Clinical consequences of Hypotension in patient with organophosphate poisoning

Objective: Aim of study is to describe the clinical consequences of hypotension in patient with organophosphate poisoning. Method: In the retrospective cohort study, we analyzed data of 66 patients with organophosphate poisoning who were treated at Bir Hospital, Nams Kathmandu. Data from those with hypotension and normal blood pressure were compared to identify significant clinical consequences. Results: All together 66 patients were enrolled in this study out of which 44(66.7%) were female and 22(33.3%) were male. After analyzing data, we found 18.2% of case with severe poisoning (ACHE < 700 U/L). Among all, 41(62%) were found to have normal blood pressure and 25(37.9%) were found to have low blood pressure. Among those with hypotension, around 56% were found to have prolonged QTc interval, p < 0.003 and there was statistically significant association between QTc prolongation and vasopressor requirement, X2(1) = 22.98, P < 0.001. Patients requiring higher dose to reach atropinization had statistically significant hypotension, P < 0.001. Those with low blood pressure were found to require more days of hospital admission, P < 0.001. Patients with hypotension were found to have severe poisoning both on the basis of POP Score severity grading, 16(64%) P <0.002 and ACHE Severity scale, 7(28%) P < 0.05. In comparison with normal blood pressure group, low blood pressure group had significantly more chance of developing complications like septic shock (2), aspiration pneumonia (5), ARDS (1) and bed sore, P = 0.002. Vasopressor requirement was significantly more among those with low blood pressure, P < 0.001. Most of hypotensive patients were needing ICU care, found to have higher WBC count P = 0.002 and lower GCS Score at admission P < 0.001. There was positive correlation between hypotension and POP Score at admission P < 0.001. Conclusion: Hypotension is a common complication in patient with organophosphate poisoning and is associated with higher POP Score, lower ACHE level, lower GCS Score, increased vasopressor requirement, more hospital stays, increasing ICU admission, more chance of developing septic shock and aspiration pneumonia.

Patients with Dipper and Nondipper High-Normal Blood Pressure Were Associated with Left Ventricular Mass

Purpose. High-normal blood pressure has been suggested to associate with target organ damage and higher left ventricular mass index (LVMI). Our aim is to find the association between people with high-normal blood pressure and their left ventricular mass index. Materials and Methods. Given a total of 181 people with office blood pressure, 24-hour ambulatory blood pressure monitoring, 35 of them are normotensive (BP < 130/85 mm Hg), and 146 people with high-normal blood pressure (BP 130–139/85–89 mm Hg), divide the high-normal blood pressure group into dipper and nondipper according to their ABPM in 24 hours. All of them were performed with echocardiography to calculate LVMI. Results. After adjusting for potential confounding factors, mean systolic blood pressure (BP) of the nondipper group is (119 + 9) mmHg in 24 h, which is significantly higher ( p < 0.05) than in the dipper group (116 + 11) mmHg, indicating the mean systolic BP is associated with the dipper type ( p < 0.05); furthermore, the higher nocturnal blood pressure is associated with the nondipper group significantly ( p < 0.05), and LVMI ((121 ± 11) g/m2) of the nondipper group is also significantly higher than in the dipper group’s LVMI ((108 ± 12) g/m2) ( p < 0.05). The multivariate linear regression analyses revealed significant and independent associations of LVMI with these factors: triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL-C), and coefficient of variation of systolic and diastolic blood pressure in 24 hours. Conclusion. After multiple relevant clinical confounding factors were adjusted, patients with dipper and nondipper high-normal blood pressure had higher LVMI. Abnormalities in circadian blood pressure variability may be associated with the left ventricular hypertrophy.

Waist Circumference Is More Closely Associated with Hypogonadism than Is Hyperglycemia, Independent of BMI in Middle-Aged Men

