Endophytic Phthalate-degrading Bacillus subtilis N-1-gfp colonizing in soil-crop system shifted indigenous bacterial community to remove di-n-butyl phthalate.

Endophytic bacteria can degrade toxic phthalate (PAEs). Nevertheless, the colonization and function of endophytic PAE-degrader in soil-crop system and their association mechanism with indigenous bacteria in PAE removal remain unknown. Here, endophytic PAE-degrader Bacillus subtilis N-1 was marked with green fluorescent protein gene. Inoculated strain N-1-gfp could well colonize in soil and rice plant exposed to di-n-butyl phthalate (DBP) as directly confirmed by confocal laser scanning microscopy and realtime PCR. Illumina high-throughput sequencing demonstrated that inoculated N-1-gfp shifted indigenous bacterial community in rhizosphere and endosphere of rice plants with significant increasing relative abundance of its affiliating genus Bacillus than non-inoculation. Strain N-1-gfp exhibited efficient DBP degradation with 99.7% removal in culture solutions, and significantly promoted DBP removal in soil-plant system. Strain N-1-gfp colonization help plant enrich specific functional bacteria (e.g., pollutant-degrading bacteria) with significant higher relative abundances and stimulated bacterial activities (e.g., pollutant degradation) compared with non-inoculation. Furthermore, strain N-1-gfp displayed strong interaction with indigenous bacteria for accelerating DBP degradation in soil, decreasing DBP accumulation in plants and promoting plant growth. This is the first report on well colonization of endophytic DBP-degrader Bacillus subtilis in soil-plant system and its bioaugmentation with indigenous bacteria for promoting DBP removal.

Diversity of endophytic bacteria in wild rice (Oryza meridionalis) and potential for promoting plant growth and degrading phthalates.

Phthalates (PAEs) accumulated in agricultural soils and rice have increased human exposure risks. Microbial degradation could efficiently reduce the residue of organic pollutants in soil and crop plants. Here, we hypothesized that endophytic bacteria from wild rice have the potential for degradation of PAEs and plant growth promoting. The endophytic bacterial community and functional diversity in wild rice (Oryza meridionalis) were analyzed for the first time, and the potential for PAE degradation and plant growth promoting by endophytes were investigated. The results of Illumina high-throughput sequencing revealed that abundant endophytes inhabited in wild rice with Proteobacteria, Bacteroidetes, Firmicutes and Actinobacteria being the dominant phyla. Endophytic bacterial diversity and complexity were confirmed by isolation and clustering of isolates. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated that endophytes exerted diverse functions such as plant growth promoting, xenobiotics biodegradation, pollution remediation and bacterial chemotaxis. Pure culture experiment showed that 30 isolated endophytic strains exhibited in vitro plant growth promoting activities, and rice plants inoculated with these strains confirmed their growth promoting abilities. Some endophytic strains were capable of efficiently degrading PAEs, with the highest removal percentage of di-n-butyl phthalate (DBP) up to 96.1% by Bacillus amyloliquefaciens strain L381 within 5 days. Synthetic community F and strain L381 rapidly removed DBP from soil (removing 91.0%-99.2% within 10 d and from rice plant slurry (removing 93.4%-99.2% within 5 d). These results confirmed the hypothesis and demonstrated the diversity of endophytic bacteria in wild rice with diverse functions, especially for plant growth promoting and removing PAEs. These multifunctional endophytic bacteria provided good alternatives to reduce PAE accumulation in crops and increase yield.

Development and validation of a nomogram to predict the risk of breast cancer-related lymphedema among Chinese breast cancer survivors.

