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Background Longitudinal serology studies can assist in analyzing kinetics of antibodies to the SARS-CoV-2 virus, helping to inform public health decision-making. Our study aims to characterize circulating antibody trends over 18 months in vaccinated participants with and without evidence of COVID-19 infection. Methods A cohort of healthcare workers employed at Boston Medical Center were followed, collecting serum samples and survey data over six timepoints from July 2020 through December 2021 (n = 527). History of SARS-CoV-2 infection, vaccination and booster status were confirmed, where possible, using electronic medical records. Serum was assessed for the qualitative detection of anti-N IgG and the semi-quantitative detection of anti-S IgG antibody levels. Piecewise regression models were utilized to characterize the antibody kinetics overtime. Results Anti-S IgG titers remained above the positivity threshold following infection and/or vaccination throughout the 18-month follow-up. Among participants with no evidence of COVID-19 infection, titers declined significantly faster in the initial 90 days post-full vaccination (β=-0.056) from December 2020 to March 2021 compared to the decline observed following booster dose uptake (β=-0.023) (p Conclusions These findings provide novel 18-month kinetics of anti-S IgG antibodies and further highlight the durability of hybrid immunity, underlining the strong humoral response stimulated by combined infection and vaccination.

Objective To examine if iron intake and genetically determined iron overload interact in predisposing to the development of childhood islet autoimmunity (IA) and type 1 diabetes (T1D). Research Design and Methods In “The Environmental Determinants of Diabetes in the Young” (TEDDY) study, 7,770 genetically high-risk children were followed from birth until the development of IA and progression to T1D. Exposures included energy-adjusted iron intake in the first 3 years of life and a genetic risk score (GRS) for increased circulating iron. Results We found a U-shaped association between iron intake and risk of GADA as first autoantibody. In subjects with GRS>=2 iron risk alleles, high iron intake was associated with increased risk of IA with insulin as first autoantibody [adjusted hazard ratio (HR, 95% CI): 1.71 (1.14; 2.58), as compared to moderate iron intake. Conclusions Iron intake may alter the risk of islet autoimmunity in children with high-risk HLA haplogenotypes.

The recently identified, globally predominant SARS-CoV-2 Omicron variant (BA.1) is highly transmissible, even in fully vaccinated individuals, and causes attenuated disease compared with other major viral variants recognized to date1–7. The Omicron spike (S) protein, with an unusually large number of mutations, is considered the major driver of these phenotypes3,8. We generated chimeric recombinant SARS-CoV-2 encoding the S gene of Omicron in the backbone of an ancestral SARS-CoV-2 isolate and compared this virus with the naturally circulating Omicron variant. The Omicron S-bearing virus robustly escapes vaccine-induced humoral immunity, mainly due to mutations in the receptor-binding motif (RBM), yet unlike naturally occurring Omicron, efficiently replicates in cell lines and primary-like distal lung cells. In K18-hACE2 mice, while Omicron causes mild, non-fatal infection, the Omicron S-carrying virus inflicts severe disease with a mortality rate of 80%. This indicates that while the vaccine escape of Omicron is defined by mutations in S, major determinants of viral pathogenicity reside outside of S.

Introduction Doctoral-level clinical chemists work in various settings including academia, healthcare systems, reference laboratories, and industry. Information regarding compensation and benefits for PhD board-certified clinical chemists is limited. The American Association for Clinical Chemistry’s (AACC) Society for Young Clinical Laboratorians (SYCL) Core Committee completed a compensation survey in February 2021 and compared that to previous survey results. Methods The 2021 salary survey was made available to all doctoral-level AACC members working in the United States and Canada. Respondents provided information on type of degree and board certification, years of experience, employment sector, geographic location, and gender. The survey collected confidential self-reported data from 208 respondents, with 187 (90%) respondents residing in the United States. Results The 25th percentile, median and 75th percentile total compensation range for 113 PhDrespondents who are certified by the American Board of Clinical Chemistry (ABCC) and residing in the United States was $150 000 to $159 000, $180 000 to $189 000, and $230 000 to $239 000, respectively. Most of the respondents worked in either an academic, hospital, or healthcare system setting, with 47% of respondents working at academic medical centers. Conclusions In this 2021 compensation survey, we continue to see an upward trend in compensation ranges for board certified PhD clinical chemists. This report is important in allowing individuals to actively advocate with prospective and current employers for equal and fair compensation as well as other benefits that impact career development and growth.

