Obtained Blood Samples Research Articles

94-LB: Measurement Accuracy of a Newly Developed Prototype System for Noninvasive Glucose Monitoring

Non-invasive glucose monitoring (NIGM) may be beneficial for people with diabetes in avoiding the need for finger pricking to obtain blood samples. The aim was to assess measurement accuracy of a prototype system for NIGM in a mixed outpatient and in-clinic setting. A total of 10 subjects with type 1 diabetes participated in the study which lasted for 27 days per subject. Subjects wore a FreeStyle Libre (FL) system on the upper arm for continuous glucose monitoring, and they performed standard blood glucose (BG) monitoring with a Contour® next ONE meter and NIGM at the thenar with the prototype system. At least 6 times per day, measurement sets were performed which consisted of 1st BG meter measurement, 1st FL scan, 1st NIGM measurement, 2nd BG meter measurement, 2nd FL scan, 2nd NIGM measurement. Results from the same measurement set were averaged by device before analysis. In-clinic sessions took place on the 6th day and on the 27th day, during which rapid glucose excursions with high and low glucose values were induced. During these sessions, measurement sets were performed every 15 minutes between 0730 and 1500. Data from 24/19 days were used for calibration of the NIGM system for the first/second group of five subjects. The data from the remaining 3/8 days (including 1 in-clinic day each) were used for independent validation of the calibration. For data from validation days, mean absolute relative differences (MARD) were calculated for FL and the NIGM prototype in comparison with the BG meter values for each subject. Outpatient MARD ranged from 8.0% to 26.2% for FL (mean: 16.1%) and from 9.1% to 26.9% for the prototype (mean: 20.2%), whereas in-clinic MARD ranged from 13.7% to 45.3% for FL (mean 23.3%) and from 19.8% to 36.0% for the prototype (mean: 25.5%). Although MARD was somewhat higher for the NIGM prototype than for FL, this proof-of-concept study showed promising results for the newly developed NIGM prototype. Disclosure G. Freckmann: Advisory Panel; Self; Abbott, Novo Nordisk Pharma GmbH, Sensile Medical AG. Consultant; Self; Metronom Health Europe, Roche Diabetes Care, Sanofi. Speaker’s Bureau; Self; Ascensia Diabetes Care, Dexcom, Inc., Ypsomed AG. S. Pleus: None. P. Wintergerst: None. D. Waldenmaier: None. N.R. Jendrike: None. M. Link: None. C. Haug: None. Funding RSP Systems A/S

Relation Between Genetic Factors and Frailty in Older Adults

ObjectivesFrailty is a geriatric syndrome that identifies individuals at higher risk of disability, institutionalization, and death. We previously reported that frailty is related to oxidative stress and cognitive impairment-related biomarkers. The aim of this study was to determine whether frailty is associated with genetic variants. DesignLongitudinal population-based cohort of 2488 community-dwelling people from Toledo, Spain, aged 65 years or older. Setting and participantsWe obtained blood samples from 78 individuals with frailty and 74 nonfrail individuals who were nonfrail (according to Fried criteria) from the Toledo Study of Healthy Ageing and extracted DNA using the Chemagic DNA blood kit. MeasuresSample genotyping was carried out by means of Axiom Exome 319 Genotyping Array (Thermo Fisher Scientific), which contains 295,988 markers [single-nucleotide polymorphisms (SNPs) and rare variants], and transferred to the GeneTitan Instrument (Affymetrix). ResultsWe found 15 SNPs (P < .001), 18 genes (P < .005), and 4 pathways (P < .05) related to cytokine-cytokine receptor interaction, natural killer cell-mediated cytotoxicity, regulation of autophagy, and renin-angiotensin system as the most strongly associated with frailty. Conclusions/ImplicationsThe specific genetic features related to energy metabolism, biological processes regulation, cognition, and inflammation highlighted by this preliminary analysis offer useful insights for finding biologically meaningful biomarkers of frailty that allow early diagnosis and treatment. Further research is needed to confirm our novel findings in a larger population. Indeed, the EU-funded FRAILOMICS research effort will address this question.

