Spike and nsp6 are key determinants of SARS-CoV-2 Omicron BA.1 attenuation.

The SARS-CoV-2 Omicron variant is more immune evasive and less virulent than other major viral variants that have so far been recognized1-12. The Omicron spike (S) protein, which has an unusually large number of mutations, is considered to be the main driver of these phenotypes. Here we generated chimeric recombinant SARS-CoV-2 encoding the S gene of Omicron (BA.1 lineage) 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 escaped vaccine-induced humoral immunity, mainly owing to mutations in the receptor-binding motif; however, unlike naturally occurring Omicron, it efficiently replicated in cell lines and primary-like distal lung cells. Similarly, in K18-hACE2 mice, although virus bearing Omicron S caused less severe disease than the ancestral virus, its virulence was not attenuated to the level of Omicron. Further investigation showed that mutating non-structural protein 6 (nsp6) in addition to the S protein was sufficient to recapitulate the attenuated phenotype of Omicron. This indicates that although the vaccine escape of Omicron is driven by mutations in S, the pathogenicity of Omicron is determined by mutations both in and outside of the S protein.

Body size and risk of multiple myeloma in the Black Women's Health Study.

BACKGROUND: Obesity is an established risk factor for multiple myeloma (MM). Relatively few prior studies, however, have evaluated associations in Black populations. METHODS: Among 55,276 participants in the Black Women's Health Study, a prospective U.S. cohort established in 1995, we confirmed 292 incident diagnoses of MM over 26 years of follow-up. Multivariable Cox proportional hazard models, adjusted for age and putative MM risk factors, were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for associations of usual body mass index (BMI), BMI at age 18, height, and waist-to-hip ratio with MM. RESULTS: Compared to women with a usual adult BMI < 25 kg/m2, the HR associated with a usual adult BMI ≥ 35 kg/m2 was 1.38 (95% CI: 0.96, 1.98). For early adult BMI, the HR comparing women with BMI ≥ 25 vs. <25 kg/m2 was 1.57 (95% CI: 1.08, 2.28). Women who were heavy in both early and later life had the highest risk compared to those who were lean at both time points (HR: 1.60; 95% CI: 1.02, 2.52). Height was also associated with the risk of MM; the HR per 10 cm was 1.21 (95% CI: 1.02, 1.43). CONCLUSIONS: These results indicate that high early adult BMI is associated with a 57% increased risk of MM in Black women and potentially highlight the importance of weight control as a preventive measure.

Vitamin D and monoclonal gammopathy of undetermined significance (MGUS) among U.S. Black women.

PURPOSE: Risk factors for monoclonal gammopathy of undetermined significance (MGUS), the asymptomatic precursor to multiple myeloma, are largely unknown. We hypothesized that low vitamin D levels might be associated with higher MGUS prevalence in a national cohort of U.S. Black women. METHODS: We screened archived serum samples (collected 2014-2017) from 3896 randomly selected participants in the Black Women's Health Study ages 50-79 for evidence of MGUS; samples had been assayed for 25-hydroxyvitamin D [25(OH)D] shortly after blood draw. We used logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI) for the association between 25(OH)D level and MGUS status, adjusting for age, body mass index, and season of blood draw. RESULTS: We identified 334 MGUS cases (8.6%) in the study population. The adjusted OR comparing women with vitamin D deficiency (25(OH)D < 20 ng/mL) to those with 25(OH)D levels ≥ 30 ng/mL was 1.27 (95% CI: 0.95, 1.72). CONCLUSION: MGUS was more prevalent among Black women with vitamin D deficiency compared to those with 25(OH)D ≥ 30 ng/mL; however, the association was not statistically significant. Future prospective studies are warranted to clarify the possible association between vitamin D and MGUS.

Prenatal smoking exposure and cardio-metabolic risk factors in adulthood: a general population study and a meta-analysis.

