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Bioaccumulation of d-Limonene in environment due to the aggrandised usage of their natural sources like citrus food wastes and industrial day to day life products has raised concern to their biotoxicity to environment biotic health. Moreover, their after-usage discharge to aquatic system has enhanced the distress of posing threat and needs attention. This study entails mechanistic and molecular evaluation of in-vivo biotoxicity of d-Limonene in zebrafish embryo models. Experimental analysis excavated the controlled concentration-dependent morphological, physiological and cellular in-vivo impact of d-Limonene in zebrafish embryos through significant changes in oxidative stress, steatosis and apoptosis regulated via 6-fold and 5-fold mRNA expression change in p53 and Sod1 genes. Computational evaluation deduced the cellular mechanism of d-limonene biotoxicity as irregularities in oxidative stress, apoptosis and steatosis due of their intrinsic interaction with metabolic proteins like Zhe1a (-4.8 Kcal/mol), Sod1(-5.3 Kcal/mol), p53, caspase3 and apoa1 leading to influential change in structural and functional integrity of the metabolic proteins. The study unravelled the measured in-vivo biotoxicity of d-Limonene at cellular and molecular level to advocate the controlled usage of d-Limonene related natural and industrial product for a sustainable environmental health., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In the modern environment conscious era, there has been a huge demand for the effective green method to fabricate biomaterials for sustained transdermal release of diltiazem hydrochloride to treat hypertension and cardiac failure. In this vein, the present study explores the amination of waste jute sourced nanocellulose (ANC) and its effect as a reinforcing filler to design electrospun polyvinyl alcohol (PVA)/chitosan based polymeric nanofibrous scaffold for drug delivery. The characterization results of FTIR (Fourier Transform Infrared Spectroscopy) confirm the successful chemical modification of nanocellulose (NCC). SEM (Scanning Electron Microscopy) results indicate the morphological modifications in ANC due to grafting. ANC enhances the mechanical properties of scaffold and sustains the release of the loaded drug to 67.89±3.39% as compared to the pure PVA/chitosan scaffold of 92.63±4.63% over a period of 72 h as shown by the results of in-vitro drug release study. Moreover, the incorporation of 0.5 % ANC improves the anti-bacterial activity against both gram-positive (97.4±4.87%, reduction in viable cells count) and gram-negative bacteria (98.5±4.93%, reduction in viable cells count). Further, the skin irritation and MTT assay authenticate the biocompatibility of the developed scaffold. The overall findings hence prove the efficacy of the engineered scaffold as a potential transdermal patch for sustained drug delivery applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A unique fluorescent molecule (ND-S) was obtained from Eosin Y in two simple yet high yielding steps (1). ND-S has special metal ion sensing ability, such that it can selectively detect toxic Hg 2+ present in very low concentration in aqueous solutions in the presence of other competing metal ions. The host-guest complexation is ratiometric and is associated with significant increase in fluorescence during the process. Isothermal titration calorimetry (ITC) experiments provided thermodynamic parameters related to interaction between ND-S and Hg 2+ . Using inductively coupled plasma mass spectrometry (ICP-MS), the Hg 2+ (aq) removal efficiency of ND-S was estimated to be 99.88%. Appreciable limit of detection (LOD = 7.4 nM) was observed. Other competing ions did not interfere with the sensing of Hg 2+ by ND-S. The effects of external stimuli (temperature and pH) were studied. Besides, the complex (ND-M), formed by 1:1 coordination of ND-S and Hg 2+ was found to be effective against the survival of Gram-positive bacteria (S. aureus and B. subtilis) with a high selectivity index. Moreover, bacterial cell death mechanism was studied systematically. Overall, we have shown the transformation of a toxic species (Hg 2+ ), extracted from polluted water by a biocompatible sensor (ND-S), into an effective and potent antibacterial agent (ND-M)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Visceral leishmaniasis (VL) is the most lethal among all leishmaniasis diseases and remains categorized as a neglected tropical disease (NTD). This study aimed to develop a peptide-based multi-epitope vaccine construct against VL using immunoinformatics methodologies. To achieve this, four distinct proteins were screened to identify peptides consisting of 9-15 amino acids with high binding affinity to toll-like receptors (TLRs), strong antigenicity, low allergenicity, and minimal toxicity. The resulting multi-epitope vaccine construct was fused in a tandem arrangement with appropriate linker peptides and exhibited superior properties related to cytotoxic T lymphocytes (CTLs), helper T lymphocytes (HTLs), and B-cell epitopes. Subsequently, a three-dimensional (3D) model of the vaccine construct was generated, refined, and validated for structural stability and immune response capabilities. Molecular docking and simulations confirmed the vaccine construct's stability and binding affinities with TLRs, with TLR4 displaying the highest binding affinity, followed by TLR2 and TLR3. Additionally, simulations predicted robust cellular and humoral antibody-mediated immune responses elicited by the designed vaccine construct. Notably, this vaccine construct includes proteins from various pathways of Leishmania donovani (LD), which have not been previously utilized in VL vaccine design. Thus, this study opens new avenues for the development of vaccines against diverse protozoan diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The estuaries provide the key pathway for travelling carbon across the land-ocean interfaces and behave as both source and sink of greenhouse gases (GHGs) in water-atmosphere systems. The sink-source characteristics of estuaries for GHGs vary spatially. The primary driving factors are adjacent ecologies (agriculture, aquaculture, etc.) and proximities to the sea. To study the sink-source characteristics of estuaries for GHGs (methane (CH 4 ), nitrous oxide (N 2 O) and carbon dioxide (CO 2 )), the water samples were collected from 53 different locations in the estuaries for estimation of dissolved GHGs concentration and air-water GHGs exchanges. The locations represent five zones (Zone I, II, III, IV and V) based on the type and degradation status of mangroves (degraded and undisturbed), anthropogenic activities, and distance from the sea. Zone I, III, V represents to the degraded mangroves far from sea, whereas, Zone II, IV surrounded by undisturbed mangroves and nearer to sea. The average dissolved CH 4 concentrations were higher in the estuaries which were adjacent to degraded mangroves (154.4 nmol L -1 ) than undisturbed mangroves (81.7 nmol L -1 ). Further, the average dissolved N 2 O concentrations were 48% higher in the estuaries nearer to degraded mangroves than that of undisturbed ones. Among the degraded mangrove sites, the dissolved CO 2 concentrations were higher at Zone I (30.1 μmol L -1 ) followed by Zone III and IV, whereas in undisturbed sites, it was higher in Zone IV (22.3 μmol L -1 ) than Zone II (17.6 μmol L -1 ). Among the 53 locations, 36, 51 and 33 locations acted as a sink (negative value of exchanges) for CH 4 , N 2 O and CO 2 , respectively. The higher sink potential for CH 4 was recorded to those estuaries adjacent to undisturbed mangroves (-791.69 μmol m -2  d -1 ) than the degraded ones (-23.18 μmol m -2  d -1 ). Similarly, the average air-water N 2 O and CO 2 exchanges were more negative in the estuaries which were nearer to undisturbed mangroves indicating higher sink potential. The pH, and salinity of the estuary water were negatively correlated with air-water CH 4 and N 2 O exchanges, whereas those were positively correlated with CO 2 exchanges. Significantly lower dissolved GHGs and air-water GHGs exchange was observed in the estuaries adjacent to the undisturbed mangrove as compared to the degraded mangrove. The reason behind higher sink behaviours of estuaries nearer to undisturbed mangroves are higher intrusion of seawater, less nutrient availability, higher salinity, low carbon contents and alkaline pH compared to estuaries adjacent to degraded mangroves and far from sea., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Immunotherapy targeting immune checkpoint proteins, such as programmed cell death ligand 1 (PD-L1), has shown impressive outcomes in many clinical trials but only 20%-40% of patients benefit from it. Utilizing Combined Positive Score (CPS) to evaluate PD-L1 expression in tumour biopsies to identify patients with the highest likelihood of responsiveness to anti-PD-1/PD-L1 therapy has been approved by the Food and Drug Administration for several solid tumour types. Current CPS workflow requires a pathologist to manually score the two-colour PD-L1 chromogenic immunohistochemistry image. Multiplex immunofluorescence (mIF) imaging reveals the expression of an increased number of immune markers in tumour biopsies and has been used extensively in immunotherapy research. Recent rapid progress of Artificial Intelligence (AI)-based imaging analysis, particularly Deep Learning, provides cost effective and high-quality solutions to healthcare. In this article, we propose an imaging pipeline that utilizes three-colour mIF images (DAPI, PD-L1, and Pan-cytokeratin) as input and predicts the CPS using AI techniques. Our novel pipeline is composed of three modules employing algorithms of image processing, machine learning, and deep learning techniques. The first module of quality check (QC) detects and removes the image regions contaminated with sectioning and staining artefacts. The QC module ensures that only image regions free of the three common artefacts are used for downstream analysis. The second module of nuclear segmentation uses deep learning to segment and count nuclei in the DAPI images wherein our specialized method can accurately separate touching nuclei. The third module of cell phenotyping calculates CPS by identifying and counting PD-L1 positive cells and tumour cells. These modules are data-efficient and require only few manual annotations for training purposes. Using tumour biopsies from a clinical trial, we found that the CPS from the AI-based models shows a high Spearman correlation (78%, p = 0.003) to the pathologist-scored CPS., Competing Interests: Declaration of competing interest None Declared., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The main objective of this paper is to improve the identification efficiency of non-linear systems using the Kalman filter (KF), which is optimised with the Artificial Electric Field (AEF) algorithm. The conventional KF suffers from the proper tuning of its parameters, which leads to a divergence problem. This issue has been solved to a great extent by the meta-heuristic AEF algorithm assisted Kalman filter (AEF-KF). This paper proposes three steps for the identification of the systems while solving the problem as mentioned above. Firstly, it converts the identification model to a measurement problem. Next, the AEF algorithm optimises the KF parameters by considering the fitness function with the KF equations. The third step is to identify the model using conventional KF algorithm with the optimised KF parameters. To evaluate the performance of the proposed method, parameter estimation error, mean squared error (MSE), fitness (FIT) percentage, statistical information and percentage improvement are considered as the performance metrics. To validate the performance of the proposed method, five distinct non-linear models are identified with the Volterra model using KF and the AEF-KF techniques under various noisy input conditions. Besides, the practical applicability of the proposed approach is also tested on two non-linear benchmark systems using experimental data sets. The obtained simulation results confirm the efficacy and robustness of the proposed identification method in terms of the convergence speed, computational time and various performance metrics as compared to KF, Kalman smoother (KS) which is optimised using different state-of-the-art evolutionary algorithms and also other existing recently reported similar types of stochastic algorithms based approaches., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 ISA. Published by Elsevier Ltd. All rights reserved.)

