Performance analysis of MIMO-STBC systems with higher coding rate using adaptive semiblind channel estimation scheme.

Semiblind channel estimation method provides the best trade-off in terms of bandwidth overhead, computational complexity and latency. The result after using multiple input multiple output (MIMO) systems shows higher data rate and longer transmit range without any requirement for additional bandwidth or transmit power. This paper presents the detailed analysis of diversity coding techniques using MIMO antenna systems. Different space time block codes (STBCs) schemes have been explored and analyzed with the proposed higher code rate. STBCs with higher code rates have been simulated for different modulation schemes using MATLAB environment and the simulated results have been compared in the semiblind environment which shows the improvement even in highly correlated antenna arrays and is found very close to the condition when channel state information (CSI) is known to the channel.

A Precise Method to Detect Post-COVID-19 Pulmonary Fibrosis Through Extreme Gradient Boosting.

The association of pulmonary fibrosis with COVID-19 patients has now been adequately acknowledged and caused a significant number of mortalities around the world. As automatic disease detection has now become a crucial assistant to clinicians to obtain fast and precise results, this study proposes an architecture based on an ensemble machine learning approach to detect COVID-19-associated pulmonary fibrosis. The paper discusses Extreme Gradient Boosting (XGBoost) and its tuned hyper-parameters to optimize the performance for the prediction of severe COVID-19 patients who developed pulmonary fibrosis after 90 days of hospital discharge. A dataset comprising Electronic Health Record (EHR) and corresponding High-resolution computed tomography (HRCT) images of chest of 1175 COVID-19 patients has been considered, which involves 725 pulmonary fibrosis cases and 450 normal lung cases. The experimental results achieved an accuracy of 98%, precision of 99% and sensitivity of 99%. The proposed model is the first in literature to help clinicians in keeping a record of severe COVID-19 cases for analyzing the risk of pulmonary fibrosis through EHRs and HRCT scans, leading to less chance of life-threatening conditions.

Carboxyl nanodiamonds inhibit melanoma tumor metastases by blocking cellular motility and invasiveness.

Carboxyl nanodiamond (cND) nanoparticles are actively internalized by B16F10 melanoma cells in culture. Treatment of B16F10 tumor cells with cNDs in vitro inhibited their ability to (i) migrate and invade through porous membranes in a transwell culture system, (ii) secrete matrix metalloproteinases (MMPs) MMP-2 and MMP-9, and (iii) express selected epithelial-mesenchymal transition markers critical for cell migration and invasion. Administration of luciferase-transfected B16F10-Luc2 melanoma cells resulted in a rapid growth of the tumor and its metastasis to different organs that could be monitored by in vivo bioluminescence imaging as well as by ex vivo BLI of the mouse organs. After tumor cells were administered intravenously in C57Bl/6 mice, administration of cNDs (50 µg i.v. every alternate day) resulted in marked suppression of the tumor growth and metastasis in different organs of mice. Subcutaneous administration of B16F10 cells resulted in robust growth of the primary tumor subcutaneously as well as its metastasis to the lungs, liver, spleen, and kidneys. Intravenous treatment with cNDs did not affect the growth of the primary tumor mass but essentially blocked the metastasis of the tumor to different organs. Histological examination of mouse organs indicated that the administration of cNDs by itself was safe and did not cause toxic changes in mouse organs. These results indicate that the cND treatment may have an antimetastatic effect on the spread of B16F10 melanoma tumor cells in mice. Further exploration of cNDs as a possible antimetastatic therapeutic agent is suggested.

A Double in vivo Biotinylation Technique for Objective Assessment of Aging and Clearance of Mouse Erythrocytes in Blood Circulation.