SUN-117 Neuroendocrine Mechanisms Responsible for the Hydromineral Control in Rats with Obesity Induced by High Fat Diet (HFD)

Besides being studied regarding cardiovascular diseases and metabolic disturbances, little is known about how obesity might modulate the hydroeletroctrolytic balance. Some studies suggest that such modulation might exist. This current study has the objective of clarify the neuroendocrine mechanisms of salt and water homeostasis in obese rats under normal and osmotic challenge conditions. Wistar male rats weighting 100 grams were used; they were divided into two groups, one submitted to high fat diet (HFD) and the other submitted to normal diet, both for 9 weeks. After that they were kept in a metabolic cage for three days in order to measure food, water and sodium intake and urinary volume. After 24 or 48 hour fluid deprivation, they were given back water, in the first hour, and water and saline (1,8% NaCl), in the second hour, determining the ingestion of both fluids. One week later, the rats were submitted again to 24 or 48 hour fluid deprivation and then euthanized to obtain blood samples and tissues. During the basal period, the obese rats drank less water and ate less food than the non-obese rats. After the 24 or 48 hour dehydration, the obese rats drank less water than the non-obese. Besides that, the dehydrated obese rats also had a reduced hematocrit, reduced plasma osmolality, and reduced levels of K⁺ in plasma, a tendency to bigger levels of Na⁺ in plasma when compared to dehydrated non-obese rats. It was possible to observe an adipose tissue mass reduction in obese dehydrated rats when compared to obese hydrated rats. Accordingly, we were able to see that obesity indeed changes the hydroelectrolytic balance and there is the hypothesis that obese rats might use fat as a water reserve through of beta oxidation of lipids, which is an issue still under investigation. Sources of Research Support: Sao Paulo Research Foudation (FAPESP) Grant #2013/09799-1; Fapesp Grant #2018/10920-3.

Time-dependent Pathologic and Inflammatory Consequences of Various Blood Sampling Techniques in Mice.

We compared 6 frequently used mouse blood-sampling methods (lateral tail incision; tail-tip amputation; sublingual, submandibular, and saphenous vein puncture; and retrobulbar sinus puncture during isoflurane anesthesia) with regard to induction of local and systemic inflammation, stomach contents, weight changes, and corticosterone levels at 6 h to 12 d after sampling. Local inflammation was assessed through histopathology and assessment of the expression of inflammation and tissue damage-related genes (S1008/9A, Cxcl2, Il1b, Nlrp3, Il6, and Il33) in sampled tissue. Systemic inflammation was assessed through quantification of plasma haptoglobin levels, measurement of blood Il1b expression, and evaluation of histopathologic changes in lung, kidney, liver, and spleen. Apart from slight, transient increases in plasma haptoglobin levels after lateral tail incision, retrobulbar sinus puncture, and saphenous vein puncture, no other signs of systemic inflammation were found. Mice subjected to retrobulbar sinus puncture, sublingual puncture, or isoflurane anesthesia only showed the highest plasma corticosterone concentrations. Retrobulbar sinus puncture had the largest effect on body weight loss. Retrobulbar sinus puncture, sublingual puncture, and submandibular puncture only showed minor and in, most cases, fastresolving inflammation. By contrast, blood sampling by lateral tail incision, tail-tip amputation, or saphenous vein puncture caused tissue damage and inflammation locally at the sampling site, which resolved more slowly compared with head-region sampling techniques, according to results from pathologic and gene expression assessments. Expression of S1008/9A, Cxcl2, Il1b, and Nlrp3 increased 10- to 1000-fold and did not return to baseline until day 6 after sampling or later and did not resolve after tail-tip amputation within the 12-d observation period. Increased expression of genes involved in inflammation and tissue repair correlated with histopathologic changes and may thus represent a quantitative supplement to histopathology. In conclusion, none of the tested methods for obtaining blood samples from mice is superior, according to simultaneous immunologic, histopathologic, and animal welfare-related parameters.

EFFECT OF EARLY FEED RESTRICTION ON BROILER PERFORMANCE, BLOOD AND CARCASS PARAMETERS UNDER SUMMER CONDITIONS.

This research work was designed to study early feed restriction effects on broiler production during Egyptian summer. One hundred ninety eight unsexed Cobb-500 broiler chickens were divided to six experimental treatments with three equal replications. All chicks were free fed starter diet during the first week. In the second week, the chicks in the control group were fed the starter diet ad libitum, but feed intake of the other experimental groups were restricted at levels of 20, 25, 30, 35 and 40%, respectively from the feed intake of the control group. At 2 weeks of age, one bird from each replicate was sacrificed to obtain blood samples. All the experimental chicks were free fed basal grower diet from 14 to 42 days-old. Broiler performance was evaluated by live weight, feed consumed and feed conversion. Carcass parameters and some blood constituents were also measured. The current results showed that feed restriction improved the marketing live body weight, feed utilization and economical efficiency of treated broiler chicks, with no negative effect on either carcass traits or blood parameters. It could be concluded that early feed restriction during the second week of life up to 35% of ad libitum feeding can enhance the marketing weight and improve the feed conversion of broilers as compared to the control treatment under summer conditions in Egypt.

