The epidemiology of non-typhoidal Salmonella gastroenteritis and Campylobacter gastroenteritis in pediatric inpatients in northern Taiwan

BackgroundCampylobacter and Non-typhoidal Salmonella (NTS) are the two most common bacterial pathogens associated with acute gastroenteritis in children. This study aims to elucidate the epidemiology of Campylobacter and NTS gastroenteritis and develop a scoring system to differentiate them.Materials and methodsThis retrospective study enrolled 886 children ≤18 years of age, hospitalized due to acute gastroenteritis with stool culture-proven Campylobacter or NTS infection from July 2012 to December 2015. Pearson's chi-square test and multivariate logistic regression were used to compare clinical manifestations and laboratory data. Receiver operating characteristic curves were plotted to evaluate the scoring system.ResultsSeasonality was found in NTS gastroenteritis from May to September, but no seasonality in Campylobacter gastroenteritis. Campylobacter jejuni and Salmonella serogroup B were the most common pathogens. The median ages were 68.2 and 18.5 months and the incidence rates of bacteremia were 0.6% and 7.1% in the Campylobacter and NTS groups, respectively. Salmonella serogroup C2 infection had the highest risk of bacteremia (OR: 5.9, 95% CI: 2.8–12.7, p < 0.001). Multivariate analysis showed significant differences in sex, age, fever, dehydration, immature WBC, CRP and Na between the two groups. A score of ≥2 points indicated Campylobacter gastroenteritis, with sensitivity 75%, specificity 77%. The positive and negative predictive values were of 73.3% and 93.9% after validation.ConclusionCampylobacter gastroenteritis is associated with older age and male sex, while NTS gastroenteritis is associated with moderate to severe dehydration and bacteremia. Salmonella serogroup C2 infection has the highest risk of bacteremia.     

Analysis of Risk Factors for Bacteremia in Children with Nontyphoidal Salmonella Gastroenteritis

To identify the risk factors for Salmonella bacteremia in infants and children with Salmonella gastroenteritis, a retrospective study of a 10-year period was conducted to evaluate 456 infants and children with culture-proven nontyphoidal Salmonella infection. Salmonella typhimurium was the most common isolate found. Among the 257 patients with gastroenteritis who had a concomitant blood culture performed, 50 exhibited bacteremia. Statistically significant differences were noted between patients with gastroenteritis and bacteremia and those without bacteremia in duration of fever ≥5 days (P<0.001; OR, 5.6; 95% CI, 2.6–12.1) and infection with group D1 Salmonella (P<0.001; OR, 6.5; 95% CI, 2.5–16.9) after adjustment for multivariate analysis. Of the 320 Salmonella strains that were serotyped, Salmonella panama was shown to be strongly associated with bacteremia (P<0.001) in children with gastroenteritis. In summary, in children with nontyphoidal Salmonella gastroenteritis, prolonged fever lasting 5 days or more and infection with a specific Salmonella serotype were risk factors closely associated with development of bacteremia. Electronic Publication     

The differential effects of 1,25‐dihydroxyvitamin D3 on Salmonella‐induced interleukin‐8 and human beta‐defensin‐2 in intestinal epithelial cells

Salmonellosis or Salmonella, one of the most common food‐borne diseases, remains a major public health problem worldwide. Intestinal epithelial cells (IECs) play an essential role in the mucosal innate immunity of the host to defend against the invasion of Salmonella by interleukin (IL)?8 and human β‐defensin‐2 (hBD‐2). Accumulated research has unravelled important roles of vitamin D in the regulation of innate immunity. Therefore, we investigated the effects of 1,25‐dihydroxyvitamin D3 (1,25D3) on Salmonella‐induced innate immunity in IECs. We demonstrate that pretreatment of 1,25D3 results in suppression of Salmonella‐induced IL‐8 but enhancement of hBD‐2, either protein secretion and mRNA expression, in IECs. Furthermore, 1,25D3 enhanced Salmonella‐induced membranous recruitment of nucleotide oligomerization domain (NOD2) and its mRNA expression and activation of protein kinase B (Akt), a downstream effector of phosphoinositide 3‐kinase (PI3K). Inhibition of the PI3K/Akt signal counteracted the suppressive effect of 1,25D3 on Salmonella‐induced IL‐8 expression, while knock‐down of NOD2 by siRNA diminished the enhanced hBD‐2 expression. These data suggest differential regulation of 1,25D3 on Salmonella‐induced IL‐8 and hBD‐2 expression in IECs via PI3K/Akt signal and NOD2 protein expression, respectively. Active vitamin D‐enhanced anti‐microbial peptide in Salmonella‐infected IECs protected the host against infection, while modulation of proinflammatory responses by active vitamin D prevented the host from the detrimental effects of overwhelming inflammation. Thus, active vitamin D‐induced innate immunity in IECs enhances the host's protective mechanism, which may provide an alternative therapy for invasive Salmonella infection.     

