Search results for: prognostic biomarker.

Authors: Marwa I. Shabayek, Ola A. Said, Hanan A. Attaia, Heba A. Awida

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Bladder carcinoma is an important worldwide health problem. Both cystoscopy and urine cytology used in detecting bladder cancer suffer from drawbacks where cystoscopy is an invasive method and urine cytology shows low sensitivity in low grade tumors. This study validates easier and less time-consuming techniques to evaluate the value of combined use of angiogenin and clusterin in comparison and combination with voided urine cytology in the detection of bladder cancer patients. This study includes malignant (bladder cancer patients, n= 50), benign (n=20), and healthy (n=20) groups. The studied groups were subjected to cystoscopic examination, detection of bilharzial antibodies, urine cytology, and estimation of urinary angiogenin and clusterin by ELISA. The overall sensitivity and specifcity were 66% and 75% for angiogenin, 70% and 82.5% for clusterin and 46% and 80% for voided urine cytology. Combined sensitivity of angiogenin and clusterin with urine cytology increased from 82 to 88%.

Keywords: angiogenin, bladder cancer, clusterin, cytology

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Authors: Prachi Abhishek Dave, Manisha Singh

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Purpose: The purpose is to study the microbiological profile, prognostic factors, and outcomes of surgery for microbial keratitis (MK) associated endophthalmitis. Methods: It is a retrospective analysis of 39 MK associated endophthalmitis cases which underwent combined PKP with VR surgery. Results: The majority (84.6%) of patients had a history of previous ocular surgery, cataract surgery being the most common (51.2%). Six patients had a history of trauma and 8 patients had corneal graft infection. The culture positivity rate was 92.3%. Organisms isolated were Streptococcal sp (20.5%), Pseudomonas (15.3%), Staphylococcal sp (12.8%) and Fungus (35.8%). Final visual acuities ranged from NPL to 20/120. Poor anatomic success was seen in 15(38.46%) eyes (9-phthisis, 6-eviscerated). Fungal cases had a poor success rate (P-0.02). Conclusion: MK associated endophthalmitis is a sight threatening ocular condition. Fungal etiology carries a poorer prognosis. Timely surgical intervention can achieve favourable anatomical and functional results.

Keywords: endophthalmitis, microbial keratitis, penetrating keratoplasty, vitreo retinal surgery

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Authors: Denis Ushakov

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Fast growing international migration as a factor of labor globalization now is one of the most important trends of world economy and determinant of social-political transformations. Study of fundamental economical reasons for international migration is relevant due to their prognostic, predictable and normative potential, which can be used in conditions of global economic non-stability. This paper analyzes role of natural-resources, financial and labor factors in economic growth of the modern states; studies relationships between stimulating role of natural resources, finance and labor with levels of modern countries’ economy development. Based on achieved results, findings about fundamental reasons of international migration; transformation of labor factor’s role in providing an economical progress of the states; efficiency of positive impact of manufacturing factors (domestic and attracted from international markets) were offered.

Keywords: international migration, migrant, labor productivity, economy efficiency of migration, migration policy

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Authors: Abdou Elhendy

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Numerous study have shown the efficacy of the percutaneous intervention (PCI) and coronary stenting in improving left ventricular function and relieving exertional angina. Furthermore, PCI remains the main line of therapy in acute myocardial infarction. Improvement of procedural techniques and new devices have resulted in an increased number of PCI in those with difficult and extensive lesions, multivessel disease as well as total occlusion. Immediate and late outcome may be compromised by acute thrombosis or the development of fibro-intimal hyperplasia. In addition, progression of coronary artery disease proximal or distal to the stent as well as in non-stented arteries is not uncommon. As a result, complications can occur, such as acute myocardial infarction, worsened heart failure or recurrence of angina. In a stent, restenosis can occur without symptoms or with atypical complaints rendering the clinical diagnosis difficult. Routine invasive angiography is not appropriate as a follow up tool due to associated risk and cost and the limited functional assessment. Exercise and pharmacologic stress testing are increasingly used to evaluate the myocardial function, perfusion and adequacy of revascularization. Information obtained by these techniques provide important clues regarding presence and severity of compromise in myocardial blood flow. Stress echocardiography can be performed in conjunction with exercise or dobutamine infusion. The diagnostic accuracy has been moderate, but the results provide excellent prognostic stratification. Adding myocardial contrast agents can improve imaging quality and allows assessment of both function and perfusion. Stress radionuclide myocardial perfusion imaging is an alternative to evaluate these patients. The extent and severity of wall motion and perfusion abnormalities observed during exercise or pharmacologic stress are predictors of survival and risk of cardiac events. According to current guidelines, stress echocardiography and radionuclide imaging are considered to have appropriate indication among patients after PCI who have cardiac symptoms and those who underwent incomplete revascularization. Stress testing is not recommended in asymptomatic patients, particularly early after revascularization, Coronary CT angiography is increasingly used and provides high sensitive for the diagnosis of coronary artery stenosis. Average sensitivity and specificity for the diagnosis of in stent stenosis in pooled data are 79% and 81%, respectively. Limitations include blooming artifacts and low feasibility in patients with small stents or thick struts. Anatomical and functional cardiac imaging modalities are corner stone for the assessment of patients after PCI and provide salient diagnostic and prognostic information. Current imaging techniques cans serve as gate keeper for coronary angiography, thus limiting the risk of invasive procedures to those who are likely to benefit from subsequent revascularization. The determination of which modality to apply requires careful identification of merits and limitation of each technique as well as the unique characteristic of each individual patient.

