ANALYSIS OF SERUM PROTEINS IN HEALTHY GIANT PANDAS (AILUROPODA MELANOLEUCA) UNDER MANAGED CARE.

Electrophoresis is a practical diagnostic tool for the identification of changes in serum protein fractions, which can be associated with a variety of diseases. Protein electrophoresis studies in Ursidae are limited, and currently no published fraction values are available for the giant panda (Ailuropoda melanoleuca). The aim of this study was to describe the serum protein fractions in the giant panda using both capillary zone electrophoresis (CZE) and standard agarose gel electrophoresis (AGE) techniques. Serum samples from nine healthy giant pandas (n = 19) were used for this study. Samples were evaluated using CZE and standard AGE. The CZE procedure successfully resolved serum proteins into seven fractions: prealbumin; albumin; and α1-, α2-, ß1-, ß2-, and γ-globulin; while AGE separated serum into only six protein fractions: prealbumin; albumin; α1-, α2-, and ß-globulins; and γ-globulin. These data will serve as a preliminary baseline for further studies and provide insight for the medical management of giant pandas.

Polyclonal hypergammaglobulinaemia: assessment, clinical interpretation, and management.

This Review outlines a practical approach to assessing and managing polyclonal hypergammaglobulinaemia in adults. Polyclonal hypergammaglobulinaemia is most commonly caused by liver disease, immune dysregulation, or inflammation, but can also provide an important diagnostic clue of rare diseases such as histiocyte disorders, autoimmune lymphoproliferative syndrome, Castleman disease, and IgG4-related disease. Causes of polyclonal hypergammaglobulinaemia can be divided into eight categories: liver disease, autoimmune disease and vasculitis, infection and inflammation, non-haematological malignancy, haematological disorders, IgG4-related disease, immunodeficiency syndromes, and iatrogenic (from immunoglobulin therapy). Measuring serum concentrations of C-reactive protein and IgG subclasses are helpful in diagnosis. IL-6-mediated inflammation, associated with persistently elevated C-reactive protein concentrations (≥30 mg/L), is an important driver of polyclonal hypergammaglobulinaemia in some cases. Although the presence of markedly elevated serum IgG4 concentrations (>5 g/L) is around 90% specific for diagnosing IgG4-related disease, mildly elevated serum IgG4 concentrations are seen in many conditions. In most cases, managing polyclonal hypergammaglobulinaemia simply involves treating the underlying condition. Rarely, however, polyclonal hypergammaglobulinaemia can lead to hyperviscosity, requiring plasmapheresis.

Evaluation of models for predicting pediatric fraction unbound in plasma for human health risk assessment.

Pediatric physiologically based pharmacokinetic (PBPK) models facilitate the prediction of PK parameters in children under specific exposure conditions. Pharmacokinetic outcomes are highly sensitive to fraction unbound in plasma (fup) as incorporated into PBPK models. Rarely is fup in children (fupchild) experimentally derived and prediction is based upon fup in adults (fupadult) as well as a ratio of plasma protein concentrations between children and adults. The objectives were to (i) evaluate protein concentration vs. age profile derived from ontogeny models, (ii) assess predictive performances of fup ontogeny models, and (iii) determine overall uncertainty in fupchild prediction resulting from a combination of quantitative structure-property relationship (QSPR) model and ontogeny models. The plasma albumin and alpha-acid glycoprotein (AAG) concentration data for pediatrics and fupchild and fupadult data were obtained from literature. The protein concentration vs. age profile derived from ontogeny models were compared to observed levels. Fupchild values were calculated according to ontogeny models using both observed and QSPR-predicted fupadult as inputs and predictive performances of ontogeny models assessed by comparing predicted fupchild to observed values. Protein concentrations vs. age profiles derived from non-linear equations were more congruent with observed albumin levels than linear or step-wise models. When observed fupadult values were used as input, the fupchild data were under-predicted with average fold error (AFE) amounts ranging 0.79-0.81 and 0.77-0.97 for albumin and AAG ontogeny models, respectively. When QSPR-predicted fupadult values were used as input, AFE of fupchild ranged 1.2-1.35 and 0.98-1.2 for albumin and AAG models, respectively. The choice of ontogeny model with respect to prediction accuracy is more important for AAG, highly bound compounds and infants. For these compounds and scenarios, experimental determination of fupchild for inclusion into a pediatric PBPK model is necessary to have confidence in PBPK model outputs.

