Multilayered synergistic regulation of phytoalexin biosynthesis by ethylene, jasmonate, and MAPK signaling pathways in Arabidopsis.

Camalexin, an indolic antimicrobial metabolite, is the major phytoalexin in Arabidopsis thaliana, and plays a crucial role in pathogen resistance. Our previous studies revealed that the Arabidopsis mitogen-activated protein kinases MPK3 and MPK6 positively regulate pathogen-induced camalexin biosynthesis via phosphoactivating the transcription factor WRKY33. Here, we report that the ethylene and jasmonate (JA) pathways act synergistically with the MPK3/MPK6-WRKY33 module at multiple levels to induce camalexin biosynthesis in Arabidopsis upon pathogen infection. The ETHYLENE RESPONSE FACTOR1 (ERF1) transcription factor integrates the ethylene and JA pathways to induce camalexin biosynthesis via directly upregulating camalexin biosynthetic genes. ERF1 also interacts with and depends on WRKY33 to upregulate camalexin biosynthetic genes, indicating that ERF1 and WRKY33 form transcriptional complexes to cooperatively activate camalexin biosynthetic genes, thereby mediating the synergy of ethylene/JA and MPK3/MPK6 signaling pathways to induce camalexin biosynthesis. Moreover, as an integrator of the ethylene and JA pathways, ERF1 also acts as a substrate of MPK3/MPK6, which phosphorylate ERF1 to increase its transactivation activity and therefore further cooperate with the ethylene/JA pathways to induce camalexin biosynthesis. Taken together, our data reveal the multilayered synergistic regulation of camalexin biosynthesis by ethylene, JA, and MPK3/MPK6 signaling pathways via ERF1 and WRKY33 transcription factors in Arabidopsis.

Treatment patterns and survival in metastatic castration-sensitive prostate cancer in the US Veterans Health Administration.

BACKGROUND: Limited real-world data exist on treatment patterns and outcomes in patients with metastatic castration-sensitive prostate cancer (mCSPC). METHODS: A retrospective cohort study was conducted, using the Veterans Health Administration claims database (April 2013-March 2018). Among 369,734 prostate cancer patients, we selected all men who developed metastases within 90 days before or after medical/surgical castration and who received androgen deprivation therapy (ADT). Patients were categorized into four cohorts: ADT-only (± <90-day nonsteroidal anti-androgen [NSAA] use), ADT + NSAA, ADT + docetaxel, and ADT + abiraterone. Main outcomes were treatment patterns, time-to-progression to metastatic castration-resistant disease, and overall survival. Multivariable analysis and sensitivity analysis were conducted. RESULTS: Of 1395 patients, 874 (63%) received ADT-only, 338 (24%) received ADT + NSAA, 108 (8%) received ADT + docetaxel, and 75 (5%) received ADT + abiraterone. Proportions on ADT-only and ADT + NSAA declined (from 66% to 60% and from 31% to 17%, respectively) over the study period, while proportions prescribed ADT + docetaxel or abiraterone increased from 3% to 9% and from 1% to 15%, respectively. Patients treated with ADT + NSAA had similar risks of castration-resistant disease (hazard ratio [HR] 1.05; 95% confidence interval [CI]: 0.87, 1.26) and overall mortality (HR 1.22; 95% CI: 0.97, 1.54) as ADT-only. CONCLUSIONS: Most patients with mCSPC initiating ADT received ADT-only or ADT + NSAA, despite the emergence of docetaxel and novel hormonal therapies. Even in the most recent period (2017 to early 2018), only 24% of men received intensified therapy with agents known to prolong survival versus ADT-only. These data in real-world clinical practice suggest substantial room for improved outcomes in patients with mCSPC.

Comparative Clinical and Economic Outcomes Associated with Warfarin Versus Apixaban in the Treatment of Patients with Venous Thromboembolism in a Large U.S. Commercial Claims Database.