Keywords: coronary artery disease, stress testing, cardiac imaging, restenosis

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Authors: M. Salvador, J. C. Martinez-Garcia, A. Moyano, M. C. Blanco-Lopez, M. Rivas

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Biosensors play a crucial role in the detection of molecules nowadays due to their advantages of user-friendliness, high selectivity, the analysis in real time and in-situ applications. Among them, Lateral Flow Immunoassays (LFIAs) are presented among technologies for point-of-care bioassays with outstanding characteristics such as affordability, portability and low-cost. They have been widely used for the detection of a vast range of biomarkers, which do not only include proteins but also nucleic acids and even whole cells. Although the LFIA has traditionally been a positive/negative test, tremendous efforts are being done to add to the method the quantifying capability based on the combination of suitable labels and a proper sensor. One of the most successful approaches involves the use of magnetic sensors for detection of magnetic labels. Bringing together the required characteristics mentioned before, our research group has developed a biosensor to detect biomolecules. Superparamagnetic nanoparticles (SPNPs) together with LFIAs play the fundamental roles. SPMNPs are detected by their interaction with a high-frequency current flowing on a printed micro track. By means of the instant and proportional variation of the impedance of this track provoked by the presence of the SPNPs, quantitative and rapid measurement of the number of particles can be obtained. This way of detection requires no external magnetic field application, which reduces the device complexity. On the other hand, the major limitations of LFIAs are that they are only qualitative or semiquantitative when traditional gold or latex nanoparticles are used as color labels. Moreover, the necessity of always-constant ambient conditions to get reproducible results, the exclusive detection of the nanoparticles on the surface of the membrane, and the short durability of the signal are drawbacks that can be advantageously overcome with the design of magnetically labeled LFIAs. The approach followed was to coat the SPIONs with a specific monoclonal antibody which targets the protein under consideration by chemical bonds. Then, a sandwich-type immunoassay was prepared by printing onto the nitrocellulose membrane strip a second antibody against a different epitope of the protein (test line) and an IgG antibody (control line). When the sample flows along the strip, the SPION-labeled proteins are immobilized at the test line, which provides magnetic signal as described before. Preliminary results using this practical combination for the detection and quantification of the Prostatic-Specific Antigen (PSA) shows the validity and consistency of the technique in the clinical range, where a PSA level of 4.0 ng/mL is the established upper normal limit. Moreover, a LOD of 0.25 ng/mL was calculated with a confident level of 3 according to the IUPAC Gold Book definition. Its versatility has also been proved with the detection of other biomolecules such as troponin I (cardiac injury biomarker) or histamine.

Keywords: biosensor, lateral flow immunoassays, point-of-care devices, superparamagnetic nanoparticles

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Authors: Chia-Feng Lu, Yuh-Jen Wang, Yu-Te Wu, Sui-Hing Yan

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This study aimed at investigating whether the functional brain networks constructed using the initial EEG (obtained when patients first visited hospital) can be correlated with the progression of cognitive decline calculated as the changes of mini-mental state examination (MMSE) scores between the latest and initial examinations. We integrated the time–frequency cross mutual information (TFCMI) method to estimate the EEG functional connectivity between cortical regions, and the network analysis based on graph theory to investigate the organization of functional networks in aMCI. Our finding suggested that higher integrated functional network with sufficient connection strengths, dense connection between local regions, and high network efficiency in processing information at the initial stage may result in a better prognosis of the subsequent cognitive functions for aMCI. In conclusion, the functional connectivity can be a useful biomarker to assist in prediction of cognitive declines in aMCI.

Keywords: cognitive decline, functional connectivity, MCI, MMSE

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Authors: Hanna Lu, Sandra S. M. Chan, Linda C. W. Lam

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Objectives: Intra-individual variability (IIV) has been considered as a biomarker of healthy ageing. However, the composite role of IIV in attention, as an impending indicator for neurocognitive disorders warrants further exploration. This study aims to investigate the IIV, as well as their relationships with attention network functions in adults with neurocognitive disorders (NCD). Methods: 36adults with NCD due to Alzheimer’s disease(NCD-AD), 31adults with NCD due to vascular disease (NCD-vascular), and 137 healthy controls were recruited. Intraindividual standard deviations (iSD) and intraindividual coefficient of variation of reaction time (ICV-RT) were used to evaluate the IIV. Results: NCD groups showed greater IIV (iSD: F= 11.803, p < 0.001; ICV-RT:F= 9.07, p < 0.001). In ROC analyses, the indices of IIV could differentiateNCD-AD (iSD: AUC value = 0.687, p= 0.001; ICV-RT: AUC value = 0.677, p= 0.001) and NCD-vascular (iSD: AUC value = 0.631, p= 0.023;ICV-RT: AUC value = 0.615, p= 0.045) from healthy controls. Moreover, the processing speed could distinguish NCD-AD from NCD-vascular (AUC value = 0.647, p= 0.040). Discussion: Intra-individual variability in attention provides a stable measure of cognitive performance, and seems to help distinguish the senior adults with different cognitive status.

Keywords: intra-individual variability, attention network, neurocognitive disorders, ageing

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Authors: Roberta Pecoraro, Elena Maria Scalisi

