Thermal performance enhancement of lauric acid using nanomaterials as composite phase change material.

In the present work, lauric acid was taken as a phase change material (PCM), and different nanoparticles (NPs) such as SiO2, TiO2, CuO, and ZnO were taken as the supporting materials. CuO NPs were prepared through the co-precipitation technique; SiO2, TiO2, and ZnO NPs were synthesized via the sol-gel technique. These NPs with different weight fractions were dispersed into molten lauric acid, individually. The variations in thermal properties (phase change temperature and latent heat for solid and liquid) of the prepared composite PCMs due to the dispersion of NPs were observed by DSC analyses. An increase in thermal conductivity of the composite PCMs was observed with the increasing weight fraction of NPs. In order to ascertain the long-term utility, a thermal reliability test was conducted on the composite PCMs with repeated heating and cooling cycles. Also, the specific heats of the pure PCM and the composite PCMs were determined as a function of temperature. Further, the experimental investigation was performed on the pure PCM and the prepared composite PCMs to assess their phase change behavior, and the test results clearly proved that the time required for the complete melting and freezing process of the composite PCMs was less when compared to pure PCM. By considering the above facts, the newly prepared composite PCMs can be recommended as a potential candidate for low-temperature solar heating applications.

Gunshot Injury to the Face With Atypical Non-linear Bullet Trajectory: A Case Report.

The disruptive effect of firearm missiles on body tissues depends on many factors. However, it mainly relates to the bullet's physical and dynamic properties and tissue-related factors. We encountered an unusual case of a gunshot injury wherein the bullet traversed the neck with an upward and non-linear trajectory with an exit from the contralateral side of the neck without damaging any vital neck organs. A 26-year-old male presented with a gunshot wound to the chin from close range. A bullet entry hole was observed on the right side of the chin, encircled by the abrasion collar, with tattoo marks around the area. The patient was conscious, with normal vital signs and no injury to the cranial nerves or aerodigestive tract. The CT imaging of the patient revealed the injury tract traversing through the muscles of the floor of the mouth to involve the left carotid and left parapharyngeal space, along with the left sternocleidomastoid muscle, with an exit hole below the left mastoid in the posterior triangle of the neck. A bullet usually travels through the body in a straight line or pathway; however, its non-linear trajectories depend on the projectile's dynamics and its interaction with the body tissues. The present case emphasizes understanding wound ballistics to know the erratic bullet trajectories in the victim's body and their interpretation, irrespective of their entry site.

Identification of proteinase 3 autoreactive CD4+T cells and their T-cell receptor repertoires in antineutrophil cytoplasmic antibody-associated vasculitis.

Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is an autoimmune disease involving autoreactivity to proteinase 3 (PR3) as demonstrated by presence of ANCAs. While autoantibodies are screened for diagnosis, autoreactive T cells and their features are less well-studied. Here, we investigated PR3-specific CD4+T cell responses and features of autoreactive T cells in patients with PR3-AAV, using a cohort of 72 patients with either active or inactive disease. Autoreactive PR3-specific CD4+T cells producing interferon γ in response to protein stimulation were found to express the G-protein coupled receptor 56 (GPR56), a cell surface marker that distinguishes T cells with cytotoxic capacity. GPR56+CD4+T cells were significantly more prominent in the blood of patients with inactive as compared to active disease, suggesting that these cells were affected by immunosuppression and/or that they migrated from the circulation to sites of organ involvement. Indeed, GPR56+CD4+T cells were identified in T-cell infiltrates of affected kidneys and an association with immunosuppressive therapy was found. Moreover, distinct TCR gene segment usage and shared (public) T cell clones were found for the PR3-reactive TCRs. Shared T cell clones were found in different patients with AAV carrying the disease-associated HLA-DP allele, demonstrating convergence of the autoreactive T cell repertoire. Thus, we identified a CD4+T cell signature in blood and in affected kidneys that display PR3 autoreactivity and associates with T cell cytotoxicity. Our data provide a basis for novel rationales for both immune monitoring and future therapeutic intervention in PR3-AAV.

A worldwide perspective on large carnivore attacks on humans.