OBJECTIVE: 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. METHODS: 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. RESULTS: 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% < I2 < 99%). However, removal of the high heterogeneity removed the associated uncertainty in the point estimate and revealed that prenatal smoking is associated with increased BMI in adulthood. There was also no evidence of publication bias in the meta-analyses. CONCLUSIONS: The findings from the meta-analyses suggested that prenatal smoking exposure is associated with an increased odds ratio of overweight, obesity, and GDM in adulthood, but not with type 2 diabetes, hypertension, waist circumference, or total cholesterol. These findings highlight the importance of abstaining from smoking by pregnant women.

Factors associated with successful vaginal birth after a cesarean section: a systematic review and meta-analysis.

BACKGROUND: Evidence for the relationship between maternal and perinatal factors and the success of vaginal birth after cesarean section (VBAC) is conflicting. We aimed to systematically analyze published data on maternal and fetal factors for successful VBAC. METHODS: A comprehensive search of Medline, Embase, and the Cumulative Index to Nursing and Allied Health Literature, from each database's inception to March 16, 2018. Observational studies, identifying women with a trial of labor after one previous low-transverse cesarean section were included. Two reviewers independently abstracted the data. Meta-analysis was performed using the random-effects model. Risk of bias was assessed by the Newcastle-Ottawa Scale. RESULTS: We included 94 eligible observational studies (239,006 pregnant women with 163,502 VBAC). Factors were associated with successful VBAC with the following odds ratios (OR;95%CI): age (0.92;0.86-0.98), obesity (0.50;0.39-0.64), diabetes (0.50;0.42-0.60), hypertensive disorders complicating pregnancy (HDCP) (0.54;0.44-0.67), Bishop score (3.77;2.17-6.53), labor induction (0.58;0.50-0.67), macrosomia (0.56;0.50-0.64), white race (1.39;1.26-1.54), previous vaginal birth before cesarean section (3.14;2.62-3.77), previous VBAC (4.71;4.33-5.12), the indications for the previous cesarean section (cephalopelvic disproportion (0.54;0.36-0.80), dystocia or failure to progress (0.54;0.41-0.70), failed induction (0.56;0.37-0.85), and fetal malpresentation (1.66;1.38-2.01)). Adjusted ORs were similar. CONCLUSIONS: Diabetes, HDCP, Bishop score, labor induction, macrosomia, age, obesity, previous vaginal birth, and the indications for the previous CS should be considered as the factors affecting the success of VBAC.

Retinoid and carotenoid status in serum and liver among patients at high-risk for liver cancer.

BACKGROUND: Approximately 2.7 million Americans are chronically infected with hepatitis C virus (HCV). HCV patients with cirrhosis form the largest group of persons at high risk for hepatocellular carcinoma (HCC). Increased oxidative stress is regarded as a major mechanism of HCV-related liver disease progression. Deficiencies in retinoid and carotenoid antioxidants may represent a major modifiable risk factor for disease progression. This study aims to identify key predictors of serum antioxidant levels in patients with HCV, to examine the relationship between retinoid/carotenoid concentrations in serum and hepatic tissue, to quantify the association between systemic measures of oxidative stress and antioxidant status, and to examine the relationship between retinoids and stellate cell activation. METHODS: Patients undergoing liver biopsy (n = 69) provided fasting blood, fresh tissue, urine and completed a diet history questionnaire. 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 UHPLC-MS-MS. Immunohistochemistry for αSMA was performed on FFPE sections and subsequently quantified via digital image analysis. Associations between urinary isoprostanes, αSMA levels, and retinoids were assessed using Spearman correlation coefficients and non-parametric tests were utilized to test differences among disease severity groups. RESULTS: There was a significant inverse association between serum retinol, lycopene, and RBP4 concentrations with fibrosis stage. Serum ß-carotene and lycopene were strongly associated with their respective tissue concentrations. There was a weak downward trend of tissue retinyl palmitate with increasing fibrosis stage. Tissue retinyl palmitate was inversely and significantly correlated with hepatic αSMA expression, a marker for hepatic stellate cell activation (r = -0.31, P < 0.02). Urinary isoprostanes levels were inversely correlated with serum retinol, ß-carotene, and RBP4. CONCLUSIONS: A decrease in serum retinol, ß-carotene, and RBP4 is associated with early stage HCV. 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. Measures of oxidative stress are associated with fibrosis stage and concurrent antioxidant depletion. Vitamin A loss is accompanied by stellate cell activation in hepatic tissue.

