Enzymes with intracellular activity have significant potential to treat diseases. Protein nanoparticles (NPs) considerably enhance intracellular delivery of enzymes. We have previously shown that a ...Salmonella effector enzyme, AvrA, delivered by NPs is capable of modulating inflammatory signals in a murine dextran sulfate sodium (DSS) colitis model. The NPs were instilled intrarectally, limiting delivery to the distal colon. Localized intestinal delivery of protein therapeutics via the oral route is a highly attractive alternative approach. However, the harsh conditions in the gastrointestinal tract can severely reduce protein function. The approach described here is to deliver therapeutic protein NPs encapsulated within gastro-protective microparticles (MPs) made from alginate and chitosan that subsequently release NPs in the small intestine and colon. A flow focusing microfluidic device was used to form alginate droplets encapsulating protein NPs. Droplets were then simultaneously crosslinked with calcium and coated with chitosan. Protein NPs encapsulated within crosslinked alginate/chitosan MPs were protected and retained their activity after incubation in simulated gastric fluid (SGF). Subsequent incubation in simulated intestinal fluid (SIF) induced release of bioactive protein NPs. Oral administration of AvrA NPs encapsulated in alginate/chitosan MPs delivered protein to intestinal epithelia and reduced clinical and histological scores of inflammation in a murine DSS-induced colitis model. Altogether, NPs in alginate/chitosan MPs are a potential oral delivery vehicle for protein therapeutics.
Projected longer-term droughts and intense floods underscore the need to store more water to manage climate extremes. Here we show how depleted aquifers have been used to store water by substituting ...surface water use for groundwater pumpage (conjunctive use, CU) or recharging groundwater with surface water (managed aquifer recharge, MAR). Unique multi-decadal monitoring from thousands of wells and regional modeling datasets for the California Central Valley and central Arizona were used to assess CU and MAR. In addition to natural reservoir capacity related to deep water tables, historical groundwater depletion further expanded aquifer storage by ∼44 km3 in the Central Valley and by ∼100 km3 in Arizona, similar to or exceeding current surface reservoir capacity by up to three times. Local river water and imported surface water, transported through 100s of km of canals, is substituted for groundwater (≤15 km3 yr−1, CU) or is used to recharge groundwater (MAR, ≤1.5 km3 yr−1) during wet years shifting to mostly groundwater pumpage during droughts. In the Central Valley, CU and MAR locally reversed historically declining water-level trends, which contrasts with simulated net regional groundwater depletion. In Arizona, CU and MAR also reversed historically declining groundwater level trends in active management areas. These rising trends contrast with current declining trends in irrigated areas that lack access to surface water to support CU or MAR. Use of depleted aquifers as reservoirs could expand with winter flood irrigation or capturing flood discharges to the Pacific (0-1.6 km3 yr−1, 2000-2014) with additional infrastructure in California. Because flexibility and expanded portfolio options translate to resilience, CU and MAR enhance drought resilience through multi-year storage, complementing shorter term surface reservoir storage, and facilitating water markets.
A variable reduction strategy (VRS) is an effective method to accelerate the optimization process of evolutionary algorithms (EAs) by simplifying the corresponding optimization problems. ...Unfortunately, the VRS is manually realized in a trial-and-error manner currently. To boost the efficiency of VRS and enable a more extensive application, we propose a variable reduction optimization problem (VROP) to represent a decision space with the smallest sets of variables. Thereafter, a heuristic rule-based automatic variable reduction algorithm (AVRA) is designed to address the VROP. AVRA sequentially reduces variables by considering several sophisticated designed heuristic rules, which search for a variable that can be utilized to represent as many variables as possible and that can be represented by the smallest set of variables possible. With AVRA, the decision variables of an optimization problem can be automatically grouped into reduced variables and core variables, where core variables can represent reduced variables and the entire decision space. During the optimization process, we only need to search the core variables to optimize the problem. Therefore, the dimensionality of the decision space can be reduced by AVRA, subsequently, simplifying the complexity of the problem and improving the search efficiency of EAs. To testify the effectiveness of AVRA, we blend AVRA with several promising EAs to solve two types of challenging problems: 1) continuous equality-constrained optimization problems and 2) nonlinear equations systems. Extensive experiments verify that an EA with AVRA outperforms the standard EA.
Colorectal cancer (CRC) is one of the main causes of morbidity and mortality due to cancer. The purpose of this in-silico study was to examine the relationship of chronic infection mechanisms caused ...by Salmonella Anti virulence agent A (AvrA) to gene mutations in the carcinogenic process of CRC.
Gene expression data on the mouse colon was obtained from the GSE22215 dataset | Gene Expression Omnibus (GEO). Adjusted p-value was calculated using Benjamini & Hochberg False Discovery Rate (FDR<0.01). Gene expression in colon adenocarcicoma tumors was obtained from The Cancer Genome Atlas's (TCGA) Genomic Data Commons (GDC) dataset containing 458 colon tumor samples.
