Evaluation of disinfectants to prevent mechanical transmission of viruses and a viroid in greenhouse tomato production

Background In recent years, a number of serious disease outbreaks caused by viruses and viroids on greenhouse tomatoes in North America have resulted in significant economic losses to growers. The objectives of this study were to evaluate the effectiveness of commercial disinfectants against mechanical transmission of these pathogens, and to select disinfectants with broad spectrum reactivity to control general virus and viroid diseases in greenhouse tomato production. Methods A total of 16 disinfectants were evaluated against Pepino mosaic virus (PepMV), Potato spindle tuber viroid (PSTVd), Tomato mosaic virus (ToMV), and Tobacco mosaic virus (TMV). The efficacy of each disinfectant to deactivate the pathogen’s infectivity was evaluated in replicate experiments from at least three independent experiments. Any infectivity that remained in the treated solutions was assessed through bioassays on susceptible tomato plants through mechanical inoculation using inocula that had been exposed with the individual disinfectant for three short time periods (0–10 sec, 30 sec and 60 sec). A positive infection on the inoculated plant was determined through symptom observation and confirmed with enzyme-linked immunosorbent assay (PepMV, ToMV, and TMV) and real-time reverse transcription-PCR (PSTVd). Experimental data were analyzed using Logistic regression and the Bayesian methodology. Results Statistical analyses using logistic regression and the Bayesian methodology indicated that two disinfectants (2% Virkon S and 10% Clorox regular bleach) were the most effective to prevent transmission of PepMV, PSTVd, ToMV, and TMV from mechanical inoculation. Lysol all-purpose cleaner (50%) and nonfat dry milk (20%) were also effective against ToMV and TMV, but with only partial effects for PepMV and PSTVd. Conclusion With the broad spectrum efficacy against three common viruses and a viroid, several disinfectants, including 2% Virkon S, 10% Clorox regular bleach and 20% nonfat dry milk, are recommend to greenhouse facilities for consideration to prevent general virus and viroid infection on tomato plants

EZH2 knockdown suppresses the growth and invasion of human inflammatory breast cancer cells.

INTRODUCTION: Inflammatory breast cancer (IBC) is the most metastatic variant of breast cancer with the poorest survival in all types of breast cancer patients and presently therapeutic targets for IBC are very limited. Enhancer of zeste homolog 2 (EZH2) is frequently expressed in human IBC and its expression positively correlates with worse clinical outcome. However, the molecular basis for EZH2 promoting IBC has not been explored. Here, we investigated the functional role of EZH2 in IBC cells by examining the effects of its knockdown on the formation of tumor spheroids and invasion of these cells in vitro and in vivo in an orthotopic xenograft model. METHODS: SUM149 and a new IBC cell line-FC-IBC-02 derived from pleural effusion fluid of an IBC patient were used in this study. Specific knockdown of EZH2 was performed using short hairpin RNA (shRNA) specific to the human EZH2 gene. Cell growth and the formation of tumor spheroids were examined in vitro. The effects of EZH2 knockdown on IBC cell migration and invasion were examined by a Boyden chamber assay. For the in vivo tumor growth studies, IBC cells were orthotopically transplanted into the mammary fat pads of immunodeficient mice. RESULTS: The results showed that EZH2 is expressed at higher levels in human IBC cell lines compared with normal human mammary epithelial cells, and the knockdown of EZH2 expression significantly suppressed cell growth and tumor spheroid formation of human IBC cells in vitro. In addition, EZH2 knockdown inhibited the migration and invasion of IBC cells. Significantly, EZH2 knockdown suppressed the angiogenesis and tumor growth of IBC cells in vivo. CONCLUSIONS: Our results provide direct evidence that EZH2 is critical for the formation of tumor spheroids and invasion of human IBC cells and could be a potential target for developing novel therapeutic strategies for human IBC

The association between the CD4/CD8 ratio and surgical site infection risk among HIV-positive adults: insights from a China hospital