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Engineered Nanoparticles (ENPs) and Nanomaterials (ENMs) concern an active research area and a sector in full expansion. They have physical-chemical characteristics and small size that improve their performance compared to common materials. Due to the increase in their production and their subsequent release into the environment, new strategies are emerging to assess risk of nanomaterials. NPs can be released into the environment through aquatic systems by human activities and exert toxicity on living organisms. We evaluated the potential toxic effect of cerium oxide (CeO2) nanoparticles because it’s used in different fields due to its peculiar properties. In order to assess nanoparticles toxicity, Fish Embryo Toxicity (FET) test was performed. Powders of CeO2 NPs supplied by the CNR-IMM of Catania are indicated as CeO2 type 1 (as-prepared) and CeO2 type 2 (modified), while CeO2 type 3 (commercial) is supplied by Sigma-Aldrich. Starting from a stock solution (0.001g/10 ml dilution water) of each type of CeO2 NPs, the other concentration solutions were obtained adding 1 ml of the stock solution to 9 ml of dilution water, leading to three different solutions of concentration (10-4, 10-5, 10-6 g/ml). All the solutions have been sonicated to avoid natural tendency of NPs to aggregate and sediment. FET test was performed according to the OECD guidelines for testing chemicals using our internal protocol procedure. A number of eight selected fertilized eggs were placed in each becher filled with 5 ml of each concentration of the three types of CeO2 NPs; control samples were incubated only with dilution water. Replication was performed for each concentration. During the exposure period, we observed four endpoints (embryo coagulation, lack of formation of somites, failure to lift the yolk bag, no heartbeat) by a stereomicroscope every 24 hours. Immunohistochemical analysis on treated larvae was performed to evaluate the expression of metallothioneins (MTs), Heat Shock Proteins 70 (HSP70) and 7-ethoxyresorufin-O-diethylase (EROD). Our results have not shown evident alterations on embryonic development because all embryos completed the development and the hatching of the eggs, started around the 48th hour after exposure, took place within the last observation at 72 hours. A good reactivity, both in the embryos and in the newly hatched larvae, was found. The presence of heartbeat has also been observed in embryos with reduced mobility confirming their viability. A higher expression of EROD biomarker was observed in the larvae exposed to the three types of CeO2, showing a clear difference with the control. A weak positivity was found for MTs biomarker in treated larvae as well as in the control. HSP70 are expressed homogeneously in all the type of nanoparticles tested but not too much greater than control. Our results are in agreement with other studies in the literature, in which the exposure of Danio rerio larvae to other metal oxide nanoparticles does not show adverse effects on survival and hatching time. Further studies are necessary to clarify the role of these NPs and also to solve conflicting opinions.

Keywords: Danio rerio, endpoints, fish embryo toxicity test, metallic nanoparticles

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Authors: Shu-Jun Li, Cheng-Cao Sun, De-Jia Li

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Recently, long noncoding RNAs (lncRNAs) have been emerged as crucial regulators of human diseases and prognostic markers in numerous of cancers, including colorectal carcinoma (CRC). Here, we identified an oncogenetic lncRNA HULC, which may promote colorectal tumorigenesis. HULC has been found to be up-regulated and acts as oncogene in gastric cancer and hepatocellular carcinoma, but its expression pattern, biological function and underlying mechanism in CRC is still undetermined. Here, we reported that HULC expression is also over-expressed in CRC, and its increased level is associated with poor prognosis and shorter survival. Knockdown of HULC impaired CRC cells proliferation, migration and invasion, facilitated cell apoptosis in vitro, and inhibited tumorigenicity of CRC cells in vivo. Mechanistically, RNA immunoprecipitation (RIP) and RNA pull-down experiment demonstrated that HULC could simultaneously interact with EZH2 to repress underlying targets NKD2 transcription. In addition, rescue experiments determined that HULC oncogenic function is partly dependent on repressing NKD2. Taken together, our findings expound how HULC over-expression endows an oncogenic function in CRC.

Keywords: long noncoding RNA, HULC, NKD2, colorectal carcinoma, proliferation, apoptosis

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Authors: Javier Enciso-Benavides, Fredy Fabian, Carlos Castaneda, Luis Alfaro, Alex Choque, Aparicio Aguilar, Javier Enciso

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Background: The use of biomarkers in breast cancer diagnosis, therapy, and prognosis has gained increasing interest. Cancer stem cells (CSCs) are a subpopulation of tumor cells that can drive tumor initiation and may cause relapse. Therefore, due to the importance of diagnosis, therapy, and prognosis, several biomarkers that characterize CSCs have been identified; however, in treatment-naïve triple-negative breast tumors, there is an urgent need to identify new biomarkers and therapeutic targets. According to this, the aim of this study was to identify serum proteins associated with cancer stem cells and pluripotency in women with triple-negative breast tumors in order to subsequently identify a biomarker for this type of breast tumor. Material and Methods: Whole blood samples from 12 women with histopathologically diagnosed triple-negative breast tumors were used after obtaining informed consent from the patient. Blood serum was obtained by conventional procedure and frozen at -80ºC. Identification of cancer stem cell-associated proteins was performed by matrix-assisted laser desorption/ionisation-assisted laser desorption/ionisation mass spectrometry (MALDI-TOF MS), protein analysis was obtained using the AB Sciex TOF/TOF™ 5800 system (AB Sciex, USA). Sequences not aligned by ProteinPilot™ software were analyzed by Protein BLAST. Results: The following proteins related to pluripotency and cancer stem cells were identified by MALDI TOF/TOF mass spectrometry: A-chain, Serpin A12 [Homo sapiens], AIEBP [Homo sapiens], Alpha-one antitrypsin, AT {internal fragment} [human, partial peptide, 20 aa] [Homo sapiens], collagen alpha 1 chain precursor variant [Homo sapiens], retinoblastoma-associated protein variant [Homo sapiens], insulin receptor, CRA_c isoform [Homo sapiens], Hydroxyisourate hydrolase [Streptomyces scopuliridis], MUCIN-6 [Macaca mulatta], Alpha-actinin-3 [Chrysochloris asiatica], Polyprotein M, CRA_d isoform, partial [Homo sapiens], Transcription factor SOX-12 [Homo sapiens]. Recommendations: The serum proteins identified in this study should be investigated in the exosome of triple-negative breast cancer stem cells and in the blood serum of women without breast cancer. Subsequently, proteins found only in the blood serum of women with triple-negative breast cancer should be identified in situ in triple-negative breast cancer tissue in order to identify a biomarker to study the evolution of this type of cancer, or that could be a therapeutic target. Conclusions: Eleven cancer stem cell-related serum proteins were identified in 12 women with triple-negative breast cancer, of which MUCIN-6, retinoblastoma-associated protein variant, transcription factor SOX-12, and collagen alpha 1 chain are the most representative and have not been studied so far in this type of breast tumor. Acknowledgement: This work was supported by Proyecto CONCYTEC–Banco Mundial “Mejoramiento y Ampliacion de los Servicios del Sistema Nacional de Ciencia Tecnología e Innovacion Tecnologica” 8682-PE (104-2018-FONDECYT-BM-IADT-AV).

