Concentration In Cells Research Articles

Design and Preparation of Lifetime-Based Dual Fluorescent/Phosphorescent Sensor of pH and Oxygen and its Exploration in Model Physiological Solutions and Cells.

In the present report, a novel dual pH-O2 sensor based on covalent conjugate of rhodamine 6G and cyclometalated iridium complex with poly(vinylpyrrolidone-block-vinyltetrazole) copolymer is reported. In model physiological solutions the sensor chromophores display independent phosphorescent and fluorescent lifetime responses onto variations in oxygen concentration and pH, respectively. Colocalization studies on Chinese hamster ovary cells demonstrate the preferential localization in endosomes and lysosomes. The fluorescent lifetime imaging microscopy-phosphorescent lifetime imaging microscopy (FLIM-PLIM) experiments show that the phosphorescent O2 sensor provides unambiguous information onto hypoxia versus normoxia cell status as well as semi-quantitative data on the oxygen concentration in cells in between these two states. However, the results of FLIM measurements indicate that dynamic lifetime interval of the sensor (≈0.5ns between pH values 5.0 and 8.0) is insufficient even for qualitative estimation of pH in living cells because half-width of lifetime distribution in the studied samples is higher than the sensor dynamic interval. Nevertheless, the variations in rhodamine emission intensity are much higher and allow rough discrimination of acidic and neutral cell conditions. Thus, the results of this study indicate that the suggested approach to the design of dual pH-O2 sensors makes possible to prepare the biocompatible and water-soluble conjugate with fast cellular uptake.

Prenatal exposure to di-n-butyl phthalate promotes RIPK1-regulated necroptosis of uroepithelial cells and induces hypospadias through the epithelial-mesenchymal transition process in newborn male rats

Maternal exposure to di-n-butyl phthalate (DBP) has been linked to the induction of hypospadias; however, the underlying mechanism remains unclear. Necroptosis is reported to be implicated in developmental malformations. This study aimed to investigate the underlying mechanism of necroptosis in the development of hypospadias. DBP was dissolved in corn oil, and pregnant rats were administered a precisely measured dose of DBP (750mg/kg/day) via gastric intubation from gestation day 14 to 18. Control rats received only corn oil. The day of birth was considered postnatal day (PND) 1. Male hypospadias rats were identified on PND 7. Genital tubercle tissues were collected and stored at −80°C for subsequent PCR analysis, cryopreserved in liquid nitrogen for western blot, or fixed in formalin for immunohistochemistry (IHC) staining. IHC staining and western blot analysis revealed increased expression of RIPK1 and necroptosis markers in genital tubercle (GT) tissue compared to the control group. Additionally, higher levels of EMT and impaired androgen receptor expression were observed in GT tissue. Exposure to increased DBP concentrations in rat primary uroepithelial cells (PUCs) led to elevated ROS production. Necroptosis markers and EMT expression levels were upregulated in PUCs following DBP incubation. Notably, treatment with DBP combined with necrostatin-1, a necroptosis inhibitor, reduced the expression of EMT markers and ROS production compared to DBP treatment alone. In vitro studies further revealed that DBP-induced necroptosis promoted the degradation of E-cadherin through the ubiquitin-proteasome pathway in PUCs. Our findings suggest that maternal exposure to DBP promotes necroptosis in uroepithelial cells by elevating ROS level and EMT status. Thus, necroptosis may play an essential role in the development of hypospadias.