Background: Yogasana improves both mental and physical health. There is sparse systematic research on the growth of yogasana practising adolescents., Objectives: This study aims to assess the differential skeletal growth pattern among pre-adolescent and adolescent girls as a result of yoga practice., Methods: A cross-sectional anthropometric study was conducted on stature (height), sitting height and leg length of 757 school-going girls (4-15 years old), divided in two groups, Yoga Group (YG) (n=380) and age matched Control Group (CG) (n=377) participants participating in recreational games other than yoga. Descriptive and inferential statistical analyses were applied. Unpaired t-test was performed for assessment of level of significance and Pearson's correlation (r) test was performed to to identify the association between growth pattern of stature and leg length at specific ages., Results: The physical growth showed an ascending trend in both Yoga group (YG) participants and control group (CG). At the onset of adolescence (10-12 years) the mean stature and leg length of YG participants were retarded (p < 0.05). Sitting height in YG was significantly (p < 0.05) low only in 10-year-olds. The similar trends were observed in stature and leg length in YG participants at 10 years (5th and 10th percentile) and 12 years (90th and 95th percentile). There was strong positive relationship between stature and leg length of YG participants (10 years, r = 0.86, p < 0.01; 11 years, r = 0.86, p < 0.01; 12 years, r = 0.72, p < 0.01). The stunted growth in YG participants during adolescence may be related to retarded growth of leg length., Conclusions: Intense yogasana practice with greater skeletal stress possibly hinders stature in adolescent girls from 10 to 12 years. This may compromise with the natural growth pattern, necessitating special care during yoga training among adolescents while selecting the type, intensity and duration of yogasanas practice., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