Single‐molecule detection of cancer mutations using a novel PCR‐LDR‐qPCR assay

Detection of low‐abundance mutations in cell‐free DNA is being used to identify early cancer and early cancer recurrence. Here, we report a new PCR‐LDR‐qPCR assay capable of detecting point mutations at a single‐molecule resolution in the presence of an excess of wild‐type DNA. Major features of the assay include selective amplification and detection of mutant DNA employing multiple nested primer‐binding regions as well as wild‐type sequence blocking oligonucleotides, prevention of carryover contamination, spatial sample dilution, and detection of multiple mutations in the same position. Our method was tested to interrogate the following common cancer somatic mutations: BRAF:c.1799T>A (p.Val600Glu), TP53:c.743G>A (p.Arg248Gln), KRAS:c.35G>C (p.Gly12Ala), KRAS:c.35G>T (p.Gly12Val), KRAS:c.35G>A (p.Gly12Asp), KRAS:c.34G>T (p.Gly12Cys), and KRAS:c.34G>A (p.Gly12Ser). The single‐well version of the assay detected 2–5 copies of these mutations, when diluted with 10,000 genome equivalents (GE) of wild‐type human genomic DNA (hgDNA) from buffy coat. A 12‐well (pixel) version of the assay was capable of single‐molecule detection of the aforementioned mutations at TP53, BRAF, and KRAS (specifically p.Gly12Val and p.Gly12Cys), mixed with 1,000–2,250 GE of wild‐type hgDNA from plasma or buffy coat. The assay described herein is highly sensitive, specific, and robust, and potentially useful in liquid biopsies.     

Effectiveness of mite‐impermeable covers: a hypothesis‐generating meta‐analysis

Asthma is a heterogeneous disease. The subject of mite allergen control has evolved into a debate dominated by a Cochrane review by Gøtzsche and Johansen (Cochrane Database of Systematic Reviews, 2008, Art. No: CD001187). A not well‐discussed aspect of that study is the selection by those authors of a univariate meta‐analysis including various interventions. This study extends the meta‐analysis by Gøtzsche and Johansen and aims to generate hypotheses on the effectiveness of various bedding interventions, including the coverage of all bedding elements. Trials were selected based on environmental criteria. The interventions were classified according to the number of barriers used. Standardized mean differences yielded the mite load, three physiological outcomes and asthma symptom scores. The influence of covariates was examined with a mixed‐effect model using the metafor package for meta‐analysis in R. Twelve trials included 1187 observations. The interventions included one barrier or product (six trials), two barriers or partial control (four trials) and three barriers or integral control (two trials). The exposure data showed considerable heterogeneity (I² = 93%). The risk of bias significantly (P = 0.04) influenced the final load, the square root of the interaction between the baseline load and the type of intervention as well (95% CI: ?0.66 to ?0.07 μg/g; P = 0.02). Changes in load showed similar tendencies. Health outcomes showed moderate to considerable heterogeneity (physiological outcomes I² = 44–94%; symptom score I² = 93%). A meta‐regression of bedding interventions indicates that integral control most significantly reduced mite load when the load was high at baseline. The number of trials was too small to allow an appropriate examination of health outcomes. Future studies are suggested to test the hypothesis that allergic patients benefit from integral control when the baseline mite load is high.     

