Drop Of Blood Research Articles

A Day Saved is a Life Saved: Direct Antimicrobial Susceptibility Testing from Positively Flagged Blood Culture Bottles and their Concordance with the Routine Method.

Sepsis is a major health problem worldwide and is associated with high morbidity and mortality with every hour delay in initiation of therapy. A conventional method of blood culture and Antimicrobial Susceptibility Testing (AST) takes around 48-72 hours. Empirical antibiotics need to be administered until the sensitivity report is made available. It has been estimated that 20-50% of the empirical antibiotics are inappropriate, resulting in prolonged hospital stays, adverse effects, and emergence of drug resistance. Additionally, this also puts an extra financial burden on both the patients and healthcare settings. Performing direct Antimicrobial Sensitivity Testing (dAST) is an important tool to reduce turn-around time (TAT) by at least 18-24 hours, thus reducing morbidity and mortality among critically ill patients. Direct AST (dAST) was performed from the positively flagged blood culture bottles received between December, 2021 to May, 2022 from Intensive Care Units (ICUs) on Mueller- Hinton Agar (MHA) using four drops of withdrawn blood. dAST was performed for six drugs: Ceftriaxone-30 μg (CTR), Piperacillin/Tazobactam-100/10 μg (PIT), Meropenem-10 μg (MRP), Ciprofloxacin-5 μg (CIP), Aztreonam-30 μg (AT), and Colistin (CL). The zone of inhibition was interpreted as per CLSI M100 ed32, 2022 guidelines. A parallel conventional method was also performed to examine for categorical agreement and disagreement. Identification was carried out using MALDI-TOF MS from the colonies that appeared on the dAST plate on the subsequent day. A total of 162 positively flagged blood culture bottles were included in the study. The majority of the Gram-negative organisms were from Enterobacterales (n=109), followed by Acinetobacter spp. (n=28) and Pseudomonas aeruginosa (n=25). Out of the 972 isolate-antimicrobial combinations, overall Categorical Agreement (CA) was seen in 936 (96.3%), whereas disagreement was observed in 36 with minor error (mE) in 21 (2.2%), major error (ME) in 7 (0.7%), and very major error (VME) in 8 (0.8%) when compared to the routine method. Categorical agreement (CA) of > 99% was seen in ceftriaxone (CTR) and ciprofloxacin (CIP). In comparison, the lowest CA was observed with meropenem (MRP) at 92%. Colistin dAST was performed using the E-strip method, and the result obtained was highly convincing, with an overall disagreement of only 1.2%. Rapid dAST from positively flagged blood culture bottles proved to significantly reduce the TAT from the time of sample collection to the first availability of antimicrobial susceptibility report with excellent categorical agreement of > 95% using the conventional disc diffusion method. Results obtained were within the acceptance criteria set by U. S. Food and Drug Administration (FDA) guidelines of > 90% categorical agreement for a new method. We were able to obtain excellent concordance for colistin using the E-strip method. Performing dAST not only saves a "day", but its proper implementation would save a "life".

LuLIPLEX: A Fast, Highly Sensitive, and Multiplexed Method for the Detection of IgE Against Major Allergens.

Diagnosis of allergies is mostly based on the patient's clinical history and allergen provocation tests. Determination of specific IgE (sIgE) profiles can be performed to support allergy diagnosis. This is commonly done invivo by the skin prick test or invitro with automated systems. Several platforms exist to quantify sIgE levels, but all these methods require access to specific instruments, often delaying the test's results. The IgE luciferase-linked immunosorbent assay (LuLISA) allows bioluminescent quantification of IgE against peanut in microliter samples, but this method awaits extension to other allergens. This study aimed to validate a new method, named LuLIPLEX, for multiplexed bioluminescent detection of sIgE against 20 major molecular allergens. Quantification of sIgE against 12 recombinant or purified food allergens and eight aeroallergens was performed by LuLIPLEX versus standard IgE detection methods (ImmunoCAP, ISAC, ALEX, or NOVEOS). Multiplexed detection of IgE against these 20 allergens was performed within 45 min using 50 μL of serum, plasma, or whole blood samples. A head-to-head comparison between LuLIPLEX and standard IgE detection methods showed a high correlation among all allergens tested. sIgE profiles in polysensitized subjects could be determined within 45 min in serum and plasma samples, as well as using a single drop of capillary blood. LuLIPLEX is a rapid and sensitive method to quantify sIgE levels against multiple allergens. Given that the test is very fast and can be performed on small and inexpensive luminometers, the IgE LuLIPLEX could allow point-of-care testing of sIgE profiles in allergic subjects.