SUMMARY: We have recently developed a new technique to objectively identify erythrocyte cohorts of defined age in mouse blood. The technique (termed double in vivo biotinylation, DIB) involves an initial biotinylation of all erythrocytes in circulation, followed after a few days by a second biotinylation, at a lower density, that labels the biotin-negative erythrocytes that have entered since the first biotinylation. The proportions of biotin(high), biotin(low), and biotin(negative) erythrocytes are enumerated by flow cytometry. The DIB technique allows us to track age-related changes on erythrocyte cohorts (Protocol A), and to simultaneously identify very young and older erythrocyte populations in the blood (Protocol B). Using this technique, we have reexamined: i) the relationship between age and buoyant density of erythrocytes, ii) erythrocyte destruction through a random removal mechanism, and iii) the expression of phosphatidylserine on aging erythrocytes. We have also used the DIB technique to study age-related changes in the expression of various markers like CD47 and CD147 and green autofluorescence in aging erythrocyte populations.

A framework for in-vivo human brain tumor detection using image augmentation and hybrid features.

Brain tumor is caused by the uncontrolled and accelerated multiplication of cells in the brain. If not treated early enough, it can lead to death. Despite multiple significant efforts and promising research outcomes, accurate segmentation and classification of tumors remain a challenge. The changes in tumor location, shape, and size make brain tumor identification extremely difficult. An Extreme Gradient Boosting (XGBoost) algorithm using is proposed in this work to classify four subtypes of brain tumor-normal, gliomas, meningiomas, and pituitary tumors. Because the dataset was limited in size, image augmentation using a conditional Generative Adversarial Network (cGAN) was used to expand the training data. Deep features, Two-Dimensional Fractional Fourier Transform (2D-FrFT) features, and geometric features are fused together to extract both global and local information from the Magnetic Resonance Imaging (MRI) scans. The model attained enhanced performance with a classification accuracy of 98.79% and sensitivity of 98.77% for the test images. In comparison to state-of-the-art algorithms employing the Kaggle brain tumor dataset, the suggested model showed a considerable improvement. The improved results show the prominence of feature fusion and establish XGBoost as an appropriate classifier for brain tumor detection in terms on testing accuracy, sensitivity and Area under receiver operating characteristic (AUROC) curve.

Relative efficacy of uptake and presentation of Mycobacterium bovis BCG antigens by type I mouse lung epithelial cells and peritoneal macrophages.

Flow cytometric studies indicated that both peritoneal macrophages (PMs) and primary lung epithelial (PLE) cells isolated from mouse lungs could take up fluorescence-tagged Mycobacterium bovis BCG. BCG uptake in both cases was significantly inhibited by cytochalasin D, indicating active internalization of BCG by these cells. Confocal microscopy data further confirmed that BCG was internalized by PLE cells. BCG sonicate antigen (sBCG) had marked toxicity toward PMs but was relatively nontoxic to PLE cells. Accordingly, BCG sonicate antigen induced a significantly higher apoptotic and necrotic response in PMs compared to that in PLE cells. Both PMs and PLE cells exposed to BCG antigens and fixed thereafter could efficiently present antigens to purified BCG-sensitized T helper cells, as assessed by the release of interleukin-2 (IL-2) and gamma interferon (IFN-γ). If, however, PLE cells were fixed before exposure to BCG, antigen presentation was abrogated, indicating that the PLE cells may in some way process the BCG antigen. A comparison of efficacies of BCG-pulsed PLE cells and PMs to present antigen at various antigen-presenting cell (APC)/T cell ratios indicated that PMs had only marginally greater APC function than that of PLE cells. Staining with specific monoclonal antibodies indicated that the cultured PLE cells used for antigen presentation essentially comprised type I epithelial cells. Our results suggest that type I lung epithelial cells may present BCG antigens to sensitized T helper cells and that their performance as APCs is comparable with that of PMs.

Quantitative assessment of elemental carbon in the lungs of never smokers, cigarette smokers, and coal miners.