β-carotene is a natural compound with immense healthcare benefits. To overcome insolubility and lack of stability which restricts its application, in this study, β-carotene from Planococcus sp. TRC1 was entrapped into formulations of chitosan‑sodium alginate microspheres (MF1, MF2 and MF3) and chitosan nanoparticles (NF1, NF2 and NF3). The maximum entrapment efficiency (%) and loading capacity (%) were 80.6 ± 4.28 and 26 ± 3.05 (MF2) and 92.1 ± 3.44 and 41.86 ± 4.65 (NF2) respectively. Korsmeyer-Peppas model showed best fit with release, revealing non-Fickian diffusion. Thermal and UV treatment exhibited higher activation energy (kJ/mol), 17.76 and 15.57 (MF2) and 37.03 and 19.33 (NF2) compared to free β-carotene (3.7 and 3.9), uncovering enhanced stability. MF2 and NF2 revealed swelling index (%) 721 ± 1.7 and 18.1 ± 1.5 (pH 6.8) and particle size 69.5 ± 3.2 μm and 92 ± 2.5 nm respectively. FESEM, FT-IR, XRD and DSC depicted spherical morphology, intactness of functional groups and masking of crystallinity. The IC 50 (μg ml -1 ) values for antioxidant and anticancer (A-549) activities were 33.1 ± 1.7, 45.1 ± 2.8, 39.3 ± 2.9 and 31.3 ± 1.7, 27.9 ± 2.4, 25.3 ± 2.2 for β-carotene, MF2 and NF2 respectively with no significant cytotoxicity on HEK-293 cells and RBCs (p > 0.05). This comparative study of microspheres and nanoparticles may allow the diverse applications of an unconventional bacterial β-carotene with promising stability and efficacies., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Alloxazine phototautomerization is believed to occur through an excited state double proton transfer (ESDPT) mechanism involving cyclic intermolecular H-bonded complexes between Alloxazine and hydroxylic solvents like water and alcohols. In AOT/alkane dispersions in the absence of any polar liquid, Alloxazine molecules reside inside the polar core of the AOT reverse micelle nanoparticles, where they involve in H-bonding with the anionic sulfonate head-groups of the AOT molecules, but are unable to generate the appropriate cyclic intermolecular H-bonded complexes conducive to ESDPT. However, tautomerization is switched on with addition of water and formation ofwater nano-droplet at the core of reverse micelle. Evidently, the Alloxazine⋅⋅⋅⋅AOT H-bonds are now replaced by Alloxazine⋅⋅⋅⋅Water H-bonds, promotingESDPT. On the other hand, Alloxazine phototautomerization is hindered in Glycerol, irrespective of whether the latter is in the bulk liquid state or in the form of a polar nano-droplet. This may be explained by steric considerations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Usage of a face mask has become mandatory in many countries after the outbreak of SARS-CoV-2, and its usefulness in combating the pandemic is a proven fact. There have been many advancements in the design of a face mask and the present treatise describes a face mask in which a simple triboelectric nanogenerator (TENG) with an electrocution layer may serve the purpose of filtration and deactivation of SARS-CoV-2. The proposed mask is designed with multilayer filters, in which the inner three layers act as a triboelectric (TE) filter and the outer one as an electrocution layer (EL). The viral particles experience a mildshock in EL due to the electric field produced between the electrocution layers by contact electrification. Four pairs of triboelectric series fabrics, i.e. polyvinylchloride (PVC)-nylon, polypropylene (PP)-polyurethane (PU), latex rubber-PU, polyimide (PI)-nylon are studied to establish the efficacy of the mask. The motional force exerted on triboelectric filter materials can produce sufficient electric power to activate EL. The proposed mask can be used by a wide range of people because of its triboelectric self-powering (harvesting mechanical energy from daily activities, e.g. breathing, talking or other facial movements) functionalities to ensure effective filtration efficiency. More importantly, it is expected to be potentially beneficial to slow down the devastating impact of COVID-19., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2020 Elsevier Ltd. All rights reserved.)