Observational Analysis of Point-of-Care Lactate Plus™ Meter in Preclinical Trauma Models

Background/Objectives: Blood lactate concentration is often used to assess systemic hypoperfusion, tissue hypoxia, and sepsis in trauma patients and serves as a prognostic indicator and marker of response to therapy. Point-of-care (POC) devices provide rapid lactate measurements with a single drop of blood. In this study, lactate values from whole blood, measured with two POC devices, Abbott i-STAT® and the Nova Biomedical Lactate (LA) Plus™ meter, are compared. Methods: An observational analysis of 760 blood samples from four preclinical trauma animal models was performed. Lactate was measured simultaneously with two POC devices (i-STAT® and the Lactate Plus™ meter). The evaluation in the form of a Bland–Altman diagram showed the agreement of the tests. Results: The Spearman correlation was 0.9180 (95% CI 0.9056 to 0.9288) for i-STAT® vs. the LA Plus™ meter. Both tests showed a significant increase in lactate after injury (p < 0.05). The i-STAT® showed a small bias compared to the LA Plus™ meter (−0.0227 ± 0.4542 with 95% limits of agreement from −0.09129 to 0.8675). The LA Plus™ meter showed slightly higher values overall (0.0227 mmol/L), but the i-STAT® showed higher results at lactate levels above 2.5 mmol/L. Conclusions: The observed similarity in performance between the two devices suggests that both are capable of reliably measuring lactate levels in a trauma situation. The LA Plus™ meter provides fast results with only one drop of blood. This study supports the prehospital use of POC devices.

Molecular detection of haemobacteria in Colombian wild birds

Colombia shows a high density and variety of bird species, making it one of the most diverse avian territories globally. Antioquia ranks among the top four provinces with the greatest variety of bird species, underscoring the importance of research efforts on the local bird fauna. Therefore, this study aimed to identify bacterial agents in the blood of wild birds from the municipality of Jardín (Antioquia, Colombia) by 16S qPCR sequencing. A descriptive cross-sectional study was conducted using non-probabilistic convenience sampling. Wild birds were captured using mist nets and blood samples were collected from each animal via puncture using sterile lancets in the brachial vein, and a drop of blood was collected on filter paper for qPCR analysis. The 16S gene in bacterial genomes was found in 13 out of 46 wild birds of the Passeriform and quasi-Passeriform orders, captured at three different locations within the study municipality at altitudes ranging from 1,665 to 2,034 m.a.s.l. Seven different bird species were recorded and four different haemobacteria were identified (i.e. Exiguobacterium spp., Escherichia coli, Stenotrophomonas spp., and Stenotrophomonas maltophilia). This study contributes to the knowledge in Colombia by identifying four different hemobacteria in wild birds. Further research is required on the health status of these birds and the attributable impacts on their populations and other related factors, including humans.

Newborn Screening Program for Cystic Fibrosis in Türkiye: Experiences from False-Negative Tests and Requirement for Optimization.