Productivity of steers of different genotypes: forecast based on interior indicators / [Produtividade de novilhos de diferentes genótipos: previsão com base em indicadores internos]

Meat productivity and quality of beef are determined by a number of factors, including pedigree traits of animals. Meat productivity is closely related to the biological patterns of their growth and development. Considering the patterns that affect meat productivity enables effective growing and fattening of livestock and obtaining commercially viable beef. To predict economically useful traits in beef cattle breeding, interior indicators can be used, as they reflect the metabolic picture of the animal's body. The research studies in physiology and biochemistry of livestock aimed at revealing the persistent mechanisms of a growing animal organism make them relevant. The article identifies a correlation between the interior indicators and the fattening indicators of three experimental groups of steers. The main forecasting factors of meat productivity indicators have been substantiated. Regression coefficients have been found and show how much the live weight varies depending on the determining factors. Meat productivity predicting procedures have been modeled with respect to the protein content in blood serum.(AU)

A produtividade e a qualidade da carne bovina são determinadas por diversos fatores, incluindo características de pedigree dos animais. A produtividade da carne tem relação íntima com os padrões biológicos de seu crescimento e desenvolvimento. A consideração dos padrões que afetam a produtividade da carne possibilita o crescimento e engorda eficazes dos rebanhos e a obtenção de carne bovina comercialmente viável. Para prever características economicamente úteis na criação de gado de corte, indicadores de interior podem ser usados, visto que refletem a imagem metabólica do corpo do animal. Tornam-se relevantes os estudos de pesquisa em fisiologia e bioquímica da pecuária com o objetivo de revelar os mecanismos persistentes de um organismo animal em crescimento. O artigo identifica uma correlação entre os indicadores internos e os indicadores de engorda de três grupos experimentais de novilhos. Os principais fatores de previsão dos indicadores de produtividade de carnes já foram comprovados. Coeficientes de regressão foram encontrados e mostram o quanto o peso vivo varia em função dos fatores determinantes. Os procedimentos de predição da produtividade da carne foram modelados em relação ao conteúdo de proteína no soro sanguíneo.(AU)

Mid-term response assessment in multiple myeloma using a texture analysis approach on dual energy-CT-derived bone marrow images - A proof of principle study.

PURPOSE: To identify textural features on dual-energy CT (DECT)-generated virtual non calcium (VNC) bone marrow images in a small group of patients with multiple myeloma undergoing systemic treatment which could potentially help for mid-term response assessment. METHODS: 44 patients (59.1 ±â€¯11.2 yr.) with multiple myeloma who underwent unenhanced whole-body reduced-dose DECT before and after systemic therapy were evaluated. All patients had current hematologic laboratory tests including serum levels of immunoglobulins, albumin, and total proteins. Using DECT post-processing, bone marrow images of the axial skeleton were reconstructed. The vertebral bodies T10-L5 were segmented for quantification of 1st order (n = 18) and 2nd order Gray Level Co-occurrence Matrix (GLCM) textural features (n = 23) based on an open-source radiomics library (Pyradiomics), which were then compared with the hematologic response category to treatment. Five patients underwent only active surveillance at intervals after previous successful therapy. RESULTS: According to hematologic diagnosis, 29 patients were classified as complete response (CR), 10 as partial response (PR) and 5 as stable disease (SD). We observed a significant drop of the 1st order textural features "10th percentile" (p = 0.009), "median" (p = 0.01), and "minimum" (p < 0.0001) after treatment, whereas the 1st order feature "range" (p = 0.0004) and the 2nd order GLCM feature "difference variance" (p = 0.007) significantly increased in patients experiencing CR. A similar trend, however, without statistical significance, could be observed in patients achieving PR after treatment. 2nd order GLCM feature "difference variance" proved to be a significant discriminator (p = 0.01) between patients with CR and PR (sensitivity 0.93, specificity 0.70) for a cut-off value of -0.28. In patients classified CR, both the mean serum protein and the beta-2 microglobulin decreased after treatment, whereas the serum albumin increased (p < 0.01). The same trend without significance could be observed in patients classified PR. CONCLUSIONS: Changes in textural features applied on VNC bone marrow images in the pre- and posttreatment settings correlate well with myeloma-specific hematologic parameters and provide complementary information for the assessment of the late effects of treatment on the bone marrow.