Keywords: triple-negative breast cancer, MALDI TOF/TOF MS, serum proteins, cancer stem cells

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Authors: Noha E. Hassan

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Little is known regarding the influence of lymphatic microvessel density (LMVD) on prognosis in endometrial cancer. Prospective study was done in tertiary education and research hospital (Shatby Alexandria university hospital) on sixty patients presented with endometrial carcinoma underwent complete surgical staging. Our aim was to assess the intratumoral and peritumoral Lymphatic microvessel density (LMVD) of endometrial carcinomas identified by immunohistochemical staining using an antibody against podoplanin and to investigate their association with classical clinicopathological factors and prognosis. The result shows that high LMVD was associated with endometroid type of tumors, lesser myometrial, adnexal, cervical and peritoneal infiltration, lower tumor grade and stage and lesser recurrent cases. There is lower lymph node involvement among cases with high intratumoral LMVD and cases of high peritumoral LMVD; that reach statistical significance only among cases of high intratumoral LMVD. No association was seen between LMVD and lymphovascular space invasion. On the other hand, low LMVD was associated with poor outcome. Finally, we can conclude that increased LMVD is associated with favorable prognosis in endometrial cancer patients.

Keywords: endometrial carcinoma, lymphatic microvessel, microvessel density, prognosis

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Authors: Yan Zeng, Li Tao, Liujun Han, Tianye Zhang, Yongan Yu, Kaijun Ma, Long Chen

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Mechanical asphyxia is one of the main cause of death; however, death by mechanical asphyxia may be difficult to prove in court, particularly in cases in which corpses exhibit no obvious signs of asphyxia. To identify a credible biomarker of asphyxia, we first examined the expression levels of all the mRNAs in human cardiac tissue specimens subjected to mechanical asphyxia and compared these expression levels with those of the corresponding mRNAs in specimens subjected to craniocerebral injury. A total of 119 differentially expressed mRNAs were selected and the expression levels of these mRNAs were examined in 44 human cardiac tissue specimens subjected to mechanical asphyxia, craniocerebral injury, hemorrhagic shock and other causes of death. We found that DUSP1 and KCNJ2 were up-regulated in tissue specimens of mechanical asphyxia compared with control tissues, with no significant correlation between age, environmental temperature and PMI, indicating that DUSP1 and KCNJ2 may associate with mechanical asphyxia-induced death and can thus serve as useful biomarkers of death by mechanical asphyxia.

Keywords: mechanical asphyxia, biomarkers, DUSP1, KCNJ2, cardiac tissue

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Authors: Rijnbout-St.James Willem, Lindner Volkhard, Scholand Mary Beth, Ashton M. Tillett, Di Gennaro Michael Jude, Smith Silvia Enrica

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Introduction: Fibrosing lung diseases are characterized by changes in the lung interstitium and are classified based on etiology: 1) environmental/exposure-related, 2) autoimmune-related, 3) sarcoidosis, 4) interstitial pneumonia, and 4) idiopathic. Among interstitial lung diseases (ILD) idiopathic forms, idiopathic pulmonary fibrosis (IPF) is the most severe. Pathogenesis of IPF is characterized by an increased presence of proinflammatory mediators, resulting in alveolar injury, where injury to alveolar epithelium precipitates an increase in collagen deposition, subsequently thickening the alveolar septum and decreasing gas exchange. Identifying biomarkers implicated in the pathogenesis of lung fibrosis is key to developing new therapies and improving the efficacy of existing therapies. The transforming growth factor-beta (TGF-B1), a mediator of tissue repair associated with WNT5A signaling, is partially responsible for fibroblast proliferation in ILD and is the target of Pirfenidone, one of the antifibrotic therapies used for patients with IPF. Canonical TGF-B signaling is mediated by the proteins SMAD 2/3, which are, in turn, indirectly regulated by Collagen Triple Helix Repeat Containing 1 (CTHRC1). In this study, we tested the following hypotheses: 1) CTHRC1 is more elevated in the ILD cohort compared to unaffected controls, and 2) CTHRC1 is differently expressed among ILD types. Material and Methods: CTHRC1 levels were measured by ELISA in 171 plasma samples from the deidentified University of Utah ILD cohort. Data represent a cohort of 131 ILD-affected participants and 40 unaffected controls. CTHRC1 samples were categorized by a pulmonologist based on affectation status and disease subtypes: IPF (n = 45), sarcoidosis (4), nonspecific interstitial pneumonia (16), hypersensitivity pneumonitis (n = 7), interstitial pneumonia (n=13), autoimmune (n = 15), other ILD - a category that includes undifferentiated ILD diagnoses (n = 31), and unaffected controls (n = 40). We conducted a single-factor ANOVA of plasma CTHRC1 levels to test whether CTHRC1 variance among affected and non-affected participants is statistically significantly different. In-silico analysis was performed with Ingenuity Pathway Analysis® to characterize the role of CTHRC1 in the pathway of lung fibrosis. Results: Statistical analyses of CTHRC1 in plasma samples indicate that the average CTHRC1 level is significantly higher in ILD-affected participants than controls, with the autoimmune ILD being higher than other ILD types, thus supporting our hypotheses. In-silico analyses show that CTHRC1 indirectly activates and phosphorylates SMAD3, which in turn cross-regulates TGF-B1. CTHRC1 also may regulate the expression and transcription of TGFB-1 via WNT5A and its regulatory relationship with CTNNB1. Conclusion: In-silico pathway analyses demonstrate that CTHRC1 may be an important biomarker in ILD. Analysis of plasma samples indicates that CTHRC1 expression is positively associated with ILD affectation, with autoimmune ILD having the highest average CTHRC1 values. While characterizing CTHRC1 levels in plasma can help to differentiate among ILD types and predict response to Pirfenidone, the extent to which plasma CTHRC1 level is a function of ILD severity or chronicity is unknown.