Large carnivores have long fascinated human societies and have profound influences on ecosystems. However, their conservation represents one of the greatest challenges of our time, particularly where attacks on humans occur. Where human recreational and/or livelihood activities overlap with large carnivore ranges, conflicts can become particularly serious. Two different scenarios are responsible for such overlap: In some regions of the world, increasing human populations lead to extended encroachment into large carnivore ranges, which are subject to increasing contraction, fragmentation, and degradation. In other regions, human and large carnivore populations are expanding, thus exacerbating conflicts, especially in those areas where these species were extirpated and are now returning. We thus face the problem of learning how to live with species that can pose serious threats to humans. We collected a total of 5,440 large carnivore (Felidae, Canidae, and Ursidae; 12 species) attacks worldwide between 1950 and 2019. The number of reported attacks increased over time, especially in lower-income countries. Most attacks (68%) resulted in human injuries, whereas 32% were fatal. Although attack scenarios varied greatly within and among species, as well as in different areas of the world, factors triggering large carnivore attacks on humans largely depend on the socioeconomic context, with people being at risk mainly during recreational activities in high-income countries and during livelihood activities in low-income countries. The specific combination of local socioeconomic and ecological factors is thus a risky mix triggering large carnivore attacks on humans, whose circumstances and frequencies cannot only be ascribed to the animal species. This also implies that effective measures to reduce large carnivore attacks must also consider the diverse local ecological and social contexts.

Correlation of angiogenic growth factors and inflammatory cytokines with the clinical phenotype of ocular tuberculosis.

PURPOSE: To determine the correlation of angiogenic growth factors and inflammatory cytokines with the clinical phenotype of ocular tuberculosis (OTB). METHODS: Vitreous fluid was analysed for cytokines in patients with OTB and non-OTB uveitis using multiplex fluorescent bead-based flow cytometric assay. The clinical phenotypes were recorded and correlated with vitreous biomarkers. RESULTS: Vitreous humour from OTB patients had elevated levels of interleukin-10 (IL-10), IL-17-A, interferon-gamma (IFN-γ), and tumour necrosis factor-alpha (TNF-α). Angiopoietin (Ang-2) levels were higher in the panuveitis phenotype. OTB posterior uveitis phenotype had relatively higher vascular endothelial growth factor (VEGF) levels and lower fibroblast growth factor (FGF) levels. Additionally, eyes with choroiditis and vasculitis had elevated levels of VEGF and Ang-2 with FGF downregulation. Both IFN-γ and IL-10 were upregulated in the choroiditis phenotype of OTB. CONCLUSION: Angiogenic growth factors and inflammatory cytokines were altered in the vitreous humour of OTB patients. IFN-γ, VEGF, and IL-10 levels are increased in choroiditis and vasculitis phenotypes. Receiver operating characteristic (ROC) curve analysis further emphasized the importance of the IFN-γ assay in the diagnosis of OTB.

Structural characterization and bioimaging of Zn2+ using meta-benziporphodimethene analogue.

'Prevention is better than cure, especially when something has no cure.' Cancer, in most patients is detected at the stage beyond which it becomes noncurative. Therefore, the early detection of cancer cells can play a crucial role in enhancing the chances of a patient's survival. In this light, we present a nonfluorescent receptor used for the detection of Zn2+ ion in MDA-MB-231 carcinoma cells that exhibits fluorescence turn-on behaviour upon binding with the metal ion. In this work, the synthesis of 11,16-bis(2,6-difluorobenzene)-6,6,21,21-tetramethyl-meta-benziporpho-6,21-dimethene and its Zn2+ chloride complex have been reported. The compounds were fully characterized using UV-visible, nuclear magnetic resonance (NMR), infrared (IR) and mass spectrometry. Furthermore, the X-ray polymorphs of a meta-benziporphodimethene analogue were added. The study of its bioimaging applications in MDA-MB-231 breast cancer cells for the detection of Zn2+ ions is reported.

Prevalence of Psychiatric Disorders in Chhattisgarh: Findings from National Mental Health Survey.

