isolates can be achieved by a combination of rapid tests. OBJECTIVE: To propose a cost-effective laboratory protocol for MTC identification. DESIGN: MTC (n = 278) was identified using microscopic morphology, two immunochromatographic assays (ICAs) (Tibilia™ and MeDiPro(®) M. tuberculosis Antigen Rapid Test) and the strand displacement amplification (SDA) method (ProbeTec), and the results were compared. RESULTS: Microscopic morphology (cord-like) had a sensitivity of 99.3%, a specificity of 84.3%, a positive predictive value (PPV) of 88.2% and a negative predictive value (NPV) of 99.1%. The overall sensitivity/specificity of the Tibilia, MeDiPro and ProbeTec assays were respectively 98.7%/98.4%, 88.0%/85.2% and 97.4%/98.4%. The PPV/NPV for Tibilia, MeDiPro and ProbeTec were respectively 98.7%/98.4%, 87.4%/85.8% and 98.7%/96.8%. Cord-like microscopy was the least expensive method and could be used for the identification of MTC. ICA offers cost-effective screening compared to the SDA method. Tibilia performed better than MeDiPro, while its diagnostic value was similar to the SDA method. CONCLUSION: We recommend a combination of microscopic morphology and Tibilia to further improve the sensitivity and PPV of MTC identification at lower cost.

Biliary papillomatosis with malignant transformation: A case report and review of the literature.

Biliary papillomatosis is a rare benign tumor that possesses a high potential for malignant transformation, and is frequently misdiagnosed. In the present report, a case of biliary papillomatosis with malignant transformation in a 63-year-old female, with a 19-year history of cholelithiasis, is presented. The patient presented with right epigastric pain and magnetic resonance imaging revealed a diffuse mass in the right liver lobe, which extended through the common bile duct. Definitive preoperative diagnosis was unable to be established; however, following surgical resection of the tumor, a diagnosis of biliary papillomatosis with malignant transformation was confirmed by pathological examination. The patient was followed up seven months later and no recurrence was detected. A supplementary review of previously published literature is additionally presented.

From SARS in 2003 to H1N1 in 2009: lessons learned from Taiwan in preparation for the next pandemic.

In anticipation of a future pandemic potentially arising from H5N1, H7N9 avian influenza or Middle East Respiratory Syndrome, and in large part in response to severe acute respiratory syndrome (SARS) in 2003, the city of Taipei, Taiwan, has developed extensive new strategies to manage pandemics. These strategies were tested during the 2009 H1N1 outbreak. This article assesses pandemic preparedness in Taipei in the wake of recent pandemic experiences in order to draw lessons relevant to the broader international public health community. Drawing on Taiwan and Taipei Centers for Disease Control data on pandemic response and control, we evaluated the effectiveness of the changes in pandemic response policies developed by these governments over time, emphasizing hospital and medical interventions with particular attention paid to Traffic Control Bundling. SARS and H1N1 2009 catalysed the Taiwan and Taipei CDCs to continuously improve and adjust their strategies for a future pandemic. These new strategies for pandemic response and control have been largely effective at providing interim pandemic containment and control, while development and implementation of an effective vaccination programme is underway. As Taipei's experiences with these cases illustrate, in mitigating moderate or severe pandemic influenza, a graduated process including Traffic Control Bundles accompanied by hospital and medical interventions, as well as school- and community-focused interventions, provides an effective interim response while awaiting vaccine development. Once a vaccine is developed, to maximize pandemic control effectiveness, it should be allocated with priority given to vulnerable groups, healthcare workers and school children.

Removal of waterborne pathogens from liver transplant unit water taps in prevention of healthcare-associated infections: a proposal for a cost-effective, proactive infection control strategy.