Since January 2015, the Cystic Fibrosis National Newborn Bloodspot Screening (CF-NBS) program has been implemented in Turkey with two samples of immune reactive trypsinogen (IRT-1/IRT-2) testing. This study aimed to evaluate the Turkish national CF screening program, which included patients referred to a tertiary pediatric pulmonology center, to ascertain the optimal cut-off values for IRT-1/IRT-2 and to identify alternative strategies for mitigating the number of late-diagnosed false-negative patients (FNPs) who initially exhibited screen negative results but were diagnosed subsequently based on clinical suspicion. The study also compared NBS-positive patients to FNPs to determine the influence of delayed diagnosis. A retrospective cohort study. Screening for CF was conducted in accordance with the national CF-NBS program within 48-72 hours of birth by collecting a few drops of heel blood on Guthrie paper. A cut-off value of 90 μg/l was accepted for the first IRT, while 70 μg/l was accepted for the second sample. Infants with elevated IRT values in both samples were referred to the CF centers for a sweat test (ST). Based on the diagnosis, the NBS-positive infants referred to our CF center for ST analysis were divided into three groups: CF; cystic fibrosis-related metabolic syndrome/cystic fibrosis screen positive, inconclusive diagnosis (CRMS/CFSPID); and false-positive NBS. In addition, the study included NBS-negative patients who initially received negative screen results but were subsequently diagnosed with CF based on clinical suspicion. Of the 227 NBS-positive infants referred within the study period, 53 (23.34%) were diagnosed with CF (true-positive NBS), 11 were classified as CRMS/CFSPID (4.84%), and 163 were classified as falsepositive NBS (71.8%). CF was diagnosed in 66 infants, 53 (80.3%) of whom were confirmed using the NBS test, while the 13 (19.7%) patients who were missed on the NBS test were diagnosed based on clinical suspicion (FNP). The study findings indicate that the IRT/IRT approach exhibited a sensitivity of 80.3% and a positive predictive value (PPV) of 23.3%. The current study is the first to analyze the NBS program for CF using data from the Western Anatolian Region of Türkiye. Due to the low sensitivity and PPV of the IRT/IRT protocol and the high proportion of false-positive infants and FNPs, the current national program is not practicable for Türkiye. False-negative results significantly delay the diagnosis and invalidate the screening objectives. It is essential to establish optimal cut-off values for IRT-1/IRT-2 or revise existing strategies to reduce the number of FNPs missed by the screening program.

High frequency of artemisinin partial resistance mutations in the Great Lakes region revealed through rapid pooled deep sequencing.

In Africa, the first Plasmodium falciparum artemisinin partial resistance mutation, was Kelch13 (K13) 561H - detected and validated at appreciable frequency in Rwanda in 2014. Surveillance to better define the extent of the emergence in Rwanda and neighboring countries is critical. We used novel liquid blood drop preservation combined with pooled sequencing to provide cost-effective rapid assessment of resistance mutation frequencies at multiple collection sites across Rwanda and neighboring regions in Uganda, Tanzania, and the Democratic Republic of the Congo (DRC). Malaria-positive samples (n=5,465) from 39 health facilities collected between May 2022 and March 2023 were sequenced in 199 pools. In Rwanda, K13 561H and 675V were detected in 90% and 65% of sites with an average frequency of 19.0% (0-54.5%) and 5.0% (0-35.5%), respectively. In Tanzania, 561H had high frequency in multiple sites. 561H appeared at 1.6% in Uganda. 561H was absent from the DRC, although 675V was seen at low frequency. Concerningly candidate mutations were observed: 441L, 449A, and 469F co-occurred with validated mutations suggesting they are arising under the same pressures. Other markers for decreased susceptibility to artemether-lumefantrine are common: P. falciparum multidrug resistance protein 1 N86 at 98.0% (63.3-100%) and 184F at 47.0% (0-94.3%) and P. falciparum chloroquine resistance transporter 76T at 14.7% (0-58.6%). Additionally, sulfadoxine-pyrimethamine-associated mutations show high frequencies. K13 mutations are rapidly expanding in the region further endangering control efforts with the potential of engendering partner drug resistance.