BACKGROUND: Venous thromboembolism (VTE), constituting deep vein thrombosis (DVT) and pulmonary embolism (PE), is a common cause of vascular-related morbidity and mortality, resulting in a significant clinical and economic burden in the United States each year. Clinical guidelines recommend that patients with DVT and PE without cancer should be initiated on anticoagulation therapy with a direct oral anticoagulant over a vitamin K antagonist. Yet there is limited real-world evidence comparing the economic burden of warfarin and apixaban in treating VTE patients in a large commercially insured population. OBJECTIVE: To compare safety and effectiveness of warfarin and apixaban and evaluate associated economic burden in treating VTE patients in a large U.S. commercial health care claims database. METHODS: The PharMetrics Plus database was used to identify oral anticoagulant (OAC)-naive patients aged ≥ 18 years who initiated apixaban or warfarin within 30 days of a qualifying VTE encounter and had continuous health plan enrollment with medical and pharmacy benefits for 6 months before treatment initiation. Apixaban initiators and warfarin initiators were matched using the propensity score matching (PSM) technique. Cox proportional hazard models were used to assess and compare the risk of major bleeding (MB), clinically relevant nonmajor (CRNM) bleeding, and recurrent VTE. Generalized linear models were used to assess and compare the all-cause health care costs. A 2-part model with bootstrapping was used to evaluate MB- and recurrent VTE-related medical costs. RESULTS: Among 25,193 prematched patients, 13,421 (53.3%) were prescribed warfarin and 11,772 (46.7%) were prescribed apixaban. After 1:1 PSM, 8,858 matched warfarin-apixaban pairs were selected with a mean follow-up of 109 days and 103 days, respectively. Warfarin was associated with a significantly higher risk of MB (HR = 1.52, 95% CI = 1.14-2.04), CRNM bleeding (HR = 1.27, 95% CI = 1017.15-1.40), and recurrent VTE (HR = 1.50, 95% CI = 1.24-1.82) compared with apixaban. Warfarin patients had significantly higher all-cause medical costs per patient per month (PPPM; $2,333 vs. $1,992; P = 0.001), MB-related costs PPPM ($112 vs. $65; P = 0.020), and recurrent VTE-related costs PPPM ($287 vs. $206; P = 0.014) compared with apixaban patients. Warfarin patients had similar all-cause total health care costs PPPM ($2,630 vs. $2,420; P = 0.051) compared with apixaban patients. CONCLUSIONS: Warfarin use was associated with a higher risk of MB, CRNM bleeding, and recurrent VTE compared with apixaban. Warfarin use was also associated with higher all-cause medical costs, MB-related medical costs, and recurrent VTE-related costs PPPM compared with apixaban. DISCLOSURES: This study was funded by Bristol Myers Squibb and Pfizer, which were also involved in the study design, as well as writing and revising of the manuscript. Guo, Rajpura, Okano, and Rosenblatt are employees of Bristol Myers Squibb. Hlavacek, Mardekian, and Russ are employees of Pfizer. Keshishian, Sah, Delinger, and Mu are employees of SIMR, LLC, which received funding from the study sponsors to conduct this study.

Corrective osteotomy for cervicothoracic deformity secondary to ankylosing spondylitis / 中华骨科杂志