Hospital water supplies often contain waterborne pathogens, which can become a reservoir for healthcare-associated infections (HAIs). We surveyed the extent of waterborne pathogen contamination in the water supply of a Liver Transplant Unit. The efficacy of point-of-use (POU) water filters was evaluated by comparative analysis in routine clinical use. Our baseline environmental surveillance showed that Legionella spp. (28%, 38/136), Pseudomonas aeruginosa (8%, 11/136), Mycobacterium spp. (87%, 118/136) and filamentous fungi (50%, 68/136) were isolated from the tap water of the Liver Transplant Unit. 28.9% of Legionella spp.-positive water samples (n = 38) showed high-level Legionella contamination (≥10(3) CFU/L). After installation of the POU water filter, none of these pathogens were found in the POU filtered water samples. Furthermore, colonizations/infections with Gram-negative bacteria determined from patient specimens were reduced by 47% during this period, even if only 27% (3/11) of the distal sites were installed with POU water filters. In conclusion, the presence of waterborne pathogens was common in the water supply of our Liver Transplant Unit. POU water filters effectively eradicated these pathogens from the water supply. Concomitantly, healthcare-associated colonization/infections declined after the POU filters were installed, indicating their potential benefit in reducing waterborne HAIs.

Spironolactone prevents dietary-induced metabolic syndrome by inhibiting PI3-K/Akt and p38MAPK signaling pathways.

OBJECTIVES: Aim of the study is to evaluate the impact of spironolactone (SPL) on indexes of metabolic syndrome (MS) and further investigate the mechanisms underlying its protective effects. METHODS: A rat model of MS was established by administering a fat- and salt-enriched diet (FS diet). The occurrence of MS was examined by measurement of blood pressure (BP), aldosterone (ALD) content, blood lipid (BL), glucose and insulin levels. Homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. Pancreatic gland tissue injury was assessed by ß-cell apoptosis. Mineralocorticoid receptor (MR) activity, phosphatidylinositol 3- kinase/Akt (PI3-K/Akt), and phosphorylation of p38MAPK (Pp38MAPK) in pancreatic gland tissue were evaluated by western blot analysis. RESULTS: SPL prevented hypertension, and dyslipidemia during MS induced by the intake of FS diet, but had no effect on K+ and Na+ disturbances. Furthermore, SPL significantly attenuated ALD and MR expression levels after FS diet. Finally, SPL inhibited phosphorylation protein kinase B (p- PKB) activation in the pancreatic gland tissue, a downstream target of PI3-K, and phosphorylation of p38MAPK pathway, critical for cellular apoptosis. CONCLUSIONS: This study demonstrates that SPL exerts a protective effect on hypertension and dyslipidemia. This protective effect may depend, at least in part, on MAPK and PI3-K pathways.

Taiwan's traffic control bundle and the elimination of nosocomial severe acute respiratory syndrome among healthcare workers.

The traffic control bundle consists of procedures designed to help prevent epidemic nosocomial infection. We retrospectively studied the serial infection control measures to determine factors most effective in preventing nosocomial infections of healthcare workers (HCWs) during the 2003 Taiwanese severe acute respiratory syndrome (SARS) epidemic. Fever screening stations, triage of fever patients, separating SARS patients from other patients, separation of entrances and passageways between patients and HCWs, and increasing hand-washing facilities all demonstrated a protective effect for HCWs (univariate analysis; P<0.05). By multiple logistic regression: (i) checkpoint alcohol dispensers for glove-on hand rubbing between zones of risk, and (ii) fever screening at the fever screen station outside the emergency department, were the significant methods effectively minimising nosocomial SARS infection of HCWs (P<0.05). The traffic control bundle should be implemented in future epidemics as a tool to achieve strict infection control measures.

Combination of molecular assay and clinical evaluation for early confirmation of tuberculosis cases.

The cost-effectiveness of the ProbeTec ET Direct TB assay (DTB) was compared with that of culture for detection of Mycobacterium tuberculosis complex in 361 acid-fast stain-positive respiratory specimens. The overall sensitivity, specificity, positive predictive value and negative predictive value of DTB were 97.7%, 86.6%, 87.2% and 97.6%, respectively. When clinical evaluation was added to DTB, the specificity and positive predictive value of DTB increased to 94.7% and 95.4%, respectively. Treatment costs of $133,521 would have been saved in this cohort if DTB, instead of culture results, had been used to eliminate 'false-positive' smear results.

Epidemiological investigation of a case of nosocomial Legionnaires' disease in Taiwan: implications for routine environmental surveillance.

An epidemiological investigation with Legionella and molecular subtyping was conducted to determine the source of a case of nosocomial Legionnaires' disease (LD) who was hospitalized in three hospitals within a month. Legionella pneumophila serogroup 3, an uncommon serogroup for infection, was isolated from the patient's sputum. Environmental surveillance revealed Legionella colonization in all three hospitals; the patient isolate matched the isolate from the first hospital by molecular typing. Culturing the hospital water supply for Legionella is a pro-active strategy for detection of nosocomial LD even in hospitals experiencing no previous cases.

