The correlation between epidurography contrast patterns and the clinical outcomes of percutaneous epidural neuroplasty (PEN) remains unclear.  To analyze the correlation between postadhesiolysis epidurography contrast patterns and the clinical outcomes of patients who undergo lumbar PEN.  This study is a retrospective analysis of 78 consecutive patients who underwent lumbar PEN between April 2012 and March 2013.  The analysis was done in the university hospital center.  The clinical outcomes of all patients were assessed before and 1, 3, 6, and 12 months after undergoing lumbar PEN. Specifically, the intensity of back and leg pain, quality of life, and procedural outcomes were evaluated using a visual analog scale (VAS), the Oswestry Disability Index (ODI), and the 12-Item Short-Form Health Survey (SF-12). P < 0.01 versus preprocedural scores). At most follow-up time points, patients exhibiting extraforaminal contrast distribution (n=22) on postadhesiolysis epidurograms exhibited a similar improvement in VAS scores and a significantly better improvement in ODI and SF-12 scores compared with patients exhibiting intracanal contrast distribution (n=56).  The VAS scores for back and leg pain, ODI score, and SF-12 score exhibited a significant improvement during the follow-up period ( Extraforaminal contrast distribution during lumbar PEN may be associated with better functional outcomes. Chronic low back pain is a common health problem with widespread socioeconomical reverberations which is experienced by most individuals at some point in their lives –3. SeverThis retrospective analysis of medical and radiographic records included 78 patients who underwent lumbar PEN procedures between April 2012 and March 2013 at a single university hospital. The study protocol was approved by the ethics committee, and informed consent was obtained from all subjects. The inclusion criterion was chronic low back pain with or without leg pain due to lumbar disc herniation and/or lumbar spinal stenosis. All patients reported a history of discogenic or radicular symptoms refractory to conservative treatments for a minimum of 6 weeks. Diagnoses were established using magnetic resonance imaging (MRI) and/or computed tomography (CT) performed before PEN. Patients with a history of spinal surgery and those with cauda equina syndrome, bleeding diathesis, associated somatic or psychiatric disease, vertebral fractures, pregnancy, and tumors or other underlying systemic diseases that could significantly affect the procedural outcomes were excluded. All procedures were performed by one of the authors (DAS) using the same procedural protocol.A standard PEN procedure was used to lyse adhesions and achieve nerve blockades in all patients, as previously described , 14. A 1Postadhesiolysis epidurography contrast patterns were defined and classified into five grades according to the system proposed by Mathis et al. : grade 1All patients were clinically evaluated before and 1, 3, 6, and 12 months after PEN by a nurse specialized in pain management and blinded to the treatment details. The intensity of leg and back pain was assessed using a subjective visual analog scale (VAS) calibrated from 0 to 10 (0 = no pain and 10 = the worst pain imaginable). For functional assessments, the Korean versions of the Oswestry Disability Index (ODI) and 12-Item Short-Form Health Survey (SF-12) were used , 23. ODIt-tests were used to assess differences in VAS, ODI, and SF-12 scores at each time point between the EF and IC groups. Chi-square tests were used to compare clinical outcomes between the two groups. Spearman's rank correlation tests were used to determine the correlation between epidurography contrast patterns and clinical outcomes. All statistical analyses were performed using the Statistical Package for the Social Sciences software . A P value of <0.05 was considered statistically significant.Age, the duration of symptoms and follow-up, and VAS, ODI, and SF-12 scores are expressed as means ± standard deviations. Student's A total of 78 patients (27 men and 51 women) were included in this study. All patients were clinically followed up for more than 11 months. The mean age was 58.1 ± 14.1 years , and the average duration of major symptoms before PEN was 7.1 ± 4.1 months . Before PEN, the VAS score for back pain, VAS score for leg pain, ODI score, and SF-12 score were 5.8 ± 2.6, 5.0 ± 3.1, 19.4 ± 7.0, and 30.0 ± 7.2, respectively .P < 0.01 when compared with preprocedural scores; The clinical outcomes of patients exhibited a significant improvement during the follow-up period. At 1, 3, 6, and 12 months after PEN, the VAS score for back pain was 2.6 ± 2.3, 3.1 ± 2.1, 3.0 ± 2.0, and 3.4 ± 2.4, respectively; the VAS score for leg pain was 1.9 ± 2.0, 2.6 ± 2.1, 2.6 ± 2.2, and 2.7 ± 2.6, respectively; the ODI score was 10.6 ± 8.0, 11.7 ± 7.4, 11.0 ± 7.3, and 11.0 ± 8.0, respectively; and the SF-12 score was 41.8 ± 9.6, 40.5 ± 9.0, 41.2 ± 9.2, and 41.2 ± 10.2, respectively (n=56), there were 19, 25, and 12 patients with grade 1, grade 2, and grade 3 patterns, respectively. In the EF group (n=22), there were 7 and 15 patients with grade 4 and grade 5 patterns, respectively. Before PEN, the VAS score for back pain was 5.7 ± 2.6 and 6.0 ± 2.4 (P=0.639), the VAS score for leg pain was 4.7 ± 3.1 and 6.0 ± 2.8 (P=0.101), the ODI score was 20.0 ± 6.9 and 17.8 ± 7.1 (P=0.222), and the SF-12 score was 29.6 ± 7.0 and 31.0 ± 7.7 (P=0.452) in the IC and EF groups, respectively, with no significant differences between groups. At most follow-up time points, the EF group exhibited a similar improvement in VAS scores and a significantly better improvement in ODI and SF-12 scores compared with the IC group (P < 0.05)). A representative case of a patient with radicular pain in the left leg who exhibited significant pain relief and functional recovery after PEN is depicted in The patients were categorized into IC and EF groups according to the contrast patterns on postadhesiolysis epidurograms. In the IC group (IC group . Thus, eEpidurograms provide anatomical information and help in confirming accurate needle placement. Spinal interventions can be safely performed using a fluoroscope and epidurograms with radiopaque dye. The aim of the present study was to evaluate the correlation between contrast patterns on postadhesiolysis epidurograms and clinical outcomes in patients who underwent lumbar PEN. We observed that PEN ameliorated the pain associated with lumbar disc herniation and/or lumbar spinal stenosis. VAS scores for leg and back pain, ODI scores, and SF-12 scores exhibited a significant improvement over the entire 12-month follow-up period. Racz et al.  reportedIn the present study, we found that the clinical outcomes of lumbar PEN were correlated with the presence of extraforaminal contrast spread on the final epidurogram obtained after adhesiolysis. Extraforaminal contrast spread is believed to be an indicator of successful adhesiolysis. The purpose of PEN is to eliminate the barriers in the epidural space that disturb drug delivery. Extraforaminal contrast spread may be related to the creation of channels around the target site through adhesiolysis, and it facilitates drug delivery to the lesion site. Exposure to the ideal amount of medication can result in better pain reduction. It can also enhance the recovery of the perineural circulation, which can improve neural congestion and decrease the associated pain. The results of our study confirmed that access to the exact position of the lesion site is associated with a better outcome. These findings suggest that mechanical adhesiolysis is meaningful only at the lesion site. Moreover, if the exact lesion site cannot be reached, both adhesiolysis and drug delivery will fail and the target area will remain untreated.In 1991, Kuslich et al.  demonstrIn the present study, there were no direct correlations between cutoff signs or filling defects and the clinical outcomes of PEN. In our practice, we cannot easily resolve cutoff signs and filling defects. Cutoff signs are a result of severe spinal stenosis, while filling defects are frequently associated with disc herniations. Currently, gross adhesions cannot be easily lysed by soft PEN. However, regardless of the presence of cutoff signs or filling defects, better outcomes were achieved with extraforaminal contrast spread. There may be microscopic adhesions that are more relevant and can be lysed by soft PEN. Park et al. also reported that there was no correlation between the degree of pain relief and epidural filling defects in patients with lumbar stenosis .This study has several limitations. These include the small sample size, the unequal groups, the short follow-up duration, and the retrospective design. Future studies should ideally include an appropriate control group. Nevertheless, the strength of our study is the use of clinical outcome measures, namely, ODI and SF-12, for assessment of the quality of life in addition to pain scores. Furthermore, the results of the study are more in line with the prognosis because the patients received a single treatment rather than repetitive treatments. With the use of epidurograms, physicians can design and plan adhesiolysis that aims at a precise target point and creates effective tunnels, which will help in predicting the prognosis.In conclusion, the findings of our study suggest that the contrast patterns on final epidurograms obtained after adhesiolysis during PEN are indicators of clinical outcomes. Specifically, extraforaminal contrast spread during PEN is associated with better functional outcomes.