PurposeIt is well known that the CD4/CD8 ratio is a special immune-inflammation marker. We aimed to explore the relationship between the CD4/CD8 ratio and the risk of surgical site infections (SSI) among human immunodeficiency virus (HIV)-positive adults undergoing orthopedic surgery.MethodsWe collected and analyzed data from 216 HIV-positive patients diagnosed with fractures at the department of orthopedics, Beijing Ditan Hospital between 2011 and 2019. The demographic, surgical, and hematological data for all patients were collected in this retrospective cohort study. We explored the risk factors for SSI using univariate and multivariate logistic regression analysis. Then, the clinical correlation between the CD4 count, CD4/CD8 ratio, and SSI was studied using multivariate logistic regression models after adjusting for potential confounders. Furthermore, the association between the CD4/CD8 ratio and SSI was evaluated on a continuous scale with restricted cubic spline (RCS) curves based on logistic regression models.ResultsA total of 23 (10.65%) patients developed SSI during the perioperative period. Patients with hepatopathy (OR=6.10, 95%CI=1.46-28.9), HIV viral load (OR=8.68, 95%CI=1.42-70.2; OR=19.4, 95%CI=3.09-179), operation time (OR=7.84, 95%CI=1.35-77.9), and CD4 count (OR=0.05, 95%CI=0.01-0.23) were risk factors for SSI (P-value < 0.05). Our study demonstrated that a linear relationship between CD4 count and surgical site infection risk. In other words, patients with lower CD4 counts had a higher risk of developing SSI. Furthermore, the relationship between CD4/CD8 ratio and SSI risk was non-linear, inverse ‘S’ shaped. The risk of SSI increased substantially when the ratio was below 0.913; above 0.913, the risk of SSI was almost unchanged. And there is a ‘threshold-saturation’ effect between them.ConclusionOur research shows the CD4/CD8 ratio could be a useful predictor and immune-inflammation marker of the risk of SSI in HIV-positive fracture patients. These results, from a Chinese hospital, support the beneficial role of immune reconstitution in HIV-positive patients prior to orthopedic surgery

The consensus guideline of perioperative antiviral therapy for AIDS patients in China based on clinical practice

The prevalence of human immunodeficiency virus (HIV) and acquired immune deficiency syndrome (AIDS) has emerged as a major public health concern in China. When patients with HIV infection undergo surgical treatment, there are two main challenges. Firstly, medical staff face a high risk of HIV infection due to occupational exposure. Secondly, the patient’s immune function is impaired, increasing the risk of opportunistic infections and postoperative complications. The surgical treatment of such patients is unique, and the risk of occupational exposure during the operation primarily depends upon the viral load of HIV/AIDS patients. Therefore, perioperative antiretroviral therapy is of paramount importance in order to standardize the perioperative antiretroviral therapy (ART) for HIV/AIDS patients. The Surgery Group of the Chinese Association of STD and AIDS Prevention and Control, in collaboration with the Treatment Association, and Surgery Group of the Chinese Medical Association of Tropical Diseases and Parasitology, has developed an expert consensus on perioperative antiretroviral therapy for HIV/AIDS patients. This consensus encompasses various aspects, including surgical risk assessment, selection of perioperative antiretroviral therapy regimens, prevention of opportunistic infections, and the crucial focus on rapid preoperative viral load reduction and immune function reconstruction for HIV/AIDS patients

Deep Sequencing of Small RNAs in Tomato for Virus and Viroid Identification and Strain Differentiation

Small RNAs (sRNA), including microRNAs (miRNA) and small interfering RNAs (siRNA), are produced abundantly in plants and animals and function in regulating gene expression or in defense against virus or viroid infection. Analysis of siRNA profiles upon virus infection in plant may allow for virus identification, strain differentiation, and de novo assembly of virus genomes. In the present study, four suspected virus-infected tomato samples collected in the U.S. and Mexico were used for sRNA library construction and deep sequencing. Each library generated between 5–7 million sRNA reads, of which more than 90% were from the tomato genome. Upon in-silico subtraction of the tomato sRNAs, the remaining highly enriched, virus-like siRNA pools were assembled with or without reference virus or viroid genomes. A complete genome was assembled for Potato spindle tuber viroid (PSTVd) using siRNA alone. In addition, a near complete virus genome (98%) also was assembled for Pepino mosaic virus (PepMV). A common mixed infection of two strains of PepMV (EU and US1), which shared 82% of genome nucleotide sequence identity, also could be differentially assembled into their respective genomes. Using de novo assembly, a novel potyvirus with less than 60% overall genome nucleotide sequence identity to other known viruses was discovered and its full genome sequence obtained. Taken together, these data suggest that the sRNA deep sequencing technology will likely become an efficient and powerful generic tool for virus identification in plants and animals

Direct Synthesis of 2‑Methylbenzofurans from Calcium Carbide and Salicylaldehyde <i>p</i>‑Tosylhydrazones

A new methodology for the construction of methyl-substituted benzofuran rings from the reactions of calcium carbide with salicylaldehyde <i>p</i>-tosylhydrazones/2-hydroxyacetophenone <i>p</i>-tosylhydrazones is described. Various 2-methylbenzofurans and 2,3-dimethylbenzofurans could be obtained in satisfactory yield by using a cuprous chloride catalyst. The advantages of this protocol include the use of a readily available and easy-to-handle acetylene source and simple workup procedure