Multiplex profiling of serum proteins in solution using barcoded antibody fragments and next generation sequencing.

The composition of serum proteins is reflecting the current health status and can, with the right tools, be used to detect early signs of disease, such as an emerging cancer. An earlier diagnosis of cancer would greatly increase the chance of an improved outcome for the patients. However, there is still an unmet need for proficient tools to decipher the information in the blood proteome, which calls for further technological development. Here, we present a proof-of-concept study that demonstrates an alternative approach for multiplexed protein profiling of serum samples in solution, using DNA barcoded scFv antibody fragments and next generation sequencing. The outcome shows high accuracy when discriminating samples derived from pancreatic cancer patients and healthy controls and represents a scalable alternative for serum analysis.

Proteome-wide assessment of diabetes mellitus in Qatari identifies IGFBP-2 as a risk factor already with early glycaemic disturbances.

BACKGROUND: Proteomics is expected to provide novel insights in the underlying pathophysiology of type 2 diabetes mellitus. In the present study, we aimed to identify and biochemically characterize proteins associated with diabetes mellitus in a Qatari population. METHODS: In a diabetes case-control study (175 cases, 164 controls; Arab, South Asian and Philippine ethnicities), we conducted a discovery study to screen 1141 blood protein levels for associations with diabetes mellitus. Additional analyses were done in controls in relation to Hb1Ac, and biochemical characterization of the main findings was performed with metabolomics (501 metabolites). We performed two-sample Mendelian Randomization to provide evidence of potential causality using data from European descent of the DIAGRAM consortium (74,124 cases of diabetes mellitus and 824,006 controls) for the identified proteins for T2D and Hb1Ac. RESULTS: After accounting for multiple testing, 30 protein levels were different (p-values<8.6e-5) between cases and controls. Of these, a higher Hb1Ac in controls was associated with a lower IGFBP-2 level (p-value = 4.1e-6). IGFBP-2 protein level was found lower among cases compared with controls across all ethnicities. In controls, IGFBP-2 was associated with 21 metabolite levels, but specifically connected to the metabolite citrulline in network analyses. We observed no evidence, however, that the association between IGFBP-2 and diabetes mellitus was causal. CONCLUSIONS: We specifically identified IGFBP-2 to be associated with diabetes mellitus, although with no evidence for causality, which was specifically connected to citrulline metabolism.

Assessment of Serological Early Biomarker Candidates for Lung Adenocarcinoma by using Multiple Reaction Monitoring-Mass Spectrometry.