Kruskal-Wallis test results have related differences in Pu-erh tea quality as well as technological processing of fruit tea to their mineral composition. In order to characterize tea elemental content, chemometric techniques such as factor analysis (FA) and cluster analysis (CA) were used. Their application allowed on differentiation of samples in view of the fermentation type, technological processing, and overall quality.This study concerns application of flame atomic absorption spectrometry (FAAS) in assessment of macro- and microelement and toxic metal levels  in dark (Pu-erh) and fruit tea leaves and their infusions. Phosphorus was also determined in the form of phosphomolybdate by spectrophotometric method. The reliability of the method was checked using three certified reference materials. The results of analysis were in agreement with the certified values, with analytical recovery ranging from 86 to 113 %. Significant correlations ( There are approximately 1500 different varieties of tea, which can be grouped by taste and color [Camellia sinensis var. assamica (L.) O. Kuntze; Theaceae) native to the Upper Mekong River Region of China’s Yunnan Province [Pu-erh refers to the tea plant  and fruit tea species are summarized in Table 3  to the ashes and evaporation to dryness on a boiling water bath. The dry residue was rewetted with 1.50 mL of 36 % HCl and heated for 1 min, covered with a watch glass. Next, the watch glass was rinsed with ultra-pure water (18 MΩ cm−1) from Milli-Q® system  above the crucible, and the resulting solution was transferred to 25-mL flasks.Tea leaves were homogenized using mill IKA® A11 Basic. About 10.0 g (±0.0001 g) of homogenized products portions was weighed and transferred to quartz crucibles. Tea samples were ashed in an electric furnace using a gradient of temperature up to 540 °C. Mineralization procedure was based on the addition of 1.50 mL 36 % HCl  and 2–3 drops of 63 % HNOIn case of infusions, 2.0 g (±0.0001 g) of materials (tea leaves) was weighed and transferred to a 250-mL beaker. Then, tea brew was prepared with 200 mL of water and infused for 5 min under the watch glass cover. Subsequently, the solution was filtered through fluted paper filter and the resulting filtrate was transferred to a quartz crucible. Then, tea infusions were evaporated to dryness on a boiling water bath. The dry residue was ashed in a furnace, following the same procedure as with the dry sample .w/v), which acts as an ionization buffer that shifts the equilibrium of ion reaction resulting in the free atomic form of the analyzed element. Magnesium and Ca were analyzed with the addition of 0.4 % w/v lanthanum oxide (lanthanum (III) oxide, Merck, Darmstadt, Germany), which is a correction buffer that connects permanently with the matrix instead of the analyzed element. Phosphorus was determined in the form of phosphomolybdate by spectrophotometric method [The concentrations of elements  were determined by atomic absorption spectrometry with an air-acetylene flame (FAAS) using the deuterium background correction. FAAS conditions are described in Table c method  using UVThe reliability of the method was checked using three certified reference materials . Digestion of these materials was performed with the same decomposition procedure used for tea samples. The recoveries of the studied elements ranged between 86 and 103 %, and the relative standard deviations (RSDs) of the test material were 0.11–14.3 % for Tea, NCS ZC73014. Tobacco INCT-OBTL-5 and INCT-PVTL-6 characterized with recoveries in the range of 87–113 % and RSDs between 0.02 and 10.3 %. All the bioelement and toxic metal measurement results in reference materials are summarized in Table Limits of detection (LODs) of the method applied for macroelements, Ca, P, Mg, K, and Na, were as follows: 0.02, 0.03, 0.02, 0.04, and 0.02 mg/100 g, respectively. LODs for microelements, Cu, Cr, Zn, Mn, Fe, Ni, and Co, in the analyzed samples were as follows: 0.009, 0.001, 0.02, 0.02, 0.01, 0.002 and 0.003 mg/100 g, respectively. LODs for toxic elements Pb and Cd amounted to 0.004 and 0.003 mg/100 g, respectively. The limits of quantification (LOQs) were defined as LOQ = 3·LOD and were equal to 0.06, 0.09, 0.06, 0.12, and 0.06 mg/100 g for the analyzed macroelements, i.e., Ca, P, Mg, K, and Na, respectively. LOQs of the method for such microelements as Cu, Cr, Zn, Mn, Fe, Ni, and Co amounted to 0.027, 0.003, 0.06, 0.06, 0.03, 0.006, and 0.009 mg/100 g, respectively. LOQs for toxic elements Pb and Cd were equal to 0.012 and 0.009 mg/100 g, respectively. The limit of detection and limit of quantification were established according to Konieczka and Namieśnik , i.e. LOIn order to validate the analytical protocol, linearity was also evaluated by determining the square correlation coefficients of the calibration curves generated by injections of standard solutions. The data concerning method’s linearity are summarized in Table The results were subjected to statistical analysis, i.e., Spearman rank correlation analysis, Kruskal-Wallis test, factor analysis (FA), and cluster analysis (CA), using Statistica 10 (StatSoft®). All selected variables were tested for normality using Shapiro-Wilk test, and normal distribution was not confirmed; thus, non-parametric tests were applied . Data weThe results concerning the analyzed elements in Pu-erh and fruit teas are listed in Table Pu-erh tea was characterized by high levels of K, P, Ca, and Mg , and these elements were more abundant in leaf tea than in tea bags. The content of Na in marketed Pu-erh tea bags (27.1 mg/100 g) was higher than in a loose form (16.2 mg/100 g), which is in agreement with results published by McKenzie et al. . SimilarThe highest K and Ca levels  were determined in teas consisting only from fruits. In case of black teas with fruit additions, the highest values among macroelements were recorded for K and P (1275 and 881 mg/100 g). Similar K values (1135 mg/100 g) were reported by McKenzie et al. . Black tDifferences in element concentration in infusions might be explained by chelation of these elements with tannic acid and tannins which exudate during the boiling of tea. Dambiec et al.  and Wróbp < 0.05, p < 0.01, p < 0.001). The most significant positive relationships (p < 0.001) in Pu-erh tea were noted for P-Zn, P-Ni, K-Ni, K-Zn, Fe-Co, Fe-Cr, Fe-Pb, Co-Cr, and Cr-Pb. In case of fruit teas, such significant positive interdependences were recorded for Mg-Fe, P-Ni, and Co-Ni.Correlation analysis was performed using the nonparametric Spearman rank test for the analyzed elements at three levels of significance , Co , Cr , Pb , and Na . Strong interdependences were also observed in samples of fruit teas and black teas with fruit additions for such elements as P , Mn , Fe , Ni , and Cd . The null hypothesis of Kruskal-Wallis test was rejected, so a post hoc test for the multiple comparisons was applied, which allowed on differences’ identification. Several elements have been identified as being dependent on the type of tea and its sort of confection. However, such interdependences were not found in case of Ca. What is interesting, it was found that Cr is strongly correlated only with the sort of confection, which can imply the influence of technological processing. All the results are presented in the Table Kruskal-Wallis test was performed for Pu-erh and fruit teas. It was found that the type of tea confection (loose or tea bag) is linked to concentrations of such elements as Fe , which was performed on raw data sets of Pu-erh and fruit teas, revealed two factors F1 and F2) that cumulatively explain 61.36 % of the total variance (F1 = 43.93 % and F2 = 17.42 %). Eigenvalues amounted to 8.59 for F1 and 6.15 for F2. It can be observed on Fig.  and F2 tSimultaneously, F2 is helpful in obtaining extra information concerning fruit and black teas with fruit addition samples, which are described by higher values of F1. Although, these samples partially overlap, but there is also observed diversification according to their type, i.e., black tea with fruit additions are characterized by Ca, and higher values of F1 and F2 Fig. .Based on the conducted analysis, it can be concluded that F1 is a factor responsible for differentiation of samples in view of the tea type (Pu-erh and fruit teas). However, factor 2 provides additional to F1 information concerning the analyzed samples, and both of them cumulatively are responsible for the diversification within the group of fruit tea, i.e., pure fruit tea from black tea with fruit additions as well as Pu-erh tea in view of its confection (loose form–tea bags) which is usually associated with its quality.Most of the authors differentiated teas in view of their type and geographical origin , 29–31. The cluster analysis (CA) was performed using Ward method and Euclidean distance on data concerning Pu-erh and fruit tea samples. Figure As can be observed, CA made it possible to differentiate tea samples in view of the tea quality and its type with respect to the chemical composition of the analyzed samples.The recommended daily intake (RDA) was calculated for Pu-erh and fruit tea infusions according to the latest available Polish  and AmerIn accordance with the EFSA  recommenNowadays, due to the increasing environment contamination, it is important to assess toxic metal and bioelement intake with food, including Pu-erh and fruit teas that consumption steadily increases. In our study, it was found that there is no health hazard associated with exposure to Cd and Pb via consumption of both kinds of the analyzed tea. However, if Ni contamination is observed, its TDI can be exceeded. Such situations can be possible in case of tea bag consumption, which as shown in statistical analysis are described mainly by heavy metals.It was also concluded that multivariate techniques constitute efficient tools helpful in differentiation of tea samples in view of their quality, type, and confection. Magnesium, Ca, K, Na, P, Mn, Cu, Fe, Zn, Co, Cr, Ni, Cd, and Pb proved to be good descriptors for diversification of the analyzed samples.


Objective. To determine the oxidative stress/inflammation behavior in patients with/without acute graft dysfunction (AGD) with Tacrolimus. Methods. Cross-sectional study, in renal transplant (RT) recipients (1-yr follow-up). Patients with AGD and without AGD were included. Serum IL-6, TNF-α, 8-isoprostanes (8-IP), and Nitric Oxide (NO) were determined by ELISA; C-reactive protein (CRP) was determined by nephelometry; lipid peroxidation products (LPO) and superoxide dismutase (SOD) were determined by colorimetry. Results. The AGD presentation was at 5.09 ± 3.07 versus 8.27 ± 3.78 months (p < 0.001); CRP >3.19 mg/L was found in 21 versus 19 in the N-AGD group (p = 0.83); TNF-α 145.53 ± 18.87 pg/mL versus 125.54 ± 15.92 pg/mL in N-AGD (p = 0.64); IL-6 2110.69 ± 350.97 pg/mL versus 1933.42 ± 235.38 pg/mL in N-AGD (p = 0.13). The LPO were higher in AGD (p = 0.014): 4.10 ± 0.69 µM versus 2.41 ± 0.29 µM; also levels of 8-IP were higher in AGD 27.47 ± 9.28 pg/mL versus 8.64 ± 1.54 pg/mL (p = 0.01). Serum levels of NO in AGD were lower 138.44 ± 19.20 µmol/L versus 190.57 ± 22.04 µmol/L in N-AGD (p = 0.042); antioxidant enzyme SOD activity was significantly diminished in AGD with 9.75 ± 0.52 U/mL versus 11.69 ± 0.55 U/mL in N-AGD (p = 0.012). Discussion. Patients with RT present with a similar state of the proinflammatory cytokines whether or not they have AGD. The patients with AGD showed deregulation of the oxidative state with increased LPO and 8-IP and decreased NO and SOD. Chronic end-stage renal disease (ESRD) is characterized by serious, irreversible kidney damage distinguished by proteinuria and a reduction in the glomerular filtration rate (GFR) , 2.Registro de Diálisis y Trasplante del Estado de Jalisco: REDTJAL) and in the State of Morelos, the number of new patients who require renal replacement therapy (RRT) is constantly increasing [Instituto Mexicano del Seguro Social: IMSS) it was $9,091.00 USD [According to the Registry of Dialysis and Transplants in the State of Jalisco  followed by the rapid succession of Tacrolimus (TAC) together with Mycophenolate Mofetil (MMF) is associated with a much lower rate of loss of the renal allograft, achieving graft survival in >90% annually . The mosα), the C-reactive protein (CRP), and the markers of OS have been reported as being significantly elevated in ESRD, with significant decreases two months after RT [Levels of the proinflammatory cytokines like interleukin 6 (IL-6), the tumor necrosis factor alpha . The ingestion of TAC for immunosuppressant therapy was considered an inclusion criterion for both groups. Excluded were the patients >55 years of age, who presented with renal comorbidities, who received a second transplant, and who were undergoing treatment with nonsteroidal anti-inflammatories, angiotensin converting enzyme (ACE) inhibitors, and antagonists of the angiotensin II receptors (ARBs), as well as recipients of transplants from perished donors. The serum levels of IL-6, TNF-α, 8-isoprostanes (8-IP), and Nitric Oxide (NO) were determined with ELISA. Nephelometry was used to determine CRP and the colorimetric method was used to determine levels of the products of lipid peroxidation (LPO)  and 4-hydroxy-alkenals) and levels of the SOD enzyme as an antioxidant.An analytical cross-sectional study was performed. Two study groups were made of RT recipients within the first year of follow-up. The first group included all patients with AGD (elevation of creatinine ≥30%) biopsy proven , in the period of Jan-2014 to Dec-2015; control group included patients without AGD (N-AGD); these patients were randomly obtained from the pool of patients who were unto protocolized graft biopsy (this is a common behavior in our setting); they were rejection-free at the time of biopsy. All patients were on triple immunosuppression scheme based on TAC, MMF, and Prednisone (considered as the most potent scheme). Both groups were first-time recipients of RT. The calculation of the sample size was determined using the formula to compare means and considering, as a variable in the determination, concentration of the superoxide dismutase enzyme (SOD), obtaining 55 patients per study group. The patients were attended to at the Department of Nephrology, Transplant Division, of the Sub-Specialties Medical Unit at the National Occidental Medical Centre of the Mexican Social Security Institute (Once blood samples were collected in two separate tubes (one with 0.1% of ethylenediaminetetraacetic (EDTA) and the other a dry tube), the plasma and serum were separated by centrifugation at 2,000 rpm for 10 minutes at room temperature. Then, the samples were stored at −80°C until processing. All of the technical readings of optical density were made with the Synergy HT (BIOTEK®) microplate reader.