Expressions of MLH1, MSH2, EPCAM, APC, and PMS2 in cases of colon adenocarcinoma tumor showed a correlation with genes that underwent changes due to Salmonella AvrA infection. Among the gens of interest, EPCAM was the gene that had the highest correlation compared to other genes (MLH1, MSH2, APC, and PMS2) (n= 514, Gene r-p value < 0.01 =22355). There were 514 genes that had a correlation with cases of AvrA infection. Tumor Necrosis Factor (TNF), which is a gene that is upregulated in AvrA infection and correlates negatively with EPCAM, had the highest BC value compared to other gens (p= 0.0000768). Survival probability showed that EPCAM was highly expressed and it can increase survival time. In addition to TNF, our study indicated that IL1B (p= 0.000419), S100A8 (p= 2.02E-05), S100A9 (p=0.000419) correlated with the gene of interest.
Late Salmonella AvrA infection affects the expression of genes involved in inflammation in colorectal cancer samples.
BACKGROUND: Colorectal cancer (CRC) is reported as the third most frequently diagnosed cancer worldwide. Salmonella infection plays a role in developing this cancer, which is chronically related to ...the avirulence protein A (AvrA) effector protein produced by the bacteria. This study was conducted to point out the effect of AvrA Salmonella on the occurrence of CRC through the regulation of TLR4/NF-κB/β-catenin/TGF-β expressions in the azoxymethane (AOM)/dextran sodium sulfate (DSS) CRC model.METHODS: A randomized control group post-test-only study was conducted using male Balb/c mice with 30-gram body weight (10-12 weeks), which were divided into three groups, namely the negative control (normal mice without any treatment), positive control (AOM/DSS-treated mice), treatment 1 (T1) (AOM/DSS-treated mice + AvrA Salmonella) groups. Colon tissue was collected and then prepared for immunohistochemistry staining using TLR4, β-catenin, NF-κB, TGF-β, and Ki67 antibodies, whereas apoptotic cells were stained using TUNEL assay.RESULTS: The expressions of TLR4, β-catenin, NF-κB, TGF-β, Ki67, and apoptosis percentage indicated significant differences among the three groups, which statistically showed p<0.05 in all observed parameters. The mean of all parameters was far more significant in the T1 group than in the negative and positive control groups.CONCLUSION: The findings revealed that AvrA Salmonella could increase the expressions of TLR4, β-catenin, NF-κB, TGF-β, and Ki67 and decrease the apoptotic percentage. Thus, AvrA Salmonella influences CRC tumorigenesis through TLR4/NF-κB/β-catenin/TGF-β and is suggested as a potential target in future preventive and curative management for CRC.KEYWORDS: AvrA Salmonella, carcinogenesis, colorectal cancer
Salmonella infection is a major public health concern, and colonization in humans can be chronic and increases the risk of cancers. Wnt signaling is a key pathway for intestinal renewal and ...development, inflammation, and tumorigenesis. In the current study, we report a novel role of Wnt1 in infection and colon cancer using cell culture models, a Salmonella-colitis colon cancer model, and human samples. In contrast to the bacteria-induced increases in Wnt2 and Wnt11, Salmonella colonization significantly reduced the level of Wnt1 in intestinal epithelial cells in vivo and in vitro. The bacterial AvrA protein is known to activate the canonical Wnt pathway. Wnt1 expression level was downregulated by AvrA-expressing Salmonella but stabilized by AvrA-deficient Salmonella in the intestine of Salmonella-colitis mice. In a chronic Salmonella-infected cancer model, the Wnt1 protein level was decreased in the AvrA+ infected group. Thus, we further assessed the functional role of Wnt1 downregulation in the inflammatory response and colorectal cancer (CRC) progression. Moreover, downregulation of Wnt1 by the Crispr-Cas9 method affected cancer cell invasion and migration. Interestingly, we found that Wnt1 was downregulated in human CRC tissue, and Wnt1 downregulation may be correlated with cancer progression. Our study provides insights into mechanisms by which enteric bacteria regulate Wnt1 expression and potentially contribute to infection-associated colon cancer.
Groundwater‐level measurements in monitoring wells or piezometers are the most common, and often the only, hydrologic measurements made at artificial recharge facilities. Measurements of gravity ...change over time provide an additional source of information about changes in groundwater storage, infiltration, and for model calibration. We demonstrate that for an artificial recharge facility with a deep groundwater table, gravity data are more sensitive to movement of water through the unsaturated zone than are groundwater levels. Groundwater levels have a delayed response to infiltration, change in a similar manner at many potential monitoring locations, and are heavily influenced by high‐frequency noise induced by pumping; in contrast, gravity changes start immediately at the onset of infiltration and are sensitive to water in the unsaturated zone. Continuous gravity data can determine infiltration rate, and the estimate is only minimally affected by uncertainty in water‐content change. Gravity data are also useful for constraining parameters in a coupled groundwater‐unsaturated zone model (Modflow‐NWT model with the Unsaturated Zone Flow (UZF) package).