PURPOSE: Plasma markers that enable diagnosis in the early stage of lung cancer is not discovered. A liquid chromatography multiple reaction monitoring-mass spectrometry (LC-MRM-MS) assay for identifying potential early marker proteins for lung adenocarcinoma is developed. EXPERIMENTAL DESIGN: LC-MRM-MS assay is used for measuring the level of 35 candidate peptides in plasma from 102 lung adenocarcinoma patients (including n = 50, 16, 24, and 12 in stage I, II, III, and IV, respectively.) and 84 healthy controls. Stable isotope labeled standard peptides are synthesized to accurately measure the amount of these proteins. RESULTS: Seven proteins are able to distinguish stage I patients from controls. These proteins are combined in to a protein marker panel which improve the sensitivity to discriminate stage I patients from controls with cross-validated area under the curve = 0.76. Besides, it is found that low expression of eukaryotic initiation factor 4A-I and high expression of lumican show significantly poor prognosis in overall survival (p = 0.012 and 0.0074, respectively), which may be used as prognostic biomarkers for lung cancer. CONCLUSIONS AND CLINICAL RELEVANCE: Proteins highlighted here may be used for early detection of lung adenocarcinoma or therapeutics development after validation in a larger cohort.

Systematic Assessment of the Effect of Internal Library in Targeted Analysis of SWATH-MS.

Targeted analysis of sequential window acquisition of all theoretical mass spectra (SWATH-MS) requires the spectral library, which can be generated by shotgun mass spectrometry (MS) or by the pseudo-spectra files directly obtained from SWATH-MS data. The external library generated by shotgun MS is employed in most SWATH-MS research. However, performance of the internal library, which is constructed by pseudo-spectra files, in the targeted analysis of SWATH-MS has not been systemically evaluated. Here, we show that up to 40% of the peptides detected by the internal library were not overlapped with those detected by the external library for most SWATH-MS data sets. However, the internal library did not identify extra phosphopeptides compared with the external library for phosphoproteomic SWATH-MS data. Therefore, the internal library should be incorporated into the external library for targeted analysis of nonphosphoproteomic SWATH-MS, given that it can significantly increase the number of peptides of SWATH-MS without requiring additional instrument measurement time.

Assessment of Serum Concentrations of Adropin, Afamin, and Neudesin in Children with Type 1 Diabetes.

INTRODUCTION: The increasing knowledge of adropin, afamin, and neudesin and the regulation of glucose metabolism and insulin resistance allows for the assessment of the differences in their concentrations between the groups with varied duration of diabetes mellitus (DM). AIM OF THE STUDY: Assessment of serum levels of adropin, afamin, and neudesin in children with type 1 diabetes, with respect to the disease duration. MATERIALS AND METHODS: The study consisted of 138 patients aged 5-18 years (M 40.58%). Children with type 1 diabetes (n = 68) were compared to the control group (n = 70). The diabetic group was divided into 4 subgroups: (I) newly diagnosed patients, after an episode of ketoacidosis (n = 14), (II) duration no longer than 5 years (n = 18), (III) 5 to 10 years (n = 27), and (IV) longer than 10 years (n = 9). Serum concentrations of adropin, afamin, and neudesin were assessed and compared between the groups of patients. The criterion for statistical significance was p < 0.05. RESULTS: The concentrations of adropin and afamin across all subgroups were lower than that in the control group, while neudesin levels were higher in diabetic patients compared to the control group. The differences were statistically significant. CONCLUSIONS: Adropin, afamin, and neudesin may play a major role in the regulation of glucose metabolism and have a significant potential as novel biomarkers to predict future metabolic disorders. However, further multicentre studies on a larger cohort of patients are necessary to specify the role of these substances in the course and treatment of type 1 diabetes.

Association of Extracellular Vesicle Protein Cargo with Race and Clinical Markers of Mortality.