Inhalation exposure to particulates such as cigarette smoke and coal dust is known to contribute to the development of chronic lung disease. The purpose of this study was to estimate the amount of elemental carbon (EC) deposits from autopsied lung samples from cigarette smokers, miners, and control subjects and explore the relationship between EC level, exposure history, and the extent of chronic lung disease. The samples comprised three subgroups representing never smokers (8), chronic cigarette smokers (26), and coal miners (6). Following the dissolution of lung tissue, the extracted EC residue was quantified using a thermal-optical transmission (TOT) carbon analyzer. Mean EC levels in the lungs of the control group were 56.68 ± 24.86 (SD) µg/g dry lung weight. Respective mean EC values in lung samples from the smokers and coal miners were 449.56 ± 320.3 µg/g and 6678.2 ± 6162 µg/g. These values were significantly higher than those obtained from the never-smoker group. EC levels in the lung and pack-years of cigarette smoking correlated significantly, as did EC levels and the severity of small airway disease. This study provides one of the first quantitative assessments of EC in human lungs from populations at high relative risk for the development of chronic lung disease.

Poly-dispersed Acid-Functionalized Single-Walled Carbon Nanotubes (AF-SWCNTs) Are Potent Inhibitor of BCG Induced Inflammatory Response in Macrophages.

The present study is focused on the modulation of Mycobacterium bovis BCG-induced inflammatory response by poly-dispersed acid-functionalized single-walled carbon nanotubes (AF-SWCNTs) in macrophages. Flow cytometric and confocal microscopy studies indicated that both BCG and AF-SWCNTs were efficiently internalized by RAW 264.7 and MH-S macrophage cell lines and were essentially localized in the cytoplasmic area. BCG-induced production of reactive oxygen species (ROS) and nitric oxide by the two cell lines was significantly inhibited by AF-SWCNTs. Using RT-PCR technique, a marked decline was observed in the expression of BCG-induced pro-inflammatory genes COX-2, iNOS, TNF-α, IL-6, and IL-1ß upon treatment with AF-SWCNTs. Results of gelatin zymography indicated that the AF-SWCNTs treatment also induced a marked decline in BCG-induced release of matrix metalloproteinases MMP-2 and MMP-9 by the two macrophage cell lines. The anti-inflammatory effect of AF-SWCNTs in downregulating BCG-induced inflammatory response was further validated in murine peritoneal macrophages. Treatment with AF-SWCNTs led to a steep decline in BCG-induced NO production in murine peritoneal macrophages in vitro as well as in vivo. Peritoneal macrophages isolated from mice treated with BCG and AF-SWCNTs had a significantly lower intracellular expression of COX-2 as compared to the peritoneal macrophages derived from mice treated with BCG alone. Taken together, our results demonstrate a potent anti-inflammatory effect of AF-SWCNTs in alleviating BCG-induced inflammatory responses in macrophages in vitro and in vivo.

Nanodiamonds inhibit scratch-wound repair in lung epithelial cell monolayers by blocking cell migration and inhibiting cell proliferation.

Proliferation and migration of lung epithelial cells following the injury to the epithelial lining of alveoli and airways in the lung are pivotal for remodeling and repair of the wound to restore normal lung function. In the present study, we examined the modulatory effect of carboxylated nanodiamonds (cNDs) on the cell division, migration, and adhesion of epithelial cells in the well-established in vitro model of wound repair and cell migration. Flow cytometry and confocal microscopy results indicated that both LA4 and A549 cells effectively internalized fluorescent carboxylated nanodiamonds (cFNDs) and the internalized nanodiamonds were essentially localized in the cytoplasmic region. Treatment with cNDs blocked the division and migration of cells to fill the scratch wound. Live cell imaging and time-lapse videography of the wound healing process indicated a significant inhibition of cell proliferation activity in cND-treated cells and blocked the wound repair process. Trans-well cell-migration assay results further support the inhibitory effect of cNDs on the cell migration process. Western blotting and immunofluorescence staining indicated that the crucial proteins involved in epithelial-mesenchymal transition (EMT) and cell migration i.e. ß-catenin, Vimentin, NM-myosin, and Focal Adhesion Kinase (FAK) were downregulated after treatment with cNDs, while the expression of E-cadherin and Claudin-1, major cell adhesion markers remained unaltered. Taken together, our results indicate that the decline in cell proliferation activity, downregulation in the expression of various crucial protein like ß-Catenin, NM-myosin, FAK, and Vimentin involved in the cell migration and unaltered expression of cell adhesion molecules E-cadherin and Claudin-1, may be the factors that contribute to the cND-mediated inhibition of EMT during the wound repair process in the monolayers of lung epithelial cells.