Key Points:
Continuous gravity data are useful to measure infiltration rate and are relatively insensitive to the change in water content
Little or no gravity change after 14 months indicates oscillatory steady state groundwater storage, with little accumulated storage
The spatial sensitivity of gravity makes it better than groundwater levels for recharge‐basin‐scale infiltration model calibration
BACKGROUND Salmonella infections manifest typically as self-limiting gastroenteritis after the consumption of contaminated food. Extra-intestinal manifestations of Salmonella infections such as ...pericarditis are rare and are usually seen in severely immunocompromised individuals. Prior case reports suggest high rates of morbidity and mortality associated with Salmonella pericarditis. Here, we present a rare case of Salmonella dublin pericarditis. CASE REPORT A 45-year-old man presented to the Emergency Department reporting chest pressure and shortness of breath. An echocardiogram showed a large pericardial effusion without tamponade physiology. Pericardial window was performed, with removal of 700 cubic centimeters of bloody fluid, with presence of fibrinous debris in the pericardial cavity. A pericardial biopsy showed chronic pericarditis, and a lymph node biopsy was negative for malignancy. Antinuclear antibody (ANA), Lyme antibodies, and human immunodeficiency virus (HIV) testing were negative. Tissue culture revealed Salmonella species. Subsequent blood cultures grew Salmonella spp. Further history-taking revealed frequent travel and recent treatment with steroids for suspected Bell's palsy. Initially, the patient was treated with ceftriaxone, which was switched to ciprofloxacin after susceptibility testing revealed ceftriaxone resistance. Final identification of the organism revealed Salmonella dublin. The patient was discharged on colchicine, ibuprofen, and a 4-week course of ciprofloxacin. Outpatient follow-up showed improvement in inflammatory markers and symptoms. CONCLUSIONS This case illustrates the rarity of Salmonella-associated pericarditis, the importance of assessing a patient's risk factors, and obtaining an extensive history when searching for an etiology of pericarditis. Investigation into why a patient was susceptible to an infection with this organism should include medication assessment and age-appropriate cancer screening. Prompt identification and treatment of the offending organism can help prevent mortality.
This report illustrates, for the first time, a case of unilateral orchitis and epididymitis in a Holstein-Friesian bull, associated with
infection (
serovar Typhimurium). A one and a half-year-old ...Holstein-Friesian bull had arrived at the Veterinary Hospital of Assiut University suffering from anorexia accompanied with persistent fever, which did not respond to oxytetracycline and flunixin meglumine injection for 15 days. Gross examination revealed left scrotal enlargement (three times its normal size), heat sensation, and induration of the testis and epididymis, which was painful on external palpation. Microbiological and pathological examinations of the left testicle, epididymis, and spleen samples were performed.
Typhimurium was recovered from the affected tissues and its critical virulence genes (
,
A and
B) were identified. Pathological examination revealed a unilateral necrotizing intratubular pyogranulomatus orchitis and epididymitis with severe peri-orchitis. In addition, splenomegaly with a firm and large whitish nodular capsular structure associated with different stages of granulomatous reaction around the white and red pulp. To the authors' knowledge, this report is the first isolation of
Typhimurium from the epididymis and testicles of a Holstein-Friesian bull. These results highlight the importance of including
Typhimurium among the health disorders associated with stressful situations in bovine with orchitis and or/epididymitis. In Egypt,
infection as being enzootic with high probability of dissemination should be considered one of genital health problems among cattle farms.
Avian salmonellosis caused by
serovar Enteritidis (
. Enteritidis) and Pullorum (
Pullorum) remains a big threat to the poultry industry and public hygiene. AvrA is an effector involved in inhibiting ...inflammation. Compared to AvrA from
. Enteritidis (SE-AvrA), the AvrA from
Pullorum (SP-AvrA) lacks ten amino acids at the C-terminal. In this study, we compared the anti-inflammatory response induced by SP-AvrA to that of SE-AvrA. Transient expression of SP-AvrA in epithelial cells resulted in significantly weaker inhibition of NF-κB pathway activation when treated with TNF-α compared to the inhibition by SE-AvrA. SP-AvrA expression in the
. Enteritidis resulted in weaker suppression of NF-κB pathway in infected HeLa cells compared to SE-AvrA expression in the cells, while SP-AvrA expressed in
Pullorum C79-13 suppressed NF-κB activation in infected HeLa and Caco 2 BBE cells to a greater extent than did SE-AvrA because of the higher expression of SP-AvrA than SE-AvrA in
Pullorum. Further analysis demonstrated that the inhibition of NF-κB pathway in
-infected cells corresponded to the downregulation of the p-JNK and Beclin-1 protein molecules. Our study reveals that AvrA modifies the anti-inflammatory response in a manner dependent on the
serotype through inhibition of NF-κB pathway.
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