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Acinetobacter baumannii is a notorious pathogen that commonly thrives in hospital environments and is responsible for numerous nosocomial infections in humans. The burgeoning multi-drug resistance leaves relatively minimal options for treating the bacterial infection, posing a significant problem and prompting the identification of new approaches for tackling the same. This motivated us to focus on non-canonical nucleic acid structures, mainly G-quadruplexes, as drug targets. G-quadruplexes have recently been gaining attention due to their involvement in multiple bacterial and viral pathogenesis. Herein, we sought to explore conserved putative G-quadruplex motifs in A. baumannii. In silico analysis revealed the presence of eight conserved motifs in genes involved in bacterial survival and pathogenesis. The biophysical and biomolecular analysis confirmed stable G-quadruplex formation by the motifs and showed a high binding affinity with the well-reported G-quadruplex binding ligand, BRACO-19. BRACO-19 exposure also decreased the growth of bacteria and downregulated the expression of G-quadruplex-harboring genes. The biofilm-forming ability of the bacteria was also affected by BRACO-19 addition. Taking all these observations into account, we have shown here for the first time the potential of G-quadruplex structures as a promising drug target in Acinetobacter baumannii , for addressing the challenges posed by this infamous pathogen. [ABSTRACT FROM AUTHOR]

The G-quadruplex structure is a highly conserved drug target for preventing infection of several human pathogens. We tried to explore G-quadruplex forming motifs as promising drug targets in the genome of Salmonella enterica that causes enteric fever in humans. Herein, we report three highly conserved G-quadruplex motifs (SE-PGQ-1, 2, and 3) in the genome of Salmonella enterica. Bioinformatics analysis inferred the presence of SE-PGQ-1 in the regulatory region of mgtA , SE-PGQ-2 in ORF of entA, and SE-PGQ-3 in the promoter region of malE and malK genes. The G-quadruplex forming sequences were confirmed by biophysical and biomolecular techniques. Cellular studies affirm the inhibitory effect of G-quadruplex specific ligands on Salmonella enterica growth. Further, PCR inhibition, reporter based assay, and RT-qPCR assays emphasize the biological relevance of G-quadruplexes in these genes. Thus, this study confirmed the presence of G-quadruplex motifs in Salmonella enterica and characterized them as a promising drug target. • Highly conserved potential G-quadruplex forming sequences (SE-PGQs) were identified Salmonella enterica. • The Acridine derivatives, BRACO-19 and 9-Aminoacridine stabilizes the SE-PGQs. • G-quadruplex regulates the expression of PGQ harboring gene in Salmonella enterica. • G-quadruplex mediated therapeutic mechanism can be used for combating Salmonella enterica infection. [ABSTRACT FROM AUTHOR]

GOLD SELEX, a novel SELEX approach has been developed that obviates the need for target immobilization for aptamer development. The approach purely relies on the affinity of the aptamers towards its target, to get detached from the gold nanoparticle (GNP) surface (weak attraction) after binding with its target. Thus, only the completely detached aptamers are selected for the next round of SELEX. This, in-process, also addresses the issue of residual binding and thus improves the sensitivity of the developed aptamers. As a proof of concept for establishing the utility of the approach for small molecules, we have developed aptamers against dichlorvos (DV), a pesticide in just 8 rounds. Using these aptamer candidates, we have developed an aptamer-NanoZyme (GNP having peroxidase mimic activity) based colorimetric assay. The developed aptamer displayed high affinity (Kd in sub micromolar range) and selectivity for DV. The developed assay could detect as low as 15 μM DV. The best-performing aptamer was also able to work in real samples like river water and commercial apple juice. The GOLD SELEX approach developed in this study, we believe, can act as a template for future SELEX strategy development and can replace the conventional SELEX strategy. [ABSTRACT FROM AUTHOR]

Despite of various advancements in biosensing, a rapid, accurate, and on-site detection of a bacterial pathogen is a real challenge due to the lack of appropriate diagnostic platforms. To address this unmet need, we herein report an aptamer-mediated tunable NanoZyme sensor for the detection of Pseudomonas aeruginosa, an infectious bacterial pathogen. Our approach exploits the inherent peroxidase-like NanoZyme activity of gold nanoparticles (GNPs) in combination with high affinity and specificity of a Pseudomonas aeruginosa-specific aptamer (F23). The presence of aptamer inhibits the inherent peroxidase-like activity of GNPs by simple adsorption on to the surface of GNPs. However, in the presence of cognate target (P. aeruginosa), owing to the high affinity for P. aeruginosa, the aptamer leaves the GNP surface, allowing GNPs to resume their peroxidase-like activity, resulting in oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB). As TMB is an electrochemically active species, we have been able to translate the NanoZyme-based method into an ultrasensitive electrochemical assay using disposable carbon screen-printed electrode. This approach is highly sensitive and allows us to rapidly detect P. aeruginosa with a low-end detection limit of ~ 60 CFU/mL in water within 10 min. This generic aptamer-NanoZyme-based electrochemical sensing strategy may, in principle, be applicable for the detection of various other bacterial pathogens. [ABSTRACT FROM AUTHOR]

Heart failure (HF) is emerging as a leading cause of death worldwide. Estimation of BNP levels is a routine diagnosis in these patients. However, in patients having high body-mass index (BMI), renal disease or in geriatric patients, BNP level is reported to be noisy and leads to incongruous conclusion. Thus, for better risk stratification among heart failure patients, it is imperative to look for a superior biomarker. In recent times, sST2 has shown promise as a biomarker. Identifying such biomarkers in peripheral blood of HF patients, need an affine and selective molecular recognition element. Thus, in the current study an aptamer (sS9_P) against sST2 was identified from an aptamer library. Systematic Evolution of Ligands through Exponential enrichment (SELEX) derived aptamer evinced role of its primer binding domains in maintaining its selectivity. This aptamer candidate demonstrated dissociation constant (Kd) in low nanomolar range, and the Limit of Detection (LOD) was ~4 ng. Circular dichroism confirms the formation of complex stem-loop like structure. The well characterized sS9_P aptamer was used in an Aptamer Linked Immobilized Sorbent Assay (ALISA) to detect sST2 level in patients' serum (n = 99). Aptamer sS9_P has shown significant discrimination to differentiate HF patients and healthy volunteers with a reasonable specificity (~83 %) with a modest sensitivity of ~64 %. While sST-2 antibody has shown poor specificity of ~44% but good sensitivity (~87%). The insight obtained from this study indicates that a combination of aptamer and antibody-based assay can be used to design a point-of-care assay for the rapid detection of HF patients in emergency settings. [ABSTRACT FROM AUTHOR]

