We have examined hepatic, genomic, and metabolic responses to dietary protein restriction in the non-pregnant Sprague-Dawley rat. Animals were pair-fed either a 6 or 24% casein-based diet for 7-10 days. At the end of the dietary period, a microarray analysis of the liver was performed, followed by validation of the genes of interest. The rates of appearance of phenylalanine, methionine, serine, and glucose and the contribution of pyruvate to serine and glucose were quantified using tracer methods. Plasma and tissue amino acid levels, enzyme activities, and metabolic intermediates were measured. Protein restriction resulted in significant differential expression of a number of genes involved in cell cycle, cell differentiation, transport, transcription, and metabolic processes. RT-PCR showed that the expression of genes involved in serine biosynthesis and fatty acid oxidation was higher, and those involved in fatty acid synthesis and urea synthesis were lower in the liver of protein-restricted animals. Free serine and glycine levels were higher and taurine levels lower in all tissues examined. Tracer isotope studies showed an ϳ50% increase in serine de novo synthesis. Pyruvate was the primary (ϳ90%) source of serine in both groups. Transmethylation of methionine was significantly higher in the protein-restricted group. This was associated with a higher S-adenosylmethionine/Sadenosylhomocysteine ratio and lower cystathione ␤-synthase and cystathionine ␥-lyase activity. Dietary isocaloric protein restriction results in profound changes in hepatic one-carbon metabolism within a short period. These may be related to high methylation demands placed on the organism and caused by possible changes in cellular osmolarity as a result of the efflux of the intracellular taurine.The metabolic and biochemical impact of qualitative and quantitative changes in dietary protein intake continues to be of interest because of their implications for public health and because of a number of studies showing a strong relationship between protein intake during pregnancy with cardiovascular function, hypertension, and glucose intolerance in the offspring (1-3). Studies in humans, primarily focused on whole body protein, nitrogen metabolism, and protein accretion, show that both high and low protein intake can impact these processes (4 -7). However, how humans adapt to protein deprivation or the biochemical mechanisms involved has not been examined. In the context of pregnancy, both high protein intake (in humans) and protein restriction in rodents have been shown to cause fetal metabolic programming and consequently altered physiological response in the offspring in adulthood (8 -11). In the rodent, dietary protein restriction during pregnancy results in growth retardation, impaired beta cell function and mass, impaired insulin sensitivity, hypertension, and other pathological responses in the offspring (1, 2). These have been associated with a change in the hypothalamic-pituitary-adrenal axis, changes in the renin-angiotensin system, and ...

Background

Background In hospitalized patients, nasopharyngeal (NP) swabs are the most common samples obtained for Respiratory Syncytial Virus (RSV) PCR testing. However, adding sputum is known to increase diagnostic yield, and saliva has been successfully used for viral respiratory infection diagnosis. We sought to compare RSV prevalence detected by PCR testing of NP swab alone versus NP swab plus saliva and sputum in adult patients hospitalized with acute respiratory illness (ARI). Methods This ongoing, prospective cohort study enrolled patients aged ≥40 years hospitalized for ARI in 4 hospitals in Louisville, Kentucky (Season 1: 27 Dec 21 – 1 Apr 22). NP swab, saliva, and sputum samples were obtained at enrollment or scavenged from standard-of-care specimens (all collected ≤3 days of admission), and PCR tested with Luminex ARIES FluA/B/RSV platform. We produced Venn diagrams of RSV positive samples by sample type for all patients and restricted to those with all 3 sample types. RSV prevalence for NP swab alone was calculated as number of patients with RSV-positive NP swabs divided by total number of patients tested. RSV prevalence by NP swab plus saliva and sputum was calculated as number of patients with RSV-positive NP swab, saliva, or sputum samples divided by total number of patients tested. Results We enrolled 653 patients and collected NP swabs (100% of patients), saliva (96%), and sputum (43% overall and 93% of the 303 sputum-producing patients). Among all patients, 28 patients tested RSV positive (Figure 1A), and when restricted to those with all 3 samples (Figure 1B), 14 tested positive. The overall cohort’s RSV prevalence by NP swab alone was 1.8% (12/653) and by NP swab plus saliva and/or sputum was 4.3% (28/653): 2.33 times higher with addition of saliva and sputum samples. Among patients with all 3 specimen types, the RSV prevalence increase was the same, and none were positive by NP swab only. Figure 1.Venn diagrams of positive RSV PCR tests (Left) A. Positive RSV PCR tests for 653 patients in overall cohort (Right) B. Positive RSV PCR tests for 275 patients with all 3 samples obtained. Conclusion RSV was most commonly detected in saliva samples. Current standard-of-care utilizing NP swab for RSV PCR testing appears to underestimate true RSV prevalence in hospitalized adult patients with ARI by more than 2-fold. Disclosures Alan Junkins, PhD, D(ABMM), Biomerieux: Advisor/Consultant Paul S. Schulz, MD, Gilead: Advisor/Consultant|Gilead: Grant/Research Support|Gilead: Honoraria|Merck: Advisor/Consultant|Merck: Grant/Research Support|Merck: Honoraria Robin Hubler, MS, Pfizer Inc.: Employee|Pfizer Inc.: Stocks/Bonds Paula Peyrani, MD, Pfizer, Inc: Employee|Pfizer, Inc: Employee|Pfizer, Inc: Stocks/Bonds|Pfizer, Inc: Stocks/Bonds Paula Peyrani, MD, Pfizer, Inc: Employee|Pfizer, Inc: Employee|Pfizer, Inc: Stocks/Bonds|Pfizer, Inc: Stocks/Bonds Qing Liu, M.S., Pfizer Inc.: I am a full time employee of Pfizer and hold Pfizer stocks Bradford J. Gessner, M.D., M.P.H., Pfizer Inc.: Employee|Pfizer Inc.: Stocks/Bonds Elizabeth Begier, M.D., M.P.H., Pfizer: Employee|Pfizer: Stocks/Bonds.