Keywords: interstitial lung disease, CTHRC1, idiopathic pulmonary fibrosis, pathway analyses

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Authors: L. S. Godlevsky, N. V. Kresyun, V. P. Martsenyuk, K. S. Shakun, T. V. Tatarchuk, K. O. Prybolovets, L. F. Kalinichenko, M. Karpinski, T. Gancarczyk

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Structures of eye bottom were extracted using multiscale texture gradient method and color characteristics of macular zone and vessels were verified in CIELAB scale. The difference of average values of L*, a* and b* coordinates of CIE (International Commision of Illumination) scale in patients with diabetes and healthy volunteers was compared. The average value of L* in diabetic patients exceeded such one in the group of practically healthy persons by 2.71 times (P < 0.05), while the value of a* index was reduced by 3.8 times when compared with control one (P < 0.05). b* index exceeded such one in the control group by 12.4 times (P < 0.05). The integrated index on color difference (ΔE) exceeded control value by 2.87 times (P < 0.05). More pronounced differences with ΔE were followed by a shorter period of MA appearance with a correlation level at -0.56 (P < 0.05). The specificity of diagnostics raised by 2.17 times (P < 0.05) and negative prognostic index exceeded such one determined with the expert method by 2.26 times (P < 0.05).

Keywords: diabetic retinopathy, multiscale texture gradient, color spectrum analysis, medical diagnostics

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Authors: J. Hidalgo de Quintana, I. Stoner, M. Tackett, G. Doran, C. Rafferty, A. Windemuth, J. Tytell, D. Pregibon

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We have developed the Multiplexed Circulating microRNA assay that allows the detection of up to 68 microRNA targets per sample. The assay combines particle­based multiplexing, using patented Firefly hydrogel particles, with single­ step RT-PCR signal. Thus, the Circulating microRNA assay leverages PCR sensitivity while eliminating the need for separate reverse transcription reactions and mitigating amplification biases introduced by target­-specific qPCR. Furthermore, the ability to multiplex targets in each well eliminates the need to split valuable samples into multiple reactions. Results from the Circulating microRNA assay are interpreted using Firefly Analysis Workbench, which allows visualization, normalization, and export of experimental data. To aid discovery and validation of biomarkers, we have generated fixed panels for Oncology, Cardiology, Neurology, Immunology, and Liver Toxicology. Here we present the data from several studies investigating circulating and tumor microRNA, showcasing the ability of the technology to sensitively and specifically detect microRNA biomarker signatures from fluid specimens.

Keywords: biomarkers, biofluids, miRNA, photolithography, flowcytometry

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Authors: Harriet Jane R. Caleja, Joel I. Ballesteros, Florian R. Del Mundo

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The dengue fever belongs to the world’s major cause of death, especially in the tropical areas. In the Philippines, the number of dengue cases during the first half of 2015 amounted to more than 50,000. In 2012, the total number of cases of dengue infection reached 132,046 of which 701 patients died. Dengue Nonstructural 1 virus (Dengue NS1 virus) is a recently discovered biomarker for the early detection of dengue virus. It is present in the serum of the dengue virus infected patients even during the earliest stages prior to the formation of dengue virus antibodies. A biosensor for the dengue detection using NS1 virus was developed for faster and accurate diagnostic tool. Biotinylated anti-dengue virus NS1 was used as the receptor for dengue virus NS1. Using the Diffractive Optics Technology (dotTM) technique, real time binding of the NS1 virus to the biotinylated anti-NS1 antibody is observed. The dot®-Avidin sensor recognizes the biotinylated anti-NS1 and this served as the capture molecule to the analyte, NS1 virus. The increase in the signal of the diffractive intensity signifies the binding of the capture and the analyte. The LOD was found to be 3.87 ng/mL while the LOQ is 12.9 ng/mL. The developed biosensor was also found to be specific for the NS1 virus.

Keywords: avidin-biotin, diffractive optics technology, immunosensor, NS1

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Authors: Neda Menbari, Ramin Mehdiabadi

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Background: breast cancer remains a critical global health issue, constituting a leading cause of cancer-related mortality in women. MicroRNAs (miRs) are natural RNA molecules that play an important role in cellular processes and regulate post-transcriptional gene expression. MiR-216b-5p is a miR that acts as a tumor suppressor. The expression levels of FoxM1 and miR-216b-5p in malignant and control cells have been evaluated by quantitative polymerase chain reaction (qPCR) technique and flow cytometry. Results: the results of this study revealed a significant downregulation of miR-216b-5p in cancerous cells compared to the control MCF-10A cells (P=0.0004). Interestingly, the expression of miR-216b-5p exhibited an inverse relationship with key clinical indicators such as tumor size, grade, and lymph node invasion. Conclusion: The study's findings showed the prognostic value of miR-216b-5p levels in breast cancer, and its reduced expression correlates with unfavorable tumor characteristics. This research recommends performing more studies on the role of FoxM1 and miR-216b-5p in breast cancer pathology which potentially paving the way for targeted therapeutic interventions.