Background Boston Medical Center (BMC) is a safety net hospital in Boston, and from the initial wave of COVID-19 there has been an overwhelming concern about the exposure of healthcare workers (HCWs) to SARS-CoV-2. Methods We conceived a study to follow a cohort of BMC HCWs, beginning in July 2020 and continuing for 15 months, collecting survey data and serum samples at approximately 3-month intervals. Serum samples were analyzed using the Abbott Architect i2000 for SARS-CoV-2 antibodies (anti-spike1-Receptor Binding Domain IgG and anti-nucleoprotein IgG). Positive anti-n IgG results were used, in addition to reverse transcription-PCR results, for identifying cases of infection. History of COVID-19 and vaccination status were confirmed, where possible, using electronic medical records. Participants were grouped according to vaccination and infection status in September 2021 for analysis of anti-s IgG trends. Results A majority of HCWs remain well above the positivity threshold for anti-spike IgG antibodies for up to 11 months post-vaccination and 15 months post-infection, regardless of combinations and permutations of vaccination and infection. Those with COVID-19 infection before vaccination had significantly higher median serum antibody concentrations in comparison to HCWs with no prior infection at each follow-up time point. Conclusions These findings further support what is known regarding the decline in serum antibody concentrations following natural infection and vaccination, adding knowledge of serum antibody levels for up to 15 months post- infection and 11 months post-vaccination.

Over 15-months we found that anti-spike RBD SARS-CoV-2 antibody concentrations follow different trends with combinations and permutations of COVID-19 infection and vaccination among healthcare workers in Boston, MA. A majority of HCWs remain well above the positivity threshold for anti-spike RBD IgG antibodies for at least 9 months following vaccination regardless of infection history. Of interest, those with COVID-19 infection before vaccination had significantly higher median serum antibody concentrations in comparison to HCWs with no prior infection at each follow-up timepoint. These findings further support what is known regarding the decline in serum antibody concentrations following natural infection and vaccination, adding knowledge of serum antibodies up to 15 months post infection and 11 months post vaccination.ImportanceBoston Medical Center (BMC) is a safety net hospital in Boston and from the initial wave of COVID-19 there has been overwhelming concern about the exposure of healthcare workers to SARS-CoV-2. We conceived a longitudinal study to assess virus exposure and trends in SARS-CoV-2 antibodies amongst healthcare workers at BMC over 15 months. We have followed HCWs through three waves of COVID-19, including the Delta variant wave from June through mid-December 2021, assessing anti-spike receptor binding domain IgG, anti-nucleocapsid IgG, and anti-spike IgM at approximately three-month intervals. Current literature largely describes antibody durability six months post vaccination. These data add to the literature by describing antibody durability and trend differences according to infection history and vaccination status. These longitudinal data contribute to a greater understanding of the ongoing COVID-19 pandemic and can help inform future research and public health decision-making regarding vaccine uptake, breakthrough infections, and overall pandemic response.

BackgroundAccurate measurement of antibodies is a necessary tool for assessing exposure to SARS-CoV-2 and facilitating understanding of the role of antibodies in immunity. Most assays are qualitative in nature and employ a threshold to determine presence of antibodies. Semi-quantitative assays are now available. Here we evaluate the semi-quantitative SARS-CoV-2 IgG II (anti-spike (S)) assay. We aim to reassess the seroprevalence using anti-S assay and subsequently compare it to the previously measured IgG (anti-nucleoprotein (N)) in health care workers at an academic medical center in Boston.Methods1743 serum samples from HCWs at Boston Medical Center were analyzed for SARS-CoV-2 anti-S IgG and IgM using the Abbott SARS-CoV-2 IgG II and□Abbott AdviseDx□SARS-CoV-2 IgM assay, respectively. Precision, linearity, positive and negative concordance with prior RT-PCR test were evaluated for anti-S IgG. Seroprevalence and its association with demographics variables was also assessed.ResultsLinearity and precision results were clinically acceptable. The positive and negative concordance for anti-S IgG with RT-PCR was 88.2% (95% CI: 79.4% - 94.2%) and 97.43% (95% CI: 95.2% - 98.8%), respectively. Overall, 126 (7.2%) of 1,743 participants were positive by anti-S IgG. Among the 1302 participants with no prior RT-PCR, 40 (3.1%) were positive for anti-S IgG antibody. The original agreement in this population with the qualitative, anti-N IgG assay was 70.6%. Upon optimizing the threshold from 1.4 to 0.49 S/CO of the anti-N IgG assay, the positive agreement of the assay increases to 84.7%.ConclusionThe anti-S IgG assay demonstrated reproducible and reliable measurements. This study highlights the presence of asymptomatic transmission among individuals with no prior history of positive RT-PCR. It also highlights the need for optimizing thresholds of the qualitative SARS-CoV-2 IgG assay for better agreement between assays by the same vendor.