Abstract Pleural tuberculosis (pTB) is diagnosed by using a composite reference standard (CRS) since microbiological methods are grossly inadequate and an accurate diagnostic test remains an unmet need. The present study aimed to evaluate the utility of Mycobacterium tuberculosis (Mtb) antigen and DNA-based tests for pTB diagnosis. Patients were classified as 'Definite TB', 'Probable TB' and 'Non-TB' disease according to the CRS. We assessed the performance of in-house antigen detection assays, namely antibody-based Enzyme-Linked ImmunoSorbent Assay (ELISA) and aptamer-based Aptamer-Linked Immobilized Sorbent Assay (ALISA), targeting Mtb HspX protein and DNA-based tests namely, Xpert MTB/RIF and in-house devR -qPCR. ROC curves were generated for the combined group of 'Definite TB' and 'Probable TB' vs. 'Non-TB' disease group and cut-off values were derived to provide specificity of ≥98%. The sensitivity of ALISA was ∼93% vs. ∼24% of ELISA (p-value ≤0.0001). devR -qPCR exhibited a sensitivity of 50% vs. ∼22% of Xpert (p-value ≤0.01). This novel aptamer-based ALISA test surpasses the sensitivity criterion and matches the specificity requirement spelt out in the 'Target product profile' for extrapulmonary tuberculosis samples by Unitaid (Sensitivity ≥80%, Specificity 98%). The superior performance of the aptamer-based ALISA test indicates its translational potential to bridge the existing gap in pTB diagnosis. Highlights • An HspX Aptamer Linked Immobilized Sorbent Assay (ALISA) is developed to detect pleural TB (pTB). • Its performance is compared with qPCR, GeneXpert and ELISA in pleural fluid samples. • HspX ALISA is superior to other tests and detects pTB with ~93% sensitivity and ~98% specificity. • ALISA demonstrates strong potential to bridge the existing gap in pTB diagnosis. [ABSTRACT FROM AUTHOR]

This review covers a broad range of well-established and novel diagnostic platforms which are currently being considered for use in commercial point-of care (POC) diagnostics utilizing aptamers instead of antibodies as the molecular recognition elements. Relevant technologies include traditional enzyme-linked colorimetric plate assays, lateral flow test strips, more exotic homogeneous “lights on” fluorescence assays, as well as new nanoparticle-based, electrochemical, electrochemiluminescence and other technologies suitable for rapid and facile POC clinical diagnostics. Advantages of using aptamers over antibodies in such POC diagnostic platforms are elucidated in each case. [ABSTRACT FROM AUTHOR]

Complex target SELEX always have been an intriguing approach to the scientific community, as it offers the potential discovery of novel biomarkers. We herein successfully performed SELEX on Bungarus caeruleus venom to develop a panel of highly affine aptamers that specifically recognizes the B. caeruleus (common krait) venom and was able to discriminate the B. caeruleus venom from Cobra, Russell's, and Saw-scaled viper's venom. The aptamers generated against the crude venom also lead to the identification of the specific component of the venom, which is β-Bungarotoxin, a toxin uniquely present in the B. caeruleus venom. The best performing aptamer candidates were used as a molecular recognition element in a paper-based device and were able to detect as low as 2 ng krait venom in human serum background. The developed aptamer-based paper device can be used for potential point-of-care venom detection applications due to its simplicity and affordability. [ABSTRACT FROM AUTHOR]

Owing to their ease in operation and fast turnaround time, lateral flow assays (LFAs) are increasingly being used as point-of-care diagnostic tests for variety of analytes. In a majority of these LFAs, antibodies are used as a molecular recognition element. Antibodies have a number of limitations such as high batch-to-batch variation, poor stability, long development time, difficulty in functionalization and need for ethical approval and cold chain. All these factors pose a great challenge to scale up the antibody-based tests. In recent years, the advent of aptamer technology has made a paradigm shift in the point-of-care diagnostics owing to the various advantages of aptamers over antibodies that favour their adaptability on a variety of sensing platforms including the lateral flow. In this review, we have highlighted the advantages of aptamers over antibodies, suitability of aptamers for lateral flow platforms, different types of aptamer-based LFAs and various labels for aptamer-based LFAs. We have also provided a summary of the applications of aptamer technology in LFAs for analytical applications. [ABSTRACT FROM AUTHOR]

Background: Pleural tuberculosis (pTB) is a diagnostic challenge because of its non-specific clinical features, lack of accurate diagnostic tools and paucibacillary nature of the disease. Methods: We, here describe the development of a novel magnetic nanoparticle antibody-conjugate and aptamer-based assay (MNp-Ab-Ap assay) targeting 4 different Mycobacterium tuberculosis ( M . tb. ) antigens (GlcB, MPT51, MPT64 and CFP-10) for pTB diagnosis. The MNp-Ab-Ap assay was developed by conjugating polyclonal antibodies on the surface of magnetic nanoparticles (MNPs) by using EDC-NHS chemistry. These conjugated MNPs were used to capture M. tb. antigens present in the pleural fluid samples. The resulting antigen-antibody complex was detected by antigen-specific 5'-biotinylated aptamers. All assays were standardized using samples of the 'Development set' (n=17) and evaluated in the 'Validation set' (n=114) in a blinded manner. Patient categorization was done using a 'Composite Reference Standard'. Assay cut-offs were determined from the 'Development set' (n=17; 'Definite & Probable' pTB; n=9 and 'Non-TB'; n=8) by calculating mean+3SD of OD 450 values of the 'Non-TB' group and applied to 'Validation set' (n=114; 'Definite' pTB; n=8, 'Probable' pTB; n=34, 'Possible' pTB; n=28 and 'Non-TB'; n=44). Results: Out of the 4 assays, MPT51-based MNp-Ab-Ap assay performed the best with 66.6% (95%CI;50.4-80.4) sensitivity and 95.4% (95%CI;85.1-99.4) specificity in the combined 'Definite and Probable' pTB group. Xpert MTB/RIF assay detected only six samples in the 'Validation set'. Binary logistic regression analysis indicated that MPT51-based MNp-Ab-Ap assay provided an incremental advantage over the existing diagnostic algorithm for pTB. Conclusions: We conclude that MPT51-based MNp-Ab-Ap assay is a novel technique that can pave the way towards rapid and accurate diagnosis of pTB., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Type VI secretion system (T6SS) is utilized by many Gram-negative bacteria to eliminate competing bacterial species and manipulate host cells. Acinetobacter baumannii ATCC 17978 utilizes T6SS at the expense of losing pAB3 plasmid to induce contact-dependent killing of competitor microbes, resulting in the loss of antibiotic resistance carried by pAB3. However, the regulatory network associated with T6SS in A. baumannii remains poorly understood. Here, we identified an Mn 2+ -dependent post-transcriptional regulation of T6SS mediated by a bonafide small RNA, AbsR28. A. baumannii utilizes MumT, an Mn 2+ -uptake inner membrane transporter, for the uptake of extracellular Mn 2+ during oxidative stress. We demonstrate that the abundance of intracellular Mn 2+ enables complementary base pairing of AbsR28- tssM mRNA (that translates to TssM, one of the vital inner membrane components of T6SS), inducing RNase E-mediated degradation of tssM mRNA and resulting in T6SS repression. Thus, AbsR28 mediates a crosstalk between MumT and T6SS in A. baumannii .IMPORTANCESmall RNAs (sRNAs) are identified as critical components within the bacterial regulatory networks involved in fine regulation of virulence-associated factors. The sRNA-mediated regulation of type VI secretion system (T6SS) in Acinetobacter baumannii was unchartered. Previously, it was demonstrated that A. baumannii ATCC 17978 cells switch from T6- to T6+ phenotype, resulting in the loss of antibiotic resistance conferred by plasmid pAB3. Furthermore, the derivatives of pAB3 found in recent clinical isolates of A. baumannii harbor expanded antibiotic resistance genes and multiple determinants for virulence factors. Hence, the loss of this plasmid for T6SS activity renders A. baumannii T6+ cells susceptible to antibiotics and compromises their virulence. Our findings show how A. baumannii tends to inactivate T6SS through an sRNA-mediated regulation that relies on Mn 2+ and retains pAB3 during infection to retain antibiotic resistance genes carried on the plasmid.