Toxicity of poly-dispersed single-walled carbon nanotubes on bone marrow derived Hematopoietic Stem and Progenitor Cells.

This study has explored the effect of acid-functionalized single-walled carbon nanotubes (AF-SWCNTs) on Hematopoietic Stem and Progenitor Cell (HSPCs) in mouse bone marrow. Administration of AF-SWCNTs induced a significant decline in the live-cell recovery from bone marrow. Lin-negative Stem cell enriched HSPCs internalized AF-SWCNTs that remained localized in cytoplasmic areas. Incubation of HSPCs with AF-SWCNTs resulted in induction of cell death, inhibition of cell cycle, and induction of reactive oxygen species (ROS) as well as the expression of Caspase 3, 7 and 9 enzymes. In vitro culture with a cytokine cocktail (SCF, GM-CSF, IL3, IL6, IL7) induced differentiation of HSPCs into lymphocytes and myeloid cells, that was inhibited in presence of AF-SWCNTs. Relative recoveries of lymphocytes specifically B lymphocytes, was significantly reduced by AF-SWCNT-treatment, whereas the relative recovery of myeloid cells remained unaltered. These results suggest that AF-SWCNTs have significant toxic effects on HSPCs and differentially suppress the ontogeny of lymphoid and myeloid cells.

Enhanced antibody response to ovalbumin coupled to poly-dispersed acid functionalized single walled carbon nanotubes.

Activated mouse B cells as compared to the resting B cells are known to internalize substantially more acid-functionalized single walled carbon nanotubes (AF-SWCNTs). It was hypothesized that the antigens coupled to AF-SWCNTs would also be taken up more efficiently by B cells. Further the enhanced uptake of the antigen by B cells may facilitate antigen presentation by B cells resulting in a better antibody response. Aim of this study was to test this hypothesis. Ovalbumin was chemically coupled to AF-SWCNTs that yielded a coupled product that had 0.08% of all carbon atoms in AF-SWCNTs occupied by ovalbumin. Coupling of ovalbumin to AF-SWCNTs was confirmed by staining the product with anti-ovalbumin antibodies. B cells incubated with ovalbumin-AF-SWCNT internalized more ovalbumin than the B cells incubated with free ovalbumin. Groups of mice were immunized subcutaneously with (a) free ovalbumin, (b) free ovalbumin and AF-SWCNTs at two different subcutaneous sites respectively on mice, and (c) ovalbumin-AF-SWCNT coupled product. In each case a primary immunization was followed by three weekly booster doses. It was found that the anti-ovalbumin antibody response assessed by ELISA, was highest in the group where ovalbumin coupled to AF-SWCNTs was used for immunization (p < 0.001). Antibody response in ovalbumin-AF-SWCNT group was comparable to the group where ovalbumin was used for immunization using complete and incomplete Freund's adjuvant (primary and secondary immunizations respectively). We propose that AF-SWCNTs could be explored as an adjuvant to improve the antibody response especially in vaccine development.

Identification of Short Exons Disunited by a Short Intron in Eukaryotic DNA Regions.

Weak codon bias in short exons and separation by a short intron induces difficulty in extracting period-3 component that marks the presence of exonic regions. The annotation task of such short exons has been addressed in the proposed model independent signal processing based method with following features: (a) DNA sequences have been mapped using multiple mapping schemes, (b) period-3 spectrums corresponding to multiple mappings have been optimized to enhance short exon-short intron discrimination, and (c) spectrums corresponding to multiple mapping schemes have been subjected to Principal Component Analysis (PCA) for identifying greater number of such short exons. A comparative study with other methods indicates improved detection of contiguous short exons disunited by a short intron. Apart from the annotation of exonic and intronic regions, the proposed algorithm can also complement the methods for the detection of alternative splicing by intron retention, as one of the characteristic feature for intron retention is the presence of two short exons flanking a short intron.