Dehydration stress response is a complex mechanism in plants involving several factors and hormone signalling pathways. RAV1 is a member of the AP2/ERF family of transcription factors that works in various developmental pathways. Here we show that downregulation of RAV1 gene expression is important for efficient dehydration stress response. Interestingly, the B3-domain transcription factor ABI3 negatively regulates RAV1 expression. In absence of ABI3, RAV1 expression increases during dehydration stress compared to control. As a part of stress response, ABI3 occupancy increases in the RAV1 promoter region. Such regulation of RAV1 gene expression seems vital as absence of RAV1 leads to reduced water loss during dehydration stress and consequently faster recovery compared to wild type. rav1 mutant seedlings show more abundant root growth under control condition and higher primary root elongation compared to wild type when subjected to dehydration stress. Mutants also exhibit enhanced ABA sensitivity compared to wild type. At the transcript level, rooting genes like NAC1, ARF16, SLR and SLR-downstream genes like ARF7, PLT3, SHR show differential expression in rav1 mutant, compared to wild type. Additionally, ethylene-responsive genes ETR1, EIN2 and ERF1 also get differentially expressed in presence and absence of RAV1 under control and stress conditions. This indicates an altered ethylene response in the rav1 mutant. All these features render rav1seedlings better equipped for responding to dehydration stress. It thus becomes evident that ABI3 mediated regulation of RAV1 gene expression is a significant part of dehydration stress signalling for efficient stress management at the molecular and morphological level., Competing Interests: Declaration of competing interest Contents of this manuscript are solely the responsibility of the authors and do not necessarily represent the official views of the funding agency., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The creation of a polymeric hydrogel from polyvinylpyrrolidone (PVP) cross-linked by Carbon Quantum Dots (CD) for the adsorption and photocatalytic degradation of both cationic and anionic dyes. PVP, an important biocompatible constituent and often surplus in cosmetic industry, was carboxylated through NaOH refluxing and covalently conjugated to surface amine functionality of CD derived from lemon juice and Cysteamine. The hybrid hydrogel was obtained from PVP-CD covalent conjugate by careful manipulation of pH and found to possess better rheological properties than only carboxylate-PVP. The monolayer physisorption of the dyes on the hydrogel was affected by hydrogen bonding, dispersion or inductive effect, and π-π interaction with the polymer backbone as well as the CD that followed pseudo-second-order kinetics. Degradation of the adsorbed dyes was instated by the unique Reactive Oxygen Species (ROS) generating ability of the CD embedded in the hydrogel matrix upon exposure to sunlight, the mechanism of which is also unveiled. The same CD-induced ROS was found to effectively annihilate both gram-positive and gram-negative bacteria in real polluted water in less than 10 min of photoexcitation of the hydrogel. The hydrogel was restored by mild acid wash that is able to perform dye adsorption and photo-degradation upto four cycles., Competing Interests: Declaration of Competing Interest The authors declare that there are no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Till date, only three techniques namely Zinc Finger Nuclease (ZFN), Transcription-Activator Like Effector Nucleases (TALEN) and Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR-Associated 9 (CRISPR-Cas9) are available for targeted genome editing. CRISPR-Cas system is very efficient, fast, easy and cheap technique for achieving knock-out gene in the cell. CRISPR-Cas9 system refurbishes the targeted genome editing approach into a more expedient and competent way, thus facilitating proficient genome editing through embattled double-strand breaks in approximately any organism and cell type. The off-target effects of CRISPR Cas system has been circumnavigated by using paired nickases. Moreover, CRISPR-Cas9 has been used effectively for numerous purposes, like knock-out of a gene, regulation of endogenous gene expression, live-cell labelling of chromosomal loci, edition of single-stranded RNA and high-throughput gene screening. The execution of the CRISPR-Cas9 system has amplified the number of accessible scientific substitutes for studying gene function, thus enabling generation of CRISPR-based disease models. Even though many mechanistic questions are left behind to be answered and the system is not yet fool-proof i.e., a number of challenges are yet to be addressed, the employment of CRISPR-Cas9-based genome engineering technologies will increase our understanding to disease processes and their treatment in the near future. In this review we have discussed the history of CRISPR-Cas9, its mechanism for genome editing and its application in animal, plant and protozoan parasites. Additionally, the pros and cons of CRISPR-Cas9 and its potential in therapeutic application have also been detailed here., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Six structurally different carbazoles (1-6) were explored as the green reducing agents for the synthesis of the fluorescent Au nanoparticles with tailor-made morphology in anionic (sodium dodecyl sulphate, SDS), cationic (cetyltrimethylammonium bromide, CTAB) and neutral (polyvinylpyrrolidone, PVP) micelle medium. Structure of the carbazoles played an important role in controlling the morphology, rate of formation and fluorescent activity of the Au nanoparticles. The Au nanoparticles formed in-situ also simultaneously catalyzed the intermolecular CC and NN couplings between the carbazoles, leading to the corresponding bis-carbazole derivatives. The free and bis-carbazole derivatives functionalized the surface of the synthesized Au nanoparticles and thereby controlling their morphology and fluorescence activity. A computational study was also made to determine the origin of the absorption and emission bands of the synthesized nanoparticles. The combined experimental and theoretical studies unraveled the nanoparticle formation process and mechanistic pathway of this green and easily implementable synthetic protocol of Au nanoparticles., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Genome-wide association studies are widely used to map genomic regions contributing to lung cancer (LC) susceptibility, but they typically do not identify the precise disease-causing genes/variants. To unveil the inherited genetic variants that cause LC, we performed focused exome-sequencing analyses on genes located in 121 genome-wide association study-identified loci previously implicated in the risk of LC, chronic obstructive pulmonary disease, pulmonary function level, and smoking behavior., Methods: Germline DNA from 260 case patients with LC and 318 controls were sequenced by utilizing VCRome 2.1 exome capture. Filtering was based on enrichment of rare and potential deleterious variants in cases (risk alleles) or controls (protective alleles). Allelic association analyses of single-variant and gene-based burden tests of multiple variants were performed. Promising candidates were tested in two independent validation studies with a total of 1773 case patients and 1123 controls., Results: We identified 48 rare variants with deleterious effects in the discovery analysis and validated 12 of the 43 candidates that were covered in the validation platforms. The top validated candidates included one well-established truncating variant, namely, BRCA2, DNA repair associated gene (BRCA2) K3326X (OR = 2.36, 95% confidence interval [CI]: 1.38-3.99), and three newly identified variations, namely, lymphotoxin beta gene (LTB) p.Leu87Phe (OR = 7.52, 95% CI: 1.01-16.56), prolyl 3-hydroxylase 2 gene (P3H2) p.Gln185His (OR = 5.39, 95% CI: 0.75-15.43), and dishevelled associated activator of morphogenesis 2 gene (DAAM2) p.Asp762Gly (OR = 0.25, 95% CI: 0.10-0.79). Burden tests revealed strong associations between zinc finger protein 93 gene (ZNF93), DAAM2, bromodomain containing 9 gene (BRD9), and the gene LTB and LC susceptibility., Conclusion: Our results extend the catalogue of regions associated with LC and highlight the importance of germline rare coding variants in LC susceptibility., (Copyright © 2018 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.)

Liver cancer is a leading cause of death related to cancer worldwide. Poly(d-l-lactide-co-glycolide) (PLGA) nanoparticles provide prolonged blood residence time and sustained drug release, desirable for cancer treatment. To achieve this, we have developed paclitaxel-loaded PLGA nanoparticles by emulsification solvent evaporation method and evaluated by in vitro and in vivo studies. The results obtained from in vitro study showed that drug loading efficiency was 84.25% with an initial burst release followed by sustained drug release. Cellular uptake and in vitro cytotoxicity of the formulated nanoparticles using HepG2, Huh-7 cancer cells and Chang liver cells were also investigated. The formulated nanoparticles showed more cytotoxic effect at lower concentration and were internalized well by HepG2 cells compared to free-drug and marketed formulation. Prolonged half-life and higher plasma and liver drug concentrations of the formulated nanoparticles were observed as compared to free drug and marketed formulation in rats. Thus, paclitaxel-loaded polymeric nanoparticle has shown its potential for the treatment of liver cancer., (Copyright © 2018 Elsevier B.V. All rights reserved.)