A-155 Development of drop-of-blood QATT heparinase and functional fibrinogen assays

Abstract Background Monitoring the blood coagulation status of patients with coagulopathy or on anticoagulant therapy is important for informed treatment decisions. Quasi-static acoustic tweezing thromboelastometry (QATT) can monitor changes in blood coagulation using a small blood drop. Here, we develop the heparinase and functional fibrinogen QATT assays. Methods Citrated whole blood (WB) samples were collected from 20 healthy volunteers using IRB-approved protocols No. 520566 and 944474. Half of the samples were used for fibrinogen, platelet count tests and the others were used for QATT measurements. For developing heparinase assay, WB samples from 4 healthy volunteers were randomly selected. For each volunteer sample, QATT experiments were performed for a WB sample, a WB sample spiked with 0.5 IU/mL heparin, and a WB sample spiked with heparin and then heparinase. For developing functional fibrinogen assay, QATT experiments were performed for a WB sample and a WB sample spiked with Cytochalasin-D (4 μM). Coagulation was triggered via intrinsic pathway. Pearson correlation coefficient (rp) or the Spearman rank correlation coefficient (rs) was used for correlation analysis. We focused on the following QATT parameters: CIT, MCF, and MCE. Results The heparinase assay effectively reversed heparinized samples (Fig. 1a). CIT of normal and heparinized samples treated with heparinase were within reference range (Fig. 1b). The effect of Cytochalasin-D was shown in Fig 1c. Strong correlation was found between MCE of Cyto-treated WB and fibrinogen level (rp = 0.75). MCE of the difference between normal and Cyto-treated WB, referred to as MCEplatelet, was also strongly correlated with platelet count test (rs = 0.76). The normal range for MCF of the functional fibrinogen assay is efficient in ruling out abnormal fibrinogen level (Fig. 1d). Conclusions QATT accurately predicted the inhibition of heparin by heparinase. QATT functional fibrinogen assay detected the contribution of both fibrinogen and platelets to the clot strength.

Antithrombotic activity of Punica granatum, L (Pomegranate): Experimental investigation and exploration of its mechanism of action on primary and secondary hemostasis in vitro and ex vivo

Hemostasis is the physiological process that stops bleeding by forming a thrombus (or blood clot). However, the inappropriate formation of a thrombus can constitute a pathophysiological event involving the same steps as those of hemostasis. Despite some adverse effects, antithrombotic therapy remains an effective means in the prevention and treatment of thrombotic diseases. In this regard, scientific research aimed at discovering new remedies of natural origin is particularly active.This study aims to evaluate the antithrombotic activity of the aqueous extract of Punica granatum peels in vivo. To understand the mechanism of action of this activity, we will investigate primary and secondary hemostasis in vitro and ex vivo. Furthermore, we will examine its antioxidant activity, toxicity, and phytochemical composition to identify the molecules responsible for the observed effects.The antithrombotic activity is assessed in vivo using the acute pulmonary thromboembolism model in mouse. Thrombosis is induced by injecting the thrombogenic solution (epinephrine + collagen) into the lateral caudal vein. Platelet aggregation, both in vitro and ex vivo, is performed on washed platelets isolated by centrifugation from rat blood. The platelet suspension is pre-incubated with the extract for 1 min. Aggregation is triggered by adding the following agonists: ADP, thrombin, collagen, and arachidonic acid. The bleeding time is measured in the absence and presence of the extract following an apical section of the mouse tail. This time corresponds to the duration of flow between the first and last drops of blood. To evaluate plasma coagulation, activated partial thromboplastin time (aPTT) and prothrombin time (PT) were measured under conditions both with and without the extract, using appropriate reagents. Acute toxicity is conducted by administering a single oral dose of the extract to mice (5 and 10 g kg-1). Possible changes in animal behavior are observed for fifteen days. The identification of polyphenols and flavonoids is performed by High Performance Liquid Chromatography (HPLC).The results obtained show that P. granatum exerted a significant preventive antithrombotic action (80 % protection) in mice. Furthermore, the extract inhibited platelet aggregation induced in vitro by all agonists in a dose-dependent manner, prolonged bleeding time, and coagulation times in rats. The results of the ex vivo study in rats confirmed these effects. We also noted that the extract did not exhibit toxicity following administration, and HPLC analysis revealed a richness of polyphenols and flavonoids in the extract.P. granatum exhibited an interesting antithrombotic activity, which could be attributed to its antiplatelet and anticoagulant actions. The phytochemical compounds identified in this extract could, at least in part, be responsible for these effects. Therefore, the demonstrated activities of this plant on hemostasis could lead to the development of more effective remedies and represent a means of prevention against thrombotic events.