The current prognostic parameters, including tumor volume, biochemistry, or immunohistochemistry, are not sufficient to reflect the properties of cancer cells that distinguish them from normal cells. Our focus is to evaluate the effects of a combination of microtubule-polymerizing Taxol and -depolymerizing colchicine on IAR20 PC1 liver cells by measuring the surface fractal dimension as a descriptor of two-dimensional vascular geometrical complexity. The fractal dimension offers a rapid means of assessing cell shape. Furthermore, we show correlations of fractal dimensions of cell contours with the latent factors from our previously employed cell shape analysis.

Introduction Nearly all existing respiratory syncytial virus (RSV) incidence estimates are based on real-time polymerase chain reaction (RT–PCR) testing of nasal or nasopharyngeal (NP) swabs. Adding testing of additional specimen types to NP swab RT–PCR increases RSV detection. However, prior studies only made pairwise comparisons and the synergistic effect of adding multiple specimen types has not been quantified. We compared RSV diagnosis by NP swab RT–PCR alone versus NP swab plus saliva, sputum, and serology. Methods This was a prospective cohort study over two study periods (27 December 2021 to 1 April 2022 and 22 August 2022 to 11 November 2022) of patients aged ≥ 40 years hospitalized for acute respiratory illness (ARI) in Louisville, KY. NP swab, saliva, and sputum specimens were collected at enrollment and PCR tested (Luminex ARIES platform). Serology specimens were obtained at acute and convalescent timepoints (enrollment and 30–60-day visit). RSV detection rate was calculated for NP swab alone and for NP swab plus all other specimen type/test. Results Among 1766 patients enrolled, 100% had NP swab, 99% saliva, 34% sputum, and 21% paired serology specimens. RSV was diagnosed in 56 (3.2%) patients by NP swab alone, and in 109 (6.2%) patients by NP swab plus additional specimens, corresponding to a 1.95 times higher rate [95% confidence interval (CI) 1.62, 2.34]. Limiting the comparison to the 150 subjects with all four specimen types available (i.e., NP swab, saliva, sputum, and serology), there was a 2.60-fold increase (95% CI 1.31, 5.17) compared to NP swab alone (3.3% versus 8.7%). Sensitivities by specimen type were: NP swab 51%, saliva 70%, sputum 72%, and serology 79%. Conclusions Diagnosis of RSV in adults was several-fold greater when additional specimen types were added to NP swab, even with a relatively low percentage of subjects with sputum and serology results available. Hospitalized RSV ARI burden estimates in adults based solely on NP swab RT–PCR should be adjusted for underestimation. Supplementary Information The online version contains supplementary material available at 10.1007/s40121-023-00805-1.