Demonstration of reciprocal diurnal variation in human serum T3 and rT3 concentration demonstrated by mass spectrometric analysis and establishment of thyroid hormone reference intervals.

Background: There has been a wide range of reference intervals proposed in previous literature for thyroid hormones due to large between-assay variability of immunoassays, as well as lack of correction for collection time. We provided the diurnal reference intervals for five thyroid hormones, namely total thyroxine (TT4), total triiodothyronine (TT3), free thyroxine (FT4), free triiodothyronine (FT3), and reverse T3 (rT3), measured in serum samples of healthy participants using a liquid chromatography/tandem mass spectrometry (LC-MS/MS) method. Methods: Couplet serum samples (a.m. and p.m.) were collected from 110 healthy females and 49 healthy males. Healthy volunteers were recruited from four participating centers between 2016 and 2018. Measurements of thyroid hormones were obtained by LC-MS/MS analysis. Results: Our study revealed significant uptrend in AM to PM FT4 ( Conclusion: When diagnosing thyroid disorders, it is important to have accurate measurement of thyroid hormones, and to acknowledge the diurnal fluctuation found, especially for FT3. Our study highlights the importance of standardization of collection times and implementation of LC-MS/MS in thyroid hormone measurement

Association Between Cost-Related Delay in Filling Prescriptions and Health Care Ratings among Medicare Advantage Recipients

Thesis (Master's)--University of Washington, 2018Patients with lower socioeconomic status frequently report not filling or delaying prescription medications due to cost (hereafter, cost-related prescription delay, CRPD). We hypothesized that CRDP would be associated with lower medical care ratings, but the effect would be confounded by socioeconomic and health factors. In the 2012 CAHPS Medicare Advantage Survey, recipients were asked to rate their medical care while reporting if they had experienced CRPD in the past 6 months. CRPD more than doubled the relative risk (RR) for poor ratings of personal doctor (RR 2.34), specialist (RR 2.14), and overall health care in the past 6 months (RR 2.40). Adjusting for sociodemographic and health status slightly reduced the RR to 1.9, but adjusting for low income subsidy and lack insurance for medications did not. Our results show that CRDP is independently associated with poor ratings of medical care irrespective of health, financial, or insurance status. Providers might improve patient ratings and thus satisfaction by explicitly discussing prescription cost and mutually forming affordable treatment plans

Correlation of Diagnostic Yield with Laboratory Parameters and Histopathological Examination in Clinically Suspected Cases of Mucormycosis in Post-COVID-19 Patients: A Single-Center Study in Ahmedabad, India

Background and objectives: Mucormycosis is a complication in post-coronavirus disease 2019 (COVID-19) patients in India. This study was done to evaluate the prognostic value of clinical, histopathologic findings, microbiological features, and biochemical parameters such as D-dimer, lactate dehydrogenase, and serum ferritin in post- COVID-19-patients with rhino-orbital mucormycosis. Methods: This retrospective observational study was carried out on biopsies taken from 50 post-COVID-19 patients suspected of mucormycosis. The biopsy specimens were processed and stained with hematoxylin and eosin, periodic acid–schiff, and Wright-Giemsa. In addition, 10–20% potassium hydroxide wet mount and culture on sabouraud dextrose agar were performed to detect Mucor. The biochemical parameters were measured using ARCHITECT ci8200 chemistry analyzer. Results: Overall, 30 cases (60%) were positive for fungal elements, and growth of Mucor spp. was found in 28 cases (56%). In histopathology, 70% of cases (n=35) showed broad, aseptate, ribbon-like hyphae with wide-angled branching diagnostic of mucormycosis. There seemed to be a site-wise overlap between the nasal/maxillary sinus and rhino-orbital/rhino-cerebral variety. There was no difference between the patients in terms of gender. The most common risk factor was diabetes mellitus (observed in 80% of cases). In patients with invasive mucormycosis, inflammatory biomarkers such as serum ferritin, serum lactate dehydrogenase, C-reactive protein, and D-dimer were greater than the normal range, whereas procalcitonin was within the reference range.    Conclusion: It can be concluded that raised metabolic markers, direct 10% KOH examination and histological features including angioinvasion as well as rhino-orbital and cerebral extension might assist doctors in diagnosis, progression, and survival rate

Proteomic Analyses of Corneal Tissue Subjected to Alkali Exposure

Fundamentally, proteins undergo changes when exposed to alkaline conditions. These changes range from aggregation, changes in solubility, elimination reactions involving the amino acid side chains, cross-linking with lipids (or other nonprotein constituents), and hydrolysis. Data presented here suggest that the hydrolytic truncation and aggregation of proteins occur as a result of alkali exposure. Proteomic analysis of alkali-exposed tissue has identified plexin D1, dynein, and other proteins. Further studies will demonstrate the feasibility of such proteins serving as biomarkers for alkali-induced damage