Differential mortality rates remain a significant health disparity in the United States, suggesting the need to investigate novel potential molecular markers associated with mortality. Extracellular vesicles (EVs), including exosomes, microvesicles and apoptotic bodies, are lipid-bound vesicles secreted by cells into the circulation. EVs mediate intercellular communication by shuttling functional signaling molecules as cargo. EV characteristics by race in the context of mortality risk factors have not been described. We isolated plasma EVs from a cross-sectional cohort of African Americans (AA) and whites and found no significant differences in EV size, distribution or concentration between race or by sex. However, EV cargo showed increased levels of phospho-p53, total p53, cleaved caspase 3, ERK1/2 and phospho-AKT in white individuals compared to AAs. phospho-IGF-1R levels were significantly higher in females compared to males. EV concentration was significantly associated with several clinical mortality risk factors: high-sensitivity C-reactive protein (hsCRP), homeostatic model assessment of insulin resistance (HOMA-IR), alkaline phosphatase, body mass index, waist circumference and pulse pressure. The association of EV proteins with mortality markers were dependent on race. These data suggest that EV cargo can differ by race and sex and is associated with mortality risk factors.

A rapid UHPLC-MS/MS screening method for the detection of the addition of porcine blood plasma to emulsion-type pork sausages.

The adulteration of meat products by the undeclared addition of commercially available blood plasma powder is quite conceivable due to low costs, high protein contents (about 70%), and advantageous functional properties. This applies particularly to pork, which has the highest meat production rate in the European Union. Evidence of this type of food fraud has been rather difficult to identify due to the lack of appropriate analytical methods, especially when adding plasma to meat of the same animal species. Consequently, a rapid UHPLC-MS/MS method for the detection of porcine blood plasma in emulsion-type pork sausages was developed. After protein extraction and tryptic digestion in a quick and simple one-pot process, species-specific marker peptides for porcine blood cell proteins (four markers) and plasma proteins (12 markers) were measured by UHPLC-MS/MS. Emulsion-type pork sausages were produced from a variety of raw materials that differed in the age or sex of the slaughtered pigs. Sausages were spiked with 0.5, 1, 1.5, 2, 3, or 5% meat substitution by one of two plasma powders, or produced as corresponding blank samples, and subjected to different thermal treatments as full or semi-preserves. Four plasma peptides were identified for the overall sample that allowed detection down to 0.7% meat substitution from the sum of their peak areas, with 5% error probability for both false positives and negatives.

Systematic assessment of antibody selectivity in plasma based on a resource of enrichment profiles.

There is a strong need for procedures that enable context and application dependent validation of antibodies. Here, we applied a magnetic bead assisted workflow and immunoprecipitation mass spectrometry (IP-MS/MS) to assess antibody selectivity for the detection of proteins in human plasma. A resource was built on 414 IP experiments using 157 antibodies (targeting 120 unique proteins) in assays with heat-treated or untreated EDTA plasma. For each protein we determined their antibody related degrees of enrichment using z-scores and their frequencies of identification across all IP assays. Out of 1,313 unique endogenous proteins, 426 proteins (33%) were detected in >20% of IPs, and these background components were mainly comprised of proteins from the complement system. For 45% (70/157) of the tested antibodies, the expected target proteins were enriched (z-score ≥ 3). Among these 70 antibodies, 59 (84%) co-enriched other proteins beside the intended target and mainly due to sequence homology or protein abundance. We also detected protein interactions in plasma, and for IGFBP2 confirmed these using several antibodies and sandwich immunoassays. The protein enrichment data with plasma provide a very useful and yet lacking resource for the assessment of antibody selectivity. Our insights will contribute to a more informed use of affinity reagents for plasma proteomics assays.

[Determination of the immunoglobulin G supply in the newborn calf]. / Bestimmung der Immunglobulin-G-Versorgung beim neugeborenen Kalb.