For the first time, a synthetic fluorescent antioxidant flavone analog was successfully anchored onto the surface of the APTES-modified mesoporous silica nanoparticles (NPs) through sulfonamide linkage. The surface chemistry and morphology of the flavone modified fluorescent silica (FMFS) NPs were studied in detail. The flavone moiety when attached onto the FMFS NP surface, imparted its characteristic fluorescence and antioxidant activities to these NPs. Moreover, the NPs are highly biocompatible as evidenced from their cytotoxicity assay on normal lung cell (L132). The fluorescence activity of these biocompatible NPs was further utilized to study their interaction with a biomolecule, BSA (Bovine Serum Albumin). It was interesting to note that the fluorescence behavior of FMFS NPs completely changed on their binding with BSA. On the other hand, the intrinsic fluorescence activity of BSA was also significantly modified due to its interaction with FMFS NPs. Thus, the sensing and detection of biomolecules like BSA in presence of FMFS NPs can be accomplished by monitoring changes in the fluorescence behavior of either FMFS NPs or BSA. Furthermore, these FMFS NPs retained their intrinsic fluorescence behavior in the cellular medium which opens up their possible use as biocompatible cell imaging agents in future., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Organogels prepared from AOT/4-chlorophenol/m-xylene are immobile in the macroscopic sense, with a well-characterized internal structure. However, the molecular level dynamics inside the gels is not too clear, although a very slow structural relaxation has been reported previously. Using a set of rod-like fluorophores, we find that the rotational mobility of a small guest molecule inside the gel can be extremely fast, indicating presence of sufficiently low-microviscosity domains. These domains consist of m-xylene solvent molecules trapped in the interstices of fiber bundles comprising columnar stacks of 4-chlorophenol surrounded by AOT molecules. However, interstitial trapping of m-xylene does retard its own dynamics, which explains the slow solvent relaxation inside the gels. Hence, the state of m-xylene in the organogel may be characterized as "bound", in contrast to the "free" state in neat m-xylene., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Introduction: The association between smoking-induced chronic obstructive pulmonary disease (COPD) and lung cancer (LC) is well documented. Recent genome-wide association studies (GWAS) have identified 28 susceptibility loci for LC, 10 for COPD, 32 for smoking behavior, and 63 for pulmonary function, totaling 107 nonoverlapping loci. Given that common variants have been found to be associated with LC in genome-wide association studies, exome sequencing of these high-priority regions has great potential to identify novel rare causal variants., Methods: To search for disease-causing rare germline mutations, we used a variation of the extreme phenotype approach to select 48 patients with sporadic LC who reported histories of heavy smoking-37 of whom also exhibited carefully documented severe COPD (in whom smoking is considered the overwhelming determinant)-and 54 unique familial LC cases from families with at least three first-degree relatives with LC (who are likely enriched for genomic effects)., Results: By focusing on exome profiles of the 107 target loci, we identified two key rare mutations. A heterozygous p.Arg696Cys variant in the coiled-coil domain containing 147 (CCDC147) gene at 10q25.1 was identified in one sporadic and two familial cases. The minor allele frequency (MAF) of this variant in the 1000 Genomes database is 0.0026. The p.Val26Met variant in the dopamine β-hydroxylase (DBH) gene at 9q34.2 was identified in two sporadic cases; the minor allele frequency of this mutation is 0.0034 according to the 1000 Genomes database. We also observed three suggestive rare mutations on 15q25.1: iron-responsive element binding protein neuronal 2 (IREB2); cholinergic receptor, nicotinic, alpha 5 (neuronal) (CHRNA5); and cholinergic receptor, nicotinic, beta 4 (CHRNB4)., Conclusions: Our results demonstrated highly disruptive risk-conferring CCDC147 and DBH mutations., (Copyright © 2015 International Association for the Study of Lung Cancer. All rights reserved.)

This paper presents an efficient approach to identify different stable and practically useful Hammerstein models as well as unstable nonlinear process along with its stable closed loop counterpart with the help of an evolutionary algorithm as Colliding Bodies Optimization (CBO) optimization algorithm. The performance measures of the CBO based optimization approach such as precision, accuracy are justified with the minimum output mean square value (MSE) which signifies that the amount of bias and variance in the output domain are also the least. It is also observed that the optimization of output MSE in the presence of outliers has resulted in a very close estimation of the output parameters consistently, which also justifies the effective general applicability of the CBO algorithm towards the system identification problem and also establishes the practical usefulness of the applied approach. Optimum values of the MSEs, computational times and statistical information of the MSEs are all found to be the superior as compared with those of the other existing similar types of stochastic algorithms based approaches reported in different recent literature, which establish the robustness and efficiency of the applied CBO based identification scheme., (Copyright © 2015 ISA. Published by Elsevier Ltd. All rights reserved.)

The surface expression of CD9 (cluster-of-differentiation antigen-9) in sperms of certain mammalian species has been attributed to its fusion with the egg and thereby dictating the fertility of species. In the present study, we investigated the association of CD9 with crossbred bull sperm quality and quantity trait was analyzed using a total of 96 Frieswal (HF × Sahiwal) crossbred. A single nucleotide polymorphism (g.358A > T) in intron 6 was significantly associated with sperm concentration (P < 0.05) and motility percentage (P < 0.01). mRNA was extracted from good (progressive motility > 50%) and motility impaired (progressive motility < 50%) bull semen. The mRNA expression and seminal plasma protein concentration of CD9 was significantly (P < 0.05) higher among good quality bull semen than motility impaired ones. Our results thus may indicate that, mutation in the intronic region may be responsible for the instability of RNA and the subsequent functional protein expression.

We studied the roles of vitamin D and its receptor, VDR, in the progression of leprosy. The majority of individuals with leprosy from Kolkata, India, with a type 1 or type 2 reaction have low levels of vitamin D3 in serum samples. Interestingly, individuals with a type 2 reaction associated with neuritis/erythema nodosum leprosum had very low VDR mRNA expression levels, ranging from 5% to 10%, compared to that of healthy control subjects; these patients also had a high bacilli index, ranging from 3+ to 5+. This is the first report to indicate that VDR expression levels may determine the complexity and severity of the progression of leprosy.

A green technique of silica nanoparticles (SiO2-NPs) formation by using a thermophilic bacterium (BKH1) as biological template is demonstrated here. SiO2-NPs are synthesized from inorganic (magnesium tri-silicate), and organic (tetraethyl orthosilicate) precursor with the help of BKH1 bacteria. BKH1 derived SiO2-NPs are subjected to Atomic Force Microscopy, Transmission Electron Microscopy, and Field Emission Scanning Electron Microscopy equipped with Energy Dispersive X-ray Analyzer to establish nanoparticle morphology. In addition, Infrared Spectroscopy reveals the presence of chemical and functional groups in SiO2-NPs samples and X-ray diffraction, the amorphous nature. The Zeta potential (ζ) reveals substantial stability of bacteria derived SiO2-NPs in the aqueous environment. Presence of two intense luminescence peaks in the UV and visible regions merits the bacteria derived SiO2-NPs for use as an optical probe in biomedical applications. This novel mode of bacteria derived SiO2-NPs formation is eco-friendly and ambient temperature synthesis approach. It avoids the complex protocol of multi-steps synthesis of silica nanoparticles, hence likely to be cost-effective. In-depth translation research is suggested for the synthesis of silica nanoparticles in large quantities using thermophilic BKH1 template., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The aim of this study was to find out the intracellular signaling transduction pathways involved in cobalt oxide nanoparticles (CoO NPs) mediated oxidative stress in vitro and in vivo system. Cobalt oxide nanoparticles released excess Co++ ions which could activated the NADPH oxidase and helps in generating the reactive oxygen species (ROS). Our results showed that CoO NPs elicited a significant (p<0.05) amount of ROS in lymphocytes. In vitro pretreatment with N-acetylene cystine had a protective role on lymphocytes death induced by CoO NPs. In vitro and in vivo results showed the elevated level of TNF-α after CoO NPs treatment. This TNF-α phosphorylated the p38 mitogen-activated protein kinase followed by activation of caspase 8 and caspase 3 which could induce cell death. This study showed that CoO NPs induced oxidative stress and activated the signaling pathway of TNF-α-caspase-8-p38-caspase-3 to primary immune cells. This study suggested that bare CoO NPs are a toxic for primary human immune cells that deals directly with human health. Surface modification or surface functionalization may open the gateway for further use of CoO NPs in different industrial use or in biomedical sciences., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