Synthetic Aperture Radar (SAR) integrated with deep learning has been widely used in several military and civilian applications, such as border patrolling, to monitor and regulate the movement of people and goods across land, air, and maritime borders. Amongst these, maritime borders confront different threats and challenges. Therefore, SAR-based ship detection becomes essential for naval surveillance in marine traffic management, oil spill detection, illegal fishing, and maritime piracy. However, the model becomes insensitive to small ships due to the wide-scale variance and uneven distribution of ship sizes in SAR images. This increases the difficulties associated with ship recognition, which triggers several false alarms. To effectively address these difficulties, the present work proposes an ensemble model (eYOLO) based on YOLOv4 and YOLOv5. The model utilizes a weighted box fusion technique to fuse the outputs of YOLOv4 and YOLOv5. Also, a generalized intersection over union loss has been adopted in eYOLO which ensures the increased generalization capability of the model with reduced scale sensitivity. The model has been developed end-to-end, and its performance has been validated against other reported results using an open-source SAR-ship dataset. The obtained results authorize the effectiveness of eYOLO in multi-scale ship detection with an F 1 score and mAP of 91.49% and 92.00%, respectively. This highlights the efficacy of eYOLO in multi-scale ship detection using SAR imagery., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

G-quadruplexes (GQs) have been primarily studied in the context of cancer and neurodegenerative pathologies. However, recent research has shifted focus to their existence and functional roles in viral genomes, revealing GQ-regulated key pathways in various human pathogenic viruses. While GQ structures have been reported in the genomes of emerging and re-emerging viruses, RNA viruses have been understudied compared to DNA viruses, including notable examples such as human immunodeficiency virus-1, hepatitis C virus, Ebola virus, Nipah virus, Zika virus, and SARS-CoV-2. The flavivirus family, comprising the Japanese encephalitis virus (JEV), poses a significant global threat due to recurring outbreaks yet lacks approved antivirals. In this study, we identified and characterized eight putative G-quadruplex-forming motifs within essential genes involved in genome replication, assembly, and internalization in the host cell, conserved across different JEV isolates. The formation and stability of these motifs were validated through a multitude of biophysical and cell-based assays. The interaction and binding affinity of these motifs with the known GQ-binding ligand BRACO-19 were supported by biophysical assays, confirming the capability of these motifs to form GQ structures. Notably, BRACO-19 also exerted antiviral properties through reduction of viral replication and infectious virus titers as well as inhibition of viral protein expression, as evaluated by the cell-based assays. This comprehensive molecular characterization of G-quadruplex structures within the JEV genome highlights their potential as promising antiviral targets for intervention strategies against JEV infection through GQ-specific ligands.

Introduction: There is no standard protocol for managing non-union of diaphyseal humerus bone, with several authors reporting their results using various techniques and methods for its management. No meta-analysis has reported the results of managing these cases with non-vascularized fibula grafting as an adjuvant for osteosynthesis., Materials and Methods: This meta-analysis was performed to estimate the pooled data for calculating the union rates in diaphyseal humerus fractures managed with non-vascularized fibula grafting. Risk of Bias was computed using the Joanna Briggs Institute appraisal tool., Results: A total of 5 studies, comprising 102 patients, were included. The pooled estimate demonstrated that 94 patients achieved bone union with intramedullary fibular strut grafting. The pooled union rate (per 100 events) was 90.59 (95 % CI, 82.86-95.04, I 2  = 0). The present meta-analysis also showed a significant improvement in DASH scores following the use of a non-vascularized fibula graft with a common effects model (SMD = 4.08; 95%CI: 3.44; 4.72; p < 0.01 I 2  = 19 %, p-value for Q test = 0.29)., Conclusion: Non-vascularized fibula grafting is an excellent adjuvant for the internal fixation of non-union diaphyseal humerus fractures. Although there is limited literature, further studies should highlight and assess the treatment of these uncommon but disabling conditions., (© 2024 Professor P K Surendran Memorial Education Foundation. Published by Elsevier B.V. All rights are reserved, including those for text and data mining, AI training, and similar technologies.)

