Chinese Wheat Mosaic Virus-Induced Gene Silencing in Monocots and Dicots at Low Temperature

Virus-induced gene silencing (VIGS) is an important tool for functional genomics studies in plants. With this method, it is possible to target most endogenous genes and downregulate the messenger RNA (mRNA) in a sequence-specific manner. Chinese wheat mosaic virus (CWMV) has a bipartite, single-strand positive RNA genome, and can infect both wheat and Nicotiana benthamiana, and the optimal temperature for systemic infection in plants is 17°C. To assess the potential of the virus as a vector for gene silencing at low temperature, a fragment of the N. benthamiana or wheat phytoene desaturase (PDS) gene was expressed from a modified CWMV RNA2 clone and the resulting photo bleaching in infected plants was used as a reporter for silencing. Downregulation of PDS mRNA was also measured by quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR). In experiments using fragments of PDS ranging from 500 to 1500 nucleotides, insert length influenced the stability and the efficiency of VIGS. The CWMV induced silencing system was also used to suppress miR165/166 and miR3134a through expression of miRNA target mimics. The relative expression levels of mature miR165/166 and miR3134a decreased whereas the transcript levels of their target genes increased. Interestingly, we also found the CWMV-induced silencing system was more efficient compare with the vector based on Barley stripe mosaic virus (BSMV) or Foxtail mosaic virus (FoMV) in wheat or the vector based on TRV in N. benthamiana at 17°C. In summary, the CWMV vector is effective in silencing endogenous genes and miRNAs at 17°C, thereby providing a powerful tool for gene function analysis in both N. benthamiana and wheat at low temperature

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Effects of Anacetrapib in Patients with Atherosclerotic Vascular Disease

BACKGROUND: Patients with atherosclerotic vascular disease remain at high risk for cardiovascular events despite effective statin-based treatment of low-density lipoprotein (LDL) cholesterol levels. The inhibition of cholesteryl ester transfer protein (CETP) by anacetrapib reduces LDL cholesterol levels and increases high-density lipoprotein (HDL) cholesterol levels. However, trials of other CETP inhibitors have shown neutral or adverse effects on cardiovascular outcomes. METHODS: We conducted a randomized, double-blind, placebo-controlled trial involving 30,449 adults with atherosclerotic vascular disease who were receiving intensive atorvastatin therapy and who had a mean LDL cholesterol level of 61 mg per deciliter (1.58 mmol per liter), a mean non-HDL cholesterol level of 92 mg per deciliter (2.38 mmol per liter), and a mean HDL cholesterol level of 40 mg per deciliter (1.03 mmol per liter). The patients were assigned to receive either 100 mg of anacetrapib once daily (15,225 patients) or matching placebo (15,224 patients). The primary outcome was the first major coronary event, a composite of coronary death, myocardial infarction, or coronary revascularization. RESULTS: During the median follow-up period of 4.1 years, the primary outcome occurred in significantly fewer patients in the anacetrapib group than in the placebo group (1640 of 15,225 patients [10.8%] vs. 1803 of 15,224 patients [11.8%]; rate ratio, 0.91; 95% confidence interval, 0.85 to 0.97; P=0.004). The relative difference in risk was similar across multiple prespecified subgroups. At the trial midpoint, the mean level of HDL cholesterol was higher by 43 mg per deciliter (1.12 mmol per liter) in the anacetrapib group than in the placebo group (a relative difference of 104%), and the mean level of non-HDL cholesterol was lower by 17 mg per deciliter (0.44 mmol per liter), a relative difference of -18%. There were no significant between-group differences in the risk of death, cancer, or other serious adverse events. CONCLUSIONS: Among patients with atherosclerotic vascular disease who were receiving intensive statin therapy, the use of anacetrapib resulted in a lower incidence of major coronary events than the use of placebo. (Funded by Merck and others; Current Controlled Trials number, ISRCTN48678192 ; ClinicalTrials.gov number, NCT01252953 ; and EudraCT number, 2010-023467-18 .)