Mushroom against Cancer: Aqueous Extract of Fomitopsis betulina in Fight against Tumors

Background/Objectives: This study investigated the anticancer potential of an aqueous extract of the fungus Fomitopsis betulina. Methods: The study assessed the effect of the extract on nine cancer cell lines, including melanoma (LM-MEL-75), lung cancer (A549), and colorectal cancer (HT29, LoVo), and four normal cell lines. The cytotoxicity of the extract was evaluated using MTT, sulforhodamine-B (SRB), and clonogenic viability assays. Additionally, the study examined the effect of the extract on plant model organisms, garden cress (Lepidium sativum) and common onion (Allium cepa), to further investigate its biological activity. Results: The assays demonstrated selective cytotoxicity of the extract toward cancer cells, while sparing normal cells. The extract induced significant cytotoxic effects at lower concentrations in lung cancer, melanoma, and colon cancer cells, showing promise as a potential anticancer agent. The results also revealed that the extract inhibited seed germination and root growth, suggesting its potential to disrupt cell cycles and induce apoptosis. Conclusions: This study highlights the therapeutic potential of F. betulina and highlights the need for further research to identify the active ingredients and mechanisms underlying its anticancer effects.

A Bis-Boron Amino Acid for Positron Emission Tomography and Boron Neutron Capture Therapy.

Trifluoroborate boronophenylalanine (BBPA) is a boron amino acid analog of 4‑boronophenylalanine (BPA) but with a trifluoroborate group (-BF3-) instead of a carboxyl group (-COOH). Clinical studies have shown that 18F-labeled BBPA ([18F]BBPA) can produce high-contrast tumor images in positron emission tomography (PET). Beyond PET imaging, BBPA is a theranostic agent for boron neutron capture therapy (BNCT). Because BBPA possesses an identical chemical structure to BNCT and PET, it can potentially predict the boron concentration for BNCT using [18F]BBPA-PET. The synthesis of BBPA was achieved by selectively fluorinating the α-aminoborate compound, taking advantage of the varying rates of solvolysis of the B-F bond. The study showcased the high-contrast [18F]BBPA-PET imaging in various tumor models, highlighting its broad applicability for both [18F]BBPA-PET and BBPA-BNCT. [18F]BBPA-PET tumor uptake remains consistent across various doses, including those used in BNCT. This enables accurate estimation of the boron concentration in tumors using [18F]BBPA-PET. With its dual boron structure, BBPA increases boron concentration in tumor cells and tumor tissues compared to BPA. Thus, less boron carrier is needed. This study introduces a new theranostic boron carrier that enhances boron accumulation in tumors, predicts boron concentration, and enhances the accuracy and effectiveness of BNCT.

A Bis‐Boron Amino Acid for Positron Emission Tomography and Boron Neutron Capture Therapy

Trifluoroborate boronophenylalanine (BBPA) is a boron amino acid analog of 4‑boronophenylalanine (BPA) but with a trifluoroborate group (‐BF3‐) instead of a carboxyl group (‐COOH). Clinical studies have shown that 18F‐labeled BBPA ([18F]BBPA) can produce high‐contrast tumor images in positron emission tomography (PET). Beyond PET imaging, BBPA is a theranostic agent for boron neutron capture therapy (BNCT). Because BBPA possesses an identical chemical structure to BNCT and PET, it can potentially predict the boron concentration for BNCT using [18F]BBPA‐PET. The synthesis of BBPA was achieved by selectively fluorinating the α‐aminoborate compound, taking advantage of the varying rates of solvolysis of the B‐F bond. The study showcased the high‐contrast [18F]BBPA‐PET imaging in various tumor models, highlighting its broad applicability for both [18F]BBPA‐PET and BBPA‐BNCT. [18F]BBPA‐PET tumor uptake remains consistent across various doses, including those used in BNCT. This enables accurate estimation of the boron concentration in tumors using [18F]BBPA‐PET. With its dual boron structure, BBPA increases boron concentration in tumor cells and tumor tissues compared to BPA. Thus, less boron carrier is needed. This study introduces a new theranostic boron carrier that enhances boron accumulation in tumors, predicts boron concentration, and enhances the accuracy and effectiveness of BNCT.