Background: Psychiatric disorders are among the leading contributors to disability in India and worldwide. The pattern, prevalence, and distribution of psychiatric disorders in the country and its regions need to be assessed to facilitate early diagnosis and treatment. No study on the epidemiology of psychiatric disorders has been conducted in the Chhattisgarh state. This paper, as part of the National Mental Health Survey (NMHS), discusses the prevalence and pattern of psychiatric disorders in Chhattisgarh state. Methods: A stratified random cluster sampling technique and random selection based on probability proportional to size (PPS) at each stage were adopted. Participants were from three selected districts of Chhattisgarh, such as Janjgir-Champa, Kabirdham, and Raipur. Adults (aged ≥18 years) residing in selected households were interviewed using Mini International Neuropsychiatric Interview (version 6.0), the Fagerstrom test for nicotine dependence, the WHO-SEARO screening questionnaire for generalized tonic-clonic seizures, and screening tools for intellectual disability and autism spectrum disorders. Results: A total of 2841 individuals were interviewed. The state's lifetime and current prevalence of psychiatric disorders for adults were 14.06% [95% confidence interval (CI) = 13.83-14.29] and 11.66% (95% CI = 11.45-11.87), respectively. Prevalence of substance use disorders, tobacco use disorders, schizophrenia and related disorders, and mood disorders was 32.4% (95% CI = 32.09-32.71), 29.86% (95% CI = 29.56-30.16), 0.8% (95% CI = 0.75-0.86), and 4.44% (95% CI = 4.31-4.58), respectively. High risk for suicide was detected in 0.28% (95% CI = 0.25-0.31). Psychiatric disorders were twice more common in males than in females. Conclusions: The study gives authentic data on the prevalence of psychiatric disorders in Chhattisgarh. This shall pave the way for policymakers and planners to design state-specific plans for dealing with mental disorders and related issues.

Lower cyst fluid carcinoembryonic antigen cutoff is helpful in the differential diagnosis of mucinous versus non-mucinous pancreatic cysts.

BACKGROUND AND AIM: Pancreatic cystic lesions (PCLs) are being diagnosed with increased frequency and have varying neoplastic potential. We conducted this multimodal, prospective study to evaluate  the role of tumor cytology and molecular markers to differentiate PCL subtypes. METHODS: Consecutive undiagnosed patients with PCLs (n = 100, mean age: 50.37 years; 41% males) were prospectively studied. Cyst fluid carcinoembryonic antigen (CEA), CA19.9, CA125, CA72.4, and vascular endothelial growth factor-alpha (VEGF-α) levels were measured by quantitative enzyme-linked immunosorbent assay (ELISA) method. Mutational analysis of the KRAS gene (exon 2, Codon 12 and 13) and GNAS gene (Exon 8, Codon 201) were performed by Sanger's sequencing. RESULTS: The mean cyst size was 4.32 ± 2.4 cm. Fluid cytology revealed definitive diagnosis in 21 (22.3%) patients. All malignant PCLs could be identified on cytology whereas 10/14 (71%) non-malignant mucinous PCLs could also be identified on cytology based on mucin staining. Among the tested tumor markers, cyst fluid CEA had the best diagnostic performance for differentiation between mucinous and non-mucinous PCLs (AUC 0.933 [95% CI 0.86-0.91]). At a cyst fluid CEA cutoff level of 45.0 ng/mL, the sensitivity, specificity, positive predictive value, and negative predictive value for differentiation between mucinous and non-mucinous cysts were 88.5%, 96.8%, 92.0%, and 95.3%, respectively (p < 0.05). KRAS and GNAS mutation had no significant diagnostic benefit in comparison to fluid cytology and CEA levels. CONCLUSIONS: Fluid CEA at a lower cutoff of 45 ng/mL is the most accurate marker to differentiate between mucinous and non-mucinous PCL. The KRAS and GNAS mutational analysis does not improve upon the diagnostic performance of fluid cytology and tumor markers.

Environmental assessment, microstructural behaviour, stress-strain characteristics, and effect of exposure to extreme temperature on sustainable concrete made with dolomite mining residues.

Mining and extraction of stones and minerals play a significant role in many countries economic growth in the world. The production of dolomite minerals in various industries in India and other countries produces vast amounts of waste in different fractions. Disposal of these types of industrial wastes in an immense quantity causes environmental pollution. The performance of dolomite mining residues on concrete properties as a fine aggregate substitute was examined. The microstructural analysis was conducted on the concrete samples to find the effect of dolomite mining residues in concrete. The stress-strain behaviour of the dolomite mining residues concrete was studied. The effect of exposure to elevated temperature and freeze-thaw on concrete properties containing dolomite mining residues was found up to 100% at 10% incremental order. The thermogravimetric analysis (TGA) and differential thermogravimetry (DTG) tests were conducted on the dolomite mining residues and concrete samples. As a test result, concrete properties influence with the incorporation of the dolomite mining residues as a substitution of river sand, but no significant effect is observed in the concrete properties containing 10% dolomite mining residues. Up to 10% of dolomite production waste can be used as a sand substitute in concrete and other applications for sustainable development.

Electroosmotic modulated unsteady squeezing flow with temperature-dependent thermal conductivity, electric and magnetic field effects.