Usability study of the qLabs® FIB: A new point-of-care system for functional fibrinogen testing

BackgroundIdentification of hypofibrinogenemia is particularly important in obstetrics since this predicts progression to severe Post-Partum Haemorrhage (PPH). ObjectivesTo evaluate the usability of qLabs® FIB at the bedside of patients with PPH. MethodsThe qLabs® FIB test was performed using a drop of whole blood from a citrated laboratory tube sampled from each parturient with PPH >1 L by a trained user. A usability survey was completed by each clinician performing the test. ResultsDuring the evaluation, 101 qLabs® FIB tests were performed. One hundred completed surveys and 58 free comments were collected. Satisfaction was achieved in 84 % of tests. Usability in emergency setting, timeliness of results, and the results display were considered positive. The qLabs® FIB results were obtained in <3 min for concentrations below 3 g/L whilst the median time to paired laboratory results was 60 (20–120) minutes. The lower a patient's fibrinogen level, the faster the qLabs® FIB result. ConclusionThe usability study was the first step in the validation process of the qLabs® FIB point of care device in obstetric settings.

Multiple oxidative modifications on hemoglobin are elevated in breast cancer patients as measured by nanoflow liquid chromatography-tandem mass spectrometry

Breast cancer is strongly connected with elevated oxidative stress. Oxidative modifications of hemoglobin can serve as biomarkers for monitoring oxidative stress status in vivo. The structure of hemoglobin modifications derived from malondialdehyde (MDA) in human blood hemoglobin exists as N-propenal and dihydropyridine (DHP). This study reports the simultaneous quantification of eleven modified peptides in hemoglobin derived from MDA and advanced histidine oxidation in 16 breast cancer patients and 16 healthy women using nanoflow liquid chromatography nanoelectrospray ionization tandem mass spectrometry. The results reveal statistically significant increases in the formation of MDA-derived N-propenal and DHP of lysine and advanced oxidation of histidine in hemoglobin of breast cancer patients with the Mann-Whitney U-test p values < 0.0001 and the AUC of ROC between 0.9277 and 1.0. Furthermore, the elevation in modified peptides is significant in patients with early stages of breast cancer. By measuring these oxidative modifications in hemoglobin from a drop of blood, the role of lipid peroxidation and oxidative stress in breast cancer can be assessed using this sensitive assay.

The Association of Dietary Micronutrient Intake and Systemic Inflammation among Patients with Type 2 Diabetes: A Cross-Sectional Study.

Inflammation contributes to the pathogenesis of type 2 diabetes (T2DM). This study sought to document how the systemic biomarkers of inflammation varied based on food choices among patients with T2DM. This cross-sectional study enrolled ambulatory patients with T2DM. Demographic and clinical information was collected. Five drops of fingerstick blood were collected using an absorbent paper device (HemaSpot HFR). C-reactive protein (CRP), serum amyloid A protein (SAA), and fibrinogen were measured using a Luminex assay. Patient-generated 7-day food diaries were analyzed using a validated food processor software. Data were analyzed by Pearson's correlation tests, linear regression and logistic regression with the significance level set at 0.05. Among the 71 participants, 43 (60.6%) were females. The average age and duration of T2DM were 64.1 ± 10.3 and 15.8 ± 9.1 years, respectively. In a simple linear regression run with selected micronutrients, iron [F (1, 53) = 5.319, p < 0.05, adj. R2 = 0.074] significantly predicted plasma CRP. This significance was lost with multiple linear regressions including age, gender, BMI, T2DM duration, T2DM complications, glycohemoglobin A1c (HbA1c) and other micronutrients. The average intake of most micronutrients by the participants was below the recommended daily intake. A higher intake of iron-rich foods was associated with higher levels of systemic inflammation in a simple linear regression model, but the association was not present after adjusting for patient factors like age, gender, BMI and T2DM-related variables. This relationship needs to be explored further given the key role of inflammation in the pathogenesis of T2DM and its associated complications.

The Flip and Fix internal limiting flap technique versus the classic temporal flap for macular hole repair.