The Utility of Soluble CD14 Subtype in Early Diagnosis of Culture-Proven Early-Onset Neonatal Sepsis and Prediction of Outcome.

This study aimed to evaluate soluble cluster of differentiation 14 subtype (sCD14-ST), also named presepsin, as an early marker for the diagnosis of culture-proven early-onset sepsis (EOS) in neonates and to assess its relation to disease severity and mortality. Out of 60 neonates with risk factors of EOS, 31 neonates were diagnosed as having culture-proven EOS. They were compared with 20 nonseptic controls. We obtained blood samples on day 1 of life for sCD14-ST measurement and sepsis screening. Blood samples were repeated on day 3 in EOS neonates. sCD14-ST was significantly higher in EOS neonates than controls (p < 0.001). Neonates who later developed septic shock had significantly higher day 1 sCD14-ST level than those who did not (p < 0.001). Furthermore, neonates who died had significantly higher day 1 sCD14-ST than survivors (p < 0.001). On day 3, there was a significant decline in sCD14-ST levels than initial levels among survivors. There was a significant positive correlation between day 1 sCD14-ST level and Tollner's sepsis severity score. sCD14-ST could be used as a powerful diagnostic and prognostic marker of EOS. Its quantitative measurement at birth could be a good predictor of sepsis severity and mortality.

Blood-Derived DNA Methylation Signatures of Crohn's Disease and Severity of Intestinal Inflammation

Crohn's disease is a relapsing and remitting inflammatory disorder with a variable clinical course. Although most patients present with an inflammatory phenotype (B1), approximately 20% of patients rapidly progress to complicated disease, which includes stricturing (B2), within 5 years. We analyzed DNA methylation patterns in blood samples of pediatric patients with Crohn's disease at diagnosis and later time points to identify changes that associate with and might contribute to disease development and progression. We obtained blood samples from 164 pediatric patients (1-17 years old) with Crohn's disease (B1 or B2) who participated in a North American study and were followed for 5 years. Participants without intestinal inflammation or symptoms served as controls (n= 74). DNA methylation patterns were analyzed in samples collected at time of diagnosis and 1-3 years later at approximately 850,000 sites. We used genetic association and the concept of Mendelian randomization to identify changes in DNA methylation patterns that might contribute to the development of or result from Crohn's disease. We identified 1189 5'-cytosine-phosphate-guanosine-3' (CpG) sites that were differentially methylated between patients with Crohn's disease (at diagnosis) and controls. Methylation changes at these sites correlated with plasma levels of C-reactive protein. A comparison of methylation profiles of DNA collected at diagnosis of Crohn's disease vs during the follow-up period showed that, during treatment, alterations identified in methylation profiles at the time of diagnosis of Crohn's disease more closely resembled patterns observed in controls, irrespective of disease progression to B2. We identified methylation changes at 3 CpG sites that might contribute to the development of Crohn's disease. Most CpG methylation changes associated with Crohn's disease disappeared with treatment of inflammation and might be a result of Crohn's disease. Methylation patterns observed in blood samples from patients with Crohn's disease accompany acute inflammation; with treatment, these change to resemble methylation patterns observed in patients without intestinal inflammation. These findings indicate that Crohn's disease-associated patterns of DNA methylation observed in blood samples are a result of the inflammatory features of the disease and are less likely to contribute to disease development or progression.

Correlation and agreement between the TEG® 5000 and the TEG® 6s during liver transplant surgery.

The TEG® 5000 and novel TEG® 6s measure the viscoelasticity of whole blood during in vitro clot formation. The two devices measure similar coagulation variables but utilize distinctly different technologies. This study aimed to determine the correlation and agreement between the thrombelastographic parameters obtained by the two devices during liver transplant surgery. We obtained blood samples at six predefined intervals during the surgery of 10 consecutive patients. Two operators proficient in the use of the TEG® 6s and TEG® 5000 systems performed thrombelastographic measurements on each sample: non-citrated TEG® 5000, citrated TEG® 5000 and citrated TEG® 6s. Agreement and correlation were assessed using Bland Altman plots and Lin's concordance correlation. There was considerable inter-device variability for the different parameters measured by the TEG® 5000 and TEG® 6s devices. Acceptable agreement was observed when results were within the normal reference ranges. However, with increasing coagulopathy, agreement was poor and results could not be considered interchangeable. Although each of the three tests appeared reliable for qualitative detection of abnormalities of clot formation during liver transplant surgery, we found their quantitative results were not interchangeable.