The present work reports the development of water-in-oil (w/o) microemulsion doped with newly designed nanocomposite comprising of gold nanoparticle (GNP) decorated single walled carbon nanotube (SWNT). This nanocomposite included cationic reverse micelle was used to boost the catalytic activity of a surface-active enzyme, Chromobacterium viscosum lipase (CV lipase). SWNT was non-covalently dispersed using cetyltrimethylammonium bromide (CTAB), cetylalaninetrimethylammonium chloride (CATAC) while GNP was synthesized by reduction of HAuCl4 with reducing/stabilizing agent trisodium citrate. Counterion exchange between cationic SWNT dispersing agent and anionic capping agent of GNP led to the formation of GNP decorated SWNT (SWNT-GNP) nanocomposite. This newly developed SWNT-GNP included CTAB reverse micelle was characterized by several microscopic and spectroscopic techniques. Interfacially located SWNT-GNP included w/o microemulsion (confirmed from biphasic and fluorescence experiment) was used as a proficient host for enhancing the catalytic activity of lipase. Lipase activity within this self-assembled soft nanocomposite improved up to 3.9-fold (second order rate constant, k2=1694±16 cm(3) g(-1) s(-1)) compared to standard CTAB reverse micelle (k2=433±7 cm(3) g(-1) s(-1)). In case of cetyltripropyl ammonium bromide (CTPAB) based reverse micelle, the observed lipase activity improved to k2=2036±11 cm(3) g(-1) s(-1) in the presence of SWNT-GNP composite. Notably, this catalytic activity of lipase within SWNT-GNP included reverse micelle was till date the highest activity found in any w/o microemulsion. The attainment of flexibility in enzyme conformation at the augmented interface was verified using circular dichroism (CD) spectroscopy., (Copyright © 2013 Elsevier B.V. All rights reserved.)

In this paper a new meta-heuristic search method, called Cat Swarm Optimization (CSO) algorithm is applied to determine the best optimal impulse response coefficients of FIR low pass, high pass, band pass and band stop filters, trying to meet the respective ideal frequency response characteristics. CSO is generated by observing the behaviour of cats and composed of two sub-models. In CSO, one can decide how many cats are used in the iteration. Every cat has its' own position composed of M dimensions, velocities for each dimension, a fitness value which represents the accommodation of the cat to the fitness function, and a flag to identify whether the cat is in seeking mode or tracing mode. The final solution would be the best position of one of the cats. CSO keeps the best solution until it reaches the end of the iteration. The results of the proposed CSO based approach have been compared to those of other well-known optimization methods such as Real Coded Genetic Algorithm (RGA), standard Particle Swarm Optimization (PSO) and Differential Evolution (DE). The CSO based results confirm the superiority of the proposed CSO for solving FIR filter design problems. The performances of the CSO based designed FIR filters have proven to be superior as compared to those obtained by RGA, conventional PSO and DE. The simulation results also demonstrate that the CSO is the best optimizer among other relevant techniques, not only in the convergence speed but also in the optimal performances of the designed filters., (Copyright © 2013 ISA. Published by Elsevier Ltd. All rights reserved.)

Developed ceria/yttria stabilized zirconia and ceria/yttria stabilized zirconia toughened alumina supported formation of apatite layer when immersed in simulated body fluid without any prior surface treatment. The formed mineral layer was confirmed as hydroxyapatite through X-ray diffraction patterns. The calcium/phosphate atomic ratio obtained from energy dispersive X-ray spectroscopy was found to be little less (Ca/P=1.5) than that of pure hydroxyapatite (Ca/P=1.7) which indicates the probability of mixed type calcium-phosphate compound formation. The achieved thickness of apatite layer was estimated through a surface profilometer and as high as ~17 μm thickness was found after 28 days of soaking. The biocompatibility of the developed materials was ensured through in vitro human osteoblast like cell (MG63) culture on ceramic discs. The morphology of attached cells was characterized through scanning electron microscopy and fluorescent microscopy which show multilayered interconnected cell growth within 8 days of culture period. Moreover, differentiation of MG63 cells was evaluated through MTT assay, total protein content and alkaline phosphatase activity., (Copyright © 2013 Elsevier B.V. All rights reserved.)

In this paper, we have demonstrated for the first time, the superb efficiency of aqueous extract of dried leaves of mahogany (Swietenia mahogani JACQ.) in the rapid synthesis of stable monometallic Au and Ag nanoparticles and also Au/Ag bimetallic alloy nanoparticles having spectacular morphologies. Our method was clean, nontoxic and environment friendly. When exposed to aqueous mahogany leaf extract, competitive reduction of Au(III) and Ag(I) ions present simultaneously in same solution leads to the production of bimetallic Au/Ag alloy nanoparticles. UV-visible spectroscopy was used to monitor the kinetics of nanoparticles formation. UV-visible spectroscopic data and TEM images revealed the formation of bimetallic Au/Ag alloy nanoparticles. Mahogany leaf extract contains various polyhydroxy limonoids which are responsible for the reduction of Au(III) and Ag(I) ions leading to the formation and stabilization of Au and Ag nanopaticles., (Copyright © 2010 Elsevier B.V. All rights reserved.)