To describe a novel technique of macular hole (MH) repair and compare it to the perfluorocarbon liquid (PFCL)-assisted temporal internal limiting membrane (ILM) flap technique. Twenty-two eyes of 22 patients with MH were randomized on a 1:1 basis into two groups. Group A cases were treated using the Flip and Fix technique, in which the single-layer ILM flap was flipped, flattened over the macula using PFCL, and fixed in place using two drops of autologous nonheparinized blood over the superonasal and inferonasal edges of the flap (away from the MH center). Group B patients had the classic temporal ILM flap technique without using the PFCL or autologous blood. All patients were followed up for a period of 6 months after the surgery. No significant difference was observed in initial and final visual acuities between Groups A and B. Intraoperative flap displacement did not occur in Group A and occurred in two eyes in Group B (18.2%). One of them showed failed MH closure and required reoperation. These two cases had a final (W shape) MH closure and a worse final vision than the remaining cases, which had either a U shape or V shape final MH closure. The study results show that the Flip and Fix technique is as effective as the PFCL-assisted temporal ILM flap technique and is associated with less ILM flap displacement risk for the repair of macular holes.

Handheld microfluidic multiple detection device for concurrent blood urea nitrogen and creatinine ratio determination using colorimetric approach

A novel microfluidic multiple detection (MMD) platform consisting of a microfluidic chip and a self-built detection system is proposed for determining the creatinine (CRE) and blood urea nitrogen (BUN) concentrations in whole blood samples. In the proposed approach, a drop of whole blood is injected into the inlet of the microfluidic chip and filtered through strips of hemofiltration paper. The plasma enters the BUN and CRE detection chambers and is absorbed by pre-placed test strips embedded with analyte-specific reagents. Secondary reagents are then introduced into the detection chambers using a finger pump, and the chip is heated to induce Berthelot and Jaffe reactions. An image of the reaction compounds is captured by a CMOS camera and transmitted to a smartphone, where the BUN and CRE concentrations are derived from predetermined calibration curves using RGB analysis software. The proposed MMD platform is demonstrated by comparing the results obtained for 43 whole blood samples collected from patients with chronic kidney disease (CKD) and health volunteers with the measurements obtained using a traditional benchtop system. The results confirm that the proposed MMD platform offers a cost-effective, convenient, and rapid technique for BUN/CRE ratio (BCR) determination in point-of-care (POC) contexts.

Multi-Biomarker Profiling for Precision Diagnosis of Lung Cancer.

Multi-biomarker analysis can enhance the accuracy of the single-biomarker analysis by reducing the errors caused by genetic and environmental differences. For this reason, multi-biomarker analysis shows higher accuracy in early and precision diagnosis. However, conventional analysis methods have limitations for multi-biomarker analysis because of their long pre-processing times, inconsistent results, and large sample requirements. To solve these, a fast and accurate precision diagnostic method is introduced for lung cancer by multi-biomarker profiling using a single drop of blood. For this, surface-enhanced Raman spectroscopic immunoassay (SERSIA) is employed for the accurate, quick, and reliable quantification of biomarkers. Then, it is checked the statistical relation of the multi-biomarkers to differentiate between healthy controls and lung cancer patients. This approach has proven effective; with 20µL of blood serum, lung cancer is diagnosed with 92% accuracy. It also accurately identifies the type and stage of cancer with 87% and 85%, respectively. These results show the importance of multi-biomarker analysis in overcoming the challenges posed by single-biomarker diagnostics. Furthermore, it markedly improves multi-biomarker-based analysis methods, illustrating its important impact on clinical diagnostics.

Clinical metabolomics: current state and prospects in Russia.

Using analytical technologies it is possible now to measure the entire diversity of molecules even in a small amount of biological samples. Metabolomic technologies simultaneously analyze thousands of low-molecular substances in a single drop of blood. Such analytical performance opens new possibilities for clinical laboratory diagnostics, still relying on the measurement of only a limited number of clinically significant substances. However, there are objective difficulties hampering introduction of metabolomics into clinical practice. The Institute of Biomedical Chemistry (IBMC), consolidating the efforts of leading scientific and medical organizations, has achieved success in this area by developing a clinical blood metabogram (CBM). CBM opens opportunities to obtain overview on the state of the body with the detailed individual metabolic characteristics of the patient. A number of scientific studies have shown that the CBM is an effective tool for monitoring the state of the body, and based on the CBM patterns (signatures), it is possible to diagnose and monitor the treatment of many diseases. Today, the CBM creation determines the current state and prospects of clinical metabolomics in Russia. This article, dedicated to the 80th anniversary of IBMC, is a review of these achievements focused on a discussion of their implementation in clinical practice.