Poly dispersed acid-functionalized single walled carbon nanotubes target activated T and B cells to suppress acute and chronic GVHD in mouse model.

Graft versus host disease (GVHD) results from hyper-activation of transplanted lymphocytes against the host antigens. Bone marrow transplantation in humans as well as some cases of blood transfusion and organ transplantation are associated with a strong GVH reaction resulting in GVHD that in many cases may be fatal. We had previously shown that poly-dispersed acid-functionalized single-walled carbon nanotubes (AF-SWCNTs) specifically target activated T and B lymphocytes and kill them. In the present study, efficacy of AF-SWCNTs to suppress the GVH reaction was tested in the mouse model. Acute GVHD was induced in mice by administering intravenously 30 or 60 million spleen cells from a parental strain (C57bl/6 mouse, MHC haplotype H-2b) to host (C57bl/6 x Balb/c) F1 mice (MHC haplotype H-2b/d)and waiting for 8-10 days. Chronic GVHD was similarly induced by administration of 30 million parent spleen cells to F1 mice and waiting for a period of 60 days. Our results demonstrate a marked decline in splenomegaly and recovery of spleen T (both CD4 and CD8) and B cells in GVHD mice treated with AF-SWCNTs. AF-SWCNTs treatment also limited T and B cell proliferation by restricting S-phage of cell cycle. Generation of anti-host cytotoxic T cells (CTLs) was also markedly suppressed by AF-SWCNT treatment of acute GVHD mice, and a significant reduction in the generation of anti-host antibodies could also be demonstrated. Taken together, our results suggest that the AF-SWCNTs can be considered as a potential therapeutic agent for treating GVHD.

Acid-functionalized single-walled carbon nanotubes alter epithelial tight junctions and enhance paracellular permeability.

Due to their unique properties, carbon nanotubes (CNTs) are being widely explored for industrial and medical applications. This has necessitated a thorough assessment of the effect of CNTs on human and animal physiology and health. Impact of CNTs on epithelial tight junctions has not been evaluated in the context of their toxic effects in many biological systems. In the present study, we examined the effect of acid functionalized single-walled carbon nanotubes (AF-SWCNTs) on the function and expression of two tight junction proteins (ZO-1 and occludin) in the Madin-Darby canine kidney (MDCK) cell line. Treatment of MDCK cells with AF-SWCNT resulted in a downregulation of tight junction proteins, decreased trans-epithelial electrical resistance (TER), increased paracellular permeability, and disruption of tight junctions. Taken together, our data demonstrate that AF-SWCNT disrupts tight junction barrier by downregulating tight junction proteins in MDCK epithelial cells.

Influence of acid functionalization on the cardiopulmonary toxicity of carbon nanotubes and carbon black particles in mice.

Engineered carbon nanotubes are being developed for a wide range of industrial and medical applications. Because of their unique properties, nanotubes can impose potentially toxic effects, particularly if they have been modified to express functionally reactive chemical groups on their surface. The present study was designed to evaluate whether acid functionalization (AF) enhanced the cardiopulmonary toxicity of single-walled carbon nanotubes (SWCNT) as well as control carbon black particles. Mice were exposed by oropharyngeal aspiration to 10 or 40 microg of saline-suspended single-walled carbon nanotubes (SWCNTs), acid-functionalized SWCNTs (AF-SWCNTs), ultrafine carbon black (UFCB), AF-UFCB, or 2 microg LPS. 24 hours later, pulmonary inflammatory responses and cardiac effects were assessed by bronchoalveolar lavage and isolated cardiac perfusion respectively, and compared to saline or LPS-instilled animals. Additional mice were assessed for histological changes in lung and heart. Instillation of 40 microg of AF-SWCNTs, UFCB and AF-UFCB increased percentage of pulmonary neutrophils. No significant effects were observed at the lower particle concentration. Sporadic clumps of particles from each treatment group were observed in the small airways and interstitial areas of the lungs according to particle dose. Patches of cellular infiltration and edema in both the small airways and in the interstitium were also observed in the high dose group. Isolated perfused hearts from mice exposed to 40 microg of AF-SWCNTs had significantly lower cardiac functional recovery, greater infarct size, and higher coronary flow rate than other particle-exposed animals and controls, and also exhibited signs of focal cardiac myofiber degeneration. No particles were detected in heart tissue under light microscopy. This study indicates that while acid functionalization increases the pulmonary toxicity of both UFCB and SWCNTs, this treatment caused cardiac effects only with the AF-carbon nanotubes. Further experiments are needed to understand the physico-chemical processes involved in this phenomenon.

