Elucidation of in Vitro Chlorinated Tyrosine Adducts in Blood Plasma as Selective Biomarkers of Chlorine Exposure

[Image: see text] Chlorine is a widely available industrial chemical and involved in a substantial number of cases of poisoning. It has also been used as a chemical warfare agent in military conflicts. To enable forensic verification, the persistent biomarkers 3-chlorotyrosine and 3,5-dichlorotyrosine in biomedical samples could be detected. An important shortfall of these biomarkers, however, is the relatively high incidence of elevated levels of chlorinated tyrosine residues in individuals with inflammatory diseases who have not been exposed to chlorine. Therefore, more reliable biomarkers are necessary to distinguish between endogenous formation and exogeneous exposure. The present study aims to develop a novel diagnostic tool for identifying site-specific chlorinated peptides as a more unambiguous indicator of exogeneous chlorine exposure. Human blood plasma was exposed in vitro to various chlorine concentrations, and the plasma proteins were subsequently digested by pronase, trypsin, or pepsin. After sample preparation, the digests were analyzed by liquid chromatography tandem mass spectrometry (LC–MS/MS) and liquid chromatography high-resolution tandem mass spectrometry (LC–HRMS/MS). In line with other studies, low levels of 3-chlorotyrosine and 3,5-dichlorotyrosine were found in blank plasma samples in this study. Therefore, 50 site-specific biomarkers were identified, which could be used as more unambiguous biomarkers for chlorine exposure. Chlorination of the peptides TY*ETTLEK, Y*KPGQTVK, Y*QQKPGQAPR, HY*EGSTVPEK, and Y*LY*EIAR could already be detected at moderate in vitro chlorine exposure levels. In addition, the latter two peptides were found to have dichlorinated fragments. Especially, Y*LY*EIAR, with a distinct chlorination pattern in the MS spectra, could potentially be used to differentiate exogeneous exposure from endogenous causes as other studies reported that this part of human serum albumin is nitrated rather than chlorinated under physiological conditions. In conclusion, trypsin digestion combined with high-resolution MS analysis of chlorinated peptides could constitute a valuable technique for the forensic verification of exposure to chlorine

Ultraviolet and optical properties of Narrow-Line Seyfert 1 galaxies

Narrow Line Seyfert 1 (NLS1) galaxies are remarkable for their extreme continuum and emission line properties which are not well understood. New results bearing on the spectroscopic characteristics of these objects are presented here, with the aim of establishing their typical ultraviolet and optical spectral behavior. We employ HST observations of 22 NLS1s, which represent a substantial improvement over previous work in terms of data quality and sample size. High signal-to-noise NLS1 composite spectra are constructed, allowing accurate measurements of the continuum shape and the strengths, ratios, and widths for lines, including weak features which are barely identifiable in other Active Galactic Nuclei (AGN) composites. We find that the NLS1 sources have redder UV-blue continua than those typically measured in other quasars and Seyferts. Objects with UV line absorption show redder spectra, suggesting that dust is important in modifying the continuum shapes. The data also permit a detailed investigation of the previously proposed link between NLS1s and z >~ 4 quasars. Direct comparison of their composite spectra, as well as a Principal Component Analysis, suggest that high-z QSOs do not show a strong preference toward NLS1 behavior.Comment: 23 pages (incl. 9 figures, 4 tables), to appear in The Publications of the Astronomical Society of the Pacifi