PURPOSE: Breast cancer-related lymphedema (BCRL) is a major long-term complication for post-surgery breast cancer survivors. Although several risk factors have been identified, lifestyle characteristics have been neglected in previous studies. The aim of this study was to develop and validate a nomogram for estimating this population's risk of developing lymphedema, taking into consideration their demographic, clinical, and personal lifestyle behaviors. METHODS: In a cross-sectional study, we collected data from 775 post-operative breast cancer survivors who had attended a follow-up session in the recent 10 years (primary cohort). Lymphedema was assessed using the Norman telephone questionnaire, self-reported by patients. Multiple logistic regression was used to identify risk factors for lymphedema, including demographic, clinical, and lifestyle-related factors. A nomogram was constructed based on those factors and was validated using a separate group of 314 breast cancer patients (validation cohort). RESULTS: The factors independently associated with lymphedema were higher body mass index (BMI), modified radical mastectomy (MRM), postsurgical infection, chemotherapy, radiotherapy, exercise of the affected arm, and the active participation in physical activity (P<0.05). The area under the curve (AUC) values of the primary and the validation cohorts were 0.721 (95% confidence interval: 0.685-0.756) and 0.702 (95% confidence interval: 0.646-0.759), respectively. CONCLUSIONS: BCRL risk factors include MRM, radiotherapy, chemotherapy, and higher BMI, while the active physical activity behavior of patients appears to be a factor against lymphedema. The nomogram incorporating the patients' clinical and lifestyle factors might be useful for predicting lymphedema in breast cancer survivors.

Prevalence and predictors of breast cancer-related arm lymphedema over a 10-year period in postoperative breast cancer patients: A cross-sectional study.

PURPOSE: Breast cancer (BC) survivors have a lifelong risk of developing lymphedema. This study investigated the prevalence of BC-related arm lymphedema among Chinese BC survivors diagnosed in the last 10 years and examined the demographic and clinical variables as well as lifestyle factors associated with lymphedema status. METHODS: In this cross-sectional study, women with BC (N = 866) who had been diagnosed and followed up in the previous 10 years were recruited from the outpatient clinic of 4 general hospitals and one cancer association in China between August 2018 and October 2019. Lymphedema status was determined using the Norman telephone questionnaire as the patient-reported occurrence of hand/lower arm/upper arm swelling. Multiple logistic regression was used to identify risk factors for lymphedema. RESULTS: The median time from BC diagnosis was 4.0 years (interquartile range, 2.0-5.0 years). 81.4% of the patients had undergone mastectomy. The prevalence of arm lymphedema among BC survivors was 49.0%. Age ≥50 years, monthly income <3000 RMB, modified radical mastectomy, postsurgical wound infection, chemotherapy, and radiotherapy were associated with an increased risk of BC-related arm lymphedema, whereas exercise of the affected arm, engagement in active physical activity, and timely reporting of symptoms of infection to a physician decreased the risk (P < 0.05). CONCLUSIONS: Arm lymphedema is a common complication for postoperative BC survivors within 10 years. It is essential to identify patients at risk of lymphedema based on demographic, clinical, and lifestyle factors and implement interventions targeting modifiable lifestyle behaviors-eg, active physical activity during the postoperative period.

A teleost bactericidal permeability-increasing protein-derived peptide that possesses a broad antibacterial spectrum and inhibits bacterial infection as well as human colon cancer cells growth.

The bactericidal permeability-increasing protein (BPI) is a multifunctional cationic protein produced by neutrophils with antibacterial, antitumor, and LPS-neutralizing properties. In teleost, a number of BPIs have been reported, but their functions are very limited. In this study, an N-terminal peptide, BO18 (with 18 amino acids), derived from rock bream (Oplegnathus fasciatus) BPI, was synthesized and investigated for its antibacterial spectrum, action mechanism, immunoregulatory property as well as the inhibition effects on bacterial invasion and human colon cancer cells growth. The results showed that BO18 was active against Gram-positive bacteria Bscillus subiilis, Micrococcus luteus, and Staphylococcus aureus, as well as Gram-negative bacteria Vibrio alginolyticus, Vibrio litoralis, Vibrio parahaemolyticus and Vibrio vulnificus. BO18 treatment facilitated the bactericidal process of erythromycin and rifampicin by enhancing the permeability of the outer membrane. During its interaction with V. alginolyticus, BO18 exerted its antibacterial activity by destroying cell membrane integrity, penetrating into the cytoplasm and binding to genomic DNA and total RNA. In vitro analysis indicated BO18 could enhance the respiratory burst ability and regulate the expression of immune related genes of macrophages. In vivo detection showed the administration of fish with BO18 before bacterial infection significantly reduced pathogen dissemination and replication in tissues. In addition, BO18 exerted a cytotoxic effect on the growth of human colon cancer cells HT-29. Together, these results add new insights into the function of teleost BPIs, and support that BO18 is a novel and broad-spectrum antibacterial peptide with potential to apply in fighting pathogenic infection in aquaculture.