With obesity and type 2 diabetes prevalence steadily increasing and no effective means in sight to support the population in obtaining and maintaining stable weight loss, there is an imminent need for pharmacological therapy to treat and prevent type 2 diabetes. Current anti-diabetic treatment is symptomatic, and very few drugs have both a strong preclinical rationale and clinical proof-of-principle as therapies targeting pathogenic processes in type 2 diabetes. The emerging appreciation of low-grade inflammation as a significant cause of insulin resistance and beta cell failure warrants exploring anti-inflammatory compounds as drug candidates. Since recent studies have demonstrated considerable phenotypic heterogeneity in the type 2 diabetic syndrome, the concept of one drug fits all is naïve, and biomarkers for the selection of type 2 diabetes subtypes for differentiated treatment based on genetic and pathogenic stratification are urgently needed. Biologics antagonizing the master pro-inflammatory cytokine interleukin-1 is one of the few principles specifically targeting low-grade inflammation in type 2 diabetes. Although early phase II studies were encouraging, subsequent underpowered studies and phase III studies designed primarily with cardiovascular endpoints have discredited the potential of anti-interleukin-1 approaches to treat the subgroup of patients that may benefit from this treatment. In this meta-analysis of 2921 individuals from eight phase I-IV studies, we demonstrate a significant overall HbA1c-lowering effect of interleukin-1 antagonism. Meta-regression analyses demonstrated a significant correlation between baseline C-reactive protein and C-peptide, and HbA1c outcome. The identification of further biomarkers for future clinical trials to define the potential of anti-interleukin-1 therapies in type 2 diabetes is urgently needed.

Objectives Severe COVID-19 results in a glucocorticoid responsive form of acute respiratory distress (ARDS)/diffuse alveolar damage (DAD). Herein we compare the immunopathology of lung tissue procured at autopsy in patients dying of SARS-CoV-2 with those dying of DAD prior to the COVID-19 pandemic. Methods Autopsy gross and microscopic features stratified by duration of illness in twelve patients who tested positive for SARS-CoV-2 viral RNA as well as seven patients dying of DAD prior to the COVID-19 pandemic were evaluated with multiplex (5-plex: CD4, CD8, CD68, CD20, AE1/AE3) and SARS-CoV immunohistochemistry to characterize the immunopathologic stages of DAD. Results We observed a distinctive pseudopalisaded histiocytic hyperplasia interposed between the exudative and proliferative phase of COVID-19 associated DAD which was most pronounced at the fourth week from symptom onset. Pulmonary macrothrombi were seen predominantly in cases with pseudopalisaded histiocytic hyperplasia and/or proliferative phase DAD. Neither pseudopalisaded histiocytic hyperplasia nor pulmonary macrothrombi were seen in non-COVID-19 DAD cases, whereas microthrombi were common in DAD regardless of etiology. Conclusion The inflammatory pattern of pseudopalisaded histiocytic hyperplasia may represent the distinctive immunopathology associated with the dexamethasone responsive form of DAD seen in severe COVID-19.

Increasing evidence suggests that autoimmunity may play a role in the pathophysiology of SARS-CoV-2 infection during both the acute and ‘long COVID’ phases of disease. However, an assessment of autoimmune antibodies in convalescent SARS-CoV-2 patients has not yet been reported.MethodologyWe compared the levels of 18 different IgG autoantibodies (AABs) between four groups: (1) unexposed pre-pandemic subjects from the general population (n = 29); (2) individuals hospitalized with acute moderate-severe COVID-19 (n = 20); (3) convalescent SARS-COV-2-infected subjects with asymptomatic to mild viral symptoms during the acute phase with samples obtained between 1.8 and 7.3 months after infection (n = 9); and (4) unexposed pre-pandemic subjects with systemic lupus erythematous (SLE) (n = 6). Total IgG and IgA levels were also measured from subjects in groups 1-3 to assess non-specific pan-B cell activation.ResultsAs expected, in multivariate analysis, AABs were detected at much higher odds in SLE subjects (5 of 6, 83%) compared to non-SLE pre-pandemic controls (11 of 29, 38%) [odds ratio (OR) 19.4,95% CI, 2.0 – 557.0, p = 0.03]. AAB detection (percentage of subjects with one or more autoantibodies) was higher in SARS-CoV-2 infected convalescent subjects (7 of 9, 78%) [OR 17.4, 95% CI, 2.0 – 287.4, p = 0.02] and subjects with acute COVID-19 (12 of 20, 60%) compared with non-SLE pre-pandemic controls, but was not statistically significant among the latter [OR 1.8,95% CI, 0.6 – 8.1, p = 0.23]. Within the convalescent subject group, AABs were detected in 5/5 with reported persistent symptoms and 2/4 without continued symptoms (p = 0.17). The multivariate computational algorithm Partial Least Squares Determinant Analysis (PLSDA) was used to determine if distinct AAB signatures distinguish subject groups 1-3. Of the 18 autoantibodies measured, anti-Beta 2-Glycoprotein, anti-Proteinase 3-ANCA, anti-Mi-2 and anti-PM/Scl-100 defined the convalescent group; anti-Proteinase 3-ANCA, anti-Mi-2, anti-Jo-1 and anti-RNP/SM defined acute COVID-19 subjects; and anti-Proteinase 3-ANCA, anti-Mi-2, anti-Jo-1, anti-Beta 2-Glycoprotein distinguished unexposed controls. The AABs defining SARS-COV-2 infected from pre-pandemic subjects are widely associated with myopathies, vasculitis, and antiphospholipid syndromes, conditions with some similarities to COVID-19. Compared to pre-pandemic non-SLE controls, subjects with acute COVID-19 had higher total IgG concentration (p-value=0.006) but convalescent subjects did not (p-value=0.08); no differences in total IgA levels were found between groups.ConclusionsOur findings support existing studies suggesting induction of immune responses to self-epitopes during acute, severe COVID-19 with evidence of general B cell hyperactivation. Also, the preponderance of AAB positivity among convalescent individuals up to seven months after infection indicates potential initiation or proliferation, and then persistence of self-reactive immunity without severe initial disease. These results underscore the importance of further investigation of autoimmunity during SARS-CoV-2 infection and its role in the onset and persistence of post-acute sequelae of COVID-19.