Background: The relationship of inflammatory bowel disease (IBD) with osteonecrosis or avascular necrosis (AVN) is uncertain., Methods: Systematic review to estimate the frequency of osteonecrosis in IBD was performed. Electronic databases were searched on 12 December 2022 to identify relevant studies. We planned to estimate the pooled prevalence of AVN in IBD, the risk in IBD when compared to the healthy population (without any chronic disease), and the impact of steroid use on osteonecrosis (IBD with and without steroid use). The risk of Bias was assessed with the Joanna Briggs Institute appraisal tool., Results: Fifteen studies including 105 154 individuals were included. The pooled rate AVN was 10.39 per 1000 patients (95% confidence interval, 4.44-24.11, I 2  = 97%). Subgroup analysis suggested that the prevalence was lower in larger studies (>1000 participants) at 3.10, 1.07; 8.98, I 2  = 98% versus 21.03, 8.69; 50.01, I 2  = 83%. The use of steroids did not seem to increase the risk of osteonecrosis in the included studies (pooled odds ratio: 1.88, 0.55-6.41, I 2  = 39%). The systematic review was limited by the absence of comparison with the control population free of chronic disease., Conclusion: IBD may be associated with a risk of osteonecrosis. Future studies should assess the risk in comparison to the healthy population and the impact of disease activity and IBD therapies on the risk., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

In order to adapt in host tissues, microbial pathogens regulate their gene expression through a variety of transcription factors. Here, we have functionally characterized Rv0792c, a HutC homolog from Mycobacterium tuberculosis. In comparison to the parental strain, a strain of M. tuberculosis with a Rv0792c mutant was compromised for survival upon exposure to oxidative stress and infection in guinea pigs. RNA sequencing analysis revealed that Rv0792c regulates the expression of genes involved in stress adaptation and virulence of M. tuberculosis. Solution small-angle X-ray scattering (SAXS) data-steered model building confirmed that the C-terminal region plays a pivotal role in dimer formation. Systematic evolution of ligands by exponential enrichment (SELEX) resulted in the identification of single-strand DNA (ssDNA) aptamers that can be used as a tool to identify small-molecule inhibitors targeting Rv0792c. Using SELEX and SAXS data-based modeling, we identified residues essential for Rv0792c's aptamer binding activity. In this study, we also identified I-OMe-Tyrphostin as an inhibitor of Rv0792c's aptamer and DNA binding activity. The identified small molecule reduced the growth of intracellular M. tuberculosis in macrophages. The present study thus provides a detailed shape-function characterization of a HutC family of transcription factor from M. tuberculosis. IMPORTANCE Prokaryotes encode a large number of GntR family transcription factors that are involved in various fundamental biological processes, including stress adaptation and pathogenesis. Here, we investigated the structural and functional role of Rv0792c, a HutC homolog from M. tuberculosis. We demonstrated that Rv0792c is essential for M. tuberculosis to adapt to oxidative stress and establish disease in guinea pigs. Using a systematic evolution of ligands by exponential enrichment (SELEX) approach, we identified ssDNA aptamers from a random ssDNA library that bound to Rv0792c protein. These aptamers were thoroughly characterized using biochemical and biophysical assays. Using SAXS, we determined the structural model of Rv0792c in both the presence and absence of the aptamers. Further, using a combination of SELEX and SAXS methodologies, we identified I-OMe-Tyrphostin as a potential inhibitor of Rv0792c. Here we provide a detailed functional characterization of a transcription factor belonging to the HutC family from M. tuberculosis.

Coronavirus disease, also known as COVID-19, is an infectious disease caused by SARS-CoV-2. It has a direct impact on the upper and lower respiratory tract and threatened the health of many people around the world. The latest statistics show that the number of people diagnosed with COVID-19 is growing exponentially. Diagnosing positive cases of COVID-19 is important for preventing further spread of the disease. Currently, Coronavirus is a serious threat to scientists, medical experts and researchers around the world from its detection to its treatment. It is currently detected using reverse transcription polymerase chain reaction (RT-PCR) analysis at the most test centers around the world. Yet, knowing the reliability of a deep learning based medical diagnosis is important for doctors to build confidence in the technology and improve treatment. The goal of this study is to develop a model that automatically identifies COVID-19 by using chest X-ray images. To achieve this, we modified the DarkCovidNet model which is based on a convolutional neural network (CNN) and plotted the experimental results for two scenarios: binary classification (COVID-19 versus No-findings) and multi-class classification (COVID-19 versus pneumonia versus No-findings). The model is trained on more than 10 thousand X-ray images and achieved an average accuracy of 99.53% and 94.18% for binary and multi-class classification, respectively. Therefore, the proposed method demonstrates the effectiveness of COVID-19 detection using X-ray images. Our model can be used to test the patient via cloud and also be used in situations where RT-PCR tests and other options aren't available., Competing Interests: Conflict of interestAll authors declare that there is no conflict of interest., (© The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2021.)

The recent SARS-CoV-2 outbreak has been declared a global health emergency. It will take years to vaccinate the whole population to protect them from this deadly virus, hence the management of SARS-CoV-2 largely depends on the widespread availability of an accurate diagnostic test. Toward addressing the unmet need of a reliable diagnostic test in the current work by utilizing the power of Systematic Evolution of Ligands by EXponential enrichment, a 44-mer G-quadruplex-forming DNA aptamer against spike trimer antigen of SARS-CoV-2 was identified. The lead aptamer candidate (S14) was characterized thoroughly for its binding, selectivity, affinity, structure, and batch-to-batch variability by utilizing various biochemical, biophysical, and in silico techniques. S14 has demonstrated a low nanomolar K D , confirming its tight binding to a spike antigen of SARS-CoV-2. S14 can detect as low as 2 nM of antigen. The clinical evaluation of S14 aptamer on nasopharyngeal swab specimens (n = 232) has displayed a highly discriminatory response between SARS-CoV-2 infected individuals from the non-infected one with a sensitivity and specificity of ∼91% and 98%, respectively. Importantly, S14 aptamer-based test has evinced a comparable performance with that of RT-PCR-based assay. Altogether, this study established the utility of aptamer technology for the detection of SARS-CoV-2., Competing Interests: The aptamer sequences reported in this study are proprietary reagents and deposited in the Indian Patent Office as a part of Indian Patent application no. 202011017852. This technology has transferred to MolBio Diagnostics Pvt. Ltd., India for scale-up and commercialization., (© 2021 The Author(s).)