Muscular involvement of extranodal natural killer/T cell lymphoma misdiagnosed as polymyositis: A case report and review of literature.

BACKGROUND: Natural killer (NK)/T cell lymphoma is a rare and highly aggressive malignant tumor, and is a special form of non-Hodgkin's lymphoma. Although extranodal involvement is frequently found in tissues such as the skin, testicular and gastrointestinal tract etc, its presence in skeletal muscle has scarcely been reported in the literature. CASE SUMMARY: We report a case of extranodal NK/T cell lymphoma with muscle swelling as the first clinical manifestation. A 42-year-old man, who initially presented with localized swelling in the double lower extremities, demonstrated gradual facial and eyelid swelling, and his imaging results showed multiple sites of muscle damage throughout the body. The final pathological results suggested NK/T cell lymphoma, and immunohistochemistry showed CD20 (-), CD3Ɛ (+), CD30 (+), CD56 (-), EBER (+), Ki67 (60%), TIA-1 (+) and CD68 (±) staining. The muscle swelling significantly improved after treatment with chemotherapy regimens. CONCLUSION: This disease is difficult to diagnose and highly invasive, and should be included in the differential diagnosis of unexplained muscle swelling.

Adjuvanted-influenza vaccination in patients infected with HIV: a systematic review and meta-analysis of immunogenicity and safety.

Adjuvanted-influenza vaccination is an efficient method for enhancing the immunogenicity of influenza split-virus vaccines for preventing influenza. However, the medical community's understanding of its performance in patients infected with HIV remains limited. To identify the advantages, we conducted a systematic review and meta-analysis with randomized controlled trials (RCTs) and cohort and case-control studies that have the immunogenicity and safety of influenza vaccines in patients infected with HIV as outcomes. We searched six different databases, and 1698 patients infected with HIV in 11 studies were included. Statistical analysis was performed to calculate the pooled standardized mean differences (SMD) or relative risk (RR) and 95% confidence interval (CI). Regarding immunogenicity, the pooled SMD of GMT (Geometric mean titer) for A/H1N1 was 0.61 (95%CI (0.40,0.82)), the pooled RR of seroconversion was 1.34 (95%CI (0.91,1.98)) for the H1N1 vaccine, 1.27(95%CI (0.64,2.52)) for the H3N2 vaccine, 1.19(95%CI (0.97,1.46)) for the B-type influenza vaccine. The pooled RR of seroprotection was 1.61 (95%CI (1.00,2.58)) for the H1N1 vaccine, 1.06 (95%CI(0.83,1.35)) for the H3N2 vaccine, and 1.13(95%CI(0.91,1.41)) for the B-type vaccine. Adjuvanted-influenza vaccination showed good general tolerability in patients infected with HIV, with the only significant increase being the rate of local pain at the injection site (RR = 2.03, 95%CI (1.06,3.86)). In conclusion, all studies evaluating injected adjuvanted influenza vaccination among patients infected with HIV showed acceptable levels of safety and immunogenicity.

Antenatal sleep quality associated with perinatal outcomes in women of advanced maternal age.