Elevated internalization and cytotoxicity of polydispersed single-walled carbon nanotubes in activated B cells can be basis for preferential depletion of activated B cells in vivo.

Uptake of polydispersed acid-functionalized single-walled carbon nanotubes (AF-SWCNTs) in resting and LPS-activated B cells was studied using fluorescence-tagged AF-SWCNTs (FAF-SWCNTs). Activated B cells internalized substantially higher amounts of FAF-SWCNTs [76.5% AF-SWCNT+ B cells, mean fluorescence intensity (MFI) 720.6] as compared to the resting B cells [39.5% AF-SWCNT+ B cells, MFI 198.5]. B cells in S and G2/M phases were found to have significantly higher uptake of FAF-SWCNTs as compared to cells in G0/G1 phase. Confocal microscopy indicated that AF-SWCNTs were essentially localized on cell membrane in resting B cells, whereas in activated B cells, AF-SWCNTs were distributed throughout the cytoplasm. Targeting of AF-SWCNTs specifically to activated B cells in vivo was examined by first administering intravenously LPS-activated B cells tagged with fluorescence tracer (CFSE) in mice, followed by FAF-SWCNTs through the same route. It was found that FAF-SWCNTs were specifically taken up by CFSE+CD19+-activated B cells (95% FAF-SWCNT+ B cells, MFI 3725) as compared to CFSE- CD19+ resting B cells (31.1% FAF-SWCNT+ B cells, MFI 428). Administration (i.v.) of LPS resulted in a significant increase in the proportion of B cell in mouse spleen that was reduced by 68% by administering AF-SWCNTs. In control mice, the corresponding decrease in B cell proportion was 49%, which was significantly lower (p < 0.005) than the decline in LPS-treated mice. These results indicate that AF-SWCNTs may have the potential as an agent for depleting activated B cells in vivo.

Isolation and quantitative estimation of diesel exhaust and carbon black particles ingested by lung epithelial cells and alveolar macrophages in vitro.

A new procedure for isolating and estimating ingested carbonaceous diesel exhaust particles (DEP) or carbon black (CB) particles by lung epithelial cells and macrophages is described. Cells were incubated with DEP or CB to examine cell-particle interaction and ingestion. After various incubation periods, the cells were separated from free extracellular DEP or CB particles by Ficoll density gradient centrifugation and dissolved in hot sodium dodecyl sulfate detergent. Insoluble DEP or CB residues were isolated by high-speed centrifugation, and the elemental carbon (EC) concentrations in the pellets were estimated by a thermal-optical-transmittance method (i.e., carbon analysis). From the EC concentration, the amount of ingested DEP or CB could be calculated. The described technique allowed the determination of the kinetics and dose dependence of DEP uptake by LA4 lung epithelial cells and MHS alveolar macrophages. Both cell types ingested DEP to a similar degree; however, the MHS macrophages took up significantly more CB than the epithelial cells. Cytochalasin D, an agent that blocks actin polymerization in the cells, inhibited approximately 80% of DEP uptake by both cell types, indicating that the process was actin-dependent in a manner similar to phagocytosis. This technique can be applied to examine the interactions between cells and particles containing EC and to study the modulation of particle uptake in diseased tissue.

A role of phosphatidylserine externalization in clearance of erythrocytes exposed to stress but not in eliminating aging populations of erythrocyte in mice.