Dietary influences on tissue concentrations of phytanic acid and AMACR expression in the benign human prostate.

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.

Kinetics of SARS-CoV-2 Serum Antibodies Through the Alpha, Delta, and Omicron Surges Among Vaccinated Health Care Workers at a Boston Hospital.

Background: Longitudinal serology studies can assist in analyzing the kinetics of antibodies to SARS-CoV-2, 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 health care workers employed at Boston Medical Center was followed to collect serum samples and survey data over 6 time points from July 2020 through December 2021 (N = 527). History of SARS-CoV-2 infection, vaccination, and booster status were confirmed, where possible, through electronic medical records. Serum was assessed for the qualitative and semiquantitative detection of IgG antibody levels (anti-nucleoprotein [anti-N] and anti-spike [anti-S], respectively). Piecewise regression models were utilized to characterize antibody kinetics over time. 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 after full vaccination (ß = -0.056) from December 2020 to March 2021 as compared with the decline observed following booster dose uptake (ß = -0.023, P < 0.001). Additionally, COVID-19 infection prior to vaccination significantly attenuated the decline of anti-S IgG when compared with no infection following vaccine uptake (P < 0.001). Lastly, fewer participants contracted Omicron when boosted (12.7%) compared to fully vaccinated (17.6%). Regardless of vaccination status, participants who were Omicron positive had lower anti-S IgG titers than those who did not test positive, but this difference was not significant. Conclusions: These findings provide novel 18-month kinetics of anti-S IgG antibodies and highlight the durability of hybrid immunity, underlining the strong humoral response stimulated by combined infection and vaccination.

Interaction Between Dietary Iron Intake and Genetically Determined Iron Overload: Risk of Islet Autoimmunity and Progression to Type 1 Diabetes in the TEDDY Study.

OBJECTIVE: To examine whether 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 GAD antibody as the first autoantibody. In children with GRS ≥2 iron risk alleles, high iron intake was associated with an increased risk of IA, with insulin as first autoantibody (adjusted hazard ratio 1.71 [95% CI 1.14; 2.58]) compared with moderate iron intake. CONCLUSIONS: Iron intake may alter the risk of IA in children with high-risk HLA haplogenotypes.

Role of spike in the pathogenic and antigenic behavior of SARS-CoV-2 BA.1 Omicron.

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 date. The Omicron spike (S) protein, with an unusually large number of mutations, is considered the major driver of these phenotypes. 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.

Fifteen-Month Follow-Up of Anti-Spike Receptor-Binding Domain SARS-CoV-2 Antibodies among Healthcare Workers in Boston, MA.

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.

Sepsis Biomarkers.

Biomarkers have been used in sepsis to assist with the diagnosis of disease as well as determining the severity of disease, that is, prognosis. These biomarkers are based on the presence of discrete molecules within the blood. Unfortunately, in 2020, a single biomarker does not have sufficient sensitivity and specificity to definitively rule in or rule out sepsis. Biomarkers have shown better performance in animal models of disease.

The 2021 PhD Board-Certified Clinical Chemist Compensation Survey.

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.

Distinctive pseudopalisaded histiocytic hyperplasia characterizes the transition of exudative to proliferative phase of diffuse alveolar damage in patients dying of COVID-19.

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. 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. 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 was seen in non-COVID-19 DAD cases, whereas microthrombi were common in DAD regardless of etiology. 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.