The Effects of a Single Freeze-Thaw Cycle on Concentrations of Nutritional, Noncommunicable Disease, and Inflammatory Biomarkers in Serum Samples

Abraham, Ransi Ann; Rana, G. C.; Agrawal, Pranjal; Johnston, Robert; Sarna, Avina; Ramesh, Sowmya; Acharya, Rajib; Khan, Nizamuddin; Porwal, Akash; Kurundkar, Sucheta Banerjee; Pandey, Arvind K.; Pullakhandam, Raghu; Nair, K. Madhavan; Kumar, Geeta Trilok; Sachdev, H. P. S.; Kapil, Umesh; Deb, Sibnath; Wagt, Arjan de; Khera, Ajay

doi:10.1055/s-0041-1726575

Cited by 6 publications

(4 citation statements)

References 21 publications

(21 reference statements)

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“…13,41 In loggerhead sea turtles, protein fractions resolved by AGE were stable over 17 years when stored at –207°C (–340.6°F) and –80°C. 26,42 Freezing at –70 to –80°C should be more stable than at –20°C, 43 but it was not assessed in this study and these ultrafreezer temperatures warrant further research in avian samples. As cryogenic preservation is recognized as the best standard practice for long-term blood samples storage, 44,45 plasma samples stored in liquid nitrogen (–180°C) were thawed after a longer time period (18 months).…”

Section: Discussionmentioning

confidence: 91%

Influence of Freezing Temperature, Freezing Duration, and Repeated Freeze/Thaw Cycles on Electrophoretic Profiles in the White Stork (Ciconia ciconia)

Thorel,

Roman,

Leclerc

2023

Journal of Avian Medicine and Surgery

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Plasma electrophoresis is an ancillary diagnostic tool in avian medicine, with agarose gel electrophoresis (AGE) and capillary zone electrophoresis (CZE) being the most common techniques. Frozen samples can be used for quantitative studies or comparative diagnostic purposes, but stability of avian plasma proteins under freezing is poorly described. To evaluate the influence of plasma freezing on electrophoretograms in white storks (Ciconia ciconia), heparin blood was sampled from 30 individuals during annual health examinations. Plasma samples were obtained after centrifugation of fresh samples and divided into aliquots. Both AGE and CZE were performed on fresh aliquots. The remaining aliquots were frozen at –20°C (–4°F) or –180°C (–292°F) and thawed following different protocols: 1 freeze/thaw cycle after 6 months at –20°C; 1, 2, 4, and 7 cycles over 12 months at –20°C; and 1 cycle after 18 months at –180°C. For both techniques, electrophoretic profiles obtained from these thawed aliquots were compared to fresh electrophoretograms. Quantitatively, significant differences (P < 0.05) in most fractions were seen from 6 months postfreezing at –20°C for both techniques. Fewer statistically significant differences were observed after 18 months under cryogenic preservation (–180°C). Qualitatively, AGE provided more repeatable and stable results than CZE over time on samples stored at –20°C, and electrophoretograms were stable after 18 months of cryogenic storage. An electromigration distortion associated with freezing was seen with CZE only. Plasma samples stored in a conventional freezer (–20°C) should not be compared to fresh plasma. For quantitative studies, cryogenic storage should be privileged.

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Section: Discussionmentioning

confidence: 91%

Influence of Freezing Temperature, Freezing Duration, and Repeated Freeze/Thaw Cycles on Electrophoretic Profiles in the White Stork (Ciconia ciconia)

Thorel,

Roman,

Leclerc

2023

Journal of Avian Medicine and Surgery

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“…They reported a 2.6% increase in vitamin D concentration after a single freeze-thaw cycle and a decrease in concentration of 4% during storage at −20 °C for up to 2 months [ 52 ]. However, Abraham et al reported a 12% reduction in vitamin D concentrations following the single freeze-thaw cycle [ 53 ]. The assays used to measure vitamin D levels also pose some potential controversies.…”

Section: Discussionmentioning

confidence: 99%

Vitamin D Status, VDR, and TLR Polymorphisms and Pulmonary Tuberculosis Epidemiology in Kazakhstan