OBJECTIVES: The increasing prevalence of advanced maternal age (AMA) coupled with poor sleep quality among pregnant women makes it important to study their association with perinatal outcomes. However, little is known about the interaction of AMA and maternal antenatal sleep on perinatal outcomes. Here, we examined whether associations between AMA and perinatal outcomes are modified by antenatal sleep quality. PARTICIPANTS: Data were collected from 446 women, with a singleton pregnancy and no pregnancy complications, who participated in the Growing Up in Singapore Towards healthy Outcomes (GUSTO) birth cohort study. MEASUREMENTS: Participants completed the Pittsburgh Sleep Quality Index (PSQI) at 26-28 weeks gestation and had perinatal outcome data collected upon delivery. Interactions between AMA and maternal sleep quality on perinatal outcomes were investigated and where significant, analyses were further stratified by maternal age. All analyses were adjusted for maternal BMI at 26-28 weeks gestation, ethnicity, and maternal education. RESULTS: Neonates of mothers of AMA and poor sleep quality (PSQI score >5) had increased odds of stay in the neonatal intensive care unit (adjusted odds ratio = 3.53, 95% CI: -1.21 to 10.27) and shorter birth length (adjusted mean difference = -1.05 cm, 95% CI: -1.82 to -0.20), as compared with women of AMA and good sleep quality (PSQI score ≤5). In women <35 years, sleep quality did not associate with perinatal outcomes. CONCLUSION: Poor sleep quality in women of AMA was associated with neonatal health outcomes. Improving maternal antenatal sleep may potentially improve perinatal outcomes in offspring of women of AMA.

Prevalence and determinants of depression in caregivers of cancer patients: A systematic review and meta-analysis.

BACKGROUND: Aim of this study was to estimate the prevalence rate of depression in cancer patient caregivers and to identify factors affecting depression and quality of life of cancer caregivers. METHODS: Relevant research articles were retrieved after literature search in several electronic databases. Random effects meta-analyses were performed to obtain pooled estimates of the prevalence rates of depression and anxiety; their respective scores, and quality of life scores. Significant relationships between depression and factors related to depression and quality of life reported in individual studies were identified. RESULTS: Thirty studies were included. Overall, 21,149 caregivers were appraised in these studies (age 52.65 years [95% CI: 49.65, 55.65]; 31.14% [28.40, 33.89] men). The prevalence of depression and anxiety were 42.30% [33.31, 51.29] % and 46.55% [35.59, 57.52], respectively. Quality of life score, as measured with Caregiver Quality of Life-Cancer scale was 64.55 [47.44, 81.66]. Patient's condition, caregiving burden, duration of caregiving, spouse caregiver, caregiver being unemployed, caregiver with chronic disease, caregiver's sleep quality, caregiver's avoidance, financial problems, and female sex were positively associated with depression whereas overall quality of life of caregiver, pre-loss grief, caregiver's education level, caregiver's age, caregiver's sense of coherence, and caregiver's bondage with patient were negatively associated with depression in caregivers. CONCLUSION: A considerably high prevalence of depression is found in cancer patient caregivers. Several factors may affect depression and their quality of life of cancer patient caregivers.

Risk Factors for Preoperative Anxiety and Depression in Patients Scheduled for Abdominal Aortic Aneurysm Repair.