Healthcare workers (HCWs) are at an increased risk of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a novel virus that causes Coronavirus Disease (COVID-19). We aim to assess the seroprevalence of SARS-CoV-2 IgG among healthcare workers and compare risk-factors between seropositive and seronegative HCWs. In this observational study, serum samples were collected from HCWs between July 13th to 26th, 2020 at Boston Medical Center (BMC). Samples were subsequently tested for SARS-CoV-2 IgG antibody using the Abbott SARS-CoV-2 IgG assay. Participants also answered a questionnaire capturing data on demographics, history of COVID-19 symptoms, occupation, infection prevention and control measures. Overall, 95 of 1743 (5.5%) participants tested positive for SARS-CoV-2 IgG. Of these, 1.8% of the participants had mild or no COVID-19 symptoms and did not require a diagnostic test. Seropositivity was not associated with gender, occupation, hand hygiene and personal protective equipment (PPE) practices amongst HCWs. However, lack of physical distancing among health care workers in work areas and break room was associated with seropositivity (p = 0.05, p = 0.003, respectively). The majority of the HCWs are negative for SARS-CoV-2 IgG. This data highlights the need to promote infection prevention measures, and the importance of distance amongst co-workers to help mitigate infection rates.

The COVID-19 pandemic has significantly impacted work, economy, and way of life. The SARS-CoV-2 virus displays unique features including widely varying symptoms and outcomes between infected individuals. Sensitive measurement of SARS-CoV-2 specific antibodies would provide new insight into virus transmission dynamics, pre-existing cross-reactive immunity, and the nuances of SARS-CoV-2 pathogenesis. To date, existing SARS-CoV-2 serology tests have limited utility due to insufficient detection of antibody levels lower than what is typically present after several days of symptoms. To measure lower quantities of SARS-CoV-2 IgM, IgG, and IgA with higher resolution than existing assays, we developed a new ELISA protocol with a distinct plate washing procedure and timed plate development via use of a standard curve. This ‘BU ELISA’ method exhibits very low signal from plasma or serum samples added to uncoated wells at as low as a 1:5 dilution. Use of this method revealed circulating SARS-CoV-2 receptor binding domain (RBD) and nucleocapsid protein (NP) reactive antibodies from blood samples drawn prior to May 2019. Of our prepandemic cohort, no SARS-CoV-2 RBD-reactive IgG antibodies were detected in subjects over 70 years of age, and SARS-CoV-2 NP-reactive antibodies were present at similar levels to infected subjects in some individuals and very low in others. Also, samples drawn in May 2020 from two individuals with no symptoms or no known virus exposure contained SARS-CoV-2 RBD-reactive antibodies at intermediate amounts compared with other subject groups (higher than pre-pandemic and lower than confirmed SARS-CoV-2 infected). The one asymptomatic SARS-CoV-2 convalescent subject in our study possessed comparable amounts of SARS-CoV-2 NP-specific IgM and IgG but drastically lower IgA than the symptomatic counterparts. Also, our assay detected positive signal from samples that gave negative results in a commercially available Lateral Flow Device (LFD) and the EUA approved Abbott IgG chemiluminescent microparticle immunoassay for SARS-CoV-2 antibody detection. We propose that this improved ELISA protocol, which is straightforward to perform, low cost, and uses readily available commercial reagents, is a useful tool to elucidate new information about SARS-CoV-2 infection and has promising implications for improved detection of all analytes measurable by this platform.