α levels were determined by ELISA, following the instructions of the kit manufacturer . First, 100 μL of diluted capture antibody was added, followed by incubation overnight at room temperature. Then, 300 μL of block buffer was added to the wells and it was incubated for 1 hour at room temperature. Serum and standards were added, followed by incubation for 2 hours at room temperature. After several washings, 100 μL of diluted detection antibody was added and incubated at room temperature for 2 hours. 100 μL diluted Avidin-HRP conjugate was added, followed by incubation for 30 minutes at room temperature. Finally, 100 μL of substrate solution was added to each well. The plate was read at a wavelength of 405 nm with correction set at 650 nm and was reported in pg/mL.TNF-Levels of the hsCRP in serum were assessed with immune nephelometry, using the BN II System . The reagents, controls SL/1 and SL/2, CardioPhase hsCRP , and N rheumatology standard were prepared according to the manufacturer's instructions. The reference curve was made, and the samples were diluted 1 : 20 and mixed with the CardioPhase hsCRP. The hsCRP levels were measured automatically by the nephelometry BN II. The levels are reported automatically in mg/L.μL of serum with 455 μL of N-methyl-2-phenylindole in acetonitrile (Reagent 1) was diluted with ferric iron in methanol. Samples were agitated, after which 105 μL 37% HCl was added, followed by incubation at 45°C for 60 minutes and centrifugation at 12,791 rpm for 10 minutes. Next, 150 μL of the supernatant was added and absorbance was measured at 586 nm. The curve pattern with known concentrations of 1,1,3,3-tetramethoxypropane in Tris-HCl was used.Plasma LPO levels were measured using the FR22 assay kit  according to the manufacturer's instructions. In this assay the chromogenic reagent reacts with MDA and 4-hydroxy-alkenals to form a stable chromophore. First, 140 μL of samples or standard was added to each well and 50 μL of 8-IP AChE tracer was added to all wells except the total activity and blank wells. 50 μL of 8-IP enzyme immunoassay antiserum was added to all wells except the total activity and blank wells. At once, 50 μL of 8-IP antiserum was added to all wells except total activity, nonspecific binding, and blank wells. The plate was covered and incubated at 4°C for 18 h and then washed 5 times with buffer. Absorbance was read at 420 nm.The immunoassay reagent kit from Cayman Chemical Company®  was used according to the manufacturer's instructions. The 8-IP assay was based on the principle of competitive binding between sample 8-IP, 8-IP acetyl cholinesterase (AChE) conjugate, and 8-IP tracer. 50 μL) was pipetted into an antibody-coated 96-well plate with 100 μL of HRP-conjugate reagent and incubated at 37°C for 1 h. The wells were then washed four times with buffer wash; 50 μL of chromogen solution A and 50 μL of chromogen solution B were added. The samples were incubated for 15 minutes at 37°C; then 50 μL of stop solution was added, and the absorbance was read at 450 nm.The levels of NO in serum were assessed by sandwich ELISA using a commercially available kit . Before performing the assay samples, reagents were kept at room temperature for 30 min. Nitric Oxide or serum  was placed, and 10 μL of the sample was added. After slow agitation, 20 µL of xanthine oxidase was added to the wells. The microplate was incubated for 20 minutes at room temperature, and the absorbency was read at a wavelength of 440 nm. Levels are reported in U/mL.Serum total SOD activity (U/mL) was determined using a kit from Cayman Chemical Company®, , following the manufacturer's protocol, to detect the O IMSS (Registration number R-2015-1301-83) and the State Research Registry (59/E-JAL/2015) put forth by the General Public Health Administration  in Jalisco, Mexico.The study was performed in accordance with the Principles of Ethics for Medical Research in Human Beings as stipulated by the Declaration of Helsinki 64th General Assembly, Fortaleza, Brazil (October 2013). Informed consent forms were signed, because it was a category III study, in agreement with the General Health Law in Mexico. The project was submitted to and approved by the local Scientific Research and Health Ethics Committee of theU test was used. The categorical variables are presented as frequencies and percentages and were analyzed with the Chi2 test. A value of p ≤ 0.05 was considered statistically significant.Continuous variables are expressed as mean ± standard deviation (SD) or standard error of the mean (SEM) and were analyzed with nonparametric tests according to the results obtained by the Kolmogorov-Smirnov test. For the comparisons between groups the Mann–Whitney p < 0.001) versus 8.27 ± 3.78 months in N-AGD (this was the time of follow-up for protocol biopsy). The age of patients with AGD was 25.39 ± 5.71 years and 28.08 ± 9.12 years in N-AGD. The male gender significantly predominated in both groups (p = 0.004): there were 48 (87%) males in the AGD group and 34 (62%) in the N-AGD group. Heights and weights between the AGD and N-AGD groups were not significantly different. Tobacco use (smoking) was present in 15 patients with AGD and in 8 N-AGD. Alcoholism was present in 14 patients with AGD and in 9 of the N-AGD. Differences in fasting glucose levels were not statistically significant. Findings of uremia were significantly increased in the AGD group with 54.78 ± 3.99 mg/dL (p < 0.001) versus the N-AGD with 36.59 ± 1.32 mg/dL, and the same behavior was found in levels of serum creatinine which was significantly higher in AGD with 1.19 ± 0.20 mg/mL (p = 0.002) versus 1.06 ± 0.27 mg/mL in N-AGD. The age of the donor was significantly higher in the AGD group with 42.54 ± 11.45 years (p < 0.001) versus 34.07 ± 10.65 years in N-AGD. The triglycerides and cholesterols were not significantly different between the groups (The AGD was developed at 5.09 ± 3.07 after transplant (e groups .p = 0.83). The average serum level of TNF-α in AGD was 145.53 ± 18.87 pg/mL versus 125.54 ± 15.92 pg/mL in N-AGD, without a significant difference (p = 0.64). Serum levels of IL-6 in AGD patients were 2110.69 ± 350.97 pg/mL versus 1933.42 ± 235.38 pg/mL in N-AGD, which was not significantly different (p = 0.13)  .p = 0.014, Mann–Whitney U test) with 4.10 ± 0.69 μM versus 2.41 ± 0.29 μM in N-AGD. A similar behavior was found in 8-IP since the average plasma level in AGD patients was also significantly elevated with 27.47 ± 9.28 pg/mL versus 8.64 ± 1.54 pg/mL in N-AGD .Serum levels of LPO in AGD were significantly elevated . The activity of the SOD enzyme was significantly diminished in the AGD group with 9.75 ± 0.52 U/mL versus 11.69 ± 0.55 U/mL in N-AGD   .From each AGD patient the histopathological diagnosis was recorded: 13% were normal; 45% had acute rejection; in 30% toxicity was found; and in 13% other diagnosis: acute tubular necrosis, and borderline changes .Renal transplantation is the RRT of choice for patients with ESRD, to provide them with better quality of life and the best survival compared to dialysis, as well as being the most cost-efficient . BesidesIn our study as in our setting, male receptors are 3 : 1 compared to female , 23; thiOne of the primary causes of later loss of the allograft is chronic graft nephropathy, which is characterized, in part, by deterioration of renal function. Registered data demonstrate that renal function in the first posttransplant year is an important predictor of the long-term result in how the RT behaves, where factors that initiate the cycle of the loss of nephrons play a predominant role. In this regard, advanced age, the masculine gender, or diabetes mellitus are considered, among other factors , 26.It has been reported that serum creatinine concentrations of ≤1.5 mg/dL at 6–12 months are associated with greater survival of the graft at 5 years. These serum creatinine results can be predictive of survival as soon as 1 month after transplantation . In patiα is a functional transmembranal homotrimeric of 26-kDa that is liberated in circulation in functional soluble form of 17-kDa. In plasma TNF-α is free or it binds with its circulating receptor [α, in general, is not present in the kidneys. After stimulation by the lipopolysaccharides, the interleukin-1α, and during inflammation, the TNF-α and its receptors are expressed in the glomerular tuft  and in the tubular cells [α were not significantly elevated (p = 0.64) compared to N-AGD.The proinflammatory cytokine TNF-receptor . The TNFar cells . In the p = 0.13). The IL-6 is a multifunctional pleiotropic proinflammatory cytokine of 26-kD molecular weight, with the ability to modulate local and systemic immunity [α, and CRP are wide studied in clinical studies regarding inflammation, all of them are predictive to mortality and other cardiovascular outcomes in RT patients; up to our knowledge this is the first evaluation of the possible association in AGD of inflammation and OS; however, it is necessary to evaluate other inflammatory markers that may play a role in AGD.The IL-6 in AGD behaved similarly with an increase that was not significant compared to the N-AGD group  that formafter RT , but theafter RT . In our after RT .The endothelial NO synthase (eNOS) is identified as a very important protective factor in renal function .p = 0.042), in detriment to the endothelial function of the RT, as well as the pathophysiological effects the NO has in relation to OS. The NO is produced from the L-arginine as a result of a catalytic reaction by the NOS enzyme. The NO can control such physiologic processes as important as blood pressure, arterial smooth muscle relaxation, platelet aggregation and adhesion, neurotransmission, and neuroendocrine secretion. It also participates in the destruction of pathogenic microorganisms and tumor cells from leukocytes and macrophages. These functions could be compromised by the diminishing of NO in AGD [2− and the activation of transcription factors that later increase the expression of inducible NOS [In our study, the levels of NO in AGD were significantly diminished compared to the N-AGD group  augments the glomerular synthesis of ROS .It is important to consider that TAC remains as the major immunosuppressant drug, and its common adverse effects include arterial hypertension, hyperlipidemia, and hyperglycemia. Pharmacological action of TAC has been previously described; the only known effect of TAC in inflammation is proinflammatory, due to the elevation of ICAM-1 and VCAM-1 dose dependent . This is ex profeso, but there is no evidence regarding the possible pharmacological interaction among TAC and statins in OS [Cardiovascular disease is the most common mortality cause after RT; statins have shown a protective lipid lowering effect, but there is no evidence regarding the possible antioxidant effect of statins  in the Rns in OS , 47. AlsIn conclusion, we found similar expression of proinflammatory cytokines in patients with and without AGD and deregulation of the OS in patients with AGD.The limitations of the present study include a small sample size and it was a cross-sectional study. We are conscious that it is convenient to provide close follow-up in patients after modifying the dose of TAC in the first year and periodic long-term follow-up.