BACKGROUND: Perioperative emotional disorders of patients underwent abdominal aortic aneurysm (AAA) repair is an emerging area of study, and preoperative mental distress of those patients remains poorly understood. The aim of this study was to investigate the prevalence and identify the risk factors of preoperative anxiety and depression in patients scheduled for AAA repair. METHODS: A total of 189 patients who underwent elective AAA repair between 2015 and 2016 were included in this study. These patients were preoperatively evaluated by Hospital Anxiety and Depression Scale (HADS). Demographics and anxiety and depression scores of the patients were documented. Logistic regression was used to identify the independent risk factors of preoperative anxiety and depression. RESULTS: A total of 150 AAA patients were included in final analysis. Of these 150 patients, 44 patients (29.3%) had borderline anxiety or clinical anxiety, and 42 patients (28.0%) were found to have borderline or clinical depression. Female (odds ratio [OR]: 2.81, 95% confidence interval [CI]: 1.08-7.26), the American Society of Anesthesiologists (ASA) Grade 3/4 (OR: 4.34, 95% CI: 1.13-16.68), higher education (OR: 1.44, 95% CI: 1.02-2.04), and abdominal or back pain (OR: 3.08, 95% CI: 1.20-7.87) were identified as significant independent risk factors of abnormal HADS-anxiety in overall patients; and higher level of education (OR: 1.87, 95% CI: 1.16-3.01) was predictive of anxiety in patients planned for endovascular aortic repair. Besides, higher body mass index (BMI) (OR: 1.18, 95% CI: 1.04-1.33) and abdominal or back pain (OR: 3.93, 95% CI: 1.70-9.11) were predictive of abnormal preoperative HADS-depression in overall patients. CONCLUSION: As for patients scheduled for AAA repair, female, higher ASA, higher level of education, and symptom may be independent risk factors for preoperative anxiety, and symptom and higher BMI may predict preoperative depression.

Syntheses, crystal structures, adsorption properties and visible photocatalytic activities of highly stable Pb-based coordination polymers constructed by 2-(2-carboxyphenyl)imidazo(4,5-f)-(1,10)phenanthroline and bridging linkers.

Five highly stable coordination polymers assembled by 2-(2-carboxyphenyl)imidazo(4,5-f)-(1,10)phenanthroline (2-HNCP) and different aromatic carboxylic acid ligands, namely, [Pb(2-NCP)(L1)]n (1), [Pb2(2-NCP)2(L2)]n·2nH2O (2), [Pb2(2-NCP)2(L2)]n (3), [Pb(2-NCP)(L3)0.5]n (4) and [Pb2(2-NCP)2(L4)]n (5), where HL1 = pyridine-4-carboxylic acid, H2L2 = 2-amino-1,4-benzenedicarboxylic acid, H2L3 = 1,4-benzenedicarboxylic acid and H2L4 = 2-hydroxy-1,4-benzenedicarboxylic acid, have been synthesized under hydrothermal conditions. Their structures have been determined by single crystal X-ray diffraction analyses and further characterized by elemental analyses and infrared spectroscopy. In 1, adjacent ladder-like chains are extended into a three-dimensional (3D) supramolecular architecture by π-π interactions. In 2, the neighboring layers are interconnected by π-π interactions to afford a 3D supramolecular architecture. 3-5 exhibit similar 3D frameworks with a Schläfli symbol of 412·63 topologies. The different auxiliary ligands and the pH value of the reaction system were discussed in regard to the formation of different structures. In addition, these five complexes present high thermal stabilities, the preferential adsorption of CO2 over N2 and excellent photocatalytic activities for dye degradation under visible light irradiation.

[Relationship between serum erythropoietin levels and brain injury in preterm infants].

OBJECTIVE: To study the relationship between the levels of erythropoietin (EPO) in serum and brain injury in preterm infants. METHODS: Three hundred and four preterm infants (gestational age: 28-34 weeks) born between October 2014 and September 2015 were enrolled in this study. Brain injury was diagnosed using cerebral ultrasound and MRI. The levels of EPO, S100 protein, neuron-specific enolase (NSE) and myelin basic protein (MBP) in serum were detected using ELISA. To compare the incidence of brain injury in different serum EPO levels in preterm infants, and the relationship between brain injury and serum EPO levels was analyzed. RESULTS: The incidence rate of brain injury in preterm infants was 41.1% (125/304). The incidence rate of brain injury in the low EPO level group was significantly higher than that in the middle-high EPO level groups (P<0.01). The serum levels of S100 protein, NSE, and MBP in the brain injury groups were significantly higher than in the control group (P<0.01). The serum EPO levels were negatively correlated with serum S100 protein concentration and NSE levels (P<0.05). According to the multiple logistic regression analysis, low gestational age, low birth weight, asphyxia, prolonged mechanical ventilation, anemia and low serum EPO levels were the risk factor for brain injury in preterm infants. CONCLUSIONS: There is a higher incidence rate of brain injury in preterm infants with lower serum EPO levels. The serum EPO levels may be correlated with brain injury in preterm infants.