A light-addressable potentiometric sensor-based extracellular calcium dynamic monitoring and imaging platform for cellular calcium channel drug evaluation

Disruption and dysregulation of cellular calcium channel function can lead to diseases such as ischemic stroke, heart failure, and arrhythmias. Corresponding calcium channel drugs typically require preliminary efficacy evaluations using in vitro models such as cells and simulated tissues before clinical testing. However, traditional detection and evaluation methods often encounter challenges in long-term continuous monitoring and lack calcium specificity. In this study, a dynamic monitoring system based on ion-sensitive membranes for light-addressable potentiometric sensor (LAPS) was developed to meet the demand for monitoring changes in extracellular calcium ion (Ca2+) concentration in live cells. The effects of Ca2+ channel agonists and blockers on 2D and 3D HL-1 cells were investigated, with changes in extracellular Ca2+ concentration reflecting cellular calcium metabolism, facilitating drug evaluation. Additionally, calcium imaging technology with optical addressing capability complemented the LAPS system's ability to perceive 3D cell morphology, enhancing its drug evaluation capabilities. This work provides a novel, label-free, specific, and stable technique for monitoring cellular calcium metabolism. It achieves both continuous monitoring at single points and custom sensing area calcium imaging, holding significant implications for drug screening and disease treatment related to human calcium homeostasis.

Chromogranin B Protects Human Umbilical Endothelial Cells against Oxidative Stress

Chromogranin B (CgB) is involved in the control of the cardiovascular system through the regulation of catecholamine release. Whether CgB can exert direct actions on the endothelium has not yet been clarified. Here, we aimed to investigate the effects of CgB on cell viability, mitochondrial membrane potential, reactive oxygen species (ROS), glutathione (GSH), nitric oxide (NO) release, and the cytosolic calcium concentration ([Ca2+]c) in human vascular endothelial cells (HUVECs) cultured under both physiological and peroxidative conditions. In HUVECs, experiments were conducted to establish the proper concentration and timing of CgB stimulation. Thereafter, specific assays were used to evaluate the response of HUVECs cultured in physiologic or oxidative stress conditions to CgB in the presence or absence of β-adrenergic receptor agonists and antagonists and intracellular pathways blockers. Analysis of cell viability, mitochondrial membrane potential, and NO release revealed that CgB was able to cause increased effects in HUVECs cultured in physiological conditions. Additionally, the same analyses performed in HUVECs cultured with H2O2, showed protective effects exerted by CgB, which was also able to counteract ROS release and maintain GSH levels. Furthermore, CgB played a dual role on the [Ca2+]c depending on the physiological or peroxidative cell culturing conditions. In conclusion, our data provide new information about the direct role of CgB in the physiological regulation of endothelial function and highlight its potential as a protective agent against peroxidative conditions, such as those found in cardiovascular diseases.

The Role of Growth Differentiation Factor-15 in the Diagnosis of Patients with Chronic Angina Undergoing Coronary Angiography

Coronary artery disease (CAD) is considered the most prevalent leading cause of myocardial ischemia and mortality worldwide and can lead to angina pectoris and myocardial infarction (MI). Growth Differentiation Factor-15 (GDF-15) is usually measured together with other biomarkers to predict all-cause mortality. During stressful conditions associated with tissue injury and inflammation like myocardial ischemia, it is released into circulation and detected at high concentration in foam cells of the atherosclerotic plaque. Its circulating levels are associated with chronic diseases rather than acute diseases mainly cardiovascular diseases (CVDs) like CAD and chronic heart failure (CHF). The aim of the current study was to detect the role of GDF-15 as a new biomarker for the diagnosis of stable angina patients with normal left ventricular ejection fraction who were presented as having stable chest pain, so that to avoid endangering them to the invasive coronary angiography. Fasting venous blood samples were taken from 90 participants who were presented as having chest pain. They were all subjected to echocardiography, electrocardiography and coronary angiography. The left ventricular ejection fraction was calculated using the biplane M mode method. GDF-15 serum level was measured by using Human GDF-15 Sandwich ELISA Kit following the manufacturer’s instructions.The current study detected a high significant difference in the serum levels of GDF-15 and serum creatinine between the patients and the control group (P≤0.01). On the other hand, there were significant differences in the serum levels of triglycerides and VLDL-C between both study groups (P≤0.05). ROC curve analysis showed that GDF-15 had AUC= 1.00, the best cut off= 254.16 with sensitivity and specificity of 100%. So, it can be concluded that GDF-15 could be used as a biomarker for the diagnosis of stable angina patients with obstructive atherosclerotic plaque and normal LV ejection fraction.