Dinuclear ruthenium(II) Schiff base complex: a first in vivo study in Swiss albino mice.

Dinuclear ruthenium(II) Schiff base complex was selected for in vivo study among many other novel metal-based compounds, because of its previously proved in vitro anticancer and antibacterial properties. The aim was to investigate the potential toxicity of this compound in animal model through biochemical and histopathological assessment. Adult Swiss albino mice of both sexes were divided into high-dose and low-dose group that received a single intraperitoneal dose of ruthenium complex (175 mg/kg and 25 mg/kg, respectively) and one control group (vehicle only). After a follow-up period of 14 days, animals were sacrificed to obtain blood samples and organs. The test compound was well tolerated in a low-dose group and did not cause any mortality. The histological findings and serum biochemistry suggested a reversible character of alterations found in vital organs of this group. However, in the high-dose group, adverse effects were more severe and indicated dose and gender-related toxicity. Mild side effects found in a low-dose group together with excellent in vitro properties, made dinuclear ruthenium(II) Schiff base complex a promising candidate for further investigation and development as anticancer and antimicrobial agent (Tab. 4, Fig. 6, Ref. 32).

Safety assessment of Chlorophytum alismifolium tuber extract (Liliaceae): Acute and sub-acute toxicity studies in Wistar rats

Objective: To explore the toxicological profile of methanol extract of Chlorophytum alismifolium (MECA) tubers in Wistar rats. Methods: MECA was subjected to acute and sub-acute studies which were conducted according to Organization for Economic Co-operation and Development (OECD 425 and 407 guidelines respectively). In the acute toxicity experiment, a limit test (5 000 mg/kg) was administered to five rats and monitored for 2 weeks. The sub-acute studies were conducted on 4 groups of rats. The first group served as control, while the 2nd, 3rd and 4th groups received MECA (150, 300 and 600 mg/kg respectively). The treatments were given orally and daily for 4 weeks. At the end of the experiment (29th day), the animals were euthanized to obtain blood samples and organs for haematological, biochemical and histological evaluations. Results: Acute toxicity study showed that the oral median lethal dose was >5 000 mg/kg. In the sub-acute studies, the results showed no significant (P>0.05) changes in the haematological, hepatic and renal indices compared to control animals. In the fourth week, a significant (P

The impact of antenatal factor XIII levels on postpartum haemorrhage: a prospective observational study.

Postpartum haemorrhage (PPH) is a leading cause of maternal mortality and morbidity. Our aim was to investigate the relationships between antenatal factor XIII (FXIII), fibrinogen levels, and blood loss at childbirth. This prospective observational study evaluated an unselected cohort of pregnant women admitted for intended vaginal deliveries of singletons at term. To determine clotting factor levels, we obtained blood samples at a maximum of three days prior to vaginal delivery. A calibrated collecting drape was used to quantify blood loss in the third stage of labour. Moderate and severe PPH were diagnosed as blood losses ≥500mL and ≥1000mL, respectively. In a multiple logistic regression analysis, we determined whether coagulation factors and their interactions could independently predict (severe) PPH. We analysed 548 vaginal deliveries that occurred during the study period. Of those, 78 (14.2%) lost ≥500mL and 18 (3.3%) lost ≥1000mL of blood. The mean pre-delivery FXIII activity in women with PPH (79.33% ± 15.5) was significantly (p < 0.001) lower than in women without PPH (86.45% ± 14.6). A receiver operating characteristic curve analysis detected antenatal FXIII cutoff levels of 83.5% and 75.5% for PPH and severe PPH, respectively. The multiple logistic regression analysis showed that FXIII alone (p < 0.001) and its interaction with fibrinogen (p = 0.03) significantly predicted PPH. FXIII was not significantly correlated with blood loss among patients with severe PPH. Our results suggested that antenatal FXIII levels may have a significant influence on PPH. The interaction between FXIII and fibrinogen might also provide slight advantages in forecasting PPH.