To achieve secure communication in heterogeneous cryptography systems, we present a heterogeneous hybrid signcryption scheme. The proposed scheme allows a sender in an identity-based cryptography system to send multi-message to multi-receiver in a certificateless cryptography system with different master keys. At the same time, all users are mapped to a distinct pseudo-identity for conditional identity privacy preservation. A trusted authority could trace the real identity when necessary. Compared with existing schemes, the proposed scheme is more practical for actual applications. In addition, the proposed scheme has indistinguishability against adaptive chosen ciphertext attacks and existential unforgeability against adaptive chosen message attacks under the random oracle model. Diverse network systems have appeared with the development of technology. These systems utilize different cryptography techniques, such as public key infrastructure (PKI), identity-based cryptography (IBC), and certificateless cryptography (CLC). A cryptographic scheme should be constructed for secure communication in heterogeneous systems. Zheng  firstly It is a practical way for large messages to use hybrid encryption perform secure communication. Hybrid encryption separates encryption into two parts: one part uses public key techniques to encrypt a one-time symmetric key, and the other part uses the symmetric key to encrypt the actual message. The public key encryption part of the algorithm is the key encapsulation mechanism (KEM), whereas the symmetric key encryption part is the data encapsulation mechanism (DEM). In 2003, a formal treatment of this paradigm originated in the work of Cramer and Shoup . Dent 1, 11 studConsidering all the above literature, it is known that none of the existing multi-recipient heterogeneous hybrid signcryption schemes for IBC to CLC. However, in today’s complex network and application environment, the information security situation is also complicated and grim. The production and collection of the mass data results in information explosion lead the network communication become more complex and low effective due to diverse system , 20and mMotivated by the above, considering with multi-PKG signcryption  and condThe rest of this paper is organized as follows: preliminary information is given in section 2. The framework and security model are presented in section 3. The heterogeneous hybrid signcryption for multi-message and multi-receiver (MHHSC) scheme is proposed in section 4. The security proof is presented in section 5. The performance evaluation of the proposed scheme is discussed in section 6. Finally, the conclusion is provided in section 7.G1 and G2 with the same prime order q, and let P is a generator of G1. A bilinear map e: G1 × G1 → G2 need satisfy the following properties:P, Q, R ∈ G1, and e = ee. Also e = eab.Bilinearity: For all P, Q ∈ G1, such that e≠1.Non-degeneracy: There exists e can be computed for P, Q ∈ G1.Computability: In this section, we describe bilinear maps and hard problems. Let consider two cyclic groups Definition 1. Given two groupsG1andG2of the same prime orderq, a bilinear mape: G1 × G1 → G2, and a generatorPofG1, the decisional bilinear Diffie-Hellman(DBDH) problem is to decide whetherT = eabcfor given  andT ∈ G2.Definition 2. Variants decisional bilinear Diffie-Hellman(VDBDH) problem is to decide whetherT = eabc−1for given  andT ∈ G2.Definition 3. Variants computational bilinear Diffie-Hellman(VCBDH) problem is to computeT = eabd−1for given .Setup: With a security parameter ℓ as the input, the PKG and KGC generate their own master key and output the system parameters params.Anony-IBC-KG: The algorithm runs by the PKG of the IBC system. With a user’s real identity RIDA and IDA,1 as the input, the algorithm generates the corresponding private key skA and pseudo-identity IDA.Anony-CLC-KG: The algorithm runs by the KGC of the CLC system. With a user’s real identity RIDBi and IDBi,1 as the input, the algorithm generates the corresponding partial private key DBi, secret key skBi, public key pkBi and pseudo-identity IDBi.Encap: Give the sender’s identity , and private key skA, receiver identity ), the algorithm outputs the encapsulation key K and encapsulation φ.Decap: Give the sender’s identity , receiver secret key, and public key , , the algorithm outputs the encapsulation key K or the symbol ⊥.MHHSC KEM consists of five algorithms:Enc: Take message M and encapsulation key K as input, the ciphertext C is then output. We denote this as C ← DEM.Enc.Dec: Take a key K and the ciphertext C as input, the message M or error symbol ⊥ is output.DEM is a symmetric encryption scheme that requires security for confidentiality and unforgeability. DEM consists of the following two algorithms:Setup, Anony-IBC-KG, and Anony-CLC-KG: Same as 3.1 MHHSC KEM.Signcrypt: Use Encap in 3.1 MHHSC KEM to obtain , use Enc in 3.2 DEM to obtain a ciphertext C, output σ ← .Unsigncrypt: Use Decap in 3.1 MHHSC KEM to obtain K, use Dec in 3.2 DEM to obtain message M, then check the equation. If it holds, receive M. Otherwise, output the symbol ⊥.The proposed MHHSC scheme consists of MHHSC KEM and DEM as follows:In the proposed scheme, the confidentiality property is defined based on the concept of indistinguishability against adaptive chosen ciphertext attacks (IND-CCA2), which considers two types of adversaries with different capabilities. A type I adversary acts as a dishonest user, whereas a type II adversary acts as a malicious KGC that can obtain the master secret key of KGC. The authenticity property is defined basis on existential unforgeability against adaptive chosen message attacks (EUF-CMA).Definition 4.  A heterogeneous hybrid signcryption scheme is said achieved IND-CCA2, if no probabilistic polynomial time adversaryA1has a non-negligible advantage in the following game:Setup: The challenger C runs the Setup algorithm and sends system parameters and public keys to A1, whereas the KGC’s master key is kept secret. Phase 1.A1 can ask several kinds of queries to the following random oracles:Partial private key query: Submit a query on IDBj. If i = 1, 2, ⋯, n), then return DBj. Otherwise, C aborts.Unsigncrypt query: Submit an unsigncrypt query under IDA, IDBj and ciphertext σ. If C runs the formal unsigncrypt algorithm and returns the answer. Otherwise, C searches the list and computes M. Then, check the equation. If holds, M is returned. Otherwise, ⊥ is output.Challenge:C decides when the Phase 1 ends. A1 selects two plaintexts M0, M1 of the same length, and IDA, IDBj to C, which wants to challenge. If C fails and aborts. A1 is not allowed to ask the partial private key of C selects b ∈ {0, 1} and runs the corresponding algorithms to obtain the ciphertext σ* transmits to A1.Phase 2.A1 can perform queries as in Phase 1. A1 cannot query the key extraction for the target identities and should not query the unsigncrypt of σ*.Guess: Finally, A1 produces a bit b′, A1 wins the game if b′ = b.Definition 5.  A heterogeneous hybrid signcryption scheme is said achieved IND-CCA2, if no probabilistic polynomial time adversaryA2has a non-negligible advantage in the following game:Setup: The challenger C runs the Setup algorithm that sends system parameters and public keys to A2. Phase 1.A2 can ask several queries to the following random oracles:Public key query: Submit a public key query on IDBj. If i = 1, 2, ⋯, n), update PK-list with , and return pkBj.Unsigncrypt query: Submit an unsigncrypt query under IDA, IDBj and ciphertext σ. If C runs the formal unsigncrypt algorithm and returns the answer. Otherwise, C searches the list and computes M. Then, check the equation. If the equation holds, return M. Otherwise, ⊥ is output.Challenge:C decides when Phase 1 ends. A2 selects two plaintexts m0, m1 of the same length, and IDA, IDBj to C, which wants to challenge. If C fails and aborts. A2 is not allowed to query for the secret key of C selects b ∈ {0, 1} and runs the corresponding algorithms to obtain the ciphertext σ* transmits to A2.Phase 2.A2 can perform queries as Phase 1. A2 cannot query the key extraction for the target identities and should not query the unsigncrypt of σ*.Guess: Finally, A2 produces a bit b′, and A2 wins the game if b′ = b.Definition 6. (Unforgeability) A heterogeneous hybrid signcryption scheme is said achieved EUF-CMA, if no probabilistic polynomial time forgerFhas a non-negligible advantage in the following game:Setup: The challenger C runs the Setup algorithm and sends system parameters and public keys to F, whereas the PKG’s master key is kept secret. Attack:F issues several kinds of queries.Private key query: Submit a query on IDA. If skA. Otherwise, C aborts.Signcrypt query: Submit a signcrypt query under IDA, σ. Otherwise, C computes σ to satisfy the equation and returns σ to F.Forgery: Finally, F outputs σ* under F wins if Unsigncrypt does not return ⊥.IDA, and n receivers IDA to send m messages to n receivers KDF in scheme denotes a key extract function in G1. Moreover, PKG and KGC can calculate pseudo-identities for users in their system, key pairs or partial private keys of all users are generated by PKG or KGC via the pseudo-identities.Setup: Let G1 and G2 be two cyclic groups with prime order q, where G1 is additive and G2 is multiplicative, and P is the generator of G1. Let e: G1 × G1 → G2 be an admissible bilinear map, a key extract function KDF: lm is the length of a key).H0: G1 → {0, 1}*, H1: {0, 1}* → G1 computes P1 = s1P, where s1 is a master secret key that only the PKG knows.PKG randomly selects H2: G1 → {0, 1}*, H3: {0, 1}* → G1, P2 = s2P, where s2 is a master secret key that only the KGC known.KGC randomly selects params = <e, P, P1, P2, G1, G2, H0, H1, H2, H3, H4, H5, KDF> and keep s1, s2 secret respectively.Public Anony-IBC-KG: Users in IBC obtain their private key as follows:A randomly selects IDA,1 = kAP and transmits  to PKG, where RIDA is the real identity of sender A. PKG calculates IDA,2 = RIDA ⊕ H0, where T denotes the valid period of this pseudo-identity. Finally, the identity os sender A is IDA = .Sender QA = H1(IDA).  is sent to A via a secure path.PKG generates a private key for IBC users as Anony-CLC-KG: Users in CLC obtain their partial private key as follows:Bi randomly selects IDBi,1 = kBiP and transmits  to KGC, where RIDBi is the real identity of receiver Bi. KGC calculates IDBi,2 = RIDBi ⊕ H2, where Ti denotes the valid period of this pseudo-identity. Finally, the identity of receiver Bi is IDBi = .Receiver QBi = H3(IDBi).  