Age dependent changes in phosphatidylserine (PS) externalization were studied in mouse erythrocytes of different age groups (range 1-55 days) by using a newly developed double in vivo biotinylation (DIB) technique. Around 3-4% of the erythrocytes freshly released in the circulation were PS(+) but this proportion fell rapidly to 1% or less and did not increase at later time points. Blocking erythrocyte clearance from the circulation by in vivo depletion of macrophages (by treatment with clodronate loaded liposomes) for up to 7 days did not result in accumulation of PS(+) erythrocytes in the circulation indicating that the low percentage of PS(+) cells within old erythrocytes (age >40 days) was not related to the clearance of PS(+) erythrocytes by macrophages. In vitro treatment with stress inducing agents like deoxyglucose or Ca(++)/calcium ionophore resulted in a marked induction of PS externalization in mouse erythrocytes and this effect was most prominent in the youngest erythrocyte population (age <10 days). Kinetics of clearance of different age groups of stress exposed erythrocytes after intravenous infusion into recipient mice indicated that the young erythrocytes were cleared at fastest rate from the circulation as compared to erythrocytes of older age groups. Within young erythrocytes exposed to stress, PS(+) erythrocytes were preferentially cleared. Taken together our results suggest that PS externalization is unlikely to have a role in the removal of old erythrocytes from blood circulation but may have a role in the clearance of stressed and damaged young erythrocytes in blood circulation.

Evidence of CD1d pathway of lipid antigen presentation in mouse primary lung epithelial cells and its up-regulation upon Mycobacterium bovis BCG infection.

Presentation of a prototype lipid antigen α-Galactosylceramide (αGC) was examined on primary epithelial cells derived from mouse lungs and on bronchoalveolar lavage (BAL) cells that essentially comprise alveolar macrophages. Presence of CD1d molecules coupled to αGC was demonstrated on both types of cells pre-treated with αGC, suggesting that both cell types are equipped to present lipid antigens. Internalization of Mycobacterium bovis Bacillus Calmette-Guérin (BCG: a prototype pathogen), a pre-requisite to the processing and presentation of protein as well as lipid antigens, was clearly demonstrated in primary lung epithelial (PLE) cells as well as BAL cells. Both PLE and BAL cells expressed CD1d molecule and a significant up-regulation of its expression occurred upon infection of these cells with BCG. Besides CD1d, the expression of other important molecules that participate in lipid antigen presentation pathway (i.e. microsomal triglyceride transfer protein (MTTP), scavenger receptor B1 (SR-B1) and Saposin) was also significantly upregulated in PLE and BAL cells upon BCG infection. In situ up-regulation of CD1d expression on lung epithelial cells was also demonstrated in the lungs of mice exposed intra-tracheally to BCG. Taken together these results suggest that lung epithelial cells may have the ability to present lipid antigens and this pathway seems to get significantly upregulated in response to BCG infection.

Age-dependent increase in green autofluorescence of blood erythrocytes.

Green auto-fluorescence (GAF) of different age groups of mouse blood erythrocytes was determined by using a double in vivo biotinylation (DIB) technique that enables delineation of circulating erythrocytes of different age groups. A significant increase in GAF was seen for erythrocytes of old age group (age in circulation more than 40 days) as compared to young erythrocytes (age less than 15 days). Erythrocytes are removed from blood circulation by macrophages in the reticulo-endothelial system and depletion of macrophages results in an increased proportion of aged erythrocytes in the blood. When mice were depleted of macrophages for 7 days by administration of clodronate loaded liposomes, the overall GAF of erythrocytes increased significantly and this increase could be ascribed to an increase in GAF of the oldest population of erythrocytes. Using the DIB technique, the GAF of a cohort of blood erythrocyte generated during a 5 day window was tracked in vivo. GAF of the defined cohort of erythrocytes remained low till 40 days of age in circulation and then increased steeply till the end of the life span of erythrocytes. Taken together our results provide evidence for an age dependent increase in the GAF of blood erythrocytes that is accentuated by depletion of macrophages. Kinetics of changes in GAF of circulating erythrocytes with age has also been defined.