Vesikari Score System (VSS) as a diagnostic tool for the etiology of acute gastroenteritis in infants and children of a Peruvian referral hospital during the first half of 2019

Propósito: Evaluar el valor del Vesikari Scoring System (VSS) como herramienta diagnóstica en la predicción de patógenos en niños con gastroenteritis aguda. Métodos: En un estudio retrospectivo, se analizó 247 historias clínicas y registros laboratoriales del Hospital de Emergencias Pediátricas (Lima, Perú), utilizando el diagnóstico de CIE-10 A00-A009 (enfermedades intestinales infecciosas). Se dividió según los patógenos detectados: no específico, viral y bacteriana. Se detectaron las bacterianas por coprocultivo y las virales por inmunoanálisis. Los casos no específicos tenían ambos resultados negativos o no hubo pruebas laboratoriales. Se ha calculado la sensibilidad, especificidad, valores predictivos negativos (VPN) y positivos (VPP), índice de probabilidad (LR) y el área bajo la curva (ROC). Resultado: El área bajo la curva ROC del VSS no fue significativo para determinar la etiología bacteriana ni viral. En el punto de corte de 9 en VSS, se analizó VPP (18.29%) y VPN (89.47%). Los leucocitos en heces tuvieron un área bajo la curva ROC de 0.8831(IC95% 0,79-0,96 y p=0.04) para etiología bacteriana, con una sensibilidad de 80.95%, especificidad de 88.24% y LR- de 0.22. Los leucocitos en heces con un punto de corte de ≥20, tuvo una precisión aceptable para diagnosticar gastroenteritis bacteriana. La prueba de moco en heces tuvo una sensibilidad de 83.33%, especificidad de 82.35% y VPP de 95.89%. Conclusión: El VSS no es una herramienta adecuada para determinar etiología de gastroenteritis viral y bacteriana. Sin embargo, se ha determinado que los leucocitos en heces con un punto de corte de ≥20 leucocitos por campo y moco en heces permite identificar la gastroenteritis aguda bacteriana.Purpose: To evaluate the Vesikari Scoring System (VSS) as a valuable diagnostic tool for predicting pathogens in children with acute gastroenteritis. Methods: In this retrospective study were analyzed 247 clinical and laboratory records of the Pediatric Emergency Hospital (Lima, Peru), with the diagnosis of ICD-10 A00-A009 (Intestinal Infectious diseases). According to the pathogens detected were divided into groups: non-specific, viral, and bacterial. Bacteria were detected by stool culture, viral pathogens by immune analysis. The non-specific group had negative on both tests, or they did not perform. We calculated sensitivity, specificity, negative and positive predictive values, negative and positive likelihood ratios, and receiver operating characteristic curves.  Results: The area under the ROC curve of VSS was not significant for viral and bacterial etiology. The cut-off point of 9 in VSS was calculated by analyzing PPV (18,29%) and NPV (89,47%). The fecal leukocyte had a ROC of 0.8831 (IC95% 0.79-0.96 and p=0.04) for bacterial group, with a sensibility of 80,95%, specificity of 88,24% and LR- of 0,22. At a cut-off point of ≥20 fecal leukocytes was an acceptable diagnostic accuracy for bacterial gastroenteritis. The stool mucus for the bacterial group had a sensibility of 83,33%, a specificity of 82,35%, and a PPV of 95,89%. Conclusion: VSS is not an adequate diagnostic tool to differentiate viral from bacterial etiology. Nevertheless, we determine that a test of fecal leukocytes, with a cut-off point of ≥20 leukocytes per field and mucus in stools, allow us to identify the bacterial etiology of gastroenteritis.Tesi

Underestimation of Radiation Doses by Compliance of Wearing Dosimeters among Fluoroscopically-Guided Interventional Medical Workers in Korea

This study aimed to estimate the level of underestimation of National Dose Registry (NDR) doses based on the workers&rsquo; dosimeter wearing compliance. In 2021, a nationwide survey of Korean medical radiation workers was conducted. A total of 989 medical workers who performed fluoroscopically-guided interventional procedures participated, and their NDR was compared with the adjusted doses by multiplying the correction factors based on the individual level of dosimeter compliance from the questionnaire. Ordinal logistic regression analysis was performed to identify the factors for low dosimeter wearing. Based on the data from the NDR, the average annual effective radiation dose was 0.95 mSv, while the compliance-adjusted dose was 1.79 mSv, yielding an 89% increase. The risks for low compliance with wearing a badge were significantly higher among doctors, professionals other than radiologists or cardiologists, workers not frequently involved in performing fluoroscopically-guided interventional procedures, and workers who did not frequently wear protective devices. This study provided quantitative information demonstrating that the NDR data may have underestimated the actual occupational radiation exposure. The underestimation of NDR doses may lead to biased risk estimates in epidemiological studies for radiation workers, and considerable attention on dosimetry wearing compliance is required to interpret and utilize NDR data

Occupational radiation exposure and morbidity of circulatory disease among diagnostic medical radiation workers in South Korea.