Correlation between the prothrombin time values obtained by point-of care prothrombin time meter and clinical laboratory in paediatric intensive care unit of a developing country

Background: Monitoring of prothrombin time (PT) is essential for adequate management of children in pediatric intensive care unit (PICU) who require serial and repeated PT measurements. In contrast to laboratory testing, point-of-care (POC) PT meters have been configured especially for bedside that are cost effective with rapid turnaround time. Our study aimed to assess its feasibility for routine use in PICU by assessing the correlation between PT values obtained through the POC PT meter and those acquired from the laboratory. Methods: A total of 200 children of either gender aged between 1 month to 12 years, admitted to PICU of Indus hospital Karachi Pakistan from February to August 2023 were analysed in this cross-sectional study. Venous blood samples were collected in PT bottles and transported to the pathology department within a time frame of 20-30 minutes for analysis. Concurrently, a drop of blood was obtained via finger prick and promptly assessed using the POC device-Coaguchek INRange system, (Roche diagnostics Ltd, USA). Pearson’s correlation was applied. Results: 200 children (96 males, 104 females) having a mean age of 6.42±3.55 years. The mean PT values with POC meter and Laboratory were 2.588±1.299 and 2.574±1.304 respectively (p=0.100). Very strong correlation of PT between POC PT meter and laboratory evaluation was observed (r=0.996, p&lt;0.001). Conclusions: The strong correlation observed between POC PT meter and laboratory supports the use of these devices for timely coagulation monitoring, potentially improving patient outcomes in PICUs.

Comparative Evaluation of Pfeiffer’s Blood Crystallization Test for Early Detection of Epithelial Dysplasia in OSMF and Healthy Individuals: A Cross-Sectional Study

Background: Unlike other malignancies, oral cancer is preceded by Oral Potentially Malignant Disorders (OPMDs). Oral Submucous Fibrosis (OSMF) is one such OPMD with malignant potential of 7.6%. The purpose of present study was to determine the efficacy of Pfeiffer’s blood Crystallization test for early detection of epithelia dysplasia in OSMF individuals. Material and methods: The study participants included were involved 50 OSMF subjects and 50 healthy controls. The blood drop collected was subjected to crystallization test and the crystal patterns formed were analyzed after 20 hours. Incisional biopsy was performed in OSMF subjects to grade the degree of epithelial dysplasia. The data obtained was subjected to statistical analysis. Results: The participant’s age in the study groups ranged up to 56 years. Difference in the crystallization pattern between diseased group and non-diseased group was significant (p-value &lt; 0.05%) with more transverse pattern in diseased group. The OSMF participants with no epithelial dysplasia showed normal crystallization pattern and maximum participants with dysplasia showed transverse pattern and the difference was statistically significant (p-value&lt;0.05%). The sensitivity and specificity of crystallization test for OSMF was found to be 91.4% and 100.0% respectively. Conclusion: Pfeiffer's Crystallization test can be considered as a less invasive and reliable screening aid for OSMF which helps clinicians to identify high‑risk individuals for malignant transformation.