Clinical utility of interleukin-18 in breast cancer patients: A pilot study

Aim: The aim of this study is to analyze the protein expression of interleukin 18 (IL-18) in patients with untreated breast cancer and further to evaluate its clinical efficacy in predicting treatment outcome. Methods: In the present study, a total of 50 untreated patients with invasive ductal carcinoma of breast were included in the study. Expression of IL-18 was studied by immunohistochemistry method. Statistical analysis was carried out using Statistical Package for Social Sciences statistical software and P ≤ 0.05 was considered statistically significant. Results: Seventy-two percent of the breast cancer patients showed the presence of cytoplasmic and/or nuclear IL-18 immunoreactivity. IL-18 expression was significantly and positively correlated with the stromal response (χ2 = 3.97, r = 0.282, P = 0.044). Further, the IL-18 immunoreactivity was significantly higher in patients with HER2 amplification as compared to luminal B (χ2 = 2.82, r = −0.523, P = 0.047) breast cancer patients. Moreover, a trend of increased IL-18 expression was observed in estrogen/progesterone receptor (ER/PR) negative patients as compared to ER/PR positive patients (χ2 = 3.41, r = −0.282, P = 0.066). Conclusion: IL-18 could be used as a potential predictive marker and guide clinicians for recommendations to newer treatment. It might serve as a potential therapeutic target to establish novel treatment approaches along with the current treatment protocol used

Multiple A2E treatments lead to melanization of rod outer segment-challenged ARPE-19 cells.

PurposeDaily phagocytosis of outer segments (OSs) and retinoid recycling by the RPE lead to the accumulation of storage bodies in the RPE containing autofluorescent lipofuscin, which consists of lipids and bisretinoids such as A2E and its oxidation products. Accumulation of A2E and its oxidation products is implicated in the pathogenesis of several retinal degenerative diseases. However, A2E accumulates in the RPE during normal aging. In this study, we used a cell model to determine the homeostatic mechanisms of RPE cells in response to A2E accumulation.MethodsTo distinguish between pathologic and normal responses of the RPE to A2E accumulation, we treated established ARPE-19 cells (cultured for 3 weeks after reaching confluence) with low micromolar amounts of A2E for several weeks. We compared the lysosomal function, lysosomal pH, degree of OS digestion, and melanization of the treated cells to untreated control cells in response to a challenge of purified rod OSs (ROSs). A2E was analyzed with high-performance liquid chromatography (HPLC); and A2E and melanin were identified with mass spectrometry.ResultsWe found that post-confluent ARPE-19 cells took up and accumulated A2E under dim light conditions. Spectral analysis of the HPLC separations and mass spectrometry showed that A2E-fed cells contained A2E and oxidized A2E (furan-A2E). A2E accumulation led to a modest increase (up to 0.25 unit) in lysosomal pH in these cells. The specific activity of cathepsin D and lysosomal acid phosphatase was reduced in the A2E-treated cells, but ROS degradation was not impaired. We found that, upon challenge with ROSs, melanin pigment was induced in the lysosomal fraction of the A2E-treated ARPE-19 cells. Thus, the ARPE-19 cells responded to the A2E treatment and ROS challenge by producing a melanin-containing lysosome fraction. We speculate that this prevents them from becoming impaired in OS processing.ConclusionsWe used a modified ARPE-19 cell model in which melanization was elicited as a response to chronic accumulation of A2E. We found that although A2E treatment led, as has been previously reported, to modest lysosomal alkalinization and lysosomal impairment of ARPE-19 cells, a potential homeostatic mechanism may involve production of a special type of lysosomes containing melanin

Multiple A2E treatments lead to melanization of rod outer segment-challenged ARPE-19 cells.

PurposeDaily phagocytosis of outer segments (OSs) and retinoid recycling by the RPE lead to the accumulation of storage bodies in the RPE containing autofluorescent lipofuscin, which consists of lipids and bisretinoids such as A2E and its oxidation products. Accumulation of A2E and its oxidation products is implicated in the pathogenesis of several retinal degenerative diseases. However, A2E accumulates in the RPE during normal aging. In this study, we used a cell model to determine the homeostatic mechanisms of RPE cells in response to A2E accumulation.MethodsTo distinguish between pathologic and normal responses of the RPE to A2E accumulation, we treated established ARPE-19 cells (cultured for 3 weeks after reaching confluence) with low micromolar amounts of A2E for several weeks. We compared the lysosomal function, lysosomal pH, degree of OS digestion, and melanization of the treated cells to untreated control cells in response to a challenge of purified rod OSs (ROSs). A2E was analyzed with high-performance liquid chromatography (HPLC); and A2E and melanin were identified with mass spectrometry.ResultsWe found that post-confluent ARPE-19 cells took up and accumulated A2E under dim light conditions. Spectral analysis of the HPLC separations and mass spectrometry showed that A2E-fed cells contained A2E and oxidized A2E (furan-A2E). A2E accumulation led to a modest increase (up to 0.25 unit) in lysosomal pH in these cells. The specific activity of cathepsin D and lysosomal acid phosphatase was reduced in the A2E-treated cells, but ROS degradation was not impaired. We found that, upon challenge with ROSs, melanin pigment was induced in the lysosomal fraction of the A2E-treated ARPE-19 cells. Thus, the ARPE-19 cells responded to the A2E treatment and ROS challenge by producing a melanin-containing lysosome fraction. We speculate that this prevents them from becoming impaired in OS processing.ConclusionsWe used a modified ARPE-19 cell model in which melanization was elicited as a response to chronic accumulation of A2E. We found that although A2E treatment led, as has been previously reported, to modest lysosomal alkalinization and lysosomal impairment of ARPE-19 cells, a potential homeostatic mechanism may involve production of a special type of lysosomes containing melanin