A sufficient supply of colostral antibodies within the first hours of life is crucial for the development and the health status in young calves. It is rational to examine the immunoglobulin uptake of single animals, but particularly on a herd basis, during herd controls and consultations. This enables economical calf rearing in accordance with animal welfare. Because of the costly, laboratory-dependent and in part time-consuming direct measurement of the absorbed immunoglobulins using radial immunodiffusion (RID) or ELISA, multiple studies attempted to develop indirect methods, which would be affordable and operational in the field. These aim to draw an inference for the absorbed quantity of colostral antibodies based on other correlated parameters. Multiple validations showed in part significant differences between various methods concerning specificity and sensitivity in comparison to the direct methods. In addition to RID and ELISA, this article presents the measurement of the γ-glutamyltransferase (GGT) activity, the determination of the total serum protein concentration using refractometry and the zinc sulphate turbidity test, and describes the advantages and disadvantages of their application. Refractory measurement and determination of the GGT activity represent a valuable alternative to a laboratory-dependent immunoglobulin G measurement. Nevertheless, there is no ideal rapid test method, such that several influencing factors have to be considered.

[Predictive value of heparin-binding protein combined with sequential organ failure assessment score in patients with septic shock].

OBJECTIVE: To explore the predictive value of heparin-binding protein (HBP) combined with sequential organ failure assessment (SOFA) score in patients with septic shock. METHODS: Seventy-eight patients with sepsis admitted to intensive care unit (ICU) of Henan Provincial People's Hospital from December 2016 to May 2017 were enrolled. Thirty healthy persons were enrolled as controls. The patient's gender, age, length of ICU stay, and blood culture results, white blood cell count (WBC), C-reactive protein (CRP), procalcitonin (PCT), blood lactate (Lac), HBP, SOFA score, acute physiology and chronic health evaluation II (APACHE II) score, organ failure and vasoactive agents usage within 24 hours of admission were recorded. The differences in the above indicators between the groups were compared, and the receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of HBP, SOFA score and their combination in patients with septic shock. RESULTS: All patients were enrolled in the final analysis, including 64 with sepsis and 14 with septic shock. Compared with the sepsis group, the proportion of patients with septic shock who were positive for blood culture, organ failure, and vasoactive agents was higher [57.1% (8/14) vs. 7.8% (5/64), 100.0% (14/14) vs. 65.6% (42/64), 100.0% (14/14) vs. 18.8% (12/64), all P < 0.01], SOFA and APACHE II scores were also higher (SOFA: 8.93±4.16 vs. 5.89±2.68, APACHE II: 22.29±4.89 vs. 15.28±5.14, both P < 0.01); however, there was no significant difference in gender, age or length of ICU stay between the two groups. Compared with the healthy control group, HBP, PCT, CRP and Lac levels were significantly increased in the sepsis group and the septic shock group. HBP in the septic shock group was significantly higher than that in the sepsis group (µg/L: 120.33±43.49 vs. 68.95±54.15, P < 0.05), but there was no significant difference in PCT, CRP or Lac between septic shock group and sepsis group [PCT (µg/L): 1.42 (0.47, 46.00) vs. 0.71 (0.19, 4.50), CRP (mg/L): 102.90±78.12 vs. 102.07±72.15, Lac (mmol/L): 1.81 (1.14, 3.65) vs. 1.59 (1.17, 2.24), all P > 0.05]. It was shown by ROC curve analysis that the area under the ROC curve (AUC) of SOFA score for predicting septic shock was 0.715 [95% confidence interval (95%CI) = 0.540-0.890, P = 0.012], and when the optimal cut-off value was 7.5, the sensitivity was 64.3%, the specificity was 76.6%. The AUC of HBP was 0.814 (95%CI = 0.714-0.913, P < 0.001), and when the optimal cut-off value was 89.43 µg/L, the sensitivity was 78.6%, the specificity was 76.6%; when the two were combined, the AUC was 0.829 (95%CI = 0.724-0.935, P < 0.001), the sensitivity was 92.9%, and the specificity was 61.9%. CONCLUSIONS: HBP can be used as a biological indicator for predicting septic shock, and the accuracy of predicting septic shock can be improved with the combination of SOFA score.

Laboratory assessment of multiple myeloma.