is sent to Bi via a secure path.KGC generates the partial private key for CLC users as Bi randomly selects the secret value skBi = xBiDBi, pkBi = xBiP.Signcrypt: A sender A signcrypts n messages mi to n receiver Bi as follows:U1 = r1P2, U2 = r1QA.Randomly selects φi = r2 ⊕ H4(Vi) and let φ = .Compute C = DEM.Enc where K = KDF(r2) and M = (m1 ⊕ R1‖m2 ⊕ R2‖⋯‖mn ⊕ Rn).Compute hi = H5.Compute Si = (r1 + hi)skA and let S = .Compute σ = ).Return ciphertext Unsigncrypt: After receiving a ciphertext σ = ), the receiver Bi decrypts σ as follows:Vi = e, Ri = e and obtain r2 = φi ⊕ H4(Vi).Compute M = DEM.Dec where K = KDF(r2). Receiver Bi recovers own message mi = (mi ⊕ Ri) ⊕ Ri.Recover hi = H5.Compute Accept the message if and only if the following equation holds:The MHHSC scheme will be discussed in this section. The proposed scheme involves four parties: PKG, KGC, sender IDU = . PKG or KGC can retrieve the real identity from any pseudo-identity by RIDU = IDU,2 ⊕ Hi for any disputed event. In addition, the pseudo-identity IDU is generated by both users and PKG or KGC. Hence, only the PKG or KGC that knows the master secret s can retrieve the real identity RIDU from IDU.Note that conditional privacy preservation for each user is mapped to a distinct pseudo-identity qHi queries to Hi for i = 1, 2, 3, 4, 5, qu queries to unsigncryption; qs queries to the signcryption; qppk queries to the partial private key; qsk queries to the secret key; qpk queries to the public key extraction; and qpkr queries to the public key replacement.In this section, we prove that the proposed IBC to CLC hybrid scheme achieves the security requirements of confidentiality and unforgeability. To demonstrate the security of our scheme, we assume that the adversary asks Theorem 1. The above MHHSC scheme is secure against adaptive chosen ciphertext attacks in the standard model assuming that the VDBDH and DBDH problems are difficult.A1 can not distinguish M. Lemma 2 proves that although adversary A2 can obtain M, it cannot distinguish message mj for IDBj.This theorem follows lemmas 1 and 2. Lemma 1 reveals that adversary Lemma 1. In the random oracle, if there is an IND-CCA2 adversaryA1has an advantageϵagainst MHHSC, then there is an algorithmCthat solves the VDBDH problem with an advantageProof. We construct a simulator C that use A1 to decide whether T = eabc−1 by providing a random instance  as the VDBDH problem. This proof consider the indistinguishability of M.Setup: At the beginning, C sets P2 = cP and proves the system parameters to the attacker A1. The target identity is Phase 1.A1 requests a number of queries. C keeps the Hi-list  and PK-list which are used to record answers to the corresponding Hi query and public key query.H3query: Input an identity IDBj. If QBj = tjP. Otherwise, calculate QBj = bP place  in the H3-list, and return QBj.H4query: If  exists in the H4-list, return h4. Otherwise, check if VDBDH oracle returns 1 when queried with the tuple . If this is the case, C returns Vj = eabc−1 and stops. Otherwise, randomly select H4-list, and return h4.Hi query: Upon receiving an Hi query, if the corresponding query exists in the Hi-list, return it to A1. Otherwise, C randomly selects an integer as the query result and returns it to A1. Meanwhile, C places the query result into the Hi-list.Partial private key query: Upon receiving a partial private key query on IDBj. If IDBj, tj, QBj) from the H3-list and sets DBj = tjc−1P return DBj. Otherwise, C aborts.Public key query: When C receives a public key query on IDBj, if there exists  in the PK-list, then C returns pkBj; otherwise, C randomly selects pkBj = xBjP, places  into the PK-list, and returns pkBj as the answer.Replace public key: When C receives a replace public key query on IDBj, C first finds  on the PK-list, then C updates the PK-list with tuple xBj = ⊥, Secret key query: When C receives a secret key query on IDBj, if C replaces public key of IDBj, then C returns ⊥. Otherwise, there exists  in the PK-list and returns xBj as answer.Unsigncrypt query: When receiving an unsigncrypt query under IDA, IDBj and ciphertext σ, if C runs the formal unsigncrypt algorithm and return the answer. Otherwise, C goes through the H4-list with  to find a value such that h4 meets the VDBDH oracle returns 1 when queried on the tuple . If such a tuple exists, return h4 and computer r2 = φj ⊕ h4, K = KDF(r2). Recover M = DEM.Dec, use H5 query to obtain hj, then check equation e = e. If holds, return M. Otherwise, output ⊥.Challenge: After the first stage, A1 outputs two plaintexts M0, M1 and IDA, IDBj to C. If C fails and aborts. Otherwise, C randomly chooses H5 query, sets T is C candidate for the VDBDH obtained from H4 query), C randomly selects K0 ∈ KMHHSC and b ∈ {0, 1} computes C* = DEM.Enc, C provides the ciphertext A1.Phase 2.A1 request a second series of queries as before.Guess: At the end of the simulation, A1 outputs a bit b′ for which the relation σ* = Signcrypt holds. If b′ = b, C outputs T = e = e = abc−1 as a solution of the VDBDH problem.qH4 + qs)qs/2k, and the probability to fail in unsigncryption queries is at most qu/2k. Note that the probability for C to not to fail in first stage is (qH3 − qppk)/qH3. Furthermore, with a probability exactly 1/(qH3 − qppk), A1 chooses to be challenged on C is Then, we assess probability. The probability to fail in signcryption queries is at most abc by providing a random instance  as the DBDH problem. This proof considers the indistinguishability of mj.Setup: At the beginning, C sets P2 = s2P and proves the system parameters to the attacker A2. The target identity is Phase 1.A2 requests a number of queries. C keeps the Hi-list  and PK-list, which are used to record answers to the corresponding Hi query and public key query.H3query: Input an identity IDBj. If QBj = tjP. Otherwise, calculates QBj = bP put  in H3-list return QBj.Hi query: Upon receiving an Hi query, if the corresponding query exists in the Hi-list, then return it to A2. Otherwise, C randomly selects an integer as the query result and returns it to A2. Meanwhile, C places the query result into the Hi-list.Public key query: Upon receiving a public key query on IDBj. If pkBj = xBjP and updates the PK-list. If pkBj = cP update the PK-list with  and return pkBj.Secret key query: When C receives a secret key query on IDBj, if IDBj, xBj, pkBj) in PK-list returns xBj.Unsigncrypt query: When receiving an unsigncrypt query under IDA, IDBj and ciphertext σ, C can compute Vj = e, obtains r2 = φj ⊕ H4(Vj), K = KDF(r2), recovers M = DEM.Dec. Then, if C fails and stops(C cannot compute Rj for skBj is only IDBj can compute). Otherwise, IDBj recovers its own message mj = (mj ⊕ Rj) ⊕ Rj. Submitting H5 query to obtain hj. Then, equation e = e is checked. If holds, mj is returned. Otherwise, output ⊥.Challenge: After the first stage, A2 outputs two plaintexts m0, m1 and IDA, IDBj to C, if C fails and abort. Otherwise, C randomly chooses H5 query, sets C* = DEM.Enc, where M* = mb ⊕ T(T is C candidate for the DBDH). Finally, C provides the ciphertext A2.Phase 2.A2 then requests a second series of queries as before.Guess: At the end of the simulation, A2 outputs a bit b′ for which believes the relation σ* = Signcrypt holds. If b′ = b, C outputs ea = abc as a solution of DBDH problem.qs/2k, and the probability to fail in unsigncryption queries is at most qu/2k. Note that the probability for C to not to fail in first stage is (qH3 − qsk)/qH3. Furthermore, with a probability exactly 1/(qH3 − qsk), A2 chooses to be challenged on C is Then, we assess probability. The probability to fail in signcryption queries is at most Theorem 2. In the random oracle model, if an EUF-CMA adversaryFhas the advantageϵagainst MHHSC, then exists an algorithmCthat solves the VCBDH problem with the advantageProof. We construct a simulator C that uses F to decide whether eabd−1 by providing a random instance  as the VCBDH problem.Setup: At the beginning, C sets P1 = dP and provides the system parameters to the attacker F. The target identity is Attack:F requests a number of queries. C keeps the Hi-lists  which are used to record answers to the corresponding Hi query.H1query: Input an identity IDA. If QA = tP is calculated. Otherwise, calculate QA = bP place  into the H1-list, and return QA.Hi query: Upon receiving a Hi query, if the corresponding query exists in the Hi-list, return it to A2. Otherwise, C randomly selects an integer as the query result and returns it to A2. Meanwhile, C places the query result into the Hi-list.Private key query: When C receives a partial private key query on IDA, if IDA, t, QA) from the H1-list and sets skA = td−1P, return skA. Otherwise, C aborts.Signcrypt query: When receiving a signcrypt query under IDA, n messages mi. If σ. Otherwise, C randomly selects U1 = xP2, Vi = e, Ri = e, φi = r2 ⊕ H4(Vi) and let φ = . Compute C = DEM.Enc where K = KDF(r2) and M = (m1 ⊕ R1‖m2 ⊕ R2‖⋯‖mn ⊕ Rn). Obtain hi from the H5 query, compute U2 = −hiQA + xP1, Si = xP, and return ciphertext σ = ).e = e holds.Equation Forge: Finally, F outputs σ* and IDA, IDBi to C. If C fails and aborts. Otherwise, by forking lemma [C selects a different hash function hi and interacts with F with the same random tape, then the adversary F can provide a different forger σ′*. We know that σ* and σ′* should satisfy the equation C derives the value of eabd−1 as e. Hence, C successfully solves the VCBDH problem.ng lemma , C selecqH4 + qs)qs/2k. With a probability of exactly 1/(qH1 − qsk), F chooses to be challenged on C is The probability of failing in signcryption queries is at most  and Fi. If the steps of computing pi(x) and Fi are not considered, Then, we compare the computational costs of scheme  with thap| = 512.To provide numerical results, we implement IBC-CLC MHHSC to measure the performance of signcryption and unsigncryption operations. Our implementation is written in C using the Pairing-Based Cryptography Library (Libpbc) . For then = 1, 10, 50, 100, 200, 500, 1000. From the figure, we can indicate that signcryption time increases with the number of recipients. However, when unsigncryption, each receiver only operates on its own message, the unsigncryption operation time is not related to the increase of the receiver. So compared with the signcryption and total operation time of the receiver for 1000, the unsigncryption operation time is 0.018, near the bottom of the axis. Therefore, we can see that our scheme can achieve more efficient communication between two systems, which have greater difference in computing power. Users in IBC can handle big data, while users in CLC only need deal with a few data, such as infrastructure-to-vehicle (I2V) communication in vehicular ad hoc networks (VANETs). Trusted authorities or road side units can be the users in IBC system, which have much more capability, and hundreds of on board units can be the users in CLC system, which ability is limited.The performing consequence of our scheme is provided in We propose a novel conditional privacy-preserving heterogeneous hybrid signcryption scheme for IBC to CLC (MHHSC), which allows to send multi-message to multi-receiver. The proposed scheme selects different master secret keys in different systems and maps a distinct pseudo-identity for each user, only the trusted authority could trace the real identity for any disputed event when necessary, which ensures conditional privacy preservation for all users in heterogeneous systems. It is definitely more practical for actual applications, such as VANETs. Moreover, we provide the formal definition and security models for the heterogeneous hybrid signcryption scheme. Proof shows that our scheme is indistinguishability against adaptive chosen ciphertext attacks and existential unforgeability against adaptive chosen message attacks, which is satisfied confidentiality and unforgeability in the random oracle model. Owing to today’s diverse and complex network system and application environment, our follow-up work could be propose a bidirectional heterogeneous signcryption scheme between IBC and CLC for multi-party user.