ObjectivesWe investigated the association between low-dose external occupational radiation exposure and circulatory disease morbidity among diagnostic medical radiation workers.MethodsA cohort of 11 500 diagnostic medical radiation workers was linked with the National Dosimetry Registry data and the National Health Insurance Service data. Relative risks (RRs) were calculated to explore the association between occupational factors and circulatory disease morbidity, and excess relative risks per 100 milligray (ERR/100 mGy) were estimated to quantify the radiation dose-response relationship.ResultsOverall, there were 2270 cases of circulatory diseases during 93 696 person-years of observation (average follow-up=8.1 years). RRs for hypertension were significantly increased for individuals who started working before 2000 compared with those who started in 2005 and later. ERR/100 mGy for all circulatory diseases was 0.14 (95% CI -0.57 to 0.99). Radiation risks of cerebrovascular diseases and ischaemic heart disease were non-significantly increased with estimates of individual cumulative doses to the heart (ERR/100 mGy=3.10 (-0.75 to 11.59) and 1.22 (-0.71 to 4.73), respectively). However, ERR estimates were generally more strongly positive for female versus male workers and for younger workers versus more than 50-year-old workers.ConclusionsThis study provides little evidence in support of a positive association between occupational radiation exposure and the overall risk of circulatory disease over a short follow-up period among medical radiation workers in South Korea. However, significantly increased RR with earlier year first worked, elevated ERR in female workers and young workers should be further followed up

Occupational radiation exposure and morbidity of circulatory disease among diagnostic medical radiation workers in South Korea.

ObjectivesWe investigated the association between low-dose external occupational radiation exposure and circulatory disease morbidity among diagnostic medical radiation workers.MethodsA cohort of 11 500 diagnostic medical radiation workers was linked with the National Dosimetry Registry data and the National Health Insurance Service data. Relative risks (RRs) were calculated to explore the association between occupational factors and circulatory disease morbidity, and excess relative risks per 100 milligray (ERR/100 mGy) were estimated to quantify the radiation dose-response relationship.ResultsOverall, there were 2270 cases of circulatory diseases during 93 696 person-years of observation (average follow-up=8.1 years). RRs for hypertension were significantly increased for individuals who started working before 2000 compared with those who started in 2005 and later. ERR/100 mGy for all circulatory diseases was 0.14 (95% CI -0.57 to 0.99). Radiation risks of cerebrovascular diseases and ischaemic heart disease were non-significantly increased with estimates of individual cumulative doses to the heart (ERR/100 mGy=3.10 (-0.75 to 11.59) and 1.22 (-0.71 to 4.73), respectively). However, ERR estimates were generally more strongly positive for female versus male workers and for younger workers versus more than 50-year-old workers.ConclusionsThis study provides little evidence in support of a positive association between occupational radiation exposure and the overall risk of circulatory disease over a short follow-up period among medical radiation workers in South Korea. However, significantly increased RR with earlier year first worked, elevated ERR in female workers and young workers should be further followed up

Risk Factors of Avulsion Fracture after Iliac Crestal Flap for Jaw Reconstruction

The iliac crestal flap is often used in maxillofacial reconstruction surgery because it has a similar bone contour to jaw bone. However, complications occur frequently in this flap, and the avulsion fracture of the anterior superior iliac spine (ASIS) occurs only in rare cases. The purpose of this study is to evaluate the risk factors affecting fractures that may occur after iliac crestal flap harvesting. In this study, we performed preoperative and postoperative CT scans of the iliac bones of 22 patients who underwent iliac crestal flaps between February 2013 and July 2019. Three of these patients had fractures within 1 month after surgery. After converting DICOM files to a three-dimensional model using Mimics software, some reference points were defined for analysis. The patients were classified into fracture and non-fracture groups. The patients’ age and sex, harvested bone, residual bone and absolute bed rest (ABR) periods were investigated. The patients’ age and sex were not associated with fracture occurrence. There was a statistically significant difference in the width of the anterior–superior part of residual bone. The fracture group showed statistically significantly shorter ABR periods. In the case of the iliac crestal flap, it was found that the factors that can affect the occurrence of fractures are the width of the residual bone and the ABR period. Iliac avulsion fracture can be prevented when the remaining bone is sufficient and the ABR period is prolonged sufficiently