Yerezhepov,

Gabdulkayum,

Akhmetova

et al. 2024

Nutrients

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Background: Tuberculosis (TB) and vitamin D deficiency remain major public health problems in Kazakhstan. Due to the high incidence of pulmonary tuberculosis in the country and based on the importance of vitamin D in the modulation of the immune response and the association of its deficiency with many health conditions, the aim of our research was to study the vitamin D status, VDR and TLR gene polymorphisms, and pulmonary tuberculosis epidemiology in Kazakhstan. Methods: A case-control study included 411 individuals diagnosed with pulmonary TB and 686 controls with no family history of pulmonary tuberculosis. Concentrations of serum vitamin D (25-(OH)D) levels were measured by electrochemiluminescence immunoassay. The gene polymorphisms were determined by real-time polymerase chain reaction (PCR) allelic discrimination assay using TaqMan probes. The association between the risk of pulmonary TB and polymorphisms was evaluated using multimodal logistic regression and assessed with the ORs, corresponding to 95% Cis, and the significance level was determined as p < 0.05. Results: 1097 individuals were recruited from 3 different regions of Kazakhstan. Biochemical data showed vitamin D deficiency (25-(OH)D < 20 ng/mL) was present in both groups, with the case group accounting for almost 95% and 43.7% in controls. Epidemiological data revealed that socioeconomic factors such as BMI < 25 kg/m2 (p < 0.001), employment (p < 0.001), diabetes (p < 0.001), and vitamin D deficiency (p < 0.001) were statistically different between case and control groups. Logistic regression analysis, adjusted by sex, age, BMI, residence, employment, smoking, alcohol consumption, and diabetes, showed that T/T polymorphism of the VDR gene (rs1544410, OR = 1.97, 95% CI: 1.04–3.72, p = 0.03) and A/A polymorphism of the TLR8 gene (rs3764880, OR = 2.44, 95% CI: 1.20–4.98, p = 0.01) were associated with a high risk of developing pulmonary tuberculosis. Conclusions: Vitamin D deficiency remains prevalent in our study cohort and is associated with TB progression. Socioeconomic determinants such as unemployment, BMI under 25 kg/m2, and diabetes are the main risk factors for the development of pulmonary TB in our study. A/A polymorphism of TLR8 (rs3764880) and T/T polymorphism (BsmI, rs1544410) of VDR genes may act as biomarkers for pulmonary tuberculosis in the Kazakh population.

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“…However, most studies used frozen plasma or serum samples. While this is the most feasible way to do discovery and early validation studies, there are differences when a capillary sample is used (54, 55) and further validation studies on the relevant clinical sample will be needed.…”

Section: Discussionmentioning

confidence: 99%

Host blood protein biomarkers to screen for Tuberculosis disease: a systematic review and meta-analysis

Gaeddert,

Glaser,

Chendi

et al. 2024

Preprint

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IntroductionNon-sputum tests are needed to improve TB diagnosis and close the diagnostic gap. The World Health Organization target product profile (TPP) for point-of-care (POC) screening tests requires minimum sensitivity 90% and specificity 70%. Our objective was to identify host blood protein biomarkers meeting TPP criteria.MethodsA systematic review was conducted and reported following PRISMA guidelines. Data extraction and quality assessment with QUADAS-2 were completed for included studies. Heterogeneity was assessed. For biomarkers reporting sensitivity and specificity in at least four studies, a random-effects meta-analysis was performed for biomarkers with similar cut-offs.ResultsWe screened 4,651 citations and included 65 studies that enrolled 16,010 participants and evaluated 156 host proteins. Most (47/65) studies enrolled adult pulmonary TB (PTB), with 15 studies in adult extra-pulmonary TB and 5 in children. Small early-stage discovery studies with case-control design were common (24/65) and had high risk of bias. For adult PTB, CRP, IP-10, NCAM-1, and SAA met TPP criteria in high-quality studies. There was a high degree of heterogeneity in biomarker cut-offs and study design. CRP at 10mg/L cut-off was meta-analyzed from 10 studies; pooled sensitivity 86% (95% CI: 80-95) and pooled specificity 67% (95% CI: 54-79). In people living with HIV (6 studies) CRP pooled sensitivity was 93% (95% CI: 90-95) and pooled specificity 59% (95% CI: 40-78).DiscussionWe identified promising biomarkers that performed well in high-quality studies. Data overall are limited and highly heterogenous. Further standardized validation across subgroups in prospective studies is needed before translating into POC assays.

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The Effects of a Single Freeze-Thaw Cycle on Concentrations of Nutritional, Noncommunicable Disease, and Inflammatory Biomarkers in Serum Samples

Cited by 6 publications

References 21 publications

Influence of Freezing Temperature, Freezing Duration, and Repeated Freeze/Thaw Cycles on Electrophoretic Profiles in the White Stork (Ciconia ciconia)

Influence of Freezing Temperature, Freezing Duration, and Repeated Freeze/Thaw Cycles on Electrophoretic Profiles in the White Stork (Ciconia ciconia)

Vitamin D Status, VDR, and TLR Polymorphisms and Pulmonary Tuberculosis Epidemiology in Kazakhstan

Host blood protein biomarkers to screen for Tuberculosis disease: a systematic review and meta-analysis

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