The UV Properties of the Narrow Line Quasar I Zwicky 1

I Zw 1 is the prototype narrow line quasar. We report here the results of our study of the UV emission of I Zw 1 using a high S/N (50-120) spectrum obtained with the HST FOS. The following main new results are obtained: 1. The Mg II and Al III doublets are partially/fully resolved. The measured doublet ratios verify theoretical predictions that the lines are thermalized in the BLR. 2. A weak associated UV absorption system is detected in N~V, and possibly also in C IV and Lya, suggesting an outflow with a velocity of 1870 km/s and velocity dispersion <300 km/s. 3. Lines from ions of increasing ionization level show increasing excess blue wing flux, and an increasing line peak velocity shift, reaching a maximum blueshift of about 2000 km/s for He II 1640. This may indicate an out-flowing component in the BLR, where the ionization level increases with velocity, and which is visible only in the approaching direction. The highest velocity part of this outflow may produce the associated UV absorption system. 4. The small C III] 1909 EW, and the small C III] 1909/Lya and C III] 1909/Si III] 1892 flux ratios indicate a typical BLR density of 10^11, i.e. about an order of magnitude larger than implied by C III] 1909 in most quasars. A BLR component of a higher density is implied by the EW and doublet ratio of the Al III 1857 doublet. 5. Prominent Fe II UV 191 emission is seen, together with weaker line emission at 1294 and 1871 A. These three features have been proposed as evidence for significant Lya pumping of the 8-10 eV levels of Fe II. 6. Significant Fe III emission is present. The Fe III UV 34 and UV 48 multiplets are clearly resolved, and Fe III UV 1, UV 47, UV 50, and UV 68 may also be present. (Shortened version)Comment: 28 pages, 1 table and 7 figures included. Uses aas2pp4.sty. Scheduled for the Astrophysical Journal November 10, 1997 issue, Vol. 48

Tacrolimus intra-patient variability is not associated with chronic active antibody mediated rejection

Background Chronic active antibody mediated rejection (c-aABMR) is a major cause of long-term kidney allograft loss. It is hypothesized that frequent sub-therapeutic exposure to immunosuppressive drugs, in particular tacrolimus (Tac), is a risk factor for the development of c-aABMR. The intra-patient variability (IPV) in Tac exposure may serve as a substitute biomarker for underexposure and/or non-adherence. In this study, the association between Tac IPV and the development of c-aABMR was investigated. Methods We retrospectively included 59 patients diagnosed with c-aABMR and compared them to 189 control patients matched for age, year of transplantation and type of kidney donor. The Tac IPV was calculated from pre-dose tacrolimus concentrations measured over a 3 year period preceding the diagnosis of c-aABMR. The mean Tac predose concentrations (C0), Tac IPV, renal allograft function and graft survival were compared between the groups. Results Tac IPV was 24.4% for the cases versus 23.6% for the controls (p = 0.47). The mean Tac C0 was comparable for the cases (5.8 ng/mL) and control patients (6.1 ng/mL, p = 0.08). Only in the c-aABMR group a significant decline in both mean Tac C0 and allograft function over the timespan of 3 years was observed (p = 0.03 and p<0.001). Additionally, in the group of c-aABMR patients a high IPV was associated with inferior graft survival (p = 0.03). Conclusions A high Tac IPV per se does not predispose to the development of c-aABMR but is associated with inferior graft survival once c-aABMR is diagnosed

Immunosuppression Has Long-Lasting Effects on Circulating Follicular Regulatory T Cells in Kidney Transplant Recipients

Background: FoxP3+ follicular regulatory T cells (Tfr) have been identified as the cell population controlling T follicular helper (Tfh) cells and B cells which, are both involved in effector immune responses against transplanted tissue. Methods: To understand the biology of Tfr cells in kidney transplant patients treated with tacrolimus and mycophenolate mofetil (MMF) combination immunosuppression, we measured circulating (c)Tfh and cTfr cells in peripheral blood by flow cytometry in n = 211 kidney transplant recipients. At the time of measurement patients were 5–7 years after transplantation. Of this cohort of patients, 23.2% (49/211) had been previously treated for rejection. Median time after anti-rejection therapy was 4.9 years (range 0.4–7 years). Age and gender matched healthy individuals served as controls. Results: While the absolute numbers of cTfh cells were comparable between kidney transplant recipients and healthy controls, the numbers of cTfr cells were 46% lower in immunosuppressed recipients (p < 0.001). More importantly, in transplanted patients, the ratio of cTfr to cTfh was decreased (median; 0.10 vs. 0.06), indicating a disruption of the balance between