Objective To report the application of C7T1extensive osteotomy and C7pedicle subtraction osteotomy(PSO) in the correction of cervicothoracic deformity secondary to ankylosing spondylitis(AS)and to investigate the efficacy and safety of the techniques.Methods Between April 2006 and August 2017,eight male patients with cervicothoracic deformity undergoing C7T1extensive osteotomy(3 cases)or C7PSO(5 cases)were retrospectively reviewed.The mean age was 31.3±14.9 years(range, 14-60 years).C2-T1kyphosis,C2-T1scoliosis and C2-T1sagittal vertical axis(SVA)were measured on the lateral cervical radio-graphs and chin-brow vertical angle(CBVA)was measured on clinical photographs preoperatively and at the final follow-up.The operative time, blood loss and complications were recorded. Results The average follow-up duration was (11.3 ± 7.9) months (range,3-48 months).In C7T1extensive osteotomy group,the mean operative time was 305 min(300-310 min)and the average blood loss was 1 250 ml(1 000-1 500 ml).Preoperative and postoperative C2-T1kyphosis were 17.0°±16.3°and-13.3°±20.2°,re-spectively.The preoperative CBVA was 20.0°±4.5°,which improved to 4.7°±5.9°at the final follow-up with a mean correction rate of 76.5%.C2-T1SVA was improved from(6.9±4.0)cm preoperatively to(3.5±1.8)cm at the final follow-up with an average correc-tion rate of 49.3%.In C7PSO group,the mean operative time was 536 min(375-730 min)and the average blood loss was 2 450 ml (700-4 200 ml).There were four patients with concomitant scoliosis and kyphosis.Preoperative and postoperative C2-T1kyphosis were 22.8°±10.5°and-13.5°±10.0°,respectively.The preoperative C2-T1scoliosis was 24.8°±12.7°,which improved to 5.0°±3.5° at the final follow-up with a mean correction rate of 79.8%.CBVA was improved from 60.5°±10.2°preoperatively to 14.3°±8.6°at the final follow-up with an average correction rate of 76.4%.C2-T1SVA was corrected to(6.4±2.5)cm at the final follow-up from(10.4 ± 4.3) cm preoperatively. One patient was presented with severe cervicothoracic scoliosis and the C2-T1scoliosis was im-proved to 10°from 33°with a 69.7% correction rate.No neurological complications,infection,or implant-related complications were observed both intraoperatively and during the follow-up period.One patient who underwent C7PSO experienced intraopera-tive subluxation of the osteomized vertebra.Fortunately,there was no neurological deficit.Solid bony fusion was observed after six-month Halo-vest immobilization.Conclusion Both C7T1extensive osteotomy and C7PSO are effective in the correction of cervico-thoracic deformity secondary to AS with acceptable complication rate.C7PSO is particularly suitable for the correction of severe and complicated biplanar cervicothoracic deformity.

Genome-Wide Identification of Klebsiella pneumoniae Fitness Genes during Lung Infection.

UNLABELLED: Klebsiella pneumoniae is an urgent public health threat because of resistance to carbapenems, antibiotics of last resort against Gram-negative bacterial infections. Despite the fact that K. pneumoniae is a leading cause of pneumonia in hospitalized patients, the bacterial factors required to cause disease are poorly understood. Insertion site sequencing combines transposon mutagenesis with high-throughput sequencing to simultaneously screen thousands of insertion mutants for fitness defects during infection. Using the recently sequenced K. pneumoniae strain KPPR1 in a well-established mouse model of pneumonia, insertion site sequencing was performed on a pool of >25,000 transposon mutants. The relative fitness requirement of each gene was ranked based on the ratio of lung to inoculum read counts and concordance between insertions in the same gene. This analysis revealed over 300 mutants with at least a 2-fold fitness defect and 69 with defects ranging from 10- to >2,000-fold. Construction of 6 isogenic mutants for use in competitive infections with the wild type confirmed their requirement for lung fitness. Critical fitness genes included those for the synthesis of branched-chain and aromatic amino acids that are essential in mice and humans, the transcriptional elongation factor RfaH, and the copper efflux pump CopA. The majority of fitness genes were conserved among reference strains representative of diverse pathotypes. These results indicate that regulation of outer membrane components and synthesis of amino acids that are essential to its host are critical for K. pneumoniae fitness in the lung. IMPORTANCE: Klebsiella pneumoniae is a bacterium that commonly causes pneumonia in patients after they are admitted to the hospital. K. pneumoniae is becoming resistant to all available antibiotics, and when these infections spread to the bloodstream, over half of patients die. Since currently available antibiotics are failing, we must discover new ways to treat these infections. In this study, we asked what genes the bacterium needs to cause an infection, since the proteins encoded by these genes could be targets for new antibiotics. We identified over 300 genes that K. pneumoniae requires to grow in a mouse model of pneumonia. Many of the genes that we identified are found in K. pneumoniae isolates from throughout the world, including antibiotic-resistant forms. If new antibiotics could be made against the proteins that these genes encode, they may be broadly effective against K. pneumoniae.