Large‐scale genomic instability in colon adenocarcinomas and correlation with patient outcome

The purpose of this study was to evaluate the association between DNA content in colon adenocarcinomas using high‐resolution image cytometry and patient outcome. Tumours from 219 patients operated for colon adenocarcinoma were analysed using high‐resolution image cytometry. Proteins involved in cell cycle propulsion (cyclins A, D1, D3 and E) and cell proliferation (c‐Myc and non‐membranous β‐catenin) have previously been reported in the same cohort and were included in this study. The results were related to disease‐free survival and to cancer‐specific death. Patients with aneuploid tumours showed shorter relapse‐free survival than patients with euploid tumours (univariate log‐rank test, p = 0.004 and multivariate Cox regression model p = 0.009, HR 0.51, 95% CI 0.31–0.84). Also the risk of death from cancer was greater in patients with aneuploid tumours (log‐rank test, p = 0.006 multivariate Cox regression model p = 0.014, HR 0.47, 95% CI 0.26–0.86). When analysing patients with Dukes stages A and B, nuclear expression of β‐catenin was highly significantly associated with both shorter relapse‐free survival (p < 0.005, HR 5.0, 95% CI 1.6–15.5) and cancer‐specific death (p = 0.036, HR 6.9, 95% CI 1.1–42.1). DNA content in colon adenocarcinomas measured by image cytometry is an independent predictor of prognosis in our patients operated for colon adenocarcinoma.     

Suppression of SPIN1‐mediated PI3K–Akt pathway by miR‐489 increases chemosensitivity in breast cancer

Drug resistance is one of the major obstacles for improving the prognosis of breast cancer patients. Increasing evidence has linked the association of aberrantly expressed microRNAs (miRNAs) with tumour development and progression as well as chemoresistance. Despite recent advances, there is still little known about the potential role and mechanism of miRNAs in breast cancer chemoresistance. Here we describe that 16 miRNAs were found to be significantly down‐regulated and 11 up‐regulated in drug‐resistant breast cancer tissues compared with drug‐sensitive tissues, using a miRNA microarray. The results also showed miR‐489 to be one of the most down‐regulated miRNAs in drug‐resistant tissues and cell lines, as confirmed by miRNA microarray screening and real‐time quantitative PCR. A decrease in miR‐489 expression was associated with chemoresistance as well as lymph node metastasis, increased tumour size, advanced pTNM stage and poor prognosis in breast cancer. Functional analysis revealed that miR‐489 increased breast cancer chemosensitivity and inhibited cell proliferation, migration and invasion, both in vitro and in vivo. Furthermore, SPIN1, VAV3, BCL2 and AKT3 were found to be direct targets of miR‐489. SPIN1 was significantly elevated in drug‐resistant and metastatic breast cancer tissues and inversely correlated with miR‐489 expression. High expression of SPIN1 was associated with higher histological grade, lymph node metastasis, advanced pTNM stage and positive progesterone receptor (PR) status. Increased SPIN1 expression enhanced cell migration and invasion, inhibited apoptosis and partially antagonized the effects of miR‐489 in breast cancer. PIK3CA, AKT, CREB1 and BCL2 in the PI3K–Akt signalling pathway, demonstrated to be elevated in drug‐resistant breast cancer tissues, were identified as downstream effectors of SPIN1. It was further found that either inhibition of SPIN1 or overexpression of miR‐489 suppressed the PI3K–Akt signalling pathway. These data indicate that miR‐489 could reverse the chemoresistance of breast cancer via the PI3K–Akt pathway by targeting SPIN1. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Association of cytotoxic T‐lymphocyte associated protein 4 gene polymorphisms with the risk and prognosis of oesophageal cancer in a high‐incidence region from northern China