Micelle-encapsulated IR783 for enhanced photothermal therapy in mouse breast cancer

BackgroundPhotothermal therapy, an emerging cancer treatment, selectively eliminates lesions using photothermal compounds that convert light into heat. IR783, a near-infrared fluorescent heptamethine cyanine dye, has been used to achieve selective hyperthermic effects in target tissues via near-infrared irradiation. To implement IR783 as a photothermal agent, IR783 biodistribution must be calibrated to achieve a constant and uniform concentration in target cells. Accordingly, we developed micelle-encapsulated IR783 (IR783 micelles) and evaluated their effectiveness as photothermal drugs. MethodsIn vitro, the photothermic effects of free IR783 and IR783 micelle solutions induced by near-infrared light irradiation were analyzed. Additionally, we investigated the mechanism of cell death mediated by photothermal therapy using free IR783 and IR783 micelles in mouse breast cancer (EMT6) cells. In vivo, the efficacy of photothermal therapy with both free IR783 and IR783 micelles was examined in EMT6-bearing mice. ResultsIn vitro, the temperature of free and micelle-encapsulated IR783 solutions increased after near-infrared irradiation. Near-infrared irradiation with free IR783 and IR783 micelles induced cytotoxicity in cancer cells by generating heat. In vivo, IR783 micelles elicited more preferential tumor tissue uptake and enhanced the antitumor effects of photothermal therapy at a lower light dose relative to free IR783. ConclusionsOverall, these results suggest that IR783 micelles could accumulate in mouse breast cancer tissues and exhibit enhanced antitumor effects when used as a photothermal therapy, with superior effects obtained at 2.1 W/cm2 (252 J/cm2) compared with that of free IR783.

MYO3B promotes cancer progression in endometrial cancer by mediating the calcium ion-RhoA/ROCK1 signaling pathway

PurposeThis study aimed to investigate the effect of MYO3B on endometrial cancer (EC) proliferation and invasion.MethodsThe expression of MYO3B in EC tissues and cells was analyzed using TCGA database, immunohistochemical staining, real-time PCR, and western blot (WB). Cell proliferation was detected by CCK8, Annexin V-APC/PI flow cytometry was used to detect apoptosis, intracellular calcium ion (Ca2+) was detected by flow cytometry with Fluo-4 AM fluorescent probe, cell migration by scratch assay, and cell invasion by Transwell assay, and the expression of proteins related to Ca2+ homeostasis and RhoA/ROCK1 signaling pathway was detected by WB and immunofluorescence staining.ResultsThe expression of MYO3B was an influential factor in EC recurrence, and the expression of MYO3B was significantly up-regulated in EC tissues and cells, but down-regulated in KLE cells, and MYO3B knockdown inhibited the proliferation, migration, and invasion ability of EC cells and promoted apoptosis, suggesting that MYO3B plays a tumor-promoting role in EC. Furthermore, MYO3B knockdown decreased Ca2+ concentration in EC cells and the RhoA/ROCK1 signaling pathway was inhibited, and the effect of MYO3B knockdown on RhoA/ROCK1 signaling was reversed by treatment with the Calmodulin agonist CALP-2, and the effects of MYO3B knockdown on cell proliferation, migration, and invasion were reversed after treatment with the RhoA agonist U-46,619.ConclusionMYO3B promotes the proliferation and migration of endometrial cancer cells via Ca2+-RhoA/ROCK1 signaling pathway. High expression of MYO3B may be a biomarker for EC metastasis.