[Construction and application of bioinformatic analysis platform for aquatic pathogen based on the MilkyWay-2 supercomputer].

As a key component of life science, bioinformatics has been widely applied in genomics, transcriptomics, and proteomics. However, the requirement of high-performance computers rather than common personal computers for constructing a bioinformatics platform significantly limited the application of bioinformatics in aquatic science. In this study, we constructed a bioinformatic analysis platform for aquatic pathogen based on the MilkyWay-2 supercomputer. The platform consisted of three functional modules, including genomic and transcriptomic sequencing data analysis, protein structure prediction, and molecular dynamics simulations. To validate the practicability of the platform, we performed bioinformatic analysis on aquatic pathogenic organisms. For example, genes of Flavobacterium johnsoniae M168 were identified and annotated via Blast searches, GO and InterPro annotations. Protein structural models for five small segments of grass carp reovirus HZ-08 were constructed by homology modeling. Molecular dynamics simulations were performed on out membrane protein A of Aeromonas hydrophila, and the changes of system temperature, total energy, root mean square deviation and conformation of the loops during equilibration were also observed. These results showed that the bioinformatic analysis platform for aquatic pathogen has been successfully built on the MilkyWay-2 supercomputer. This study will provide insights into the construction of bioinformatic analysis platform for other subjects.

Effect of high pressure processing on rheological and structural properties of milk-gelatin mixtures.

There is an increasing demand to tailor the functional properties of mixed biopolymer systems that find application in dairy food products. The effect of static high pressure processing (HPP), up to 600MPa for 15min at room temperature, on milk-gelatin mixtures with different solid concentrations (5%, 10%, 15% and 20% w/w milk solid and 0.6% w/w gelatin) was investigated. The viscosity remarkably increased in mixtures prepared with high milk solid concentration (15% and 20% w/w) following HPP at 300MPa, whereas HPP at 600MPa caused a decline in viscosity. This was due to ruptured aggregates and phase separation as confirmed by confocal laser scanning microscopy. Molecular bonding of the milk-gelatin mixtures due to HPP was shown by Fourier-transform infrared spectra, particularly within the regions of 1610-1690 and 1480-1575cm(-1), which reflect the vibrational bands of amide I and amide II, respectively.

S100A4 siRNA inhibits human pancreatic cancer cell invasion in vitro.

OBJECTIVE: Pancreatic cancer is one of the most deadly cancers, which is characterized by its high metastatic potential. S100A4 is a major prometastatic protein involved in tumor invasion and metastasis which precise role in pancreatic cancer has not been fully investigated. We knocked down the S100A4 gene in the Bxpc-3 pancreatic cancer cell line via RNA interference to study the changes in cell behavior. METHODS: Real-time polymerase chain reaction and western blotting were used to detect mRNA and protein expression levels of S100A4, matrix metalloproteinase (MMP)-2, E-cadherin and thrombospondin (TSP)-1. Transwell chambers were used to detect the migration and invasion abilities; a cell adhesion assay was used to detect adhesion ability; colony forming efficiency was used to detect cell proliferation; flow cytometry was used to detect apoptosis. RESULTS: S100A4 mRNA expression was reduced to 17% after transfection with S100A4-siRNA, and protein expression had a similar trend. mRNA and protein expression of MMP-2 was reduced and that of E-cadherin and TSP-1 was elevated, indicating that S100A4 affects their expression. S100A4-silenced cells exhibited a marked decrease in migration and invasiveness and increased adhesion, whereas overall proliferation and apoptosis were not overtly altered. CONCLUSION: S100A4 and its downstream factors play important roles in pancreatic cancer invasion, and silencing A100A4 can significantly contain the invasiveness of pancreatic cancer.