BackgroundA subset of COVID-19 patients exhibit clinical features of cytokine storm. However, clinicopathologic features diagnostic of hemophagocytic lymphohistiocytosis (HLH) have not been reported. Pathologic studies to date have largely focused on the pulmonary finding of diffuse alveolar damage (DAD). To this aim, we study the reticuloendothelial organs of four consecutive patients dying of COVID-19 and correlate with clinical and laboratory parameters to detect HLH.MethodsAutopsies restricted to chest and abdomen were performed on four patients who succumbed to COVID-19. Spleen, liver, and multiple pulmonary hilar/mediastinal lymph nodes were sampled in all cases. Bone marrow was obtained by rib squeeze in a subset of cases. Routine H&E staining as well as immunohistochemical staining for CD163 was performed to detect hemophagocytosis. Clinical and laboratory results from pre-mortem blood samples were used to calculate H-scores.FindingsAll four cases demonstrated DAD within the lungs. Three of the four cases had histologic evidence of hemophagocytosis within pulmonary hilar/mediastinal lymph nodes. One case showed hemophagocytosis in the spleen but none showed hemophagocytosis in liver or bone marrow. Lymphophagocytosis was the predominant form of hemophagocytosis observed. One patient showed diagnostic features of HLH with an H-score of 217 while a second patient was likely HLH with a partial H-score of 145 due to missing triglyceride level. Both patients exhibited high fever and early onset rise in serum ferritin; however, neither bicytopenia, pancytopenia, nor hypofibrinogenemia were observed in either. The remaining two patients had H-scores of 131 and 96.InterpretationThis is the first report of SARS-CoV-2 associated HLH. Identification of HLH in a subset of patients with severe COVID-19 will inform clinical trials of therapeutic strategies.

Background Urine immunoassays are frequently employed for methadone screening because they are relatively inexpensive and widely available. However, immunoassays are notoriously prone to false positives. We report that the use of vortioxetine (Trintellix® in the USA and Canada, Brintellix® worldwide) could cause false positives in the Roche KIMS Methadone II Urine immunoassay (MDN2). Methods We performed a spiking study using a parent drug vortioxetine concentration of 7500 ng/ml. Results Urine specimens from seven patients on typical vortioxetine doses tested positive for methadone in the Roche assay but negative for methadone in a confirmatory (GC/MS) assay and two other immunoassay platforms. Because of the pharmacokinetics of vortioxetine and the high cross-reactivity of a metabolite in the MDN2 assay, routine use of the drug could cause false positives even without detectible parent drug in the urine. Conclusions Vortioxetine is commonly prescribed for mood disorders, which have high prevalence in patients treated for opioid addiction. For that reason, it is important that clinicians are aware of this interference.

Background SARS-CoV-2 continues to spread globally, including in limited resource settings. It is therefore important to derive general case definitions that can be useful and accurate in the absence of timely test results. We aim to validate the World Health Organization (WHO) case definition, a symptom-screening tool currently used to identify SARS-CoV-2 cases in a cohort of symptomatic health care providers (HCP) who completed a symptom survey interview and received a PCR test at Boston Medical Center (BMC) between March 13, 2020 and May 5, 2020. Methods We classified each HCP as a probable or not probable case of SARS-CoV-2 based on the WHO case definition. Using PCR test as gold standard, we computed the sensitivity and specificity of the WHO case definition. We used a stepwise logistic regression model on all PCR-tested HCP to identify symptoms predictive of PCR positivity. Results Of 328 included HCP, 109 (33.2%) were PCR positive, 213 (64.9%) negative, and 6 (1.8%) had indeterminate test result. The sensitivity and specificity of the WHO case definition were 65.1% and 74.6%, respectively. The positive predictive value was 56.8% and the negative predictive value was 80.7%. Symptoms found to be predictive of PCR positivity were fever, headache, loss of smell and/or loss of taste, and muscle ache/joint pain. Sore throat was found to be predictive of PCR negativity. The area under the curve using the final model was 0.8412. All statistically significant symptoms included in the final model, were also included in the WHO case definition. Conclusion In our largely symptomatic HCP cohort, our model yielded similar symptoms to those identified in the WHO probable case definition. As seen in similar studies, it is unlikely that further adjustment will improve the performance of a SARS-CoV-2 case definition. However, it is concerning that 35% (38/109) of PCR positive SARS-CoV-2 HCP would have been classified as not probable cases by the WHO definition, given that this definition does not even include asymptomatic cases. This is further evidence for global building of laboratory capacity and development of affordable diagnostics to improve global pandemic control. Disclosures All Authors: No reported disclosures

Additional file 20: Table S3. Newcastle-Ottawa quality assessment (DOCX 62 kb)

A meta-analysis of the association of iron overload with gestational diabetes mellitus (GDM) may inform the health debate. We performed a meta-analysis investigating the association of iron biomarkers and dietary iron exposure with GDM. We identified 33 eligible studies (N = 44,110) published in 2001-2017. The standardized mean differences (SMD) in women who had GDM compared to pregnant women without were 0.25 mu g/dL (95% CI: 0.001-0.50) for iron, 1.54 ng/mL (0.56-2.53) for ferritin, 1.05% (0.02 to 2.08) for transferrin saturation, and 0.81 g/dL (0.40-1.22) for hemoglobin. Adjusted odds ratio for GDM were 1.58 (95% CI: 1.20-2.08) for ferritin, 1.30 (1.01-1.67) for hemoglobin, and 1.48 (1.29-1.69) for dietary heme intake. We did not find any differences in TIBC or transferrin concentration in women with and without GDM. We also did not find any association of increased transferrin receptor or increased intake of total dietary iron, non-heme iron or supplemental iron, with increased odds ratios for GDM. Considerable heterogeneity was present among the studies (0-99%), but no evidence of publication bias. Accumulating evidence suggests that circulating and dietary iron biomarkers among pregnant women are associated with GDM, but the results should be interpreted with caution due to the high heterogeneity of analyses. Randomized trials investigating the benefits of iron reduction in women at high risk for GDM are warranted.