The protective effects of WIN55212-2 for lung injury in paraquat poisoned mice

Objective To explore the protective effects of cannabinoid analogues WIN55212-2 on paraquat poisoned mice. Methods Totally 35 healthy male C57BL/6 mice were randomly(random number) divided into four groups: PQ group (paraquat poisoned, n=10), WIN 1 mg group (PQ+WIN55212-2 1 mg n=10), WIN 2 mg group (PQ+WIN55212-2 2 mg, n=10), control group (n=5). The PQ poisoned animal models were established in the PQ group, WIN 1 mg group and WIN 2 mg group by intraperitoneally injection of paraquat with a concentration of 20 mg/kg. Intraperitoneal injection of WIN55212-2 (containing Tween 80 cosolvent) at the concentration of 1 mg/kg and 2 mg/kg was performed 1 h before PQ exposure in the two interfered groups. Equivalent volume of saline was given to the control group. WIN55212-2 was injected twice a week from the second week. In the acute phase (14 d), 5 mice were randomly sacrificed in the PQ group, WIN 1 mg group and WIN 2 mg group, and 3 mice were sacrificed in the control group to obtain blood sample, bronchoalveolar lavage fluid (BALF) and lung tissue. All the remaining mice were executed on day 28, and the tissue samples were collected as mentioned above. HE staining and Masson staining were performed to observe the changes of lung tissues after PQ poisoning. Changes of TNF-α, IL-6 and TGF-β in plasma and BALF were measured by ELISA. Results In the acute phase, the pathological sections of lung tissues in the PQ group, WIN 1 mg group and WIN 2 mg group showed diffuse inflammation, which was improved after the intervention of WIN5522-2, especially in the WIN 1 mg group. IL-6 levels of BALF in the PQ group, WIN 1 mg group, WIN 2 mg group and the control group were (1 024.77±124.74)U/L, (620.48±99.76)U/L, (823.29±157.88)U/L, and (180.42±20.22)U/L, respectively. IL-6 levels in the WIN 1 mg group and the WIN 2 mg group were statistically lower than those in the PQ group (P=0.021, P=0.016). However, no difference was found between the two intervention groups(P=0.114). The similar condition was also found in TNF-α in BALF and plasma. In the chronic phase, mice in the PQ group, WIN 1 mg group and WIN 2 mg group showed fibrosis in tissue by HE and Masson staining, and the inflammatory condition was improved after the intervention of WIN5522-2, which was more obvious in the WIN 1 mg group. In BALF, TNF-α level was (321.64±50.54)U/L, (260.23±48.19)U/L, (278.89±29.40)U/L, (89.76 ± 10.87)U/L in the PQ group, WIN 1 mg group, WIN 2 mg group and the control group. Differences were found between the WIN 1 mg group and the control group and the WIN 2 mg group. Similar differences were also observed in plasma TNF-α, but not in TGF-β. Conclusions A small dose of WIN55212-2 can improve the general condition of PQ poisoning mice, and reduce the inflammatory and fibrosis-related cytokines levels in PQ poisoning mice. Key words: Paraquat; Mice model; WIN55212-2; Cannabis receptor agonist; Acute lung injury; Chronic lung fibrosis

High Prevalence of MERS-CoV Infection in Camel Workers in Saudi Arabia.

Middle East respiratory syndrome (MERS), a highly lethal respiratory disease caused by a novel coronavirus (MERS-CoV), is an emerging disease with high potential for epidemic spread. It has been listed by the WHO and the Coalition for Epidemic Preparedness Innovations (CEPI) as an important target for vaccine development. While initially the majority of MERS cases were hospital acquired, continued emergence of MERS is attributed to community acquisition, with camels likely being the direct or indirect source. However, the majority of patients do not describe camel exposure, making the route of transmission unclear. Here, using sensitive immunological assays and a cohort of camel workers (CWs) with well-documented camel exposure, we show that approximately 50% of camel workers (CWs) in the Kingdom of Saudi Arabia (KSA) and 0% of controls were previously infected. We obtained blood samples from 30 camel herders, truck drivers, and handlers with well-documented camel exposure and from healthy donors, and measured MERS-CoV-specific enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay (IFA), and neutralizing antibody titers, as well as T cell responses. Totals of 16/30 CWs and 0/30 healthy control donors were seropositive by MERS-CoV-specific ELISA and/or neutralizing antibody titer, and an additional four CWs were seronegative but contained virus-specific T cells in their blood. Although virus transmission from CWs has not been formally demonstrated, a possible explanation for repeated MERS outbreaks is that CWs develop mild disease and then transmit the virus to uninfected individuals. Infection of some of these individuals, such as those with comorbidities, results in severe disease and in the episodic appearance of patients with MERS.IMPORTANCE The Middle East respiratory syndrome (MERS) is a coronavirus (CoV)-mediated respiratory disease. Virus transmission occurs within health care settings, but cases also appear sporadically in the community. Camels are believed to be the source for community-acquired cases, but most patients do not have camel exposure. Here, we assessed whether camel workers (CWs) with high rates of exposure to camel nasal and oral secretions had evidence of MERS-CoV infection. The results indicate that a high percentage of CWs were positive for virus-specific immune responses but had no history of significant respiratory disease. Thus, a possible explanation for repeated MERS outbreaks is that CWs develop mild or subclinical disease. These CWs then transmit the virus to uninfected individuals, some of whom are highly susceptible, develop severe disease, and are detected as primary MERS cases in the community.