A novel approach has been developed for the detection of  56   kDa tissue - specific antigen  (TSA) gene of  Orientia tsutsugamushi  a causative agent of scrub typhus disease. The approach was developed by immobilization of 5' NH2 labeled ssDNA probe selective to 56   kDa   TSA  gene, to the surface of AuNPs/CNF modified screen-printed electrode. An electrochemical response was recorded with single stranded genomic DNA (ssDNA) of O. tsutsugamushi isolated from patient sample, using cyclic voltammetry and electrochemical impedance spectroscopy. The electrode surface was characterized by Field-Emission Scanning electron microscope (FE-SEM), Fourier Transform Infrared Spectroscopy (FTIR) and Raman Spectroscopy at each step of fabrication. The DNA biosensor shows optimum response within 50-60 s at room temperature (25 ± 3 °C). The sensor shows higher sensitivity [7849 (µA/cm 2 )/ng DNA], fast response time (60 s), wider linear range (0.04-2.6 ng) with limit of detection of 0.02 ng/µl of ssDNA sample., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest in the publication., (© King Abdulaziz City for Science and Technology 2020.)

Defining target product profiles (TPPs) for aptamer-based diagnostics is crucial to the success or failure of aptamer businesses or products. A well-conceived TPP will place the aptamer in an assay for a target against which antibodies are ill-suited or have difficulty detecting the analyte, such as some highly related proteins or poorly immunogenic small molecule haptens. Strong TPPs can also take advantage of the unique nucleic acid nature of aptamers, to produce assays with longer shelf life or special chemical properties and ability to be modified versus protein-based antibodies. The following chapter reviews the essence of well-conceived TPPs especially with respect to aptamer targets for diagnostics and illustrates several examples of commercial aptamer diagnostic success. Graphical Abstract.

The successful management of tuberculosis (TB) requires efficient diagnosis and treatment. Further, the increasing prevalence of drug-resistant TB highlights the urgent need to develop novel inhibitors against both drug-susceptible and drug-resistant forms of disease. Malate synthase (MS), an enzyme of the glyoxylate pathway, plays a vital role in mycobacterial persistence, and therefore it is considered as an attractive target for novel anti-TB drug development. Recent studies have also ascribed an adhesin function to MS and established it as a potent diagnostic biomarker. In this study, a panel of Mycobacterium tuberculosis (Mtb) MS-specific single-stranded DNA aptamers was identified by Systematic Evolution of Ligands by EXponential enrichment (SELEX). The best-performing G-quadruplex-forming 44-mer aptamer, MS10, was optimized post-SELEX to generate an 11-mer aptamer, MS10-Trunc. This aptamer was characterized by various biochemical, biophysical, and in silico techniques. Its theranostic activity toward Mtb was established using enzyme inhibition, host cell binding, and invasion assays. MS10-Trunc aptamer exhibited high affinity for MS (equilibrium dissociation constant [K D ] ∼19 pM) and displayed robust inhibition of MS enzyme activity with IC 50 of 251.1 nM and inhibitor constant (K i ) of 230 nM. This aptamer blocked mycobacterial entry into host cells by binding to surface-associated MS. In addition, we have also demonstrated its application in the detection of tuberculous meningitis (TBM) in patients with sensitivity and specificity each of >97%., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)

G-quadruplex structure forming motifs are among the most studied evolutionarily conserved drug targets that are present throughout the genome of different organisms and susceptible to influencing various biological processes. Here we report highly conserved potential G-quadruplex motifs (PGQs) in three essential genes (espK, espB, and cyp51) among 160 strains of the Mycobacterium tuberculosis genome. Products of these genes are involved in pathways that are responsible for virulence determination of bacteria inside the host cell and its survival by maintaining membrane fluidity. The espK and espB genes are essential players that prevent the formation of mature phagolysosome and antigen presentation by host macrophages. The cyp51 is another PGQ-possessing gene involved in sterol biosynthesis pathway and membrane formation. In the present study, we revealed the formation of stable intramolecular parallel G-quadruplex structures by Mycobacterium PGQs using a combination of techniques (NMR, circular dichroism [CD], and gel electrophoresis). Next, isothermal titration calorimetry (ITC) and CD melting analysis demonstrated that a well-known G-quadruplex ligand, TMPyP4, binds to and stabilizes these PGQ motifs. Finally, polymerase inhibition and qRT-PCR assays highlight the biological relevance of PGQ-possessing genes in this pathogen and demonstrate that G-quadruplexes are potential drug targets for the development of effective anti-tuberculosis therapeutics., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

The entry and survival of Mycobacterium tuberculosis (Mtb) within host cells is orchestrated partly by an essential histone-like protein HupB (Rv2986c). Despite being an essential drug target, the lack of structural information has impeded the development of inhibitors targeting the indispensable and multifunctional C-terminal domain (CTD) of HupB. To bypass the requirement for structural information in the classical drug discovery route, we generated a panel of DNA aptamers against HupB protein through systemic evolution of ligands by exponential (SELEX) enrichment. Two G-quadruplex-forming high-affinity aptamers (HupB-4T and HupB-13T) were identified, each of which bound two distinct sites on full-length HupB, with an estimated K D of ∼1.72 μM and ∼0.17 μM, respectively, for the high-affinity sites. While HupB-4T robustly inhibited DNA-binding activity of HupB in vitro, both the aptamers recognized surface-located HupB and significantly blocked Mtb entry into THP-1 monocytic cells (p < 0.0001). In summary, DNA aptamers generated in this study block DNA-binding activity of HupB, inhibit virulent Mtb infection in host cells, and demonstrate aptamers to be inhibitors of HupB functions. This study also illustrates the utility of SELEX in developing inhibitors against essential targets for whom structural information is not available., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)