Prenatal smoking exposure is associated with obesity and other cardio-metabolic risk factors in children, but no previous meta-analysis has been conducted in adults. We investigated the association of prenatal smoking exposure in the Danish General Suburban Population Study (GESUS) with BMI, waist circumference, total cholesterol, type 2 diabetes, gestational type 2 diabetes, and hypertension in adulthood. We subsequently performed a meta-analysis, adding published studies investigating the association between prenatal smoking and the risk of cardio-metabolic outcomes among individuals at least 18 years of age. We included 19 eligible observational studies with various cardio-metabolic outcomes (N = 24,201–308,981 adults). In individuals exposed to prenatal smoking, the pooled random effects adjusted odds ratio were 1.35 (95% CI: 1.16–1.56) for being overweight, 1.46 (1.39–1.54) for being obese, 1.07 (0.89–1.29) for type 2 diabetes, 1.17 (0.92–1.48) for hypertension, and 1.38 (1.19–1.61) for gestational diabetes mellitus (GDM), compared with no exposure. The standardized means in waist circumference, total cholesterol, diastolic, and systolic blood pressure were not different in individuals exposed vs. not exposed to prenatal smoking. Heterogeneity was moderate to high (51%

BACKGROUND Alpha-methylacyl-CoA racemase (AMACR) is an enzyme involved in fatty acid metabolism that is markedly over-expressed in virtually all prostate cancers (PCa), relative to benign tissue. One of AMACR's primary substrates, phytanic acid, is derived predominately from red meat and dairy product consumption. Epidemiological evidence suggests links between dairy/red meat intake, as well as phytanic acid levels, and elevated PCa risk. This study investigates the relationships among dietary intake, serum and tissue concentrations of phytanic acid, and AMACR expression (mRNA and protein) in the histologically benign human prostate. METHODS Men undergoing radical prostatectomy for the treatment of localized disease provided a food frequency questionnaire (n = 68), fasting blood (n = 35), benign fresh frozen prostate tissue (n = 26), and formalin-fixed paraffin-embedded (FFPE) sections (n = 67). Serum and tissue phytanic acid concentrations were obtained by gas chromatography–mass spectrometry. We extracted RNA from epithelial cells using laser capture microdissection and quantified mRNA expression of AMACR and other genes involved in the peroxisomal phytanic acid metabolism pathway via qRT-PCR. Immunohistochemistry for AMACR was performed on FFPE sections and subsequently quantified via digital image analysis. Associations between diet, serum, and tissue phytanic acid levels, as well as AMACR and other gene expression levels were assessed by partial Spearman correlation coefficients. RESULTS High-fat dairy intake was the strongest predictor of circulating phytanic acid concentrations (r = 0.35, P = 0.04). Tissue phytanic acid concentrations were not associated with any dietary sources and were only weakly correlated with serum levels (r = 0.29, P = 0.15). AMACR gene expression was not associated with serum phytanic acid (r = 0.13, P = 0.47), prostatic phytanic acid concentrations (r = 0.03, P = 0.88), or AMACR protein expression (r = −0.16, P = 0.20). CONCLUSIONS Our data underscore the complexity of the relationship between AMACR and its substrates and do not support the unifying hypothesis that excess levels of dietary phytanic acid are responsible for both the overexpression of AMACR in prostate cancer and the potential association between PCa risk and intake of dairy foods and red meat. Prostate 75:200–210, 2015. © 2014 Wiley Periodicals, Inc.

A 17-year-old boy with a past medical history of substance abuse (marijuana and alcohol) was presented to the emergency department with extreme agitation and suicidal statements. His mother claimed he consumed 4 glasses of hard liquor with caffeine shortly before presenting to the hospital. His urine toxicology screen was negative for amphetamines, barbiturates, benzodiazepines, cocaine, opiates, and tricyclic antidepressants. However, his urine toxicology screen was positive for cannabinoids. Other laboratory results …

Approximately 2 years ago, I started my training in clinical chemistry at Boston Children's Hospital. A colleague at the hospital once said something that will always stay with me: “let the data tell you.” With a steep learning curve ahead of me and a finite number of hours in a day, I found myself getting up at 5:30 am to go to the gym to have the only hour of sanity throughout my day. As an avid long distance runner, I use my FiBit HR to monitor my heart rate to obsessively assess my progress. After the morning respite, I usually have challenging days on the mass spectrometer …