Lower mitochondrial dysfunction in survivor septic patients with mitochondrial DNA haplogroup JT

ObjectiveThe comparison on mitochondrial function between severe septic patients and healthy control subjects according to mitochondrial deoxyribonucleic acid (mtDNA) haplogroup has not been previously reported; and this was the objective of the current study. MethodsProspective, multicenter, observational study. We obtained blood samples from 198 severe septic patients at days 1, 4 and 8 of severe sepsis diagnosis and from 96 sex- and age-matched healthy controls to determine mtDNA haplogroup and platelet respiratory complex IV (CIV) specific activity. The endpoint of the study was 30-day mortality. ResultsWe included 198 severe septic patients (38 with mtDNA haplogroup JT and 160 with mtDNA haplogroup non-JT) and 96 healthy control subjects (16 with mtDNA haplogroup JT and 80 with mtDNA haplogroup non-JT). We have no found statistically significant differences in platelet CIV specific activity between healthy controls and survivor severe septic patients with mtDNA haplogroup JT at days 1, 4 and 8 of severe sepsis diagnosis; and the remaining severe septic patients showed lower platelet CIV specific activity than healthy controls with the same mtDNA haplogroup. ConclusionsThe new finding of our study was that survivor severe septic patients and healthy controls with mtDNA haplogroup JT showed no different platelet Civ specific activity.

1467P - Efficacy and safety of nivolumab for cytotoxic chemotherapy unfit patients with advanced non-small cell lung cancer: A phase II study

Background: This single-center phase II study aimed to examine the efficacy and feasibility of nivolumab for patients with advanced non-small cell lung cancer (NSCLC) who were unsuitable for single-agent chemotherapy or targeted agents because of their poor performance status (PS). Methods: In this study, we enrolled previously treated patients with advanced NSCLC with poor PS of 2–4 without any indication of cytotoxic chemotherapy or targeted therapies. All enrolled patients received nivolumab (3 mg/kg, every 2 weeks) until progression or unacceptable toxicities. In addition, we prospectively obtained peripheral blood mononuclear cells (PBMC) and plasma from patients after obtaining their informed consent before treatment. Results: Between April 2016 and December 2017, we enrolled 33 patients with NSCLC and poor PS, including 23 patients with PS 3–4. In this study, the response rate was 29.0% (90% confidence interval (CI), 17.7%–43.7%), and the disease control rate was 41.9% in 31 evaluable patients. PS improvement rate was 29.0%. In addition, the median progression-free survival (mPFS), median overall survival (mOS), and 1-year survival rates were 1.5 (95% CI, 1.0–2.7) months, 3.8 (95% CI, 2.4–16.3) months, and 37.6%, respectively. Among patients harboring EGFR-mutations (mut), the mPFS was 2.6 and 1.1 months in EGFR-wild type (wt) and EGFR-mut (p = 0.015), respectively, and the mOS was 9.3 and 1.9 months in EGFR-wt and EGFR-mut (p = 0.029), respectively. During the study period, treatment-related deaths were observed in 2 patients (6.1%). We obtained blood samples from 75% of the enrolled patients (data will be updated in the meeting). Conclusions: To the best of our knowledge, this is the first trial to investigate the efficacy of immune checkpoints in patients with advanced NSCLC with poor PS. Although the response rate was similar to those with good PS, shorter survival was observed. Excluding patients harboring driver mutations may enhance treatment efficacy to show a survival benefit even for poor PS patients. Nevertheless, the biomarker investigation warrants focus on the PBMC analysis. Clinical trial identification: Clinical trial registration: UMIN000020855/UMIN000021734. Legal entity responsible for the study: Tokyo Metropolitan Komagome Hospital. Funding: Has not received any funding. Disclosure: Y. Okuma: Research Funding: Chugai Pharma (Inst). Y. Hosomi: Honoraria: AstraZeneca; Boehringer Ingelheim Japan; Bristol-Myers Squibb Japan; Chugai Pharma; Eli Lilly Japan; Ono Pharmaceutical; Taiho Pharmaceutical; Research Funding: Chugai Pharma (Inst); Eli Lilly Japan (Inst); Ono Pharmaceutical. N. Yamamoto: Honoraria: AstraZeneca; Bristol-Myers Squibb Japan, Chugai Pharma, Eli Lilly Japan, Ono Pharmaceutical, Pfizer; Research Funding: Astellas Pharma (Inst), Bayer (Inst), Boehringer Ingelheim (Inst), Chugai Pharma (Inst), Daiichi Sankyo (Inst), Eisai (Inst), Kyowa Hakko Kirin (Inst), Eli Lilly Japan (Inst), Novartis (Inst), Pfizer (Inst), Quintiles (Inst), Taiho Pharmaceutical (Inst), Takeda (Inst). All other authors have declared no conflicts of interest.