Laboratory testing plays an essential role in the diagnosis and management of patients with multiple myeloma. A variety of chemistry and molecular assays are routinely used to monitor patient progress, response to treatment and relapse. Here, we have reviewed current literature and core guidelines on the details of laboratory testing in myeloma-related investigations. This includes the use and value of protein electrophoresis, serum free light chain and cytogenetic testing. Furthermore, we discuss other traditional chemistry assays essential to myeloma investigation, and potential interferences that may arise due to the disease nature of myeloma, that is, the presence of a monoclonal immunoglobulin. Finally, we discuss the importance of communication in protein electrophoresis results, where laboratorians are required to relate clinically relevant myeloma-relevant information to the ordering physician on the background of a complex pattern of serum or urine proteins. Laboratory testing in myeloma-related investigation relies on several traditional chemistry assays. However, we anticipate new tests and technologies to become available in the future with improved analytical sensitivity, as well as improved clinical sensitivity in identifying patients who are at high risk of progression to multiple myeloma.

Immunoelectrophoresis: A Method with Many Faces.

Immunoelectrophoresis (IEP) was the first practical method that combined electrophoresis and immunoprecipitation for identifying and characterizing proteins within complex mixtures. Over the years, IEP has been extended to include a variety of techniques and, as a general name, has been applied to virtually any technique that involves electrophoresis and antigen-antibody precipitin reaction for proteins. Because of the diversity in technical details of different IEP versions, the method described here deals only with classic IEP. Although it requires some manual expertise, IEP is versatile, relatively easy to customize, and economical with no need for expensive instrumentation. Further, it can discern identity, partial identity, and nonidentity of the proteins. Any low-viscosity body fluid specimen or, possibly, culture fluid and tissue extract could be tested with IEP if proper antibodies are available. With these attributes, classic IEP remains a valuable tool for clinical diagnostic testing, purity checking of biochemical and pharmaceutical products, and research.

Critical assessment of different methods for quantitative measurement of metallodrug-protein associations.

Quantitative screening for potential drug-protein binding is an essential step in developing novel metal-based anticancer drugs. ICP-MS approaches are at the core of this task; however, many applications lack in the capability of large-scale high-throughput screenings and proper validation. In this work, we critically discuss the analytical figures of merit and the potential method-based quantitative differences applying four different ICP-MS strategies to ex vivo drug-serum incubations. Two candidate drugs, more specifically, two Pt(IV) complexes with known differences of binding affinity towards serum proteins were selected. The study integrated centrifugal ultrafiltration followed by flow injection analysis, turbulent flow chromatography (TFC), and size exclusion chromatography (SEC), all combined with inductively coupled plasma-mass spectrometry (ICP-MS). As a novelty, for the first time, UHPLC SEC-ICP-MS was implemented to enable rapid protein separation to be performed within a few minutes at > 90% column recovery for protein adducts and small molecules. Graphical abstract Quantitative screening for potential drug-protein binding is an essential step in developingnovel metal-based anticancer drugs.

Health status assessment of traumatic injury freshwater turtles.

A group of injured yellow-bellied sliders (Trachemys scripta) and river cooters (Pseudemys concinna) were evaluated for a variety of health values at presentation to the NC State Turtle Rescue Team and prior to release. An i-STAT Portable Clinical Analyzer and CG8+ cartridges were used to determine venous blood gas and biochemical values, the packed cell volume (PCV) and total protein were evaluated using hematocrit tubes and high speed centrifugation, and a differential WBC percentage was determined manually with Diff-Quick stained blood smear slides. Forty-six turtles were sampled on presentation and twenty-three of those were sampled again prior to release. Blood values were analyzed for significant differences between samples collected at presentation and prior to release, as well as differences between surviving and non-surviving turtles. Five variables were identified as significantly different between presenting and recuperated samples: pH, pCO2, Glu, % heterophils, and % eosinophils. When comparing samples between turtles that survived versus those that did not, two variables were identified as being significant prognostic indicators; lactate and PCV. Identification of these significant variables can aid in determining patient prognosis and triage therapy for injured aquatic turtles.