This study was conducted on 94 Frieswal (5/8 Holstein Friesian 3/8 Sahiwal) crossbred bulls of three different grades, categorized based on their semen freezability visualising Group 1 (consistently freezable semen producer bulls, N = 11), Group 2 (inconsistent freezable, N = 16) and Group 3 (Non freezable, N = 67). Each group was further divided into two classes that is young (up to 30 months) and adult (31 to 70 months) bulls depending upon their age. Sperm morphology was studied by using the eosin-nigrosin staining technique. Bulls age significantly (P < 0.01) affected semen quality and sperm morphology. In adult bulls, semen volume, mass activity and sperm concentration were 36%, 17.56% and 19.6%, respectively, higher than young. Initial progressive motility (%) and livability showed significant (P < 0.01) improvement with the advancement of age (43.37 ± 1.21 and 67.71 ± 1.11, respectively, in young; 53.02 ± 1.11 and 74.17 ± 1.03, respectively, in adult). In young bulls, sperm head, mid piece, tail abnormality and total abnormal sperm percent (12.38 ± 0.92, 4.87 ± 0.24, 11.01 ± 0.60 and 28.26 ± 1.34, respectively) were 1.85, 1.27, 1.20 and 1.44 folds higher than that of their mature stage (6.69 ± 0.64, 3.82 ± 0.32, 9.14 ± 0.64 and 19.66 ± 1.31, respectively). Significant reduction (P < 0.01) in micro cephalic sperm, free heads, bent mid piece, looped mid piece and proximal protoplasmic droplets were observed at mature age as compared with their younger stage. In bulls of consistent freezing category, abnormal sperm heads significantly decreased from 4.40 ± 0.31% to 3.28 ± 0.02% on maturity. Similarly, in inconsistent freezing grade bulls sperm head abnormality (9.28 ± 0.75% to 5.13 ± 1.20%) and total abnormal sperm percent (24.89 ± 1.43 to 18.73 ± 3.40) was decreased over the age. On the contrary, in non-freezing category bulls' sperm morphology did not show significant (P > 0.05) improvement with age advancement, rather some abnormalities like long slender head, under developed/deformed head, abaxial implantation of mid piece, double mid piece, stump tail and distal protoplasmic droplets tend to increased significantly (P < 0.05) with age of bulls. Results indicated that in potential Frieswal bulls semen quality and sperm morphology were improved from young to mature stage, where as, in poor quality (non-freezing) semen producer bulls neither the morphology nor the semen quality showed any improvement with maturity. It was recommended that crossbred bulls producing more than 25% morphologically abnormal sperms in young age (below 30 months) along with poor progressive motility (<50%) and low sperm concentration (<1000 million/ml) need immediate culling with out any expectation of further improvement in semen quality with age advancement.

In an attempt to find natural, environmentally benign, green-chemical agents for the synthesis of metal nanoparticles, we have demonstrated for the first time the excellent efficiency of ethanol and water extracts of a natural, non-toxic material, Indian propolis and two of its chemical constituents, pinocembrin and galangin in the rapid synthesis of stable Ag and Au nanoparticles having wide spectrum of fascinating morphologies. Both of these two extracts were found to be extremely efficient in the synthesis of Ag and Au nanoparticles under alkaline condition. For a given metal ion precursor, the kinetics of particle synthesis were remarkably similar in all the cases, as it is evident from the absorption spectra monitored over time. Moreover they exhibited similar redox behavior under alkaline condition (pH approximately 10.62). The efficiency of the ethanol and water extracts of Indian propolis towards Ag and Au nanoparticles synthesis was compared with that of naturally occurring hydroxyflavonoids, pinocembrin and galangin isolated from Indian propolis; which are equally efficient in the rapid synthesis of Ag and Au nanoparticles and stabilization of the resultant particles.

We explored the application of three different aqueous solutions derived from Black Tea leaf extracts in the synthesis of Au and Ag nanoparticles. The plain tea leaf broth, as well as that containing the ethyl acetate extract of tea leaves, were found to be extremely efficient, leading to rapid formation of stable nanoparticles of various shapes: spheres, trapezoids, prisms and rods. For a given metal ion precursor, the kinetics of particle synthesis were remarkably similar in these two solutions, as evidenced from their absorption spectroscopy monitored over time. Moreover, they exhibited similar redox behavior. In contrast, with the other solution, containing the dichloromethane (CH(2)Cl(2)) extract of tea leaves, we failed to detect any nanoparticle generation under similar reaction conditions. Our results suggest that the reduction of metal ions and stabilization of the resultant particles in the first two solutions involved the same class of biomolecules. We identified these biomolecules as the tea polyphenols, including flavonoids, which were present in comparable amounts in both the tea leaf broth and ethyl acetate extract, but are absent in the CH(2)Cl(2) extract of tea leaves. The efficiency of the tea leaf extracts towards Au and Ag nanoparticle synthesis were compared with that of a naturally occurring hydroxyflavonoid, quercetin.

Noncovalent protein delivery into cells via peptide carriers is an emerging concept. Only a handful of such peptides are known. To address various limitations associated with protein delivery for therapeutic purposes, a greater number of different delivery peptides would be required. No general method exists for creating such peptides. By combining a sequence of 16 lysine residues (K16) with the signal peptide (SP) sequence of Kaposi's fibroblast growth factor (K-FGF), we have synthesized a peptide (K16SP) that efficiently and noncovalently delivers functionally intact proteins (immunoglobulin G molecules, beta-galactosidase, and green fluorescent protein) into mammalian cells. The peptides K16 and SP each alone did not show any noncovalent protein-carrying capacity. K16SP appears to be nontoxic to cells and three to four times more efficient than a commercially available peptide reagent. Our approach offers proof-of-concept of a general strategy for creating a diverse array of peptide carriers for eventual therapeutic applications.

Purpose: Aqueous decoction of Chenopodium album seeds (CAD) was assessed for its sperm-immobilizing and contraceptive efficacy in laboratory mammals., Method: Spermicidal efficacy was evaluated in vitro by a modified Sander-Cramer test. The mode of spermicidal action was assessed by (a) supravital and double fluoroprobe staining of sperm, (b) hypoosmotic swelling tests and (c) transmission electron microscopy. Contraceptive efficacy was evaluated by intrauterine and vaginal application of CAD in rats and rabbits, respectively, followed by their mating and evaluation of pregnancy outcomes., Results: The minimum effective concentration of CAD that induced instantaneous immobilization of rat spermatozoa in vitro was 2 mg/mL. The mechanism of CAD action involved disintegration of sperm plasma membrane and dissolution of acrosomal cap causing sperm death. Fertilization of oocytes and establishment of implantation were prevented in the uterine horn that was administered with CAD, while these events occurred unhindered in the untreated contralateral side. In rabbit, intravaginal application of CAD significantly blocked the establishment of pregnancy., Conclusion: CAD possesses appreciable spermicidal potential, which may be explored as an effector constituent of vaginal contraceptive.

Three steroidal saponins, racemosides A (1), B (2) and C (3), were isolated from the methanolic extract of the fruits of Asparagus racemosus, and characterized as (25S)-5beta-spirostan-3beta-ol-3-O-{beta-D- glucopyranosyl (1-->6)-[alpha-L-rhamnopyranosyl (1-->6)-beta-D-glucopyranosyl (1-->4)]-beta-D-glucopyranoside}, (25S)-5beta-spirostan-3beta-ol-3-O-alpha-L-rhamnopyranosyl (1-->6)-beta-D-glucopyranosyl (1-->6)-beta-D-glucopyranoside and (25S)-5beta-spirostan-3beta-ol-3-O-{alpha-L-rhamnopyranosyl-(1-->6)-[alpha-L-rhamnopyranosyl (1-->4)]-beta-D-glucopyranoside}, respectively, by spectrometric analysis and some chemical strategies.