Three-dimensional microfluidic paper-based analytical device fabricated by cutting technique for highly sensitive and specific colorimetric detection of triglycerides in whole blood utilizing a mixed indicator system

In this study, we developed a three-dimensional microfluidic paper-based analytical device (3D-µPAD) for quantifying triglycerides (TGs) in whole blood samples. This method provides an alternative to conventional TGs detection and offers several advantages. The 3D-µPAD was fabricated using an in-house computer-controlled X-Y knife plotter, eliminating the need for hydrophobic reagents. This approach results in a more rapid, simple, highly reproducible, and cost-effective mass production process. To separate plasma from a drop of whole blood, the LF1 membrane was incorporated as a sample zone on the 3D-µPAD. The hydrolysis of TGs was catalyzed by lipase, producing H+ ions detected by a phenol red indicator read against cyan ink to reduce background interference. A spectrophotometer (i1 Pro 3 mini) measured the green color intensity in the 3D-µPAD’s detection zone, which was proportional to the TGs concentration and quantified within 10 min. Results were consistent with clinical methods, with a detection limit of 0.182 mmol L−1 in the 0.1–6 mmol L 0207B1 TGs concentration range. Using mixed indicators successfully reduced background interference and enhanced sensitivity for TGs determination. The 3D-µPAD demonstrated a storage stability of approximately 30 days. Our 3D-µPAD prototype offers a rapid, cost-effective, and user-friendly approach for pre-clinical TGs diagnostics, expanding the potential for point of care whole blood sample analysis.

Assess human blood uranium levels of some Iraqi companies

The goal of this study is to measure the uranium concentration levels in the blood of Iraqi workers employed in certain government companies. Assessing the initial level of uranium toxicity in their blood and the possibility of health problems occurring. 184 blood samples from Iraqi government companies and the control group were collected in this study. A solid-state nuclear track detector (CR-39) was used to measure the amount of uranium present. Two drops of blood (100 μl) were placed on CR-39. The CR-39 was irradiated with a thermal neutron using the fission-track technique (241Am-9Be) to determine the uranium concentration in blood samples. The statistical analysis is carried out using the Origin Lab 2024 version. The results show the average of uranium concentration at all locations has a higher level compared to the control group. The blood samples from workers at the phosphate company had the highest amount (1.021 ± 0.050 μg/l), compared to samples from other factories. This result confirms that there is a connection between the concentration of uranium and phosphate substances. The results suggest that there is a slight increase in uranium levels that is related to both age and years of employment.

First Report on Clinical Feasibility of Dried Blood Spot Technique for Hemoglobin Estimation in Cholistani Cattle

Background: The dried blood spot (DBS) technique using filter papers has revolutionized the conventional blood sampling techniques through ease of blood collection, storage and transport. Various analytes (such as hormones, antigens, antibodies and hematochemical attributes) are being estimated through DBS globally. However, this technique has not yet been implied in Pakistan. This research work is the first of its kind regarding hemoglobin (Hb) estimation in Cholistani cattle (n=63) blood through DBS technique using filter paper.Methods: Three methods of Hb estimation were implied in the present study viz. through veterinary hematology analyzer (HbA), and two indirect cyanmethemoglobin methods (HbIC and HbICX) using measured (20µL) and unmeasured blood drops on the filter paper, respectively.Results: Results revealed that HbA and HbIC were non-significantly (P≥0.05) different from each other, being different from HbICX (P≤0.05). The HbICX gave overestimated values of Hb as compared to HbA and HbIC. The sensitivity, specificity, positive predictive value, and negative predictive value for HbIC were higher being 86.1%, 88.3%, 88.0%, and 86.0%, respectively as compared to the lower values of 45.0%, 12.0%, 12.0% and 45.0%, for HbICX. Bland and Altman test revealed a better level of agreement between HbA and HbIC. Around the mean difference line, there was no proportional bias in data distribution (Mean= -0.16, 95% CI= 0.34 to -0.67). Similarly, measures attained through Cronbach alpha and intraclass correlation coefficient between HbA and HbIC were higher being 0.703 and 0.825 values for single and average, respectively, as compared to the values of 0.200 and 0.333 between HbA and HbICX.Conclusion: It is concluded that the indirect cyanmethemoglobin method for Hb estimation is reliable and accurate for cattle blood, if a measured quantity of blood drop is taken on a filter paper. We recommend this DBS technique for Hb estimation in cattle blood for resource-poor settings and for livestock herds being reared distantly from the laboratories. For future, it is recommended that DBS technique with various other modifications and for other hematochemical biomarkers may be validated for livestock blood.Keywords: Dried blood spot; Hemoglobin; Cholistani cattle