Tuberculous meningitis (TBM) is the most severe manifestation of tuberculosis and its diagnosis remains a challenge even today due to the lack of an adequate test. HspX antigen of Mycobacterium tuberculosis was previously established as a reliable diagnostic biomarker for TBM in an ELISA test format using anti-HspX polyclonal antibodies. Towards overcoming the limitations of batch-to-batch variation and challenges of scalability in antibody generation, we utilized Systematic Evolution of Ligands by EXponential enrichment (SELEX) to develop high affinity DNA aptamers against HspX as an alternative diagnostic reagent. Post-SELEX optimization of the best-performing aptamer candidate, H63, established its derivative H63 SL-2 M6 to be superior to its parent. Aptamer H63 SL-2 M6 displayed a specific and high affinity interaction with HspX (K d ∼9.0 × 10 -8  M). In an Aptamer Linked Immobilized Sorbent Assay (ALISA), H63 SL-2 M6 significantly differentiated between cerebrospinal fluid specimens from TBM and non-TBM subjects (n = 87, ***p < 0.0001) with ∼100% sensitivity and ∼91% specificity. Notably, ALISA exhibited comparable performance with previously reported antibody-based ELISA and qPCR. Altogether, our findings establish the utility of HspX aptamer for the reliable diagnosis of TBM and pave the way for developing an aptamer-based point-of-care test for TBM., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Background: The data regarding Valproate and its influence on serum folate and homocysteine levels are conflicting. The aim of this study was to evaluate whether differences exist in homocysteine, folate, and vitamin B12 levels in children receiving Valproate., Methods: A total of 55 newly diagnosed epileptic children with ages ranging from 2 to 15 years were enrolled at the start of study but after 3 months follow up, the total sample size finally was only 50 epileptic children. 5 children dropped out of study due to poor follow up. 50 age and gender matched healthy control subjects were also studied on enrollment at the start of study. Serum homocysteine levels were analyzed by enzyme immunoassay method using the kits provided by Axis-Shield Diagnostics Ltd (Dundee DD2 1XA, United Kingdom). Serum folate and serum vitamin B12 were estimated by Competitive Chemiluminescent Enzyme Immunoassay method., Results: The serum homocysteine level in epileptic children was found to be significantly increased after Valproate monotherapy as compared to before therapy. Moreover, a highly significant decrease was observed in the levels of serum folate in epileptic children after Valproate monotherapy as compared to before therapy. But a non significant difference was observed in serum vitamin B12 levels in epileptic children before and after Valproate monotherapy., Conclusions: Thus, we conclude that there is a significant increase in the levels of homocysteine and a significant decrease in the concentration of serum folate while vitamin B12 decreases non-significantly after Valproate monotherapy. The atherogenic effect of increased serum homocysteine level is well established; the patients under Valproate monotherapy should be monitored for possible atherogenic effects. Considering the above observation and results of children undergoing Valproate monotherapy, these children should be screened for levels of serum homocysteine, folate, and vitamin B12 and treated when their levels are found to be disturbed.

Background: Oxidative stress has been considered to be a pathogenic factor of diabetic complications including nephropathy. There are many controversies and limited studies regarding the antioxidant enzymes in diabetic nephropathy., Aim: This study was to evaluate the levels of antioxidant enzymes and lipid peroxidation in Type-2 Diabetes Mellitus (DM) patients with and without nephropathy., Materials and Methods: The study included 90 age and sex matched subjects. Blood samples of all subjects were analyzed for all biochemical and oxidative stress parameters., Results: The malondialdehyde (MDA) levels and catalase (CAT) activity were significantly increased and reduced glutathione (GSH) levels and activities of glutathione peroxidase (GPx) and glutathione reductase (GR) were significantly decreased in Type-2 DM with and without nephropathy as compared to controls and also in Type-2 DM with nephropathy as compared to Type-2 DM without nephropathy. There were an excellent positive correlation of glycohemoglobin (HbA1c) with MDA and a good negative correlation of GPx with GSH in controls. There were positive correlations of GR, CAT, and superoxide dismutase (SOD) with MDA in Type-2 diabetes patients with nephropathy., Conclusions: Intensity of oxidative stress in Type-2 diabetic patients with nephropathy is greater when compared with Type-2 diabetic patients without nephropathy as compared to the controls.

Background: Periodontal disease is closely related to type 2 diabetes and is an important complication of diabetes. There are few studies about the relationship the glycohemoglobin levels with severity of periodontitis in non-diabetic population. We therefore planned this study to evaluate the glycohemoglobin levels with severity of periodontitis in non-diabetic population., Methods: This study was conducted on 50 age and gender matched subjects in each of the three groups (according to the grades of mobility in periodontitis), a total of 150 non-diabetic periodontitis patients (Grade 1, Grade 2, and Grade 3 mobility) and 50 non-diabetic periodontitis patients with Grade 0 mobility (controls), in collaboration with the Department of Periodontics of Dental College and Department of Biochemistry, PGIMS, Rohtak, Haryana. After obtaining informed consent, fasting venous blood samples of all the non-diabetic periodontitis patients of all grades were collected aseptically for HbA1c, plasma glucose, and serum C-reactive protein (CRP) estimation., Results: A total of 150 non-diabetic periodontitis patients (Grade 1, Grade 2, and Grade 3 mobility) and 50 age and gender matched controls participated in the study. There was no significant difference in fasting plasma glucose and postprandial plasma glucose in non-diabetic periodontitis patients with Grade 1, Grade 2, and Grade 3 mobility as compared to controls, non-diabetic periodontitis patients with Grade 1 mobility as compared to Grade 2, non-diabetic periodontitis patients with Grade 1 mobility as compared to Grade 3 and non-diabetic periodontitis patients with Grade 2 mobility as compared to Grade 3. Glycohemoglobin and serum C-reactive protein levels were significantly increased in non-diabetic periodontitis patients with Grade 1, Grade 2, and Grade 3 mobility as compared to controls, non-diabetic periodontitis patients with Grade 1 mobility as compared to Grade 3 and non-diabetic periodontitis patients with Grade 2 mobility as compared to Grade 3. The difference of serum C-reactive protein levels were significant. However, glycohemoglobin levels were non-significant between non-diabetic periodontitis patients with Grade 1 and Grade 2 mobility., Conclusions: The evidence of association between periodontitis and increased glycohemoglobin increases attention to the diagnosis and treatment of periodontitis, consequently improving the patient's oral health and prevention of occurrence in future diabetes. An understanding of these correlations is important to allow dental health care providers to inform patients with periodontitis of their increased risks and to counsel such patients to seek additional medical assessment or intervention as indicated.

Background: Individuals with obesity and abdominal adiposity are at higher risk for hyperinsulinaemia, insulin resistance, and diabetes. This study was, therefore, designed to evaluate the association of both generalized and regional obesity with metabolic variables and biochemical indices., Methods: 200 confirmed patients of type-2diabetes of either gender were studied., Results: A statistically significant degree of dyslipidemia was depicted in obese class-II subjects; however, females had a lower degree of dyslipidemia as compared to male subjects with statistically significant results only for HDL-C. Further, multiple logistic regression analysis revealed that BMI is a stronger predictor of FPG and HbA1c as compared to WHR., Conclusions: Higher plasma glucose levels were depicted at a lower BMI, which turned out to be stronger predictor of glycemic control as compared to WHR. Moreover, BMI, WHR and male gender was significantly correlated with the metabolic parameters and even much more pronounced association with BMI.