Protective Effect of Coenzymes Complex Combined with Vitamin E on Liver and Kidney Injury Induced by Neonatal Hyperbilirubinemia / 天津医药

Objective To study the effect of coenzymes complex combined with vitamin E on liver and kidney inju-ry induced by neonatal hyperbilirubinemia. Methods One hundred and fifty full-term neonatals with hyperbilirubinemia were chosen as observation groups, who were divided into mild, moderate and severe groups according bilirubin. Forty five healthy full-term newborns in the same period, who are either healthy or with physiological jaundice, were selected as con-trol group. Serum total bilirubin, gamma-glutamyl transferase (γ-GGT) , Malondialdehyde (MDA) and Cys-C were measured within 24 hours of hospital admission.The observation groups were randomly divided into regular and combined treatment groups. Coenzymes complex combined with vitamin E were given in addition to regular method to combined group while reg-ular group only received regular methods.All above biochemical indexes were tested in the 7th day after medication admin-istration. Results Serum MDA were higher in all observation groups (mild, moderate and severe groups) than in control group (P＜0.05);but the levels ofγ-GGT and Cys-C increased in moderate and severe groups compared with control group (P＜0.05). There were positive correlations (P＜0.05) between levels of serum total bilirubin with MDA,γ-GGT and Cys-C. Positive linear correlation were found between MDA with γ-GGT and Cys-C(P＜0.01). After early intervention,γ-GGT, Cys-C and MDA declined with drop of bilirubin level. This is more prominent and faster in a in combined treatment group than regular group (P＜0.05).Conclusion In hyperbilirubinemia newborns, lipid peroxidation activated by bilirubin may lead to damages of liver and kidney. Coenzymes combined with vitamin E have protective effect to these damages.

K83 site affects PICK1 PDZ binding ability / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the role of 83 site in interaction of GluR2 C-terminal and PICK1 PDZ domain.</p><p><b>METHODS</b>Docking structure of PICK1 PDZ domain with GluR2 C terminal PDZ binding motif was built with computer software. After K83 site was substituted by other amino acid, the structure and binding energy were recalculated; meanwhile, site specific mutants were constructed using wild type full length cDNA as template. Mutants were co-transfected with GluR2 into HEK293T cells. After staining, the distribution of PICK1 and GluR2 were observed under confocal microscope.</p><p><b>RESULTS</b>Wild type PICK1 and GluR2 formed many co-clusters in HEK293T cells as reported by other research groups; but different K83 mutant had different distribution in HEK293T cells.</p><p><b>CONCLUSION</b>The K83 site in PDZ domain of PICK1 is important for the interaction between PICK1 and GluR2. Altering lysine will probably change the hydrophobic interactions, the hydrogen bonds or the electrostatic interactions formed between PICK1 PDZ domain and GluR2 C terminal; accordingly, that will change the binding capacity between PICK1 and GluR2 in varying degrees.</p>

High expression of thymosin alpha 1 by injecting recombinant PVX vector into the tomato fruit / 生物工程学报

For expression of foreign genes in plant, plant virus vector provides many advantages, such as high expression level, short expression period and wider plant hosts. In the present study, we report the expression of thymosin alpha 1 (Talpha1) in tomato fruits by potato virus X (PVX) vector. Talpha1 gene fragment from plasmid pGEM-T containing Talpha1 gene was cloned into plant virus vector pGR107 and the resulting pGR107-Talpha1 plasmid was confirmed by digestion with Sal I and Cla I. To express the Talpha1 protein, Agrobacterium tumefaciens GV3101 transformed with pGR107-Talpha1 was directly injected into tomato fruits through the fruit stylar apex at different developmental stages. The ELISA results showed that Talpha1 protein was expressed successfully in fruits, and the highest expression level was obtained from 2.5-3 week-old tomato fruits inoculated by bacterium at 1.0 OD600 density.