Vamifeport: Monography of the First Oral Ferroportin Inhibitor.

Over the last few years, several mechanisms that are involved in congenital diseases characterized by ineffective erythropoiesis have been described. Therefore, multiple new target drugs are being developed in preclinical models against the main regulators of normal erythropoiesis. Above all, the key mechanism that regulates systemic iron homeostasis, represented by the hepcidin-ferroportin axis, is considered to be the target for new therapies. The main hypothesis is that iron restriction, through blocking ferroportin (the unique iron transporter in mammals) in such diseases, ameliorates erythropoiesis. The action of vamifeport is different from the currently approved drugs in this setting since it acts straight on the ferroportin-hepcidin axis. The data presented in the sickle cell disease (SCD) Townes mouse model showed a preclinical proof-of-concept for the efficacy of oral ferroportin inhibitor. Vamifeport reduced hemoglobin concentration in red blood cells (RBCs) and diminished intravascular hemolysis and inflammation, improving hemodynamics and preventing vascular occlusive crises. On this basis, clinical trials were commenced in patients with SCD, non-transfusion-dependent (NTD) thalassemia and transfusion-dependent (TD) thalassemia. Preliminary data in NTD thalassemic patients also confirm the safety and efficacy in decreasing iron level. In conclusion, vamifeport represents a new option in the panorama of drugs targeting the hepcidin-ferroportin axis, but its efficacy is still under investigation as a single agent.

Catalytically distinct IDH1 mutants tune phenotype severity in tumor models.

Mutations in isocitrate dehydrogenase 1 (IDH1) impart a neomorphic reaction that produces D-2-hydroxyglutarate (D2HG), which can inhibit DNA demethylases to drive tumorigenesis. Mutations affect residue R132 and display distinct catalytic profiles for D2HG production. We show that catalytic efficiency of D2HG production is greater in IDH1 R132Q than R132H mutants, and expression of R132Q in cellular and xenograft models leads to higher D2HG concentrations in cells, tumors, and sera compared to R132H. Though expression of IDH1 R132Q leads to hypermethylation in DNA damage pathways, DNA hypomethylation is more notable when compared to R132H expression. Transcriptome analysis shows increased expression of many pro-tumor pathways upon expression of IDH1 R132Q versus R132H, including transcripts of EGFR and PI3K signaling pathways. Thus, IDH1 mutants appear to modulate D2HG levels via altered catalysis, resulting in distinct epigenetic and transcriptomic consequences where higher D2HG levels appear to be associated with more aggressive tumors.

Mesoporous PtIr alloy single-particle: A novel SECM-tip for in situ monitoring NO of single-cell

As a vital factor in cell metabolism, nitric oxide (NO) is associated with nitrosative stress and subsequent inflammations and diseases. In situ, real-time NO monitoring is challenging due to its relative trace concentration and fast diffusion in cell. Scanning electrochemical microscopy (SECM) is suited uniquely for single-cell analysis, and its electrochemical response to targets can be further enhanced by improving the interfacial properties of its tip. Here, an ultramicroelectrodes (UMEs) modification strategy based on bimetallic single-particle was proposed for the first time. This mesoporous platinum/iridium alloy single-particle (mPtIr SP) interface using micelle-assisted electrodeposition was characterized by scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). And the nucleation kinetic progress which can be defined as "oil-in-water-like" electrodeposition was discussed in detail. The high sensitivity (203.86 μA/μM·cm2) and good selectivity for NO detection benefits from the high catalysis of the PtIr alloy and the high mass transfer properties of the porous interface. In particular, this novel UME can real-time monitor NO release from a single MCF-7 cell stimulated by perfluorooctanoic acid (PFOA), providing new ideas for contaminant toxicity assessment, health diagnostics, and disease treatment.