Introduction. To evaluate whether waist circumference (WC) or hyperglycemia is more closely associated with hypogonadism in middle-aged men. Research Design and Methods. This cross-sectional study analyzed male participants under 65 years old from the MJ Health Screening Center in Taiwan from 2007 to 2016. Basic patient characteristics with relevant parameters were obtained. We used the chi-square test to perform a correlation analysis for HbA1c and WC between participants with and without hypogonadism. A one-way ANOVA with post hoc Scheffe’s method was applied to compare the mean testosterone (T) among the HbAlc and WC groups (normal blood sugar with normal WC (NBSNW), abnormal blood sugar with normal WC (ABSNW), normal blood sugar with abnormal WC (NBSAW), and abnormal blood sugar with abnormal waist circumference (ABSAW)). Results. The 5,680 participants were divided into two groups based on the presence ( n = 599 ) or absence of hypogonadism ( n = 5,081 ), which was defined as total testosterone TT < 300 ng / dL . The mean TT of group NBSAW ( 443.71 ± 220.59 ng / dl ) was significantly lower than that of group ABSNW ( 506.64 ± 191.08 ng / dl , p < 0.001 ). Moreover, the mean TT of group ABSAW ( 398.89 ± 146.24 ng / dl ) was significantly lower than that of group ABSNW ( 506.64 ± 191.08 ng / dl , p < 0.001 ). The ORs after adjusting for BMI, TG, HDL, SBP, and DBP were statistically significant when comparing NBSAW vs. NBSNW ( OR = 2.846 ; 95 % CI = 2.266 – 3.575 ; p < 0.001 ), ABSNW vs. NDNW ( OR = 1.693 ; 95 % CI = 1.309 – 2.189 ; p < 0.001 ), and ABSAW vs. NBSNW ( OR = 4.613 ; 95 % CI = 3.634 – 5.856 ; p < 0.001 ). Conclusion. The current study showed that WC should be the risk factor that is more closely associated with hypogonadism than hyperglycemia in middle-aged men.

Serum Uric Acid to HDL-Cholesterol Ratio is Associated with New-Onset NAFLD in Lean Chinese People with Normal Blood Lipid Levels: a Longitudinal Cohort Study

Background: Nonalcoholic fatty liver disease (NAFLD) is commonly occurred in the non-obese individuals. The serum uric acid (UA) to high-density lipoprotein cholesterol (HDL-c) ratio (UHR) is considered as a predictive factor of NAFLD. However, it is still difficult to confirm the correlation in lean Chinese people with normal blood lipid levels. It is aimed to analyze the correlation of UHR with NAFLD among lean Chinese population without dyslipidemia and compared UHR with other predictors in this study.Methods: 9838 lean people without NAFLD were included in a retrospective cohort study. NAFLD was diagnosed by liver ultrasound.Results: A total of 9,838 lean patients with normal blood lipid levels were included in the final study. During the five-year follow-up period, the overall prevalence of NAFLD was 8.7%. across the quintile 1, 2, 3, 4 and 5 of UHR, the prevalence of NAFLD among lean patients was increased from 2.4%, 5%, 7.9%, 10.3% to 17.8%. After adjustment for age, markers of liver and kidney function, gender and metabolic indicators, multivariate cox proportional hazard regression analysis demonstrated that the hazard ratio(HR) was 1.99(1.43-2.73) in highest UHR (quintile 5) compared with lowest UHR(quintile 1). The area under the curve(AUC) of UHR (0.690) was higher than in UA (0.666) and HDL-c (0.636), which showed that the predictive ability of the UHR to new-onset NAFLD was better than serum uric acid and HDL-c. Even within the normal range of UA and HDL-c levels, UHR was independently associated with NAFLD, and HR (95% confidence interva, 95%CI) for NAFLD in Quintile 5 of UHR was 6.74(4.32-10.53). Compared with other significant predictors, the AUC value of UHR(0.67) was similar to that of low-density lipoprotein cholesterol(LDL-c)/HDL-c ratio(0.68), non-high-density lipoprotein cholesterol (NHDL-c)/HDL-c ratio(0.68) and alanine aminotransferase (ALT)/ aspartate aminotransferase (AST) ratio(0.7), and superior to that of LDL-c (0.63), remnant cholesterol (RC,0.59), albumin(ALB)/alkaline phosphatase(ALP) ratio(0.61). The sensitivity of UHR (70.5%) was the highest among all indicators. In the subgroup of ALT, the AUC of UHR was 0.70, which was the highest among all predictors in subjects with ALT< 40. For subjects with elevated ALT levels (ALT > 40 U/L), there was no statistical significance among RC (P=0.441), ALB/ALP (P=0.419) and ALT/AST(P=0.159). In contrast, UHR's performance in predicting NAFLD was meaningful and reliable (AUC=0.61, p<0.001).Conclusions: UHR serve as an inexpensive and reliable predictor of NAFLD in lean Chinese people with normal blood lipid levels. It can be used to identify people at high risk of NAFLD.