Raman Scattered He II λ\lambda 6545 Line in the Symbiotic Star V1016 Cygni

We present a spectrum of the symbiotic star V1016 Cyg observed with the 3.6 m Canada-France-Hawaii Telescope, in order to illustrate a method to measure the covering factor of the neutral scattering region around the giant component with respect to the hot emission region around the white dwarf component. In the spectrum, we find broad wings around H$\alpha$ and a broad emission feature around 6545${\rm \AA}$ that is blended with the [N II]$\lambda$ 6548 line. These two features are proposed to be formed by Raman scattering by atomic hydrogen, where the incident radiation is proposed to be UV continuum radiation around Ly$\beta$ in the former case and He II $\lambda$ 1025 emission line arising from $n=6\to n=2$ transitions for the latter feature. We remove the H$\alpha$ wings by a template Raman scattering wing profile and subtract the [N II] $\lambda$ 6548 line using the 3 times stronger [N II] $\lambda$ 6583 feature in order to isolate the He II Raman scattered 6545 \AA line. We obtain the flux ratio $F_{6545}/F_{6560}=0.24$ of the He II $\lambda$ 6560 emission line and the 6545 \AA feature for V1016 Cyg. Under the assumption that the He II emission from this object is isotropic, this ratio is converted to the ratio $\Phi_{6545}/\Phi_{1025}=0.17$ of the number of the incident photons and that of the scattered photons. This implies that the scattering region with H I column density $N_{HI}\ge 10^{20}{\rm cm^{-2}}$ covers 17 per cent of the emission region. By combining the presumed binary period $\sim 100$ yrs of this system we infer that a significant fraction of the slow stellar wind from the Mira component is ionized and that the scattering region around the Mira extends a few tens of AU, which is closely associated with the mass loss process of the Mira component.Comment: 12 pages, 6 figures, accepted for publication in Ap

Multiwavelength Monitoring of the Narrow-Line Seyfert 1 Galaxy Akn 564. II. Ultraviolet Continuum and Emission-line Variability

We present results of an intensive two-month campaign of approximately daily spectrophotometric monitoring of the narrow-line Seyfert 1 galaxy Akn 564 with HST. The fractional variability amplitude of the continuum variations between 1365-3000 A is ~6%, about a factor 3 less than that found in typical Seyfert 1 galaxies over a similar period of time. However, large amplitude, short time-scale flaring behavior is evident, with trough-to-peak flux changes of about 18% in approximately 3 days. We present evidence for wavelength-dependent continuum time delays, with the variations at 3000 A lagging behind those at 1365 A by about 1 day. These delays may be interpreted as evidence for a stratified continuum reprocessing region, possibly an accretion-disk structure. The Lyman-alpha 1216 emission-line exhibits flux variations of about 1% amplitude.Comment: 27 pages, 14 figures. Accepted by Astrophysical Journa

Level of Digitization in Dutch Hospitals and the Lengths of Stay of Patients with Colorectal Cancer

A substantial amount of research has been published on the association between the use of electronic medical records (EMRs) and quality outcomes in U.S. hospitals, while limited research has focused on the Western European experience. The purpose of this study is to explore the association between the use of EMR technologies in Dutch hospitals and length of stay after colorectal cancer surgery. Two data sets were leveraged for this study; the HIMSS Analytics Electronic Medical Record AdoptionModel (EMRAMSM) and the Dutch surgical colorectal audit (DSCA). The HIMSS Analytics EMRAM score was used to define a Dutch hospital's electronic medical records (EMR) capabilities while the DSCA was used to profile colorectal surgery quality outcomes (specifically total length of stay (LOS) in the hospital and the LOS in ICU). A total of 73 hospitals with a valid EMRAM score and associated DSCA patients (n = 30.358) during the study period (2012-2014) were included in the comparative set. A multivariate regression method was used to test differences adjusted for case mix, year of surgery, surgical technique and for complications, as well as stratifying for academic affiliated hospitals and general hospitals. A significant negative association was observed to exist between the total LOS (relative median LOS 0,974, CI 95% 0.959-0,989) of patients treated in advanced EMR hospitals (high EMRAM score cohort) versus patients treated at less advanced EMR care settings, once the data was adjusted for the case mix, year of surgery and type of surgery (laparoscopy or laparotomy). Adjusting for complications in a subgroup of general hospitals (n = 39) yielded essentially the same results (relative median LOS 0,934, CI 95% 0,9150,954). No consistent significant associations were found with respect to LOS on the ICU. The findings of this study suggest advanced EMR capabilities support a healthcare provider's efforts to achieve desired quality outcomes and efficiency in Western European hospitals