Efficacy of point-of-entry copper--silver ionisation system in eradicating Legionella pneumophila in a tropical tertiary care hospital: implications for hospitals contaminated with Legionella in both hot and cold water.

A medical centre in Southern Taiwan experienced an outbreak of nosocomial Legionnaires' disease, with the water distribution system thought to be the source of the infection. Even after two superheats and flush, the rate of Legionella positivity in distal sites in hospital wards and intensive care units (ICUs) was 14% and 66%, respectively. Copper-silver ionisation was therefore implemented in an attempt to control Legionella colonisation in both hot- and cold-water systems. Environmental cultures and ion concentration testing were performed to evaluate the efficacy of ionisation. When the system was activated, no significant change in rate of Legionella positivity in the hospital wards (20% vs baseline of 30%) and ICUs (28% vs baseline of 34%) of the test buildings over a three-month period was found, although all Legionella positivity rates were below 30%, an arbitrary target for Legionnaires' disease prevention. When ion concentrations were increased from month 4 to month 7, however, the rate of Legionella positivity decreased significantly to 5% (mean) in hospital wards (P=0.037) and 16% (mean) in ICUs (P=0.037). Legionella positivity was further reduced to 0% in hospital wards and 5% (mean) in ICUs while 50% sites were still positive for Legionella in a control building. Although Legionella was not completely eradicated during the study period, no culture- or urine-confirmed hospital-acquired Legionnaires' disease was reported. Ionisation was effective in controlling Legionella for both hot and cold water, and may be an attractive alternative as a point-of-entry systematic disinfection solution for Legionella.

Effect of flow regimes on the presence of Legionella within the biofilm of a model plumbing system.

AIMS: Stagnation is widely believed to predispose water systems to colonization by Legionella. A model plumbing system was constructed to determine the effect of flow regimes on the presence of Legionella within microbial biofilms. METHODS AND RESULTS: The plumbing model contained three parallel pipes where turbulent, laminar and stagnant flow regimes were established. Four sets of experiments were carried out with Reynolds number from 10,000 to 40,000 and from 355 to 2,000 in turbulent and laminar pipes, respectively. Legionella counts recovered from biofilm and planktonic water samples of the three sampling pipes were compared with to determine the effect of flow regime on the presence of Legionella. Significantly higher colony counts of Legionella were recovered from the biofilm of the pipe with turbulent flow compared with the pipe with laminar flow. The lowest counts were in the pipe with stagnant flow. CONCLUSIONS: We were unable to demonstrate that stagnant conditions promoted Legionella colonization. SIGNIFICANCE AND IMPACT OF THE STUDY: Plumbing modifications to remove areas of stagnation including deadlegs are widely recommended, but these modifications are tedious and expensive to perform. Controlled studies in large buildings are needed to validate this unproved hypothesis.

Using an integrated infection control strategy during outbreak control to minimize nosocomial infection of severe acute respiratory syndrome among healthcare workers.

Healthcare workers (HCWs) are at risk of acquiring severe acute respiratory syndrome (SARS) while caring for SARS patients. Personal protective equipment and negative pressure isolation rooms (NPIRs) have not been completely successful in protecting HCWs. We introduced an innovative, integrated infection control strategy involving triaging patients using barriers, zones of risk, and extensive installation of alcohol dispensers for glove-on hand rubbing. This integrated infection control approach was implemented at a SARS designated hospital ('study hospital') where NPIRs were not available. The number of HCWs who contracted SARS in the study hospital was compared with the number of HCWs who contracted SARS in 86 Taiwan hospitals that did not use the integrated infection control strategy. Two HCWs contracted SARS in the study hospital (0.03 cases/bed) compared with 93 HCWs in the other hospitals (0.13 cases/bed) during the same three-week period. Our strategy appeared to be effective in reducing the incidence of HCWs contracting SARS. The advantages included rapid implementation without NPIRs, flexibility to transfer patients, and re-inforcement for HCWs to comply with infection control procedures, especially handwashing. The efficacy and low cost are major advantages, especially in countries with large populations at risk and fewer economic resources.