FISH By Imaging Flow Cytometry in CLL for Diagnosis and MRD Assessment

Chronic lymphocytic leukaemia is a genetically heterogeneous disease with treatment and prognosis varying based on chromosomal abnormalities. These are detectable in up to 80% of cases when tested on the nuclei of interphase cells by fluorescence in situ hybridisation (FISH). Despite the clinical importance of FISH in management, as only up to 200 nuclei are generally assessed, it is not suitable for minimal residual disease (MRD) assessment. Since clinical decisions are based on detection thresholds of 10 -4, MRD assays are restricted to flow cytometry and molecular based assessment. Here we have explored the utility of a cutting-edge automated imaging flow cytometry method that incorporates cell immunophenotype and FISH ("immuno-flowFISH") to detect chromosomal abnormalities in CLL. Aims: Our aim was to determine the capability of immuno-flowFISH using imaging flow cytometry to detect del(17p) and +12 in CLL, and, the lowest limit of detection. We hypothesized that this integrated automated immuno-flowFISH method would be suitable for MRD assessment of CLL. Methods: Blood from 75 patients with CLL, at diagnosis or on therapy, was analysed. For MRD studies, cells from the CI cell line were spiked into normal blood at concentrations of 0.001 - 10%. After red cell lysis, samples were incubated with CD3, CD5 and CD19 fluorophore-conjugated antibodies (fluorophores: BV480, BV605, AF647). Following fixation and membrane permeabilization, DNA was denatured at 78 oC for 5 mins. FISH probes to 17p12 and centromeres of chromosomes 12 and 17 were added and hybridized for 24 hours at 42 oC. Nuclei were then stained with SYTOX AADvanced and up to 600,000 cells acquired on the Amnis ® ImageStream ®XMk II imaging flow cytometer. Digital images (x60 objective) and quantitative data derived from computational algorithms (IDEAS software) were used to assess FISH signals overlying cell nuclei. IDEAS was then used to assess the number and percent CD19/CD5-positive CLL cells with FISH abnormalities. Results: Between 10,000 and 600,000 cells (mean 60,000) were acquired. CLL (CD19/CD5-positive) and T- (CD3/CD5-positive) cells could be clearly identified by their immunophenotype and assessed individually for probe signals. FISH signals were identifiable on the digital images as specific "spots" overlying the SYTOX AADvanced nuclear stain. The IDEAS software could enumerate the number of FISH spots per cell and this was confirmed by quantitative mean channel fluorescence intensity for each probe. A chromosome 12 or 17 abnormality was detected in 23 of the 75 CLL cases. Of these, 10 cases had only one 17p signal (but 2 for the centromere of chromosome 17), indicative of del(17p). Del(17p) was detected in 2-35% of CD19/CD5-positive cells (i.e. 0.4-23% or 270-35,441 of all cells), the lowest seen in a patient on cytoreductive therapy. In 13/75 cases, there were 3 FISH signals for CEP12, consistent with trisomy 12 (+12) in 0.1-46% of all cells analysed; the lowest number of 0.1% was when 26 out of 26,000 cells analysed were CD19/CD5-positive and had +12. We also performed multi-FISH, incorporating CEP12, CEP17 and 17p probes together with the CD3, CD5 and CD19 antibodies. This required 7-fluorophores (antibodies, probes and nucleus) and confirmed the ability to detect del(17p) and chromosome 12 copy number simultaneously in a single analysis. Spiking of CI CLL cells into normal blood demonstrated that +12 could be detected to a lowest limit of 10 -5. In all analyses, CLL cells had normal diploid spots for the control CEP17 probe, and the CD3/CD5-positive T cells had dual signals for CEP12, CEP17 and 17p12 probes on numerical analysis and on digital imagery. Conclusion: This study of confirms that high-throughput automated imaging flow cytometry, integrating FISH and immunophenotyping, can detect chromosomal defects in CLL. The lowest limit of detection for a FISH-detectable abnormality was 10 -5. This high sensitivity and specificity exceeds current clinical practice (10 -4), and was achieved through the analysis of many thousands of cells and positive identification of CLL cells based on their phenotype. This immuno-flowFISH method does not require any prior cell separation and is automated. It adds a new dimension to chromosomal analysis in CLL, both at diagnosis and for MRD monitoring. The high precision and specificity of immuno-flowFISH illustrates that this has a real place as a new MRD assessment tool for CLL. Disclosures No relevant conflicts of interest to declare.