Effects of Confined Microenvironments with Protein Coating, Nanotopography, and TGF-β Inhibitor on Nasopharyngeal Carcinoma Cell Migration through Channels.

Distant metastasis is the primary cause of unsuccessful treatment in nasopharyngeal carcinoma (NPC), suggesting the crucial need to comprehend this process. A tumor related to NPC does not have flat surfaces, but consists of confined microenvironments, proteins, and surface topography. To mimic the complex microenvironment, three-dimensional platforms with microwells and connecting channels were designed and developed with a fibronectin (FN) coating or nanohole topography. The potential of the transforming growth factor-β (TGF-β) inhibitor (galunisertib) for treating NPC was also investigated using the proposed platform. Our results demonstrated an increased traversing probability of NPC43 cells through channels with an FN coating, which correlated with enhanced cell motility and dispersion. Conversely, the presence of nanohole topography patterned on the platform bottom and the TGF-β inhibitor led to a reduced cell traversing probability and decreased cell motility, likely due to the decrease in the F-actin concentration in NPC43 cells. This study highlights the significant impact of confinement levels, surface proteins, nanotopography, and the TGF-β inhibitor on the metastatic probability of cancer cells, providing valuable insights for the development of novel treatment therapies for NPC. The developed platforms proved to be useful tools for evaluating the metastatic potential of cells and are applicable for drug screening.

Shipment of Glycerolized RBC Segments for Red Cell Concentrate Compatibility Testing.

Background: Red cell concentrate (RCC) cryopreservation allows for long-term storage of RCCs with rare phenotypes. Currently, tubing segments are not produced for these frozen units. Pre-transfusion compatibility testing therefore requires thawing and deglycerolization of the whole unit. A study was conducted to demonstrate the feasibility of using segments for compatibility testing, including circumstances where segments would require shipment to a reference laboratory. Study Design and Methods: RCCs produced using the red cell filtration method from citrate-phosphate-dextrose whole blood collections were glycerolized (40%) at day 21 post-collection and segments were generated prior to freezing. Room temperature (RT, 18°C-20°C) or water bath (WB, 37°C) thawing of segments was performed prior to storage at RT or at refrigerated temperatures (cold, 1°C -6°C) for 0, 24, 48, or 72 hours followed by deglycerolization and hemolysis testing. Additional segments were thawed and shipped in temperature-controlled containers at either RT or 1°C -10°C for antibody screening. Results: Hemolysis and RBC recovery results did not show significant differences over the storage period or between thawing and storage conditions. RBC recovery ranged from 46% to 64%. Hemoglobin (Hb) recovery ranged from 56% to 96%; for RT-thawed segments, recovery was significantly higher at 24 hours and lower at 72 hours for RT storage compared with cold storage. WB-thawed, cold-stored segments had higher Hb recoveries at 48 hours. Phenotype assessment was successful for all segments regardless of thawing method or shipping condition. Discussion: The shipment of thawed segments containing glycerolized red cells is feasible for the purpose of conducting pretransfusion phenotype evaluations or pretransfusion compatibility checks.

Tree Rings Mercury Controlled by Atmospheric Gaseous Elemental Mercury and Tree Physiology.

Tree rings are an emerging atmospheric mercury (Hg) archive. Questions have arisen, though, regarding their mechanistic controls and reliability. Here, we report contrasting tree-ring Hg records in three collocated conifer species: Norway spruce (Picea abies), Scots pine (Pinus sylvestris), and European larch (Larix decidua), which are from a remote boreal forest. Centennial atmospheric Hg trends at the site, derived from varved lake sediments, peats, and atmospheric monitoring, indicated a steady rise from the 1800s, peaking in the 1970s, and then declining. Prior to ca. 2005, larch and spruce tree rings reproduced the peak in the atmospheric Hg trend, while pine tree rings peaked in the 1930s, likely due to the prolonged sapwood period and ambiguity in the heartwood-sapwood boundary of pine. Since ca. 2005, tree rings from all species showed increasing Hg concentrations in the physiologically active outer rings despite declining atmospheric Hg concentrations. The good agreement between Hg and nitrogen concentrations in active tree-ring cells indicates a similar transport mechanism and cautions against their applicability as atmospheric Hg archives. Our results suggest that tree-ring Hg records are controlled by atmospheric Hg and tree physiology. We provide recommendations for using tree-ring Hg archives that take tree physiology into account.