A technologist questioned an undetectable glucose concentration in an arterial blood gas sample. Investigation revealed that it was a penile sample collected from a 31-year-old man with erectile dysfunction. The patient presented to the emergency department with priapism of 72-h duration. A penile blood gas sample was subsequently sent to the lab (sample 1). Following phenylephrine treatment, additional penile blood gas …

The prostate gland contains a heterogeneous milieu of stromal, epithelial, neuroendocrine and immune cell types. Healthy prostate is comprised of fibromuscular stroma surrounding discrete epithelial-lined secretory lumens and a very small population of immune and neuroendocrine cells. In contrast, areas of prostate cancer have increased dysplastic luminal epithelium with greatly reduced or absent stromal population. Given the profound difference between stromal and epithelial cell types, it is imperative to separate the cell types for any type of downstream molecular analysis. Despite this knowledge, the bulk of gene expression studies compare benign prostate to cancer without micro-dissection, leading to stromal bias in the benign samples. Laser-capture micro-dissection (LCM) is an effective method to physically separate different cell types from a specimen section. The goal of this protocol is to show that RNA can be successfully isolated from LCM-collected human prostatic epithelium and used for downstream gene expression studies such as RT-qPCR and RNAseq.

Hypertension is a well-established risk factor for cardiovascular disease (CVD).2 Assessment of lipids, physical activity, alcohol consumption, and other CVD risk factors in this patient population is an essential component of disease management. The use of nonfasting samples for lipid panels has been a matter of ongoing debate. In 2009, Denmark was the first to use nonfasting lipid panels for routine testing based on recommendations from the Danish Society for Clinical Biochemistry. Although the 2013 American College of Cardiology/American Heart Association guidelines preferred fasting samples for routine lipid panels, the choice of specimen type was left to the clinician's discretion. Recently, the European Atherosclerosis Society (EAS) and European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) made a strong …

Monitoring for metabolic sequelae of antipsychotic medications is inconsistent in clinical settings. In this study, frequency of such monitoring in 40 individuals experiencing a first episode of psychosis was analyzed according to the setting in which they received treatment (i.e., inpatient unit, outpatient clinic, or metabolic screening clinic). The traditional outpatient clinic was the least likely of the three settings to monitor blood glucose, blood pressure, weight, and waist circumference. In any setting, blood lipids were measured in only 2 of the 40 patients. Reasons for these findings and recommendations for reducing barriers to screening are presented.

Approximately 3.2 million Americans are chronically infected with Hepatitis C, forming the largest group of persons at high risk for HCC, with an annual incidence of 3-4%. Increased oxidative stress is regarded as the major mechanism of progression in chronic liver disease (CLD) and subsequent HCC. Hepatic iron overload facilitates the production of reactive oxygen species (ROS), thereby inducing liver damage and fibrosis. We have previously shown that retinoid and carotenoid depletion occurs early in disease and may represent a major modifiable risk. This study aims to determine, in CLD patients and controls, whether retinoid levels modify the association between serum iron concentrations and: (a) oxidative stress biomarkers in urine (b) tissue changes associated with hepatic stellate cell (HSC) activation, and (c) CLD progression. Patients with HCV undergoing liver biopsy (n = 69) provided fasting blood, fresh tissue, urine and completed a diet history questionnaire (DHQ). Serum and questionnaire data from healthy volunteers (n = 11), normal liver tissue from public repositories and patients without liver disease (n = 11) were also collected. Urinary isoprostanes, serum and tissue retinoid concentrations were obtained by LC-MS-MS. Serum iron, ferritin and transferrin were quantitated. Immunohistochemistry for αSMA was performed on FFPE sections and subsequently quantified via digital image analysis. Associations between urinary isoprostanes, αSMA levels, serum iron, serum and tissue retinoids were assessed using Spearman correlation coefficients and non-parametric tests were utilized to test differences among disease severity groups. There was a significant increasing trend for serum transferrin levels with progressive fibrosis, but not for serum iron and ferritin concentrations. All subjects had transferrin saturation within the normal range. Serum β-carotene and lycopene were inversely associated with serum ferritin concentrations (r = -0.29, P < 0.01; r = -0.29, P < 0.01) however, this relationship was not observed for other serum iron measurements. Serum iron measurements were not associated with hepatic retinoids/carotenoids, αSMA expression or urinary isoprostanes. Tissue retinyl palmitate was inversely and significantly correlated with hepatic αSMA expression (r = -0.3, P < 0.02). Serum iron overload was not evident in this population even though a depletion of serum retinoids/carotenoids is observed. The inverse correlation observed between serum ferritin and retinoids/carotenoids suggests a possible joint effect in CLD patients. Our data provide confirmation that hepatic retinoid loss is accompanied by HSC activation, a key player in fibrosis. Further evaluation of hepatic iron status is warranted to assess the relationships between iron and retinoids in HCV. Citation Format: Yachana Kataria, Ryan Deaton, Erika Enk, Milita Petrauskaite, Scott Cotler, Donald Jensen, Peter Gann. Joint relationship between iron and retinoids in patients at high-risk for liver cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2821. doi:10.1158/1538-7445.AM2015-2821