The most important anti‐tumour immune response is mediated by T lymphocytes. Cytotoxic T lymphocyte‐associated protein 4 (CTLA4) plays a critical role in the immune surveillance against tumours as an inhibitory immune checkpoint molecule of T‐cell activation. This study was designed to explore the association of CTLA4 polymorphisms with the susceptibility to oesophageal squamous cell carcinoma (ESCC) and prognosis of patients with ESCC in a high‐incidence population from northern China. CTLA4 rs5742909 C/T and rs231775 G/A single nucleotide polymorphisms (SNPs) were genotyped using polymerase chain reaction–ligase detection reaction (PCR‐LDR) method in 577 ESCC patients and 580 controls. Upper gastrointestinal cancer family history increased the risk of ESCC (the sex‐, age‐ and smoking status‐adjusted OR = 1.383, 95%CI = 1.094–1.749). The genotype frequencies of these two SNPs in the patients with ESCC were similar to that in the controls. Survival analyses were conducted in 204 patients with ESCC with five‐year survival information. The mean survival time of ESCC patients with rs231775 SNP A/A genotype in age over 60 years group was 23.2 months, significantly shorter than that of those with G/G genotype (47.3 months). The A/A genotype was associated with increased death risk of patients with ESCC older than 60 years (adjusted HR = 4.544, 95%CI = 1.913–10.790). CTLA4 rs231775 SNP might be used as genetic marker of worse prognosis for patients with ESCC over 60 years in a high‐incidence population from northern China.     

Improved multiparametric MRI discrimination between low‐risk prostate cancer and benign tissues in a small cohort of 5α‐reductase inhibitor treated individuals as compared with an untreated cohort

The purpose of this study was to determine whether 5α‐reductase inhibitors (5‐ARIs) affect the discrimination between low‐grade prostate cancer and benign tissues on multiparametric MRI (mpMRI). Twenty men with biopsy‐proven Gleason 3 + 3 prostate cancer and 3 T mpMRI were studied. Ten patients (Tx) had been receiving 5‐ARIs for at least a year at scan time. Ten untreated patients (Un) were matched to the treated cohort. For each subject two regions of interest representing cancerous and benign tissues were drawn within the peripheral zone of each prostate, MR measures evaluated, and cancer contrast versus benign (contrast = (MR_Tumor ? MR_Healthy)/MR_Healthy) calculated. Decreased cancer contrast was noted on T₂‐weighted images: 0.4 (Un) versus 0.3 (Tx). However, for functional MR measures, a better separation of cancerous and benign tissues was observed in the treated group. Cancer contrast on high‐b diffusion‐weighted imaging (DWI) was 0.61 (Un) versus 0.99 (Tx). Logistic regression analysis yielded higher AUC (area under the curve) values for distinguishing cancerous from benign regions in treated subjects on high‐b DWI (0.71 (Un), 0.94 (Tx)), maximal enhancement slope (0.95 (Un), 1 (Tx)), peak enhancement (0.84 (Un), 0.93 (Tx)), washout slope (0.78 (Un), 0.99 (Tx)), K ^trans (0.9 (Un), 1 (Tx)), and combined measures (0.86 (Un), 0.99 (Tx)). Coefficients of variation for MR measures were lower in benign and cancerous tissues in the treated group compared with the untreated group. This study's results suggest an increase in homogeneity of benign and malignant peripheral zone prostatic tissues with 5‐ARI exposure, observed as reduced variability of MR measures after treatment. Cancer discrimination was lower with T₂‐weighted imaging, but was higher with functional MR measures in a 5‐ARI‐treated cohort compared with controls.     

Specific and Cross‐reactive Plasmablast Response in Humans after Primary and Secondary Immunization with Vi Capsular Polysaccharide Typhoid Vaccine

Secondary immunization with polysaccharide vaccines may imply a risk of hyporesponsiveness. Despite the wide use of typhoid Vi capsular polysaccharide vaccine, its potential tendency to hyporesponsiveness has been inadequately addressed. While previous studies have explored serum antibody responses, we applied a more sensitive approach, a single‐cell assay for circulating plasmablasts, to compare primary and secondary responses. Twelve subjects received primary and booster doses of the Vi vaccine (Typherix^®) at 30‐ to 37‐month intervals. Plasmablasts specific to the Vi or typhoidal O antigens or cross‐reactive with paratyphoid and non‐typhoidal Salmonella strains were identified as antibody‐secreting cells (ASC) with ELISPOT. Before vaccinations, none had plasmablasts specific to the antigens tested. Twelve of 12 subjects showed a Vi‐specific response after primary, but only eight of 12 after booster vaccination. All responded to typhoidal O‐9,12 antigen after both immunizations. The geometric mean of plasmablasts specific to the Vi antigen was 59 (95% CI 24–119) and 1 (0‐54) IgA + IgG + IgM‐ASC/10⁶ peripheral blood mononuclear cell (PBMC) after primary and booster immunizations, respectively, and 20 (9–49) and 56 (29–103) to the O‐9,12 antigen. We detected 1 (0–28) and 17 (6–36) ASC/10⁶ PBMC cross‐reactive with Salmonella Paratyphi A; 3 (0–30) and 22 (8–48) with S. Paratyphi B; 3 (0–29) and 18 (7–47) with S. Paratyphi C; 19 (10‐34) and 51 (26‐94) with Salmonella Enteritidis; and 1 (0–35) and 23 (9–52) with Salmonella Typhimurium, respectively. One‐third of the vaccinees, although responding to the O‐9,12 antigen, failed to respond to the Vi antigen after booster immunization, suggesting hyporesponsiveness in part of the vaccinees. The findings warrant further investigation.     