Characterization of a critical role for CFTR chloride channels in cardioprotection against ischemia/reperfusion injury.

AIM: To further characterize the functional role of cystic fibrosis transmembrane conductance regulator (CFTR) in early and late (second window) ischemic preconditioning (IPC)- and postconditioning (POC)-mediated cardioprotection against ischemia/reperfusion (I/R) injury. METHODS: CFTR knockout (CFTR(-/-)) mice and age- and gender-matched wild-type (CFTR(+/+)) and heterozygous (CFTR(+/-)) mice were used. In in vivo studies, the animals were subjected to a 30-min coronary occlusion followed by a 40-min reperfusion. In ex vivo (isolate heart) studies, a 45-min global ischemia was applied. To evaluate apoptosis, the level of activated caspase 3 and TdT-mediated dUTP-X nick end labeling (TUNEL) were examined. RESULTS: In the in vivo I/R models, early IPC significantly reduced the myocardial infarct size in wild-type (CFTR(+/+)) (from 40.4% ± 5.3% to 10.4% ± 2.0%, n=8, P<0.001) and heterozygous (CFTR(+/-)) littermates (from 39.4% ± 2.4% to 15.4% ± 5.1%, n=6, P<0.001) but failed to protect CFTR knockout (CFTR(-/-)) mice from I/R induced myocardial infarction (46.9% ± 6.2% vs 55.5% ± 7.8%, n=6, P>0.5). Similar results were observed in the in vivo late IPC experiments. Furthermore, in both in vivo and ex vivo I/R models, POC significantly reduced myocardial infarction in wild-type mice, but not in CFTR knockout mice. In ex vivo I/R models, targeted inactivation of CFTR gene abolished the protective effects of IPC against I/R-induced apoptosis. CONCLUSION: These results provide compelling evidence for a critical role for CFTR Cl(-) channels in IPC- and POC-mediated cardioprotection against I/R-induced myocardial injury.

Peptidoglycan recognition proteins kill bacteria by activating protein-sensing two-component systems.

Mammalian peptidoglycan recognition proteins (PGRPs), similar to antimicrobial lectins, bind the bacterial cell wall and kill bacteria through an unknown mechanism. We show that PGRPs enter the Gram-positive cell wall at the site of daughter cell separation during cell division. In Bacillus subtilis, PGRPs activate the CssR-CssS two-component system that detects and disposes of misfolded proteins that are usually exported out of bacterial cells. This activation results in membrane depolarization, cessation of intracellular peptidoglycan, protein, RNA and DNA synthesis, and production of hydroxyl radicals, which are responsible for bacterial death. PGRPs also bind the outer membrane of Escherichia coli and activate the functionally homologous CpxA-CpxR two-component system, which kills the bacteria. We exclude other potential bactericidal mechanisms, including inhibition of extracellular peptidoglycan synthesis, hydrolysis of peptidoglycan and membrane permeabilization. Thus, we reveal a previously unknown mechanism by which innate immunity proteins that bind the cell wall or outer membrane exploit the bacterial stress defense response to kill bacteria.

In Vitro Synergy of Biochanin A and Ciprofloxacin against Clinical Isolates of Staphylococcus aureus.

Many clinical isolates of Staphylococcus aureus (S. aureus) are resistant to numerous antimicrobials, including the fluoroquinolones (FQs). Flavonoids such as biochanin A (BCA) are compounds that are naturally present in fruits, vegetables, and plant-derived beverages. The goal of this investigation was to study the possible synergy between the antimicrobial agents BCA and ciprofloxacin (CPFX) when used in combination; CPFX was chosen as a representative FQ compound. We used S. aureus strain ATCC 25923 and 11 fluoroquinolone (FQ)-resistant methicillin-resistant S. aureus (MRSA) strains. Results from the drug susceptibility testing and checkerboard assays show that the minimum inhibitory concentration (MIC) of BCA ranged from 64 µg/mL to 512 µg/mL. When BCA was combined with CPFX, the fractional inhibitory concentration index (FICI) data showed that there was synergy in all 12 of the S. aureus strains tested. No antagonistic activity was observed in any of the strains tested. The results of time-kill tests and agar diffusion tests confirm that there was synergy between BCA and CPFX against S. aureus strains. These results suggest that BCA can be combined with FQs to produce a powerful antimicrobial agent.