We planned to determine the association of body mass index (BMI) with diabetes mellitus (DM) and impaired fasting glucose (IFG) in Chinese pulmonary tuberculosis (PTB) patients.3,505 newly-diagnosed PTB patients registered in PTB clinics in Linyi of China between September 2010 and March 2013 were enrolled. DM and IFG were identified based on fasting plasma glucose levels. ROC analysis was used to predict the ability of screening of BMI for DM and IFG in PTB patients.2, patients with DM and IFG had significantly increased trends when BMI ≥ 24.0 kg/m2, and aORs were 2.28 (95%CI 1.44–3.60) and 1.30 (95%CI 1.04–1.64), respectively. After adjustment for age, gender, and educational level, there was an increased odd in BMI ≥ 23.41 kg/m2 for IFG, and a decreased odd in BMI < 19.82 kg/m2 for DM (p < 0.05). The cut-offs of BMI for screening IFG and DM in PTB patients were 22.22 kg/m2 (AUC 0.56) and 22.34 kg/m2 (AUC 0.59).Compared with 18.5–23.9 kg/mIn PTB patients, BMI is significantly associated with IFG and DM. However, the predictive power of BMI was not sufficient, so it may only be a limited screening tool for DM and IFG among PTB patients in China. The co-morbidity of diabetes mellitus (DM) and pulmonary tuberculosis (PTB) represents a double burden with significant public health implications , 2. GlobAlso, patients with impaired fasting glucose (IFG) are more prone to progress to the DM stage -approximately 8.8% per year in China , 7- in tWe designed this cross-sectional study using primary data to explore the association of BMI with DM or IFG in PTB patients in China, and to determine the optimal BMI cut off value for prediction of DM or IFG in Chinese adult patients with PTB.A total of 3,505 PTB patients, 18-45 years of age, were selected from counties of Linyi in Shandong province, China. All the PTB patients were newly-diagnosed and registered for Directly Observed Treatment, Short Course (DOTS) in PTB clinics between September 2010 and March 2013, and also were diagnosed in the PTB clinic of each county by X-ray and sputum smear examination. All participants have written informed consent.DM was diagnosed by World Health Organization criteria based on the fasting plasma glucose (FPG) level . The diagnosis of IFG was based on criteria for the classification of glucose tolerance based on the FPG level, which was defined as a FPG range from 5.6–6.9 mmol/L . The participants fasted for 8–10 h before the blood testing.2) was calculated using the following formula: BMI = weight (kg)/height (m)2. Underweight, normal weight, overweight, and obese categories were defined using the modified criteria for the Chinese population; the BMI cut-off values were 18.5 kg/m2, 24.0 kg/m2, and 28.0 kg/m2, respectively ), gender, educational level, BMI . A P value <0.05 was considered statistically significant.SPSS version 17.0  was used for statistical analyses. The characteristics of PTB patients in the DM, IFG and normal FPG groups and inter-quartile range (IQR) of BMI were compared and analyzed. The mean and standard deviation for continuous variables, such as age, BMI, and blood lipid content, and proportions for categorical variables, including the prevalence of DM or IFG, the percentage of age group, gender, and educational level, are reported. One-way analysis of variance was used to test continuous variables. A chi-square test was used to compare categorical variables. Multinomial logistic regression analyses were performed, and the variables for inclusion in the multivariate model were chosen based on plausibility and variables with P < 0.05). In the PTB + IFG group, 65.2% were male, which was more than the PTB group . The percentage of patients with a high school or higher educational level in the PTB + DM and PTB + IFG groups were lower than the PTB group (P < 0.05). BMI means and the proportion of overweight or obese patients in the PTB + DM and PTB + IFG groups were higher than the PTB group (P < 0.05). The means of TG and TCHO were 1.6 mmol/L and 4.8 mmol/L in the PTB + DM group, respectively, which were higher than the PTB + IFG and PTB groups (P < 0.05).A total of 3 505 PTB patients were included in our study (18–45 years of age), among whom 96 and 529 had co-existing DM and IFG, respectively. The mean age was 31 · 8 years for all PTB patients, and the mean age in the PTB + DM group was the highest (P < 0.05). Overweight or obese patients had significantly higher odds ratios of DM and IFG . In addition, male gender was positively associated with the IFG .Multivariate logistic regression analysis of DM or IFG in PTB patients is shown in Table 2; Q2, 19.82 kg/m2; Q3, 21.45 kg/m2; and Q4, 23.41 kg/m2 ; the prevalence in the patients 31–45 years of age and males were also the same trends.The participants were divided into four groups based on the BMI baseline quartiles, as follows: Q1, <19.82 kg/m2), the percentage in the PTB group was somewhat higher; at higher BMI, it was lower than the PTB + IFG group and PTB + DM group.The percentages of different BMI levels among the PTB + DM, PTB + IFG, and PTB groups are shown in Fig. The univariate and multivariate odds ratios for IFG and DM based on BMI quartiles are shown in Table 2 in all patients and 22.23 kg/m2 in males; and the PPV, NPV, PLR and NLR were 0.70, 0.38, 1.17 and 0.81 for all patients and 0.73, 0.38, 1.36 and 0.80 for males, respectively. With respect to DM, the area under the ROC curve was 0.59 for all PTB patients, 0.58 for patients 31–45 years of age, and 0.61 for males. The optimal BMI cut-off for DM was 22.34 kg/m2 in all patients, 22.53 kg/m2 in patients 31–45 years of age, and 22.41 kg/m2 in males; and the PPV, NPV, PLR and NLR were 0.77, 0.42, 1.58 and 0.65 for all patients, 0.78, 0.39, 1.45 and 0.64 for 31–45 years of age, 0.78, 0.43, 1.66 and 0.60 for males, respectively.The results of ROC analysis showed that BMI might be a weak potential predictor for IFG or DM in PTB patients Table . With re2 (Q4). Until now the optimal BMI cut off point for IFG and DM screening among PTB patients has not been estimated in China. Based on the current study, the optimal BMI cut-offs for predicting DM and IFG were 22.22 kg/m2 and 22.34 kg/m2, respectively.In this survey of Chinese adults (18–45 years of age), we showed that overweight and obesity were positively associated with DM and IFG in PTB patients. The prevalence of DM and IFG was higher when the BMI was ≥ 23.41 kg/m2 was shown in a previous meta-analysis [2) compared to normal weight patients was estimated to be 3.2 (95% CI = 3.1–3.3) [After adjusted for the associated factors, including young age, high educational level, and excess weight, BMI was still a risk factor for DM or IFG in PTB patients. In a previous cohort study , the incanalysis . The rel3.1–3.3) . Meanwhi3.1–3.3) . Thus, o2 range was negatively associated with DM in PTB patients, and a BMI > 23.41 kg/m2 was positively associated with IFG. Moreover, for a BMI ≥ 23.41 kg/m2, the prevalence of DM and IFG in PTB patients was much higher in our study. A cross-sectional study carried out in Spain showed that the prevalence of DM in overweight or obese patients was 23.6%, and the higher the BMI, the higher the prevalence of DM [2 (Q1), the prevalence of DM in PTB patients presented slightly decreased, although the differences were not significant. This finding might be attributable to the impact of PTB. Abundant epidemiologic evidence has indicated that low BMI is a risk factor for PTB, and the biological mechanism underlying the relationship has been well-described by induced impairment of cellular immunity [In the model adjusted by all of the relevant risk factors, a BMI in the 19.82–21.45 kg/mimmunity . In addi2 for men and 24.0 kg/m2 for women [2 for DM and ≥ 22.0 kg/m2 for glucose tolerance abnormalities were better anthropometric indices [We were able to establish BMI cut-off values for DM and IFG in PTB patients in the current study. Although BMI might not be the strongest risk factor to screen undiagnosed DM compared with age, waist circumference, and a family history of DM , it is aor women . A cross indices ; thus, B indices , and in  indices .There were some limitations in our study. First, there were some limitations in the generalizability of our findings: the age range and only Chinese participants. The age range in our survey was 18–45 years, which comprised young and middle-aged adults in China. Because there were no elderly in our study, which comprise a high-risk group and have a higher prevalence than young and middle-aged adults , the preIn conclusion, the present study revealed that BMI might be a limited screening tool for DM and IFG in PTB patients in Chinese young and middle-aged people. However, the predictive power of BMI was still poor. This study is only a prelude to the upcoming research in co-existing infectious diseases and chronic non-communicable diseases, especially in Chinese populations. Further studies are warranted to verify the effects of nutritional status on the co-morbidity of these two diseases.


Non-linear behaviors of a single neuron described by Fitzhugh-Nagumo (FHN) neuron model, with external electromagnetic radiation considered, is investigated. It is discovered that with external electromagnetic radiation in form of a cosine function, the mode selection of membrane potential occurs among periodic, quasi-periodic, and chaotic motions as increasing the frequency of external transmembrane current, which is selected as a sinusoidal function. When the frequency is small or large enough, periodic, and quasi-periodic motions are captured alternatively. Otherwise, when frequency is in interval 0.778 < ω < 2.208, chaotic motion characterizes the main behavior type. The mechanism of mode transition from quasi-periodic to chaotic motion is also observed when varying the amplitude of external electromagnetic radiation. The frequency apparently plays a more important role in determining the system behavior. Dynamic behaviors of single neuron or neural network are essential to understanding complex behaviors in brain or even serious diseases in nervous system. On account of complexity of nervous system, hundreds of equations should be set up to describe neuronal behavior precisely. However, main properties like electrical activities in neurons catch the attention of most researchers, and based on which, several models are established to physically study mode selection of neurons. For example, four-variable Hodgkin-Huxley (HH) equations are usually used to investigate the main properties of neurons via membrane potential  neuron model shows its validity on oscillatory dynamic behavior in a neuron .k0ρ(φ)u. The memory conductance ρ(φ) of a memristor controlled by magnetic flux, which is often described byThe additive magnetic flux is induced by changing the distribution of ionic concentration of lectrolytes. The evolution of magnetic flux across the membrane is described by the third equation in Equation (1), which has great effects on membrane potential u in first equation in Equation (1) by a induced current denoted by the last term v is slow variable for current and magnetic flux across the membrane. The physical meaning of other parameters are present in the reference  = . Values of other parameters are listed here, a = 0.15, μ1 = 0.2, μ2 = 0.3, ε = 0.002, α = 0.1, β = 0.2, I0 = 0.6, k0 = −1, k1 = 0.2, k2 = 1.0.The second I0 could lead to transformation of mode of electrical activities. However, in this section we focus on ω as our control parameter in system. Ma  (Duan et al., We plot the semi-Poincáre diagram with 0 < ω < 3.0, which is shown in Figure Hopf bifurcation plays the key role in motion type selection. Irreducible frequency introduced into system by Hopf bifurcation leads the system from periodic motion to quasi-periodic motion or from quasi-periodic to chaos. In order to specify the statement, we also provide the frequency spectrum of second variable for the three different motion type when the parameter ω is closed to each other. We select ω = 0.754, ω = 0.755, and ω = 0.78 for periodic, quasi-periodic, and chaotic motion types. One, two, and three irreducible frequencies are discovered in frequency spectrum diagram (see Figure ext = A cos 2πft. The amplitude A is also an important control parameter, which decides the state of system when other parameters are fixed. We still use semi-Poincáre diagram to describe the scenario as varying A during 0 < A < 1.0. The situation of A > 1.0 will not shown in this paper because the the system stay in equilibrium state, which means all the variables remain constants.External electromagnetic radiation is chosen as a cosine function φ14, respectively.Three semi-Poincáre diagrams are plotted as ω = 0.618, ω = 0.4, and ω = 0.6, corresponding to periodic, quasi-periodic, and chaotic motion when A is fixed in 0.1, shown in Figures A < 1.0, periodic motion is discovered, but is is interrupted by quasi-periodic motion several times, when ω = 0.618 (Figure A < 1.0 (Figure In large span of 0 < 8 Figure . When A 0 Figure . SimilarA < 0.0019, however, three closed orbits appear when A is >0.0019. The new