miR‐29a inhibition normalizes HuR over‐expression and aberrant AU‐rich mRNA stability in invasive cancer

The activities of RNA‐binding proteins are perturbed in several pathological conditions, including cancer. These proteins include tristetraprolin (TTP, ZFP36) and HuR (ELAVL1), which respectively promote the decay or stability of adenylate‐uridylate‐rich (AU‐rich) mRNAs. Here, we demonstrated that increased stabilization and subsequent over‐expression of HuR mRNA were coupled to TTP deficiency. These findings were observed in breast cancer cell lines with an invasive phenotype and were further confirmed in ZFP36‐knockout mouse fibroblasts. We show that TTP–HuR imbalance correlated with increased expression of AU‐rich element (ARE) mRNAs that code for cancer invasion genes. The microRNA miR‐29a was abundant in invasive breast cancer cells when compared to non‐tumourigenic cell types. When normal breast cells were treated with miR‐29a, HuR mRNA and protein expression were up‐regulated. MiR‐29a recognized a seed target in the TTP 3′ UTR and a cell‐permeable miR‐29a inhibitor increased TTP activity towards HuR 3′ UTR. This led to HuR mRNA destabilization and restoration of the aberrant TTP–HuR axis. Subsequently, the cancer invasion factors uPA, MMP‐1 and MMP‐13, and cell invasiveness, were decreased. The TTP:HuR mRNA ratios were also perturbed in samples from invasive breast cancer patients when compared with normal tissues, and were associated with invasion gene expression. This study demonstrates that an aberrant ARE‐mediated pathway in invasive cancer can be normalized by targeting the aberrant and functionally coupled TTP–HuR axis, indicating a potential therapeutic approach. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Beneficial metabolic phenotypes caused by loss‐of‐function APOC3 mutations

Loss‐of‐function mutations in APOC3, triglycerides, and coronary disease. TG & HDL Working Group of the NHLBI Exome Sequencing Project (2014) N Engl J Med 371:22–31 Loss‐of‐function mutations in APOC3 and risk of ischemic vascular disease. Jørgensen et al. (2014) N Engl J Med 371:32–41     

Sleep disorders and an increased risk of Parkinson's disease in individuals with non‐apnea sleep disorders: a population‐based cohort study

Sleep disorders are common non‐motor symptoms in patients with Parkinson's disease. Our study aims to explore the relationship between non‐apnea sleep disorders and future Parkinson's disease. This is a cohort study using a nationwide database. The participants were recruited from the Taiwan National Health Insurance Research Database between 2000 and 2003. A total of 91 273 adult patients who had non‐apnea sleep disorders without pre‐existing Parkinson's disease were enrolled. An age‐, gender‐, income‐, urbanization‐ and Charlson comorbidity index score‐matched control cohort consisting of 91 273 participants was selected for comparison. The two cohorts were followed for the occurrence of Parkinson's disease, death or until the end of 2010, whichever came first. The Kaplan–Meier analyses revealed patients with non‐apnea sleep disorders tended to develop Parkinson's disease (log‐rank test, P < 0.001). After a multivariate adjustment in a Cox regression model, non‐apnea sleep disorders was an independent risk factor for the development of Parkinson's disease [crude hazard ratio: 1.63, 95% confidence interval (CI): 1.54–1.73, P < 0.001; adjusted hazard ratio: 1.18, 95% CI: 1.11–1.26, P < 0.001]. In the subgroup analysis, patients with chronic insomnia (lasting more than 3 months) had the greatest risk (crude hazard ratio: 2.91, 95% CI: 2.59–3.26, P < 0.001; adjusted hazard ratio: 1.37, 95% CI: 1.21–1.55, P < 0.001). In conclusion, this study revealed that non‐apnea sleep disorders, especially chronic insomnia, are associated with a higher risk for future Parkinson's disease.     