Keywords: breast cancer, gene expression, FOXM1, microRNA

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Authors: Asha Dukkipati

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Cognitive neuroscience explores neural functioning and aberrant brain activity during cognitive and perceptual tasks. Neurocinematics is a subfield of cognitive neuroscience that observes neural responses of individuals watching a film to see similarities and differences between individuals. This method is typically used for commercial use, allowing directors and filmmakers to produce better visuals and increasing their results in the box office. However, neurocinematics is increasingly becoming a common tool for neuroscientists interested in studying similar patterns of brain activity across viewers outside of the film industry. In this review, it argue that neurocinematics provides an easy, naturalistic approach for studying and diagnosing learning disorders. While the neural underpinnings of developmental learning disorders are traditionally assessed with well-established methods like EEG and fMRI that target particular cognitive domains, such as simple visual and attention tasks, there is initial evidence and theoretical background in support of neurocinematics as a biomarker for learning differences. By using ADHD, dyslexia, and autism as case studies, this literature review discusses the potential advantages of neurocinematics as a new tool for learning disorders research.

Keywords: behavioral and social sciences, neuroscience, neurocinematics, biomarkers, neurobehavioral disorders

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Authors: Chen Wang, Chun Liang

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Microsatellite instability (MSI) is characterized by high degree of polymorphism in microsatellite (MS) length due to a deficiency in mismatch repair (MMR) system. MSI is associated with several tumor types and its status can be considered as an important indicator for tumor prognostic. Conventional clinical diagnosis of MSI examines PCR products of a panel of MS markers using electrophoresis (MSI-PCR) which is laborious, time consuming, and less reliable. MSIpred, a python 2 package for automatic classification of MSI was released by this study. It computes important somatic mutation features from files in mutation annotation format (MAF) generated from paired tumor-normal exome sequencing data, subsequently using these to predict tumor MSI status with a support vector machine (SVM) classifier trained by MAF files of 1074 tumors belonging to four types. Evaluation of MSIpred on an independent 358-tumor test set achieved overall accuracy of over 98% and area under receiver operating characteristic (ROC) curve of 0.967. These results indicated that MSIpred is a robust pan-cancer MSI classification tool and can serve as a complementary diagnostic to MSI-PCR in MSI diagnosis.

Keywords: microsatellite instability, pan-cancer classification, somatic mutation, support vector machine

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Authors: Shima Mahmoudi, Setareh Mamishi, Babak Pourakbari, Majid Marjani

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In the last years, the potential role of distinct T-cell subsets as biomarkers of active tuberculosis TB and/or latent tuberculosis infection (LTBI) has been studied. The aim of this study was to investigate the potential role of interleukin-2 (IL-2) in whole blood stimulated with M. tuberculosis-specific antigens in the QuantiFERON-TB Gold In Tube (QFT-G-IT) for the discrimination of active and latent tuberculosis. After 72-h of stimulation by antigens from the QFT-G-IT assay, IL-2 secretion was quantitated in supernatants by using ELISA (Mabtech AB, Sweden). Observing the level of IL-2 released after 72-h of incubation, we found that the level of IL-2 were significantly higher in LTBI group than in patients with active TB infection or control group (P value=0.019, Kruskal–Wallis test). The discrimination performance (assessed by the area under ROC curve) between LTBI and patients with active TB was 0.816 (95%CI: 0.72-0.97). Maximum discrimination was reached at a cut-off of 13.9 pg/mL for IL-2 following stimulation with 82% sensitivity and 86% specificity. In conclusion, although cytokine analysis has greatly contributed to the understanding of TB pathogenesis, data on cytokine profiles that might distinguish progression from latency of TB infection are scarce and even controversial. Our data indicate that the concomitant evaluation of IFN- γ and IL-2 could be instrumental in discriminating of active and latent TB infection.

Keywords: interleukin-2, discrimination, active TB, latent TB

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Authors: Alex Kizuk

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This paper studies the trope or meme of the Siren in terms of what long-standing cultural continuities can be found in college classrooms today. Those who have raised children may remember reading from Hans Christian Anderson's 'The Little Mermaid' (1836), not to mention regaling them with colorful Disneyesque versions when they were younger. Though Anderson tempered the darker first ending of the story to give the little mermaid more agency in her salvation—a prognostic developed in Disney adaptations—nonetheless, the tale pivots on an image of a 'heavenly realm' that the mermaid may eventually come to know or comprehend as a beloved woman on dry land. Only after 300 years, however, may she hope to see that 'which lives forever' and 'rises through thin air, up to the shining stars. Just as [sea-people] rise through the water to see the lands on earth.' What students today can see in this example is a trope of the agonistic soul in a hard-won disembarkation at a harbour of knowledge--where the seeker after truth may come to know through persistence (300 years)—all that is good and true concerning human life. This paper discusses several such examples from the Great Books and popular culture to suggest that teaching in the world of the 21st century could do worse than accede to some such perennial seeking.

Keywords: the Great Books, tradition, popular culture, 21st century directions in teaching

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Authors: Arif İsmailov, Naila Hasanova, Gunay Orujalieva

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Homocysteine (HCY) is an indicator of prognostic value in monitoring regenerative processes in osteoporosis and osteoporotic fractures. The osteoporosis is known to be a serious health and economic problem, especially for women in the postmenopausal period. The study was carried out on patients 45-83 years old divided into 3 groups: group I – 14 patients with osteoporosis , group II – 15 patients with non-osteoporotic fractures, group III – 25 patients with osteoporotic fractures. The control group consisted of practically healthy 14 people. A blood sample was taken at 3 stages to monitor the dynamics of HCY level: on the 1st day before treatment, on the 10th day of treatment and 1 month after it. Blood levels of Hcy were determined at a wavelength of 450 nm by the ELİSA(Cloud Clone Corp.Elisa kits,USA). The statistical evaluation was performed by using SPSS 26.0 program (IBM SPSS Inc., USA).The results showed that on the 1st day before the treatment HCY concentration was statistically increased 2.7 times(PU = 0.108) in group I, 5.6 times (PU <0.001) in group II and 6.5 times (PU <0.001) in group III compared to the control group. Thus, the average value of HCY in group I was 1.76 ± 0.56 μg/ml; in group II – 3.57 ± 0.62 μg/ml; in group III – 4.2 ± 0.50 μg/ml. HCY level increases more sharply after fractures,especially in osteoporotic patients. In treatment period Vitamin D plays an important role in synthesis of the Cystathionine β‐synthase enzyme, which regulates HCY metabolism. Increased Hcy levels could lead to an increase in the risk of fracture through the interference in collagen cross-linking.