Retrospective Analysis of Transfusion-Related Adverse Reactions: A 15-Month Study of a Single Center’s Experience

The aim of this study was to evaluate transfusion-related adverse reactions (TRARs). In this study, all adverse reactions (ARs) related to blood/blood product transfusions conducted between 01.01.2022 and 31.03.2023 at the Health Sciences University Türkiye, Adana City Training and Research Hospital were evaluated. In total, 97,926 records of blood and blood component transfusions were evaluated during the study period. The distribution of blood components used was as follows: 57,066 (58.2%) red blood cell concentrates, 27,345 (28%) fresh frozen plasma, 12,282 (12.5%) pooled platelet concentrates, 564 (0.6%) apheresis platelet concentrates, and 669 (0.7%) cryoprecipitates. In total, 40 AR reports were associated with transfusions. The probability levels of the relationship degrees of reactions for these 40 cases were as follows: 2 cases; not likely (5%); 32 cases; likely (80%); 2 cases; highly likely (5%); and 4 cases, unassessable (10%). All unwanted reactions were acute, and there were no delayed reactions. No transfusion reaction (TR) leading to death occurred. Of the patients who developed reactions, 60% (n=24) were female, and 40% (n=16) were male. The ages of patients with unwanted reactions ranged from 2 to 86 years, with a median age of 33. Among the cases with unwanted reactions, 8 were children (20%) and 32 were adults (80%). In our study, the frequency of allergic TR was 8.1 per 100,000 children and 32.6 per 100,000 adults. A statistically significant difference in the distribution of blood component types among cases based on the types of unwanted reaction was observed (p=0.003). In this retrospective evaluation of 15 months of data from a single center, 97,926 blood component transfusions were performed, and the prevalence of TRAR was 40.8 per 100,000 blood components.

Decision Variables for the Use of Radioactive Iodine in Patients with Thyroid Cancer at Intermediate Risk of Recurrence.

The use of radioactive iodine (RAI) after total thyroidectomy for patients at the American Thyroid Association (ATA) who are at intermediate risk of recurrence is controversial. This is due to the lack of prospective randomized trials proving a benefit to recurrence or survival of RAI therapy in this group. In the absence of such evidence, clinicians struggle to recommend for or against this therapeutic approach which frequently results in overtreatment. This review describes key elements in the decision-making process that help clinicians more comprehensively evaluate the need for RAI therapy in patients with thyroid cancer at intermediate risk of recurrence. A clear definition of the purpose of RAI therapy should be conveyed to patients. In this sense, adjuvant RAI therapy intends to decrease recurrence, and ablation therapy is used to facilitate surveillance. Better stratification of the intermediate risk category into a low-intermediate subgroup and an intermediate-high-risk subgroup results in less heterogeneity and a more precise prediction of recurrence risk. The evaluation of post-operative thyroglobulin levels may prevent the overtreatment of low-intermediate-risk patients when their thyroglobulin level is <2.5 ng/mL. the integration of tumor genomics (when available) alongside pathologic features can enhance the ability of the clinician to predict iodine concentration in thyroid cancer cells. Finally, a detailed consideration of the adverse effects of RAI, patients' comorbidities, and patient preferences will result in a patient-centered personalized approach. Systematic examination of these variables will ultimately provide a framework for making more educated decisions on the use of RAI in patients at intermediate risk of recurrence that will prevent overtreatment and minimize harm.