orbit is growing lager when we increase A. The process of growth of the new orbit is stopped by a Hopf bifurcation and system enters the chaotic state when A is >0.0832. The growth process of orbit in Poincáre section is shown in Figure It is worth noting that Ruelle-Takens route to chaos also exists when varying the parameter A. Take the scenario shown in Figure Ist, and another comes from the external electromagnetic radiation φext.In this paper, we study dynamical behavior of a single neuron system driven by two plane waves. One of them is provided by external current denoted by ext fixed, system state is chosen from periodic, quasi-periodic, and chaotic motion. Specifically, system behavior switches back and forth between periodic and quasi-periodic when 0 < ω < 0.778. In the interval of 0.778 < ω < 2.208, only chaotic motion is fond. But when ω is beyond 2.208, the system alternates again between periodic and quasi-periodic motions and the chance of finding quasi-periodic motion is much more than 0 < ω < 0.778.With frequency of electromagnetic radiation φWe also study the non-linear behavior of system varying amplitude of electromagnetic radiation A at three ω values. Very different scenarios are displayed. Periodic, quasi-periodic, or chaotic motion type characterize the scenario at different ω value, respectively.ext and amplitude of electromagnetic radiation A. However, it seems ω plays a much more important role in controlling the system state. In summary, system behavior is very sensitive to frequency of external force, which might be related to the ratio of frequencies of two external plane waves.The route to chaos, induced by Hopf bifurcation, is discovered varying both frequency ω of electromagnetic radiation φAll authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The simple detection of DCP by “turn-on” fluorescence property of the chemosensor makes it unique for easy and rapid  The simple organophosphates, present in pesticides eventually become more popular chemical weapon due to its very simple method of manufacturing, availability, and low cost along with its dispensability. Despite of its common use as a pesticide, various analogues of such organophosphates are found to be very potent nerve agent, which irreversibly damage functions of nerve cells. Among all the highly toxic, volatile nerve agents, Sarin (GB), Soman (GD) and Tabun (GA) are of most common. If the nerve agents are being inhaled or absorbed through skin, their reactive phosphate group irreversibly react with the hydroxyl group of cellular acetylcholinesterase, which is responsible for breaking down the acetylcholine neurotransmitter, leads to its inactivation. The fatal consequences are neurological imbalance at cholinergic synapses, failure of several organs, paralysis of central nervous system and rapid death5. On 4th April 2017, Khan Shaykhun, a city of Syria faced the destructive activities of suspected chemical warfare with nerve agent Sarin6. For public safety concern and national security, a reliable, facile and reactive detection method of such nerve agents is of great global interest towards the development of practical ‘in field’ devices.1995, The terrorist attack on Tokyo subway introduces the whole world with a new threat to mankind- Chemical Warfare Agents (CWAs)10, fluorometry17, interferometry18, electrochemistry22, surface acoustic wave (SAW) devices24, photoacoustic spectroscopy25, moleculer imprinting coupled with lanthanide luminescence and fibre optics28, enzymatic assay30, and gas chromatography-mass spectrometry31 have been explored to detect substantial number of CWAs. But the problem arises when these existing methods show one or more drawbacks such as low response and sensitivity, limited selectivity, non-specificity and non-portability, difficulties in real time detection, false positive readings and operational complexity. The chemosensors mediated detection is an extremely convenient and simple method based on optical outputs such as color change or change in fluorescence intensity after the exposure to the target analytes35. Chemosensors make such detection method attractive and widely acceptable by fulfilling the key features of a practical sensing system i.e. high selectivity, sensitivity, specificity, portability, ability of real time sensing along with cost effectiveness37. Implementation of a reliable chemosensor is a highly challenging but it brings a considerable importance for their wide application in medical, environmental and biological field. Generally, diethylchlorophosphate (DCP) is used as mimic of the nerve agent Sarin. DCP is much less toxic than Sarin but its mode of action is similar.Till today, several detection methods, such as- colorimetryARC, 3-aminophenol based rhodamine conjugate. This chemosensor is simple and efficient in showing fluorescence signal in presence of DCP in aqueous medium both in in vivo and in vitro systems. There are reports that described recognition of deadly and relevant molecules like DCP and other organophosphates. A comparative assessment of our chemosensor has been provided with that of the published one . Characterization of the probe ARC has been done by 1H NMR, 13C NMR, 31P NMR and MS analysis . Within a few seconds in contact with DCP, ARC gives a prompt fluorescence response with a full signal emission maxima at 585 nm (λex = 520 nm) - a 100 fold increased fluorescence signal (Фfs = 0.58) compared to its own  - a nonreactive analogue of DCP; some common metal ions such as Zn2+, Cu2+, Co2+, Hg2+; reactive oxygen species (ROS), i.e. NaOCl, tBuOOH, H2O2; common acid chlorides, i.e. acetyl chloride, benzoyl chloride; and common pesticides, i.e. benzene hexachloride (BHC), phorate and chlorothalonil  e.g. diethylcyanophosphonate (DCNP), diethyl(1-phenylethyl)phosphonate (DPEP), diethyl(methylthiomethyl)phosphonate (DMTMP), diethyl-(2-oxopropyl)phosphonate (DOPP), Ethyl methylphosphonate (EMP), Diethyl methylphosphonate (DEMP) by UV-vis and fluorescence method Figs S and S7. Ps Fig. S. We haveil Fig. S. From thARC and DCP showed that the reaction is complete within 20 seconds  over the chemosensor ARC (1 µM) in aqueous medium  at room temperature. The pseudo-first-order rate constant (k′) has been calculated as k′ = 0.172 s−1  has been calculated according to the equation38:t and Fmax are the fluorescence intensities at 585 nm (λex = 520 nm) at time ‘t’ and the maximum value obtained after the reaction is complete, respectively, and k′ is the observed pseudo-first order rate constant.A kinetic investigation of the reaction between ds Fig. S. We then−1 Fig. S. This raARC to DCP has also been successfully established in gaseous phase, as our probe can satisfy an important role of a convenient chemosensor kit. We did the experiment using a Whatman-41 filter paper strip immersed in ARC, keeping inside in a glass chamber. A rapid “turn-on” fluorescence response was noticed after adding a few drops of DCP in that glass chamber . We therefore maintained the pH of ARC at 7 while doing all experiments. However, while adding DCP at neutral pH the probe shows its deep pink color even at pH 8  method using the Gaussian 09 program  than that of the chemosensor ARC (4.61 eV)  Fig. S. The maiDCP from Catfish brain through the fluorescence microscopic imaging upon binding with ARC in Catfish brain. To test the efficiency of binding of DCP to ARC and subsequent fluorescence emission, fish brain, more specifically the slices from optic tectum– the region which have earlier been shown to possess high level of acetylcholinesterase activity41, were used in the present study. As shown in Fig. ARC alone does not produce detectable fluorescence signal, while incubation of ARC followed by DCP, promoted a robust increase in fluorescence emission indicating specificity of interaction. The experiment has also been done in reverse way, i.e. first incubation of DCP followed by ARC which gives the same result as earlier. These also suggest successful penetration of ARC and DCP into the Catfish brain tissue.Furthermore, we attempted to visualize ARC in A549 human cancer cell line  was carried out using a micromass Q-TOF MicroTM instrument by using methanol as a solvent. Fluorescence spectra were recorded on a Perkin Elmer Model LS 55 spectrophotometer. UV spectra were recorded on a SHIMADZU UV-3101PC spectrophotometer. The following abbreviations are used to describe spin multiplicities in 1H NMR spectra: s = singlet; d = doublet; t = triplet; m = multiplet.Rhodamine B, phosphorus oxychloride and 3-amino phenol were purchased from Sigma-Aldrich Pvt. Ltd. (India). Unless mentioned otherwise, materials were obtained from commercial suppliers and were used without further purification. Solvents were dried according to the standard procedures. Elix Millipore water was used throughout all experiments. 3 and allowed to reflux for 24 h under a dinitrogen blanket. The excess amount of POCl3 was removed in a rotary evaporator. The acid chloride was dried under vacuum and used for the next step without further purification. This was dissolved in dry acetonitrile (100 mL) then 3-amino phenol  followed by triethylamine (5 mL) were added slowly. Once the addition was complete, the resulting mixture was heated to reflux for 24 h. The reaction mixture was then concentrated in a rotary evaporator under low pressure and then extracted with dichloromethane. After drying it over anhydrous Na2SO4, the organic layer was evaporated completely. The residue was purified by column chromatography with the eluent chloroform: ethyl acetate  to get the product ARC with 74% yield : d (ppm) = 9.34 , 7.84–7.86 , 7.51–7.55 , 7.00–7.02 , 6.88–6.92 , 6.47–6.52 , 6.35–6.40 , 6.280–6.286 , 6.19–6.20 . 13C NMR : d (ppm) = 167.18, 157.54, 154.15, 152.65, 148.76, 138.42, 133.74, 129.95, 129.29, 128.82, 124.16, 123.26, 116.78, 114.14, 113.75, 108.55, 106.18, 97.72, 44.07, 12.87. HRMS (TOF MS): : Calcd. for C34H35N3O3: 533.27. Found: m/z = 534.27 . 1H, 13C NMR and HRMS spectra are shown in Figs SRhodamine B 4.80 g, 10 mmol) was taken in a 250 mL RB flask containing 100 mL freshly distilled POCl0 mmol waARC (1 × 10–6 M) was prepared in water-acetonitrile . 1 × 10−5 M DCP solution was prepared in Millipore water. All experiments were carried out in aqueous medium at neutral pH. During titration, each time a 1 × 10−6 M solution of ARC was filled each time in a quartz optical cell with 1 cm optical path length and DCP stock solution was added into the quartz optical cell gradually by using a micropipette. For all fluorescence measurements, excitations were provided at 520 nm, and emissions were collected from 540 to 640 nm.A stock solution of ARC and DCP has been done by measuring the fluorescence spectra after mixing ARC and DCP in a cubic 4-sided quartz cell of 3 ml. The reaction was carried out at room temperature under the excess amount of DCP  and the reaction was expected to reach 100% conversion of chemosensor ARC to ARC-DCP complex. ARC and DCP solution of concentration 1 µM and 10 µM, respectively, were prepared in aqueous medium and mixed to investigate the kinetic. The excitation wavelength was 520 nm.The kinetic study of the reaction between 2 polystyrene, filter-capped tissue culture flask in an atmosphere of 5% CO2 and 95% air at 37 °C in CO2 incubator. When the cells reached the logarithmic phase, the cell density was adjusted to 1.0 × 105 per/well in culture media. The cells were then used to inoculate in a glass bottom dish, with 1.0 mL (1.0 × 104 cells) of cell suspension in each dish. After cell adhesion, culture medium was removed. The cell layer was rinsed twice with phosphate buffered saline (PBS) (pH 7.0), and then treated with ARC or ARC and DCP according to the experimental need.A549 cell  lines were prepared from continuous culture in Dulbecco’s Modified Eagle’s Medium  supplemented with 10% fetal bovine serum (Invitrogen), penicillin (100 μg/mL), and streptomycin (100 μg/mL). Cells were initially propagated in 75 cm4 A549 cells in 1000 μL of medium, were seeded on sterile 35 mm glass bottom culture dish , and incubated at 37 °C in a CO2 incubator for 10 hours. Then cells were washed with 500 μL DMEM followed by incubation with ARC (1 µM) dissolved in 1000 μL DMEM at 37 °C for 1 h in a CO2 incubator and cells were washed thrice with phosphate buffered saline (PBS) (pH 7.0) to remove excess ARC observed under an Olympus IX81 microscope equipped with a FV1000 confocal system using 1003 oil immersion Plan Apo (N.A. 1.45) objectives. Images obtained through section scanning were analyzed by fluorochrome filter POPO-3 with excitation at 534 nm monochromatic laser beams, and emission spectra were integrated over the range 580 nm (single channel). The cells were again incubated with DCP (10 µM) for 10 min and excess DCP was washed thrice with PBS (pH 7.0) followed by observations under microscope. For all images, the confocal microscope settings, such as transmission density, and scan speed, were held constant to compare the relative intensity of intracellular fluorescence. All images were captured using 2% lesser light.For confocal imaging studies, 1 × 10In vitro studies established the ability of the chemosensor ARC to detect DCP in biological system. Human cell A549 (ATCC No CCL-185) were used as models. However, to materialize this objective, it is a prerequisite to assess the cytotoxic effect of ARC and ARC-DCP complex on live cells. The well-established MTT assay was adopted to study cytotoxicity of above mentioned complexes at varying concentrations. A cytotoxicity measurement for each experiment shows that the chemosensor ARC does not have any toxicity on the tested cells and ARC-DCP complex does not exert any significant adverse effect on cell viability at tested concentrations.Clarias batrachus , average body weight 65.48 ± 3.91 g, were maintained in laboratory condition as described earlier43. All the experimental protocols were approved by the Institutional Animal Ethical Committee (Reg. no.: 1819/GO/Ere/S/15/CPCSEA) of Visva-Bharati University, Santiniketan, India. We also confirm that all the experiments were performed in accordance with the relevant guidelines and regulations. During autopsy, fishes were killed by decapitation; brain was dissected out and placed immediately in phosphate buffer saline, PBS . Catfish brain, specifically the optic tectum  region was sliced using sharp blades and incubated in 6 well culture plate , either in PBS alone or supplemented with the ARC (1 µM) for 15 min in a humidified chamber at 23 ± 20 C. Brain slices were washed thrice with PBS, probed with the guest (DCP: 10 µM), for 5 min with gentle shaking (40 rpm), washed again to remove unbound guest and observed under an inverted fluorescent microscope  using HI PLAN I 10 × /0.22 PH1 objective and ROHD filter (excitation and emission ranges at 520 nm and 580 nm respectively). Images were acquired through Leica EC4 Digital Camera and DFC 3000 G and processed using LAS X software.Live specimens of adult male catfish, Supplementary Information