Keywords: homocysteine, osteoporosis, osteoporotic fractures, Vitamin D

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Authors: Madiha Liaqat, Rehan Ahmed Khan, Shahid Kamal

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Multiple imputation with delta adjustment is a versatile and transparent technique for addressing univariate missing data in the presence of various missing mechanisms. This approach allows for the exploration of sensitivity to the missing-at-random (MAR) assumption. In this review, we outline the delta-adjustment procedure and illustrate its application for assessing the sensitivity to deviations from the MAR assumption. By examining diverse missingness scenarios and conducting sensitivity analyses, we gain valuable insights into the implications of missing data on our analyses, enhancing the reliability of our study's conclusions. In our study, we focused on assessing logPSA, a continuous biomarker in incomplete prostate cancer data, to examine the robustness of conclusions against plausible departures from the MAR assumption. We introduced several approaches for conducting sensitivity analyses, illustrating their application within the pattern mixture model (PMM) under the delta adjustment framework. This proposed approach effectively handles missing data, particularly loss to follow-up.

Keywords: loss to follow-up, incomplete response, multiple imputation, sensitivity analysis, prostate cancer

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Authors: J. Bernasovská, R. Lohajová Behulová, E. Petrejčiková, I. Boroňová, I. Bernasovský

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Y chromosome microdeletions are the most common genetic cause of male infertility and screening for these microdeletions in azoospermic or severely oligospermic men is now standard practice. Analysis of the Y chromosome in men with azoospermia or severe oligozoospermia has resulted in the identification of three regions in the euchromatic part of the long arm of the human Y chromosome (Yq11) that are frequently deleted in men with otherwise unexplained spermatogenic failure. PCR analysis of microdeletions in the AZFa, AZFb and AZFc regions of the human Y chromosome is an important screening tool. The aim of this study was to analyse the type of microdeletions in men with fertility disorders in Slovakia. We evaluated 227 patients with azoospermia and with normal karyotype. All patient samples were analyzed cytogenetically. For PCR amplification of sequence-tagged sites (STS) of the AZFa, AZFb and AZFc regions of the Y chromosome was used Devyser AZF set. Fluorescently labeled primers for all markers in one multiplex PCR reaction were used and for automated visualization and identification of the STS markers we used genetic analyzer ABi 3500xl (Life Technologies). We reported 13 cases of deletions in the AZF region 5,73%. Particular types of deletions were recorded in each region AZFa,b,c .The presence of microdeletions in the AZFc region was the most frequent. The study confirmed that percentage of microdeletions in the AZF region is low in Slovak azoospermic patients, but important from a prognostic view.

Keywords: AZF, male infertility, microdeletions, Y chromosome

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Authors: Vandana Singh, Subash Singh

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Introduction: Oral squamous cell carcinoma is the most common malignant tumor of the oral cavity. Normally the death of cell and the growth are active processes and depend not only on external factors but also on the expression of genes like Bcl-2, which activate and inhibit apoptosis. The term Bcl-2 is an acronym for B-cell lymphoma/ leukemia -2 genes. Objectives: An attempt was made to evaluate Bcl-2 oncoprotein expression in patients with oral precancer and cancer and to assess possible correlation between Bcl-2 oncoprotein expression and clinicopathological features of oral precancer and cancer. Material and Methods: This is a selective prospective clinical and immunohistochemical study. Clinicopathological examination is correlated with immunohistochemical findings. The immunolocalization of Bcl-2 protein is performed using the labeled streptavidin biotin (LSAB) method. To visualize the reaction, 3, 3-diaminobenzidine (DAB) is used. Results: Bcl-2 expression was positive in 11 [36.66 %, low Bcl-2 expression 3 (10.00 %), moderate Bcl-2 expression 7 (23.33 %), and high Bcl-2 expression 1 (3.33 %)] oral cancer cases and in 14 [87.50 %, low expression 8 (50 %), moderate expression 6 (37.50 %)] precancer cases. Conclusion: On the basis of the results of our study we conclude that positive Bcl-2 expression may be an indicator of poor prognosis in oral cancer and precancer. Relevance: It has been reported that there is deregulation of Bcl-2 expression during progression from oral epithelial dysplasia to squamous cell carcinoma. It can be used for revealing progression of epithelial dysplasia to malignancy and as a prognostic marker in oral precancer and cancer.

Keywords: BcL-2, immunohistochemistry, oral cancer, oral precancer

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Authors: Zahra Salehi Shahrbabaki, Zahra Chavoshzadeh, Fahimeh Abdollahimajd, Samin Sharafian, Tolue Mahdavi, Mahnaz Jamee

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Background: Inborn errors of immunity (IEIs) are monogenic diseases of the immune the system with broad clinical manifestations. Despite the increasing genetic advancements, the diagnosis of IEIs still leans on clinical diagnosis. Dermatologic manifestations are observed in a large number of IEI patients and can lead to proper approach, prompt intervention and improved prognosis. Methods: This cross-sectional study was carried out between 2018 and 2020 on IEIs at a Children's tertiary care center in Tehran, Iran. Demographic details (including age, sex, and parental consanguinity), age at onset of symptoms and family history of IEI with were recorded. Results :212 patients were included. Cutaneous findings were reported in (95 ,44.8%) patients. and 61 of 95 (64.2%) reported skin lesions as the first clinical presentation. Skin infection (69, 72.6%) was the most frequent cutaneous manifestation, followed by an eczematous rash (24, 25 %). Conclusions: Skin manifestations are common feature in IEI patients and can be readily recognizable by healthcare providers. This study tried to provide information on prognostic consequences.