Next‐generation‐sequencing‐based risk stratification and identification of new genes involved in structural and sequence variations in near haploid lymphoblastic leukemia

Near haploidy (23–29 chromosomes) is a numerical cytogenetic aberration in childhood acute lymphoblastic leukemia (ALL) associated with particularly poor outcome. In contrast, high hyperdiploidy (51–67 chromosomes) has a favorable prognosis. Correct classification and appropriate risk stratification of near haploidy is frequently hampered by the presence of apparently high hyperdiploid clones that arise by endoreduplication of the original near haploid clone. We evaluated next‐generation‐sequencing (NGS) to distinguish between “high hyperdiploid” leukemic clones of near haploid and true high hyperdiploid origin. Five high hyperdiploid ALL cases and the “high hyperdiploid” cell line MHH‐CALL‐2, derived from a near haploid clone, were tested for uniparental isodisomy. NGS showed that all disomic chromosomes of MHH‐CALL‐2, but none of the patients, were of uniparental origin, thus reliably discriminating these subtypes. Whole‐exome‐ and whole‐genome‐sequencing of MHH‐CALL‐2 revealed homozygous non‐synonymous coding mutations predicted to be deleterious for the protein function of 63 genes, among them known cancer‐associated genes, such as FANCA, NF1, TCF7L2, CARD11, EP400, histone demethylases, and transferases (KDM6B, KDM1A, PRDM11). Only eight of these were also, but heterozygously, mutated in the high hyperdiploid patients. Structural variations in MHH‐CALL‐2 include a homozygous deletion (MTAP/CDKN2A/CDKN2B/ANRIL), a homozygous inversion (NCKAP5), and an unbalanced translocation (FAM189A1). Together, the sequence variations provide MHH‐CALL‐2 with capabilities typically acquired during cancer development, e.g., loss of cell cycle control, enhanced proliferation, lack of DNA repair, cell death evasion, and disturbance of epigenetic gene regulation. Poorer prognosis of near haploid ALL most likely results from full penetrance of a large array of detrimental homozygous mutations. © 2013 Wiley Periodicals, Inc.     

Direct visualization of endogenous Salmonella‐specific B cells reveals a marked delay in clonal expansion and germinal center development

CD4⁺ T cells and B cells are both essential for acquired immunity to Salmonella infection. It is well established that Salmonella inhibit host CD4⁺ T‐cell responses, but a corresponding inhibitory effect on B cells is less well defined. Here, we utilize an Ag tetramer and pull‐down enrichment strategy to directly visualize OVA‐specific B cells in mice, as they respond to infection with Salmonella‐OVA. Surprisingly, OVA‐specific B‐cell expansion and germinal center formation was not detected until bacteria were cleared from the host. Furthermore, Salmonella infection also actively inhibited both B‐ and T‐cell responses to the same coinjected Ag but this did not require the presence of iNOS. The Salmonella Pathogenicity Island 2 (SPI2) locus has been shown to be responsible for inhibition of Salmonella‐specific CD4⁺ T‐cell responses, and an examination of SPI2‐deficient bacteria demonstrated a recovery in B‐cell expansion in infected mice. Together, these data suggest that Salmonella can simultaneously inhibit host B‐ and T‐cell responses using SPI2‐dependent mechanisms.     