Bioguided-fractionation of the methanol extract of the leaves of Careya arborea led to isolation of a triterpenoid saponin, designated arborenin, and characterized as 3-O-beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl-2 alpha,3beta-dihydroxy-taraxast-20-en-28-oic acid (1), together with desacylescin III (2). The structures were determined on the basis of extensive 2D NMR spectroscopic analysis. The saponin showed in vitro antileishmanial activity against Leishmania donovani (strain AG 83).

Tumor-induced immunosuppression often leads to failure in cancer therapy. Here, in an attempt to understand the course of tumor-dependent immunosuppression in young adult murine model, we found that in Ehrlich's ascites carcinoma (EAC) bearing mice, CD4(+) and CD8(+) populations of peripheral blood were depleted within first week of tumor inoculation. However, there was a rise in these populations at the end of second week only to fall back severely at the end of third week. These pulsating changes were also reflected in spleen. Interestingly, in thymus, production of CD4(+) and CD8(+) increased during first two weeks of tumor inoculation indicating the effort of thymus to replenish these populations in peripheral blood and spleen in response to their initial depletion, restricting tumor growth in between first and second weeks. However, at third week, due to (a) block in thymocyte maturation leading to increase in CD4(-)8(-) and decrease in CD4(+)8(+), (b) inhibition in formation of functional isotypes, and (c) thymocyte apoptosis, thymic reinforcement was stalled. Further investigation for the underlying mechanism of such thymic atrophy revealed down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bax, resulting in decreased Bcl-2/Bax ratio thereby inducing apoptosis. Above findings accounted for the significant decrease in CD4(+) and CD8(+) of peripheral blood and spleen by the end of third week culminating in total collapse in the fight back mechanism of host and uncontrolled growth of tumor. All these results signify the importance of thymus in modulating the immune status of the host during tumor development.

Semen was collected from six adult (3.5-7-year-old) Murrah buffalo bulls at weekly intervals for 1 year and evaluated for routine parameters, motion characteristics, reactivity in hypoosmotic solution, and acrosomal and other morphological abnormalities of the spermatozoa. The overall motility (MOT), straight line velocity (VSL), curvilinear velocity (VCL), linearity (LIN), lateral head displacement (ALH) and average path velocity (VAP) were 66.85+/-2.79%, 26.58+/-0.24 and 107.07+/-1.47 microm/s, 26.91+/-0.01%, 11.19+/-0.09 and 61.78+/-2.79 microm/s, respectively. Significant seasonal variation was observed in sperm kinematics and hypoosmotic swelling (HOS) reactivity. Except for LIN, the mean values of sperm dynamics were higher during summer and rainy season and significantly lower in winter season. Sperm kinematics showed significant (P<0.01) positive correlation (r=0.25-0.60) with plasmallemal integrity. Ejaculates with less than 50% HOS-reactive spermatozoa had significantly lowered MOT, VSL, VCL and VAP as compared to the ejaculates with >50% HOS-positive spermatozoa. No significant difference was observed in sperm kinematics among the ejaculates having 50-70% and >70% HOS-reactive spermatozoa. The trend of motion dynamics of the spermatozoa with respect to HOS reactivity was similar in all the three seasons (summer, rainy and winter). The results indicate that ejaculates having more than 50% of HOS-reactive sperm show a higher magnitude of sperm kinematics compared to ejaculates having less than 50% HOS-positive spermatozoa.

The equilibrium denaturation of tetrameric soybean agglutinin (SBA) in urea and guanidine hydrochloride (GdnHCl) has been examined by steady-state fluorescence and size-exclusion chromatography. The denaturation of SBA reveals two distinct and separable transitions: dissociation (native tetramer<-->tertiary monomer) and unfolding (tertiary monomer<-->unfolded monomer). The urea denaturation curves of N-dimethyl and acetyl derivatives of SBA are also similar to unmodified lectin but the midpoints, [D](1/2), are shifted to lower denaturant concentrations. The free energy of stabilization of tertiary structure (DeltaG(u,aq)) of SBA is estimated to be 4.5-4.6 kcal mol(-1), which shows a decrease by approximately 10-15% for both N-dimethyl SBA and acetyl-SBA. The free energy term (DeltaG(d, aq)) for the relative stability of the quaternary structure of SBA and its derivatives shows that the decrease in stability relative to SBA occurs by <10% for N-dimethyl SBA while for acetyl-SBA, this occurs by approximately 30%. However, the m values depicting the dependence of free energy on denaturant concentration for SBA and its derivatives are similar for dissociation as well as unfolding, which suggest similar denaturation pathways of unmodified and modified SBA.

Certain oligomannose-type glycopeptides have been previously shown to be bivalent for binding to concanavalin A and capable of precipitating the lectin by forming homogeneous cross-linked lattices [L. Bhattacharyya, M. I. Khan, and C.F. Brewer, Biochemistry, 27 (1988) 8762-8767]. In the present study, the effect of protein environment on the binding properties of an oligomannose-type oligosaccharide has been examined through quantitative precipitation analysis of the interactions of concanavalin A (Con A) with the soybean (Glycine max) agglutinin (SBA), which is a tetrameric glycoprotein possessing a single Man9-oligomannose chain per monomer. The results showed that SBA forms two different types of cross-linked complexes with tetrameric Con A, depending on the relative ratio of the two molecules in solution. At a concentration of one equivalent or less, SBA forms a 1:1 complex with Con A. At concentrations exceeding one equivalent, SBA forms a 2:1 complex with Con A. However, SBA forms only 1:1 cross-linked complexes with dimeric forms of Con A, such as acetyl- and succinyl-Con A. The results demonstrated that the total valency of the carbohydrate of SBA is a function of both the quaternary structure of Con A, as well as the relative ratio of SBA to Con A. In addition, the individual Man9-oligosaccharide, which as a glycopeptide is bivalent for binding to Con A, expresses univalency when present on the protein matrix of SBA.

Macroglobulin was purified from the eggwhite of the marine turtle (Caretta caretta Linn.) by gel filtration through Sephadex G-200 and Sepharose 4B. It was characterized by physical techniques including sedimentation velocity, diffusion and viscosity. Its molecular weight (Mr) was determined as 724,000 with four subunits of equal molecular weight. A large amount of water was hydrodynamically associated with the macroglobulin. It inhibited the activities of trypsin and papain and did not cross-react with human alpha 2-macroglobulin. Its amino acid composition was similar to that of human alpha 2-macroglobulin. Results suggest that turtle eggwhite macroglobulin is a homologous but distinct protein from human alpha 2-macroglobulin.