Approximately 3.2 million Americans are chronically infected with hepatitis C (HCV), forming the largest group at high-risk for hepatocellular carcinoma (HCC). Oxidative stress due to chronic inflammation impels the progression of chronic liver disease (CLD) & subsequent HCC. Deficiencies in dietary antioxidants such as retinoids and carotenoids may represent a modifiable risk factor for HCC progression. This study aims to identify key predictors of serum antioxidant levels in patients with CLD, to quantify the association between systemic measures of oxidative stress and antioxidant status, and to examine the relationship between retinoid/carotenoid concentrations in serum and hepatic tissue. Patients with HCV undergoing liver biopsy (n=69) provided fasting blood, fresh tissue, urine, and responses to a diet history questionnaire (DHQ). We also collected serum and questionnaire data from healthy volunteers (n=11), and normal liver tissue from public repositories and patients without liver disease (n=11). Serum and tissue retinoid concentrations were obtained by mass spectroscopy. Serum retinol binding protein 4 (RBP4) was quantified by ELISA. Urine 8OHdG and F2α-isoprostanes were determined by mass spectroscopy. Dietary retinoid intake was estimated by DHQ. Associations between diet, serum and tissue retinoids, and urinary oxidative stress markers were assessed by Spearman correlation coefficients. Dietary β-carotene and lutein were associated with their respective serum concentrations (r = 0.24, p = 0.05; r = 0.33, p < 0.01). There was a significant downward trend for serum retinol, lycopene and RBP4 concentrations with progressive fibrosis; mean serum concentrations were lower in subjects with chronic HCV but no fibrosis compared to controls. Urinary isoprostanes levels were inversely correlated with serum retinol (r= -0.22, p = 0.05), β-carotene (r= -0.22 p = 0.05), and RBP4 (r= -0.25 p = 0.03). Urinary 8OHDG did not correlate with any serum retinoids or carotenoids. Serum β-carotene and lycopene were strongly associated with the respective tissue concentrations (r = 0.56, p Depletion of serum retinol, β-carotene, and RBP4 worsens with liver fibrosis severity. RBP4 might be a better marker for vitamin A status due to its stronger association with disease severity. Retinoid and carotenoid levels decline as disease progresses, and our data suggest that this decline occurs early in the disease process, even before fibrosis is apparent. In HCV patients, decreases in retinoid/carotenoid levels are accompanied by an increase in urinary F2α-isoprostanes, an indicator of lipid peroxidation due to oxidative stress. Dietary or chemopreventive strategies for replenishing antioxidant levels should be studied further. Citation Format: Yachana Kataria, Erika Enk Rueter, Ryan Deaton, Donald Jensen, Scott Cotler, Peter Gann. Retinoid and carotenoid depletion in patients at high-risk for liver cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2144. doi:10.1158/1538-7445.AM2014-2144

Alpha-methylacyl-CoA racemase (AMACR) is an enzyme that is overexpressed in prostate cancer, with lower levels present in normal prostatic tissues. AMACR is involved in the metabolism of phytanic acid - a branched chain saturated fatty acid that has been linked with elevated risks of prostate cancer in several epidemiologic studies. Although considerable research has been done on AMACR as a clinical biomarker in prostate cancer, no study has evaluated the possible link between phytanic acid and AMACR gene expression in the normal human prostate, and thus the possibility that AMACR plays a causal role in prostate carcinogenesis. Fasting blood and histologcally benign prostate tissue samples were obtained from 31 men undergoing radical prostatectomy for the treatment of localized disease. Concentrations of phytanic acid in serum and tissues were determined by isotope dilution gas chromatography-mass spectrometry. RNA was extracted from epithelial cells isolated from fresh-frozen prostate tissues by laser capture microdissection, and AMACR mRNA levels were quantified using real time polymerase chain reaction. Tissue phytanic acid concentrations were not significantly correlated with serum levels nor with prostatic AMACR mRNA levels. Notably, AMACR was significantly inversely correlated with body mass index (Spearman correlation coefficient (r)= –0.43, p=0.02) and directly correlated with Gleason score (r=0.35, p=0.05). Stratification by body mass index revealed an unexpected inverse correlation between wAMACR gene expression levels and phytanic acid concentrations in the prostate (r= –0.95, p= 0.001) in obese men. In conclusion, blood and tissue levels of phytanic acid were not correlated, which suggests that blood measures should not be utilized as a surrogate measure of this fatty acid in the target organ. Contrary to expectation, tissue phytanic acid levels were significantly inversely associated with AMACR gene expression levels in the prostate, but only among obese men.These findings require confirmation in other, larger studies. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4480. doi:1538-7445.AM2012-4480

Clinical assay results for the study population by fibrosis stage - median (interquartile range). (DOCX 38 kb)

Clinical assay results for the study population by fibrosis stage - median (interquartile range). (DOCX 38 kb)