Cytokines in relation to hCG are significantly altered in asymptomatic women with miscarriage \u2013 a pilot study

BackgroundSpontaneous abortion is one of the most common complications in early pregnancy. A preventive test to identify women who will experience a miscarriage, even before first symptoms occur, is not established. Activation of maternal immunological tolerance seems to be essential for early fetal development and various cytokines have been described in different stages of pregnancy. Therefore, we aimed to investigate if chemokine levels at the time of pregnancy testing relative to human Choriogonadotropin (hCG) are altered in patients who will experience a miscarriage in this pregnancy.MethodsWe obtained blood samples from 39 women. Dependent on the follow-up, patients with a positive pregnancy test were subsequently divided in two groups: ongoing pregnancy (n = 22) and miscarriage (n = 17) in this pregnancy. Immunological and endocrine profiling of maternal plasma at the time of pregnancy testing (5th week of gestation) was performed for each group at the time of pregnancy test using Multiplex and ELISA analysis.ResultshCG was significantly decreased in patients with abortion whereas levels of IL-1ra, MIP-1a and TNF-alpha were significantly increased. GCSF/ IL-1ra-ratio was 1.66-fold increased in patients with ongoing pregnancy. TGF-beta /MIP1a-ratio was significantly 3.45-times higher in patients with miscarriage. Comparing patients with ongoing pregnancy to patients experiencing a miscarriage, we could demonstrate significant alterations of the ratios MIP1a/hCG, IL-1ra/hCG, TNFalpha/hCG, MCP1/hCG, IL-6/hCG, TPO/hCG and TGF-beta1/hCG. The strongest effects were seen for the ratio MIP1a/hCG, IL-1ra/hCG and TNFalpha/hCG.ConclusionsWe have shown that cytokines in relation to hCG after 4 weeks of gestation are significantly altered in women with miscarriage, promising potential as a prognostic biomarker.

Is waist circumference more strongly associated with metabolic risk factors than waist-to-height ratio in Asians?

Differential distribution of fats can vary among ethnic groups and thus have varying effects on metabolic risk. Measuring metabolic risk of individuals using simple anthropometric measurements is essential to replace current invasive methods of obtaining blood samples. Waist-to-height ratio (WHtR) has been advocated as the best simple anthropometric measurement, but, because of the high visceral fat of Asians, there has been speculation as to the possibility of using only waist circumference (WC) to measure metabolic risk. The aim of this study was to compare the performance of WC and WHtR in terms of their association with measures of obesity and metabolic risk factors (e.g., homeostasis model assessment for insulin resistance, low-density lipoprotein, triacylglycerol, and ratio of triacylglycerol to high-density lipoprotein) and to obtain an optimal cutoff value for one anthropometric measurement. The study was performed on healthy Asian Chinese (N = 527) men (n = 209) and women (n = 318) who participated in a cross-sectional study conducted at the Clinical Nutrition Research Centre located in Singapore. Association of WC and WHtR with metabolic risk factors was obtained using Spearman's rank correlation coefficient. Optimal cutoff value was obtained using receiver operating characteristic curve. WC and WHtR performed equally well in both sexes in terms of their strength of association between metabolic risk factors. Receiver operating characteristic curve analysis showed that 73.5 cm (in women) and 82.5 cm (in men) were the optimal WC cutoff values to identify insulin resistance. It is suggested that WC is a simpler anthropometric measurement that has strong association with an individual's metabolic risk level.