Background: In type 2 diabetic patients, persistence of hyperglycemia has been reported as a cause of increased production of oxygen free radicals (FR), which leads to oxidative stress (OS) and becomes the main factor for predisposition to the cardiovascular complications in diabetes. Diabetic postmenopausal women are prone to cardiovascular disease due to reduced production of estrogen which is a potent antioxidant and prevents oxidative stress (OS) in body. The study is being aimed to find out the status of antioxidant enzymes (AOEs) and malondialdehyde (MDA) in post-menopausal diabetic women., Methods: The study was conducted with a total of 70 cases, which included 35 Type 2 diabetic post-menopausal females (45 - 60 years) with diabetic CVD complication as the study group and 35 age matched type 2 diabetic postmenopausal females without CVD complication., Results: All diabetic post menopausal females with CVD had significantly higher levels of fasting plasma glucose (FPG), postprandial plasma glucose (PPPG), total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C), catalase (CAT), and malondialdehyde (MDA) and significantly lower levels of HDL-C, reduced glutathione (GSH), glutathione reductase (GR), glutathione peroxidase (GPx), and superoxide dismutase (SOD) as compared to the levels of control subjects., Conclusions: During menopause, reduced production of estrogen causes hypertriglyceridemia, hypercholesterolemia, and hyperlipoproteinemia whose oxidation causes the accumulation of FR in the cell, which precipitates OS. Also, type 2 diabetic subjects with CVD poor glycemic control and impaired AOEs result in increased oxidative injury by failure of protective mechanisms, which further leads to oxidative stress.

Background: Individuals with obesity and abdominal adiposity are at higher risk for hypeinsulinaemia, insulin resistance, and diabetes. This study was therefore designed to investigate the relationship of obesity with oxidative stress and the role of abdominal adiposity on obesity induced oxidative stress, and further to explore the possible mechanism of obesity associated metabolic syndrome., Methods: A total of 150 subjects (120 men and 30 women), aged 17-26 years of both genders, were studied. Body Mass Index and Waist-to-Hip Ratio were taken as a measure of generalized obesity and abdominal adiposity. The biochemical tests done included fasting blood glucose (FBG), lipid profile parameters, serum malondialdehyde (as a biomarker of oxidative stress), and serum adiponectin., Results: The concentration of serum malondialdehyde (MDA) increased with increasing levels of BMI (as per the NIH classification), which was found to be non-significant statistically in overweight subjects while obese class-I and class-II subjects exhibited a statistically significant (p < 0.001) higher level of serum malondialdehyde as compared to normal-weight subjects. Furthermore, according to the present study groups, on comparison with normal-weight subjects (Group-I), obese subjects with abdominal adiposity (Class-2) had statistically significant (p < 0.001) higher serum concentration of malondialdehyde while a non-significant difference was observed in obese subjects without abdominal adiposity (Class-1). Even within the subset of obese subjects, a statistically significant (p < 0.001) difference was depicted, suggesting the role of abdominal adiposity. Karl Pearson coefficient of correlation revealed a statistically significant negative correlation of malondialdehyde with adiponectin (r = - 0.587; p < 0.001)., Conclusions: Obese subjects exhibit increased systemic oxidative stress, which is enhanced when obesity is associated with abdominal adiposity and, moreover, increased oxidative stress is associated with adiponectin deficiency.

Background: For many years, several studies have demonstrated a relationship between insulin like growth factor-1 (IGF-1), thyroid hormones, and various malignancies. IGF-1 plays an important role in tumor proliferation in various malignancies. The relationship between IGF-1 and thyroid hormones is complex and not fully understood. Therefore we planned to evaluate the level of IGF-1 and thyroid hormones in patients of acute leukemia., Methods: The present study included 25 patients with acute leukemia (Acute Myeloid Leukemia, n = 16; Acute Lymphoid Leukemia n = 9, mean age 28.16 years). 25 age and gender matched healthy individuals were taken as control (mean age 27.17 years). In all the subjects, serum IGF-1 was measured by enzyme linked immunosorbent assay (ELISA), serum total triiodothyronine (T3), thyroxine (T4) by radioimmunoassay (RIA), thyroid stimulating hormone (TSH) by immunoradiometric assay (IRMA), and free T3 (FT3) and free T4 (FT4) by chemilluminiscence. These tests were done before starting of chemotherapy and either 6 to 8 weeks after chemotherapy or at the time of remission, whichever was earlier., Results: At the time of diagnosis, patients with acute leukemia showed a significantly increased level of IGF-1 as compared to controls (198.32 +/- 67.55 vs 160.64 +/- 45.96; p < 0.01). After 6 to 8 weeks of chemotherapy, patients with acute leukemia showed a significant decrease in the level of IGF-1 compared to the baseline values (198.32 +/- 67.55 vs 155.6 +/- 45.96; p < 0.01). Though FT3, FT4, total T3, and total T4 values in these patients were within the normal range, these values were still significantly higher compared to controls. TSH levels were significantly lower in patients at the time of presentation and the levels increased after chemotherapy., Conclusions: The estimation of IGF-1 and thyroid hormones may be helpful in assessing the disease activity and predicting the response of chemotherapy.

Background: Subclinical hypothyroidism (SH) represents the mildest form of thyroid hormone deficiency and may be associated with adverse consequences [Subclinical hypothyroidism was defined as a TSH level > 4.0 mIU/L and a normal free thyroxine level 0.6-1.8 ng/dL]. The identification of patients with subclinical hypothyroidism having an increased cardiovascular risk (CVR) is important. The aim of the study was to evaluate atherosclerotic risk factors in patients with subclinical hypothyroidism., Methods: Forty patients with subclinical hypothyroidism and forty healthy euthyroid controls, age and gender matched were included in the study. Serum total triiodothyronine (T3), thyroxine (T4), TSH, free T3 (FT3) and free T4 (FT4) were measured by enzyme linked immunosorbent assay (ELISA). Atherosclerotic risk factors measured were high sensitivity-CRP (hs-CRP), Lipoprotein (a) [Lp (a)] and lipid parameters. Lipid parameters (triglycerides, total cholesterol and high density lipoprotein cholesterol) were analysed by enzymatic colorimetric, endpoint method whereas the hs-CRP and Lp (a) were measured by quantitative latex turbidimetric method., Results: Patients with subclinical hypothyroidism had significantly higher levels of serum hs-CRP, Lp (a), total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) when compared to same parameters of controls. Further, a significant positive correlation was observed between TSH and hs-CRP, Lp (a), LDL-C and TC in subjects with subclinical hypothyroidism. However, TG levels showed no significant correlation with TSH levels., Conclusions: We concluded that the SH patients presented increased concentration of some CVR factors. The potential benefits of diagnosis and treatment of subclinical hypothyroidism may have possible advantages firstly by preventing the progression to overt hypothyroidism and secondly decrease the risk of death from cardiovascular disease (CVD) by starting appropriate therapy to improve lipid parameters. Further research is needed on subclinical hypothyroidism and the associated atherosclerotic risk factors.