Differential timing of antibody‐mediated phagocytosis and cell‐free killing of invasive African Salmonella allows immune evasion

Nontyphoidal Salmonellae commonly cause fatal bacteraemia in African children lacking anti‐Salmonella antibodies. These are facultative intracellular bacteria capable of cell‐free and intracellular survival within macrophages. To better understand the relationship between extracellular and intracellular infection in blood and general mechanisms of Ab‐related protection against Salmonella, we used human blood and sera to measure kinetics of Ab and complement deposition, serum‐mediated bactericidal killing and phagocytosis of invasive African Salmonella enterica serovar Typhimurium D23580. Binding of antibodies peaked by 30 s, but C3 deposition lagged behind, peaking after 2–4 min. C5b‐9 deposition was undetectable until between 2 and 6 min and peaked after 10 min, after which time an increase in serum‐mediated killing occurred. In contrast, intracellular, opsonized Salmonellae were readily detectable within 5 min. By 10 min, around half of monocytes and most neutrophils contained bacteria. The same kinetics of serum‐mediated killing and phagocytosis were observed with S. enterica Typhimurium laboratory strain SL1344, and the S. enterica Enteritidis African invasive isolate D24954 and laboratory strain PT4. The differential kinetics between cell‐free killing and phagocytosis of invasive nontyphoidal Salmonella allows these bacteria to escape the blood and establish intracellular infection before they are killed by the membrane attack complex.     

Synaptopodin‐2 suppresses metastasis of triple‐negative breast cancer via inhibition of YAP/TAZ activity

Triple‐negative breast cancer (TNBC) represents the most aggressive subtype of breast cancer, with a high incidence of distant metastasis; however, the underlying mechanism for this frequent recurrence remains unclear. Herein, we show that synaptopodin‐2 (SYNPO2), a putative tumour suppressor in aggressive cancer, is frequently downregulated in TNBC by methylation of the promoter of SYNPO2. Low expression levels of SYNPO2 correlated significantly with 5‐year metastatic relapse, and predicted poorer prognosis in breast cancer patients. Reintroduction of SYNPO2 inhibited the invasion and spontaneous metastasis of TNBC cells in vivo. Strikingly, downregulation of SYNPO2 is essential for the maintenance of stem cell‐like properties in TNBC cells, leading to efficient distant colonization and metastasis outgrowth. Moreover, we demonstrate that SYNPO2 inhibits the activities of YAP and TAZ by stabilizing LATS2 protein, and transduction of YAP‐S127A abrogates the repressive role of SYNPO2 in metastasis. Finally, immunohistochemical (IHC) analysis of breast cancer patient specimens indicated that the SYNPO2–LATS2–YAP axis is clinically relevant. These findings uncover a suppressive role of SYNPO2 in TNBC metastasis via inhibition of YAP/TAZ, and suggest that SYNPO2 might provide a potential prognosis marker and novel therapeutic strategy. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Prognostic Evaluation of CapG,Gelsolin, P‐gp,GSTP1, and Topo‐II Proteins in Non‐Small Cell Lung Cancer

Metastasis and multidrug resistance (MDR) are the main reasons for the poor prognosis of non‐small cell lung cancer (NSCLC) patients. The use of biomarkers may contribute to a more accurate prediction of tumor metastasis, a better response to chemotherapy, and better patient survival. Gelsolin‐like actin‐capping protein (CapG) and gelsolin have been identified as playing important roles in tumor invasion and metastasis. Permeability glycoprotein (P‐gp), glutathione S‐transferase pi (GSTP1), and topoisomerase‐II (Topo‐II) are proteins that are closely related to MDR. In this study, we assessed the prognostic significance of CapG and gelsolin (both markers of tumor motility), and of P‐gp, GSTP1, and Topo‐II (markers of MDR) in NSCLC patients. One hundred and twenty‐one patients with pathologically confirmed, resectable NSCLC were included in the study. The expression levels of the five kinds of proteins mentioned above were determined by immunohistochemistry (IHC). The correlation between the clinical characteristics and IHC findings were analyzed. Expression of CapG, gelsolin, and P‐gp was found to be associated with an increased risk of death (Hazard Ratio (HR) = 2.799, 95% Confidence Interval (CI) = 1.2705–6.169, P = 0.011; HR = 3.968, 95% CI = 1.811–8.693, P = 0.001; HR = 3.251, 95% CI = 1.456–7.260, P = 0.004, respectively), whereas expression of GSTP1 and Topo‐II was not. These results suggest that higher tumor motility and MDR may be important in NSCLC prognosis. Anat Rec, 2012. © 2011 Wiley Periodicals, Inc.