Continued expansion of tick-borne pathogens: Tick-borne encephalitis virus complex and Anaplasma phagocytophilum in Denmark

Tick-borne encephalitis virus (TBEV) is a tick-transmitted flavivirus within the tick-borne encephalitis (TBE) complex. The TBE complex is represented by both TBEV and louping ill virus (LIV) in Denmark. Anaplasma phagocytophilum is also transmitted by ticks and is believed to play an essential role in facilitating and aggravating LIV infection in sheep.This study aimed to describe the distribution of TBE complex viruses in Denmark, to establish the possible emergence of new foci and their association with the distribution of A. phagocytophilum. We performed a nationwide seroprevalence study of TBE complex viruses using roe deer (Capreolus capreolus) as sentinels and determined the prevalence of A. phagocytophilum in roe deer. Danish hunters obtained blood samples from roe deer during the hunting season of 2013-14. The samples were examined for TBEV-specific antibodies by virus neutralization tests (NT). A. phagocytophilum infection was assessed by specific real-time-PCR.The overall seroprevalence of the TBE complex viruses in roe deer was 6.9% (51/736). The positive samples were primarily obtained from a known TBE endemic foci and risk areas identified in previous sentinel studies. However, new TBE complex risk areas were also identified. The overall prevalence of A. phagocytophilum was 94.0% (173 PCR-positive of 184 roe deer), which is twice the rate observed ten years ago.These results point to an expansion of these tick-borne diseases geographically and within reservoir populations and, therefore, rationalize the use of sentinel models to monitor changes in transmission of tick-borne diseases and development of new risk areas. We found no association between TBE complex-positive roe deer and the prevalence of A. phagocytophilum, as almost all roe deer were infected. Based on our findings we encourage health care providers to be attentive to tick-borne illnesses such as TBE when treating patients with compatible symptoms.

Systemic Oxygen Transport with Rest, Exercise, and Hypoxia: A Comparison of Humans, Rats, and Mice.

The objective of this article is to compare and contrast the known characteristics of the systemic O2 transport of humans, rats, and mice at rest and during exercise in normoxia and hypoxia. This analysis should help understand when rodent O2 transport findings can-and cannot-be applied to human responses to similar conditions. The O2 -transport system was analyzed as composed of four linked conductances: ventilation, alveolo-capillary diffusion, circulatory convection, and tissue capillary-cell diffusion. While the mechanisms of O2 transport are similar in the three species, the quantitative differences are naturally large. There are abundant data on total O2 consumption and on ventilatory and pulmonary diffusive conductances under resting conditions in the three species; however, there is much less available information on pulmonary gas exchange, circulatory O2 convection, and tissue O2 diffusion in mice. The scarcity of data largely derives from the difficulty of obtaining blood samples in these small animals and highlights the need for additional research in this area. In spite of the large quantitative differences in absolute and mass-specific O2 flux, available evidence indicates that resting alveolar and arterial and venous blood PO2 values under normoxia are similar in the three species. Additionally, at least in rats, alveolar and arterial blood PO2 under hypoxia and exercise remain closer to the resting values than those observed in humans. This is achieved by a greater ventilatory response, coupled with a closer value of arterial to alveolar PO2 , suggesting a greater efficacy of gas exchange in the rats. © 2018 American Physiological Society. Compr Physiol 8:1537-1573, 2018.

An unexpected diversity of trypanosomatids in fecal samples of great apes.

Charismatic great apes have been used widely and effectively as flagship species in conservation campaigns for decades. These iconic representatives of their ecosystems could also play a role as reservoirs of several zoonotic diseases. Recently it was demonstrated that African great apes can host Leishmania parasites (Kinetoplastea: Trypanosomatidae). Given that this finding raised a strong negative reaction from leishmania experts and the subsequent discussion did not lead to a clear resolution, we decided to analyze wild gorilla (Gorilla gorilla gorilla) and chimpanzee (Pan troglodytes troglodytes) fecal samples collected from the same area in Cameroon as in the original study. Fecal samples, used to circumvent the difficulties and ethics involved in obtaining blood samples from endangered wild apes, were screened by three different PCR assays for detection of Leishmania DNA. We did not detect any leishmania parasites in analyzed feces; however, sequencing of SSU rRNA revealed an unexpected diversity of free-living bodonids (Kinetoplastea: Bodonidae) and parasitic trypanosomatids (Kinetoplastea: Trypanosomatidae) other than Leishmania. A single detected Phytomonas species, found in chimpanzee feces, most likely originated from animal plant food. On the other hand, the presence of four free-living bodonid species and four parasitic insect monoxenous trypanosomatid, including two possible new species of the genus Herpetomonas, could be explained as ex post contamination of feces either from the environment or from flies (Diptera: Brachycera).