Keywords: primary immuno deficiency, inborn errror of metabolism, skin manifestation, skin infection

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Authors: Ghada E. Esheba, Ghufran Kheshaifaty, Kholoud Al-Harbi, Wafa'a Al-Harbi, Ala'a Al-Orabi, Moayad Turkistani

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Krukenberg tumor is a rare metastatic ovarian carcinoma that usually occurs in female between 30 - 40 year old and rarely seen after menopause. Stomach is the most common primary site. Histopathological features of krukenberg tumors appear as diffuse stromal proliferation, mucus-production, and numerous signet-cells and these tumors spread mostly by lymphatic route. Treatment and prognostic factors are not well established. This study describes a 34 year old female with a unilateral ovarian mass discovered accidentally during cesarean section delivery and it was misdiagnosed as luteoma of pregnancy, but histopathological examination showed a diffuse infiltration of the ovary and omentum by signet ring cells. These findings were not correlated with luteoma of pregnancy or any other types of primary ovarian tumors like surface epithelial tumor, sex cord stromal tumor or germ cell tumor. However, after the analysis of immunohistochemical results (negative CK7, positive CK20 and CDX-2), the finding was the diagnostic of metastatic krukenberg tumor. Two weeks later, the patient was evaluated and a large gastric tumor was found in her stomach and she underwent gastrectomy.

Keywords: CK7, CK20, CDX-2, Krukenburg tumor, metastatic ovarian tumor

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Authors: Saurav Dhawan, Sanchit Bansal

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Background T-MACS is a computer-based decision aid that ‘rules in’ and ‘rules out’ ACS using a combination of the presence or absence of six clinical features with only one biomarker measured on arrival: hs-cTnT. T-MACS had 99.3% negative predictive value and 98.7% sensitivity for ACS, ‘ruling out’ ACS in 40% of patients while ‘ruling in’ 5% at the highest risk. We aim at benchmarking the use of T-MACS which could help to conserve healthcare resources, facilitate early discharges, and ensure safe practice. Methodology Randomized retrospective data collection (n=300) was done from ED electronic records across 3 hospital sites within MFT over a period of 2 months. Data was analysed and compared by percentage for the usage of T-MACS, number of admissions/discharges, and in days for length of stay in hospital. Results MRI A&E had the maximum compliance with the use of T-MACS in the trust at 66%, with minimum admissions (44%) and an average length of stay of 1.825 days. NMG A&E had an extremely low compliance rate (8 %), with 75% admission and 3.387 days as the average length of stay. WYT A&E had no TMACS recorded, with a maximum of 79% admissions and the longest average length of stay at 5.07 days. Conclusion All three hospital sites had a RAG rating of ‘RED’ as per the compliance levels. The assurance level was calculated as ‘Very Limited’ across all sites. There was a positive correlation observed between compliance with TMACS and direct discharges from ED, thereby reducing the average length of stay for patients in the hospital.

Keywords: ACS, discharges, ED, T-MACS

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Authors: Ramji Kalidoss, Snekhalatha Umapathy, V. Dhinakaran, J. M. Mathana

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Chemi-resistive sensors used in breathalyzers have become a hotspot between the international breath research communities. These sensors exhibit a significant change in its resistance depending on the temperature it gets heated thus demanding high power leading to non-portable instrumentation. In this work, numerical simulation to identify the suitable combination of substrate and heater profile using COMSOL multiphysics was studied. Ni-Cr and Pt-100 joule resistive heater with various profiles were studied beneath the square and circular alumina substrates. The temperature distribution was uniform throughout the square substrate with the meander shaped pt100 heater with 48 mW power consumption for 200 oC. Moreover, this heater profile induced minimal stress on the substrate with 0.5 mm thick. A novel Graphene based ternary metal oxide nanocomposite (GO/SnO2/TiO2) was coated on the optimized substrate and heater to elucidate the response of diabetes biomarker (acetone). The sensor exhibited superior gas sensing performance towards acetone in the exhaled breath concentration range for diabetes (0.25 – 3 ppm). These results indicated the importance of substrate and heater properties along with sensing material for low power portable breathalyzers.

Keywords: Breath Analysis, Chemical Sensors, Diabetes Mellitus, Graphene Nanocomposites, Heater, Substrate

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Authors: Kazeem Ajasa Badmus, Zulkifli Idrus, Goh Yong Meng, Kamalludin Mamat-Hamidi

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This study was aimed at evaluating the roles played by early age in performance, organs weights, meat quality traits, and telomere length integrity. One hundred male Cobb 500® broiler chickens were grouped into ten replicates of ten chickens each. Growth performance, measurement of telomere length, weights of organs, and meat quality traits were determined on days 14, 28, and 42 of the experiment. There were significant (p < 0.05) differences obtained in the chicken growth performance across ages. Telomere length of blood, muscle, liver, and heart on day 14 were significantly (p < 0.05) shorter than telomere length obtained on days 28 and 42 of the age. Weights of organs on day 14 were significantly (p < 0.05) higher than those obtained on days 28 and 42. In this study, birds slaughtered on day 14 presented the highest (p < 0.05) pH, drip loss, redness, and yellowness. They, however, showed lower (p < 0.05) cooking loss, shear force, and lightness. There was a significant association between age, telomere length, and meat quality traits. It is therefore concluded that telomere length attrition is associated with early age metabolic activities and could be used to measure chicks' welfare.

Keywords: age, telomere length, organ weights, meat quality

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