Negative enrichment of circulating tumor cells from unmanipulated whole blood with a 3D printed device

Reliable and routine isolation of circulating tumor cells (CTCs) from peripheral blood would allow effective monitoring of the disease and guide the development of personalized treatments. Negative enrichment of CTCs by depleting normal blood cells ensures against a biased selection of a subpopulation and allows the assay to be applied on different tumor types. Here, we report an additively manufactured microfluidic device that can negatively enrich viable CTCs from clinically-relevant volumes of unmanipulated whole blood samples. Our device depletes nucleated blood cells based on their surface antigens and the smaller anucleated cells based on their size. Enriched CTCs are made available off the device in suspension making our technique compatible with standard immunocytochemical, molecular and functional assays. Our device could achieve a ~ 2.34-log depletion by capturing > 99.5% of white blood cells from 10 mL of whole blood while recovering > 90% of spiked tumor cells. Furthermore, we demonstrated the capability of the device to isolate CTCs from blood samples collected from patients (n = 15) with prostate and pancreatic cancers in a pilot study. A universal CTC assay that can differentiate tumor cells from normal blood cells with the specificity of clinically established membrane antigens yet require no label has the potential to enable routine blood-based tumor biopsies at the point-of-care.

Double Product as a Predictor of Coronary Artery Disease in Males with Normal Blood Pressure

Aim: This study investigates whether in males with normal blood pressure that underwent exercise stress test Double Product (DB) or Double Product Ratios to workload (DPR) are self-sufficient in predicting the presence of coronary artery disease (CAD). Method: 78 male patients with normal blood pressure went through bicycle exercise stress test (EST) and within four weeks they underwent coronary angiography. 34 of them resulted with normal coronary arteries and 44 of them were diagnosed with one or multiple vessel CAD. Pressure rate double product was calculated in rest, in the first stage of the exercise test, in the peak of exercise, and also in the second, fourth, and sixth minutes of recovery of the stress test. Also, the ratios of pressure rate double product with the workload, which reflects the relative growth of double product to the workload, were calculated. The results were compared between the two groups, the one with normal coronary arteries, and those with abnormal coronary angiography. Results: DP in rest and in the first stage of the exercise are not significantly different for normotensive males with or without CAD, while the peak DB value of the exercise, as well as the econd and sixth minutes of the recovery are significantly lower for the group of patients with CAD. The starting peak workloads were significantly lower in the patients' group with CAD (p=0.0002 and p 0.0001). On the other hand, the double product to workload ratio at the first stage was significantly lower in males with normal blood pressure and CAD. Significant lower DPR are detected also in the fourth and sixth minutes of recovery. Conclusion: The values of DP in the peak exercise and at the second and sixth minutes of the recovery phase are significantly lower in normotensive males with abnormal coronary angiography compared to those with normal coronary arteries, and these findings can be used to detect the presence of CAD despite the ECG changes and they are not affected by the baseline characteristics of the patients. The ratios between DP and workload at the first stage, and in the fourth and sixth minutes of recovery are significantly lower in normotensive male patients with coronary artery disease compared to those without CAD and can be independent predictive parameters for the disease.

CAR T cells targeting CD99 as an approach to eradicate T-cell acute lymphoblastic leukemia without normal blood cells toxicity

CAR T cell therapy has shown dramatic clinical success in relapsed or refractory B-ALL and other hematological malignancies. However, the loss of specific antigens, cell fratricide, T cell aplasia, and normal T cell separation are challenges in treating T cell leukemia/lymphoma with CAR T therapy. CD99 is a promising antigen to target T-ALL and AML as it is strongly expressed on the majority of T-ALL and AML. Here, we isolated a low-affinity CD99 (12E7) antibody, which specifically recognizes leukemia cells over normal blood cells. Moreover, T cells transduced with an anti-CD99-specific CAR that contained the 12E7 scFv expanded with minor fratricide and without normal blood cells toxicity. We observed that our anti-CD99 CAR T cells showed robust cytotoxicity specifically against CD99+ T-ALL cell lines and primary tumor cells in vitro and significantly prolonged cell line-derived xenografts (CDXs) or patient-derived xenografts (PDXs) models survival in vivo. Together, our results demonstrate that anti-CD99 CAR T cells could specifically recognize and efficiently eliminate CD99+ leukemia cells.