The high level of bicarbonate may add to the pollution load during discharge. We therefore suggest a new method for bicarbonate-rich wastewater remediation in acidic soil. In our laboratory irrigation experiments, wastewater irrigation efficiently increased the pH and decreased the exchangeable aluminum in the acidic soil. Furthermore, the wastewater method efficiently remediated the entire soil body, while lime application remediated only a portion of the topsoil. Wastewater irrigation also improved soil fertility .The swine industry in China is experiencing a wastewater crisis. In this work, we found that swine wastewaters were particularly high in bicarbonate (1.52–9.25 g/L, mean = 5.68 g/L,  However, the Chinese swine industry is experiencing a wastewater disposal crisis owing to the launching of the “Environmental Protection Storm” by the Chinese government. In Zhejiang Province alone, about 45 thousand farms have been forced to close since 20132.China is the largest swine producer in the world3 and high concentrations of chemical oxygen demand4 and ammonia5 in swine wastewater pose an economic and technical challenge regarding treatment. For example, 6.0 billion tons of swine wastewater are discharged annually in China without appropriate disposal4. Wastewater reuse is an emerging strategy that can confer a variety of environmental benefits6. Generally, water and nutrients are regarded as the only useful wastewater components. Pollutants and nutrients have been the focus of previous investigations on swine wastewater composition8. In recent years, bicarbonate in wastewater has been recognized to facilitate algal blooms9 and cause toxicity in some aquatic organisms11. However, if swine wastewater with high ammonia concentrations also possesses sufficiently high levels of bicarbonate, it can be suitable for acidic soil remediation for the following two reasons. (1)The bicarbonate in wastewater neutralizes the protons in the soil  exchange:The large volumeEquation  after thHerein, we investigated the characteristics of swine wastewater from 30 swine farms in Zhejiang Province and explored the feasibility of applying swine wastewater for acidic soil remediation.n = 42; see Table n = 42; see Table Bicarbonate concentrations were particularly high  and SL2 (3–6 cm), but wastewater application was effective in all layers  values were 327.4 mg/kg and 782.6 mg/kg for NH4-N and P, respectively. These results indicate that the acidic soil efficiently retained the N and P present in the wastewater.Both the NHerm Fig.  for the 12. Therefore, swine wastewater is often high in both biosolids and dissolved organic matter, which are rich in carbon  was reported to be 142 mg/L13, while the mean concentration in 77 rivers in North America, South America, Asia, Africa, Europe, and Oceania was 146 mg/L11. Therefore, the bicarbonate concentration in the swine wastewater is ten times higher than that in typical freshwater systems. This high level of bicarbonate could add to the pollution load of the discharged wastewater.Swine wastewater consists primarily of residual drinking water, rinse water, urine, solid manure, and residual feed. A fattening pig can generate 8.0 kg of wastewater daily, as well as 2.0 kg of solid manure and 3.3 kg of urineon Table . Organicns Table . Additio14, occupying approximately half of the total arable land area16. Cropland soil acidification has intensified in recent decades17. Al toxicity is the dominant factor limiting crop growth in acid soil16. Currently, liming is the most common and practical strategy to combat soil acidification and acidity. In contrast to the easily released H+ in acidic soil, Al3+ in the exchange complex18 can only be released by Ca2+ replacement when lime is in contact with soil. Therefore, the remediation efficiency for Al toxicity is dependent on the degree of lime contact in the soil, which is determined by the liming rate and degree of lime mixing in the soil. As a) soil has substantial spatial heterogeneity because of its complicated composition and large variability in particle size, and b) lime has a very low mobility in soil, the degree of lime mixing in soil can be low. In particular, the degree of lime mixing in soil can be far lower in field situations than under experimental conditions. The results of this study highlighted the low degree of lime contact in soil, and indicated that liming could limit the quality of remediation. In the field, lime may only remediate a portion of the topsoil, while wastewater can efficiently remediate the entire soil body. Swine wastewater irrigation is simple and convenient, can contribute to solving the wastewater discharge problem in swine production, and help remediate acid soil, resulting in soil of high quality. Moreover, Swine wastewater irrigation can be practiced at a cost as low as that of wastewater distribution. A 10,000-head swine farm can generate approximately 20,000 tons of wastewater annually, which can be applied at a rate of 0.8 L/kg to approximately 6 ha of acidic soil for remediation of the top 30-cm soil layer. Furthermore, the high concentrations of organic carbon and nutrients  in wastewater could be beneficial for soil fertility. In particular, P addition and pH adjustment because of wastewater irrigation could substantially increase the P availability, as acidic soil is generally deficient in P. Generally, there are very low concentrations of persistent contaminants . Soil remediation with suitable wastewater is a novel strategy than can drive a range of benefits.We suggest that the environmental protection authorities should adopt a policy encouraging swine wastewater utilization instead of closing swine farms to eliminate swine wastewater generation.Acidic soils are widely distributed in China and other countiesA total of 1 L of swine wastewater  was collected at each farm in an acid-rinsed polyethylene bottle between November 2016 and July 2017. For the wastewater samples from farms #17 and #19, the sample volume was 25 L each. Fresh swine urine, drinking water, and rinse water were also sampled at farm #17 with an acid-rinsed polyethylene bottle. All samples were transported within 12 h in an icebox to the laboratory, separated into 250 mL subsample aliquots in acid-rinsed polyethylene bottles, and stored at −4 °C.4-N, NO3-N, PO4-P, and heavy metal concentrations according to the Chinese State Environmental Protection Agency Standard Methods13. The wastewater samples were first centrifuged at 7,000 rpm for 2 min, after which the collected supernatant was filtered through a 0.45 μm cellulose membrane. The filtrates were analyzed for HCO3−, Cl−, and SO42− , DOC , NH4-N (using the Nash-reagent spectrophotometric method), NO3-N (using the phenoldisulfonic acid method), and PO4-P (using the molybdenum–antimony anti-spectrophotometric method). In addition, K, Ca, Mg, Fe, Cu, Zn, Al, Mn, Cd, Pb, Ni, As, Cr, Co, Sr, and Sn concentrations were determined via inductively coupled plasma mass spectrometry . The solid suspension (SS) was determined by filtering 100 mL of wastewater through a pre-weighed Whatman filtration paper, and then weighing the filtration paper and the solid particles after drying to a constant weight at 7 °C. The C and N contents of the SS were determined by a CHNOS elemental analyzer  after the dried SS was ground and passed through a 0.149 mm sieve.Wastewater pH was measured with a PHB-4 model pH meter . Wastewater samples were also analyzed for bicarbonate, dissolved organic carbon (DOC), NH3, AR grade) at rates of 0, 0.6, 1.2, 2.4, and 4.8 g/kg, and irrigating with deionized water at rates of 0, 0.4, 0.8, 1.2, and 1.6 L/kg, respectively; (2) wastewater irrigation using the sample from farm #17: irrigating using the wastewater at rates of 0, 0.4, 0.8, 1.2, and 1.6 L/kg; and (3) wastewater irrigation using the sample from farm #19: irrigating using wastewater at rates of 0, 0.4, 0.8, 1.2, and 1.6 L/kg. Irrigations consisted of intervals in which 0.4 L/kg of irrigant was added at a time after air-drying of the soil. The soil was stirred vigorously during irrigation. The experimental design was randomized, and three replicates were performed for each sample.Irrigation experiment 1 was conducted at our lab between February and April 2017. The room was kept a constant temperature of 25 °C. The acidic soil consisted of inceptisol obtained from a tea farm in Hangzhou, China. The soil was air-dried, ground, and passed through a 2 mm sieve. A 1.2 kg soil sample was placed in a polyethylene pot . Three treatments were applied. (1) Liming: applying lime  at a rate of 4.8 g/kg, and irrigating with deionized water at a rate of 2.4 L/kg; and (2) wastewater application using the sample from farm #19: irrigating with the wastewater at a rate of 2.4 L/kg. Irrigations consisted of intervals in which 0.4 L/kg of irrigant was added at a time after air-drying of the soil. The experimental design was randomized, and three replicates were performed for each sample. After air-drying of the irrigated soil, the soil pH was measured in each 3 cm layer from the top downward.Irrigation experiment 2 was conducted at our lab between March and April 2017. This irrigation experiment employed polyethylene columns with diameters of 9.0 cm and heights of 30.0 cm. The same soil used in irrigation experiment 1 was used in experiment 2. Two treatments were applied. (1) Liming: applying lime .Soil exchangeable Al and Ca were determined using a KCl extraction method. The KCl concentration was 1 mol/L, and the soil-to-liquid ratio was 1:10. After mixing the soil and the KCl solution, the mixture was agitated on a reciprocating shaker at 120 rpm for 30 minutes. After a 30-minute centrifugation period, the supernatant was filtered through a 0.22 μm pore-size Millipore filter for Al and Ca measurement. Al and Ca concentrations were determined via inductively coupled plasma mass spectrometry .4-N and PO4-P. The soil adsorption capacity for NH4-N and PO4-P was calculated based on the concentrations before and after the adsorption equilibrium. The adsorption isotherms were simulated with the Langmuir equation in Origin 9.0 (OriginLab Corporation).The soil adsorption experiment was conducted at our lab in July 2017. A 1.50 g sample of the same acidic soil used in the irrigation experiments was placed in a 50 mL centrifuge tube with either 10.0 mL of the original wastewater sampled at farm #17 in July 2017 or wastewater diluted at ratios of 1:2, 1:4, 1:6, 1:8, and 1:10. The tubes were shaken at 25 ± l °C for 24 h and then centrifuged at 7,000 rpm for 2 min, after which the collected supernatant was filtered through a 0.45 μm cellulose membrane. The filtrates were analyzed for NHThe calcium conversion efficiency  is defined as the percent of applied Ca converted to soil exchangeable Ca:Zero-Al pH is defined as the soil pH when exchangeable Al decreases to zero with the application of a remediation agent.P ≤ 0.05. Statistical analyses were performed in SPSS, version 16.0 .The means and standard deviations of the data were calculated using Microsoft Excel. A t-test was used to compare the paired means. Differences were considered significant at Supplementary Information