Alloreactive T cells to assess acute rejection risk in kidney transplant recipients

Background.Memory T cells are important mediators of transplant rejection but are not routinely measured before or after kidney transplantation. The aims of this study were as follows: (1) validate whether pretransplant donor-reactive memory T cells are reliable predictors of acute rejection (AR) (2) determine whether donor-reactive memory T cells can distinguish AR from other causes of transplant dysfunction. Methods.Samples from 103 consecutive kidney transplant recipients (2018-2019) were obtained pretransplantation and at time of for-cause biopsy sampling within 6 mo of transplantation. The number of donor-reactive interferon gamma (IFN-gamma) and interleukin (IL)-21-producing memory T cells was analyzed by enzyme-linked immunosorbent spot (ELISPOT) assay. Results.Of the 63 patients who underwent a biopsy, 25 had a biopsy-proven acute rejection (BPAR; 22 aTCMR and 3 aAMR), 19 had a presumed rejection, and 19 had no rejection. Receiver operating characteristic analysis showed that the pretransplant IFN-gamma ELISPOT assay distinguished between patients who later developed BPAR and patients who remained rejection-free (area under the curve [AUC] 0.73; sensitivity 96% and specificity 41%). Both the IFN-gamma and IL-21 assays were able to discriminate BPAR from other causes of transplant dysfunction (AUC 0.81; sensitivity 87% and specificity 76% and AUC 0.81; sensitivity 93% and specificity 68%, respectively). Conclusions.This study validates that a high number of donor-reactive memory T cells before transplantation is associated with the development of AR after transplantation. Furthermore, it demonstrates that the IFN-gamma and IL-21 ELISPOT assays are able to discriminate between patients with AR and patients without AR at the time of biopsy sampling.Personalised Therapeutic

Intraoperative inducibility of atrial fibrillation does not predict early postoperative atrial fibrillation

Background--Early postoperative atrial fibrillation (EPoAF) is associated with thromboembolic events, prolonged hospitalization, and development of late PoAF (LPoAF). It is, however, unknown if EPoAF can be predicted by intraoperative AF inducibility. The aims of this study are therefore to explore (1) the value of intraoperative inducibility of AF for development of both EPoAF and LPoAF and (2) the predictive value of de novo EPoAF for recurrence of LPoAF. Methods and Results--Patients (N=496, 75% male) undergoing cardiothoracic surgery for coronary and/or valvular heart disease were included. AF induction was attempted by atrial pacing, before extracorporeal circulation. All patients were on continuous rhythm monitoring until discharge to detect EPoAF. During a follow-up period of 2 years, LPoAF was detected by ECGs and Holter recordings. Sustained AF was inducible in 56% of patients. There was no difference in patients with or without AF before surgery (P=0.159), or between different types of surgery (P=0.687). In patients without a history of AF, incidence of EPoAF and LPoAF was 37% and 2%, respectively. EPoAF recurred in 58% patients with preoperative AF, 53% developed LPoAF. There were no correlations between intraoperative inducibility and EPoAF or LPoAF (P > 0.05). EPoAF was not correlated with LPoAF in patients without a history of AF (P=0.116), in contrast to patients with AF before surgery (P < 0.001). Conclusions--Intraoperative AF inducibility does not predict development of either EPoAF or LPoAF. In patients with AF before surgery, EPoAF is correlated with LPoAF recurrences. This correlation is absent in patients without AF before surgery