Background: Increasing evidence in both experimental and clinical studies suggests that free radical mediated oxidative stress plays a major role in the pathogenesis of both types of diabetes mellitus. Proteins and lipids are among the prime targets for oxidative stress. In this study we evaluated oxidative stress in Type 1 Diabetes Mellitus (insulin dependent diabetes mellitus, IDDM) patients by estimating lipid peroxidation and the effect of vitamin E on oxidative stress and metabolic parameters., Methods: A total of 40 children (20 Type 1 Diabetes Mellitus patients + 20 healthy controls) were examined in the study. Oxidative stress parameters malondialdehyde (MDA), antioxidants, reduced glutathione (GSH), vitamin E and metabolic parameters were studied. All the type 1 Diabetes Mellitus patients were supplemented with 600 mg/daily vitamin E for three months. After three months of supplementation all the parameters mentioned above were studied again., Results: Reduced glutathione and vitamin E levels were lower and malondialdehyde levels were higher in Type 1 Diabetes Mellitus patients compared to healthy controls (p < 0.05). After supplementation with vitamin E in diabetic patients a significant decrease (p < 0.05) in MDA levels and significant increase in GSH (p < 0.05) and vitamin E (p < 0.05) levels were found. A negative correlation between MDA and vitamin E, between MDA and GSH and a positive correlation between vitamin E and GSH was found. Significant changes were not observed in metabolic parameters in Type 1 Diabetes Mellitus patients after vitamin E supplementation (p > 0.05)., Conclusions: Vitamin E ameliorates oxidative stress in Type 1 Diabetes Mellitus patients and improves antioxidant defense system. However, vitamin E does not have any advantage for metabolic parameters.

Background: Carcinomas are considered to be associated with increased cellular proliferation under antigenic stimulation and inflammation. Therefore, the markers of increased cellular turnover such as adenosine deaminase and uric acid were estimated quantitatively along with markers of inflammation such as C-reactive protein in squamous cell carcinoma of head and neck region., Methods: The levels of adenosine deaminase, uric acid, and C-reactive protein were estimated in 50 patients of squamous cell carcinoma of head and neck region before starting any treatment and compared with their corresponding levels in 30 healthy age and gender matched controls. The patients were divided into four groups depending on their staging and their results were compared statistically and the coefficient of correlation was calculated using Pearson's formula., Results: The levels of adenosine deaminase, uric acid, and C-reactive protein were found to be significantly higher in patients of head and neck cancers as compared to the levels in controls (p < 0.001). The levels were also observed to rise with staging. A positive correlation was observed between adenosine deaminase and uric acid (r = 0.743, p < 0.001), adenosine deaminase and C-reactive protein (r = 0.648, p < 0.001) and also between C-reactive protein and uric acid (r = 0.712, p < 0.001)., Conclusions: Thus, estimation of adenosine deaminase, uric acid, and C-reactive protein can help in making the diagnosis and assessing the severity of disease in patients of head and neck carcinoma.

Background: High levels of glycated hemoglobin (HbA(1c)) have been associated with Coronary Vascular Diseases (CVD) in diabetic patients. Recent studies have reported no association between elevated glycated hemoglobin (HbA(1c)) and incident cardiovascular disease (CVD) among women without diabetes. There are many controversial studies on topics such as "Glycated hemoglobin levels (HbA(1c)) have been associated with cardiovascular diseases (CVD) in the non-diabetic patients". Therefore, we planned this study., Methods: The present study was conducted on 50 age matched controls and 50 clinically diagnosed non-diabetic CVD patients of either gender. The study included 50 patients with myocardial infarction (MI) admitted to the ICCU ward of J.L.N. Medical College and Hospital, Ajmer (Rajasthan). The following information was recorded from admission sheets of non-diabetic CVD patients of either gender: history of diabetes, hypertension, and cigarette smoking; demographic indices; coronary heart disease and diabetes mellitus treatment; serum cholesterol; serum triglycerides (TG); high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C); fasting and non-fasting blood glucose levels and Glycated haemoglobin levels (HbA(1c)). Glycosylated hemoglobin (HbA(1c)) was measured by latex agglutination inhibition assay., Results and Conclusions: The HbA(1c) levels in healthy controls (n = 50) and non-diabetic CVD subjects (n = 50) observed were 4.32 +/- 0.34% and 5.80 +/- 0.20%, respectively. HbA(1c) levels in these subjects were significantly higher than controls (p < 0.001). The HbA(1c) levels in non-diabetic CVD patients are higher in comparison to controls.

Background: Intrauterine growth restriction (IUGR) is a common diagnosis in obstetrics and carries an increased risk of prenatal mortality and morbidity. It is associated with short and long term negative outcome in fetuses, infants, and children. The aim of the study was the determination of serum homocysteine along with folic acid and vitamin B12 concentration in pregnancies complicated with IUGR (intrauterine growth restriction)., Methods: The study was performed in a group of 180 pregnant subjects in the third trimester. Fasting (overnight, at least 12 hour) blood samples were collected from 150 pregnant patients with IUGR and 30 normal pregnant women. Serum total homocysteine (tHcy), serum folic acid, and serum vitamin B12 levels were measured., Results: Mean serum concentration of total homocysteine in the group of pregnant patients with IUGR was 11.14 +/- 4.05 microM/L whereas in the group of normal pregnant women it was 7.42 +/- 2.93 microM/L. The serum folic acid level in the group of pregnant patients with IUGR was 10.24 +/- 3.91 ng/mL while in the group of normal pregnant women it was 15.20 +/- 3.41 ng/mL. The serum vitamin B12 level in the group of pregnant patients with IUGR was 146.99 +/- 43.51 pg/mL where as in the group of normal pregnant women it was 171.96 +/- 25.75 pg/mL. The differences were statistically significant., Conclusions: The study revealed that increasing serum homocysteine levels in pregnancies complicated with IUGR were accompanied by decreasing levels of serum folic acid and vitamin B12. Treatment with folic acid and vitamin B12 could improve fetal weight in IUGR pregnant patients with increased homocysteine levels.