Influence of boron on carrot cell wall structure and its resistance to fracture.

Plant cell wall structure integrity and associated tissue mechanical properties is one of key determinants for the perceived texture of plant-based foods. Carrots (Daucus carota) were used to investigate the effect of mineral supply of boron (B) and/or calcium (Ca), during plant growth, on the plant cell wall structure and mechanical properties of matured root tissues. Five commercial cultivars of carrots, Kuroda (orange), Dragon Purple, Kuttiger White, Yellow, and Nutri-Red, were cultivated under controlled glasshouse conditions over two seasons. Significant increases in the accumulation of B and Ca were found for all cultivars of carrots when additional B and Ca were included in the nutrient feeding solutions throughout the plant growth period. Elevated levels of B in carrot root tissue reduced the uptake of Ca and other mineral nutrients and enhanced plant cell wall structural integrity, its resistance to fracture, and the weight and size (both diameter and length) of carrots. Although higher amounts of Ca were accumulated in the plant materials, the additional supply of Ca did not have a significant effect on the mechanical properties of mature plant tissues or on the uptake of B by the plant. The results suggest that B cross-linking of pectin (rhamnogalacturonan II) has a greater influence on mature tissue mechanical properties than Ca cross-linking of pectin (homogalacturonan) when supplied during plant growth.

Effects of allicin on CYP2C19 and CYP3A4 activity in healthy volunteers with different CYP2C19 genotypes.

OBJECTIVE: To investigate the interaction between allicin and omeprazole and to observe the effects of allicin on CYP2C19 and CYP3A4 activity in healthy Chinese male volunteers with different CYP2C19 genotypes. METHODS: Eighteen subjects (six CYP2C19*1/CYP2C19*1, four CYP2C19*1/CYP2C19*2, two CYP2C19*1/ CYP2C19*3, and six CYP2C19*2/ CYP2C19*2) were enrolled in a two-phase randomized crossover trial. In each phase, all subjects received placebo or a 180 mg allicin capsule once daily for 14 consecutive days. The pharmacokinetics of omeprazole (20 mg orally on day 15) was determined for up to 12 h following administration by high-performance liquid chromatography. RESULTS: In carriers of the CYP2C19*1/CYP2C19*1 and CYP2C19*1/CYP2C19*2 or *3 genotype, allicin treatment increased the peak plasma concentration (C(max)) of omeprazole by 49.7 +/- 7.2 (p < 0.001) and 54.2 +/- 9.2% (p < 0.001), and increased the area under the plasma time-concentration curve (AUC(0-infinity)) of omeprazole by 48.1 +/- 9.0 (p = 0.001) and 73.6 +/- 26.7% (p < 0.001), respectively. The ratio of AUC(0-infinity) of 5-hydroxyomeprazole to omeprazole (a marker for CYP2C19 activity) decreased significantly (p < 0.001 and p = 0.001, respectively). However, no pharmacokinetic parameters were significantly changed by allicin in CYP2C19*2/CYP2C19*2. The C(max) and AUC(0-infinity) of omeprazole sulfone were unchanged in all three genotypes. CONCLUSIONS: Allicin reduced the metabolism of omeprazole by inhibiting CYP2C19 activity in individuals with the CYP2C19*1/CYP2C19*1 and CYP2C19*1/CYP2C19*2 or *3 genotypes, but not in those with the CYP2C19*2/ CYP2C19*2 genotype. Allicin did not significantly affect the activity of CYP3A4 in all subjects.