Categorization of the Inevitable Causes of Blood and Components Wastage Detected in South Indian Urban Blood Centre

Background: Since there is no artificial substitute for human blood, it plays a vital role in the patient management in this era of modern medicine. Therefore the donated blood from a healthy donor is precious and must not be wasted without proper utilization. Blood wastage is a universal phenomenon which needs a careful approach with strict guidelines. The study aims at categorizing the various inevitable causes of wastage with an attempt to determine ways of preventing the loss due to non-utilization. Materials and Methods: A retrospective – cross sectional study was conducted in a newly started Stand-alone Blood Centre in South Indian Urban city between June, 2022 and June 2024.All the components including Whole Blood (WB), Packed Red Blood Cell Concentrate (PRBC), Platelet concentrate (PC) and Fresh Frozen Plasma (FFP) were included in the study. The reasons for blood and component wastage was noted down from the various Registers of the Blood Centre and all the data was analyzed using Microsoft Excel Spread Sheet. Results: A total of 2456 units of blood were collected. 5284 components were prepared from these which includes 42 units of Whole Blood, 2397 Packed Red Blood cells(PRBC), 2393 Fresh frozen Plasma(FFP), 452 Platelet Concentrate(PC).Overall discard rate was found to be 11.90%. PC discard rate was highest (49.34%) followed by WBU (19.04%), PRBC (13.01%) AND FFP (3.59%). The commonest cause for the discarding was expiry of the shelf life of blood bags (74.72%) followed by damaged bags (9.69%) and TTI’s (6.76%). Conclusion: Blood and its components wastage is an inevitable reality which needs to be approached as upmost priority. Zero wastage of blood and components may be targeted by recommendations like appropriate training of the blood bank staff, keeping negative group donors on reserve, strictly following FIFO policy while dispensing blood, whole blood unit collection may be minimized, encouraging Single Donor Platelets (SDP) by apheresis instead of Random Donor Platelets (RDP) preparation, Blood Utilization Committee to be made mandatory and permission for reciprocal sharing of blood components among blood banks.

Distinct In Vitro Differentiation Protocols Differentially Affect Cytotoxicity Induced by Heavy Metals in Human Neuroblastoma SH-SY5Y Cells.

The SH-SY5Y cell line is widely used in neurotoxicity studies. However, the effects of inducing cell differentiation on the cytotoxic effects of heavy metals are unclear. Therefore, we investigated the effects of mercuric chloride (HgCl2), cadmium chloride (CdCl2), arsenic trioxide (As2O3), and methylmercury (MeHg) on SH-SY5Y cells differentiated in the presence of insulin-like growth factor-I (IGF-I) or all-trans retinoic acid (ATRA). Neurite outgrowth with distinct changes in neuronal marker expression, phenotype, and cell cycle was induced in SH-SY5Y cells by IGF-I treatment for 1day or ATRA treatment for up to 7days. The cytotoxic effects of HgCl2 decreased at lower concentrations and increased at higher concentrations in both IGF-I- and ATRA-differentiated cells compared with those in undifferentiated cells. Differentiation with IGF-I, but not with ATRA, increased the cytotoxic effects of CdCl2. Decreased cytotoxic effects of As2O3 and MeHg were observed at lower concentrations in IGF-I-differentiated cells, whereas increased cytotoxic effects of As2O3 and MeHg were observed at higher concentrations in ATRA-differentiated cells. Changes in the cytotoxic effects of heavy metals were observed even after 1day of ATRA exposure in SH-SY5Y cells. Our results demonstrate that the differentiation of SH-SY5Y cells by IGF-I and ATRA induces different cellular characteristics, resulting in diverse changes in sensitivity to heavy metals, which depend not only on the differentiation agents and treatment time but also on the heavy metal species and concentration.