Modern lubrication systems are increasingly deploying smart (functional) materials. These respond to various external stimuli including electrical and magnetic fields, acoustics, light etc. Motivated by such developments, in the present article unsteady electro-magnetohydrodynamics squeezing flow and heat transfer in a smart ionic viscous fluid intercalated between parallel plates with zeta potential effects is examined. The proposed mathematical model of problem is formulated as a system of partial differential equations (continuity, momenta and energy). Viscous dissipation and variable thermal conductivity effects are included. Axial electrical distribution is also addressed. The governing equations are converted into ordinary differential equations via similarity transformations and then solved numerically with MATLAB software. The transport phenomena are scrutinized for both when the plates move apart or when they approach each other. Also, the impact of different parameters such squeezing number, variable thermal conductivity parameter, Prandtl number, Hartmann number, Eckert number, zeta potential parameter, electric field parameter and electroosmosis parameter on the axial velocity and fluid temperature are analysed. For varied intensities of applied plate motion, the electro-viscous effects derived from electric double-capacity flow field distortions are thoroughly studied. It has been shown that the results from the current model differ significantly from those achieved by using a standard Poisson-Boltzmann equation model. Axial velocity acceleration is induced with negative squeeze number (plates approaching,S< 0) in comparison to that of positive squeeze number (plates separating,S> 0). Velocity enhances with increasing electroosmosis parameter and zeta potential parameter. With rising values of zeta potential and electroosmosis parameter, there is a decrease in temperatures forUe> 0 for both approaching i.e. squeezing plates (S< 0) and separating (S> 0) cases. The simulations provide novel insights into smart squeezing lubrication with thermal effects and also a solid benchmark for further computational fluid dynamics investigations.

TLR9 signalling activation via direct ligation and its functional consequences in CD4 + T cells.

CpG Oligodeoxynucleotides (ODNs) are established TLR9 ligands; however, their functional responses in CD4+ T cells are believed to be independent of TLR9 and MyD88. We studied ligand-receptor interactions of ODN 2216 and TLR9 in human CD4+ T cells and assessed their consequences in terms of TLR9 signalling and cell phenotype. We demonstrated that the uptake of ODN 2216, a synthetic TLR9 agonist, is controlled by TLR9 signalling molecules and results in an increase in the expression of TLR9 signalling molecules, regulated via a feedback mechanism. Next, the uptake of ODN 2216 resulted in TLR9 signalling dependent but MyD88 independent increase in expression of TGF-ß. Finally, ODN 2216 treated CD4+ T cells showed an anti-inflammatory phenotype that was similar to Th3 type of regulatory T cells. These Th3-like cells were able to suppress the proliferation of untreated CD4+ T cells. Collectively, our results demonstrate a direct and interdependent relationship between ODN 2216 uptake and TLR9 signalling in CD4+ T cells. Our findings thus pave the way for future research to explore direct modulation of adaptive immune cells, using innate immune ligands, to subvert exaggerated inflammatory responses.

Analysis of energy storage materials for developments in solar cookers.

Solar energy is accessible freely and can be utilized for many household and industrial applications. The consumption of solar energy for cooking applications has found significant success. Various innovations have been employed in facilitating cooking during off-sunshine hours. Thermal energy storage helps in overcoming the fluctuations in the supply of energy required for cooking during different time periods of the day. This study focuses on the different types of thermal energy storage mediums that are currently utilized in solar cooking. Primarily, oils and pebbles are most commonly used as sensible heat storage (SHS) while organic phase change materials (PCMs) are used as latent heat thermal energy storage materials (LHTES). The properties and performances of various SHS and latent heat storage (LHS) mediums have been compared for their suitable utilization. SHS materials are cost-effective but have lower thermal gradient compared to LHTES materials. The energy storage capability of LHTES is high while degradation with the increasing number of charging and discharging cycles is also considerable. The melting point should be close to the utilization temperature for being used as LHTES as thermal diffusivity of the materials greatly influences the performance of solar cookers. The cooking time is lower for solar cooking systems equipped with energy storage compared to non-equipped cooking systems. It is recognized that the use of energy storage has been proved as a huge advantage to solar cooking systems, however, the design, and heat transfer characteristics of the cooking vessel along with the storage material type and volume must be optimized in order to make this technology more influential.

In vitro and ex vivo functional characterization of human HLA-DRB1∗04 restricted T cell receptors.

Recent advances in single-cell sequencing technologies enable the generation of large-scale data sets of paired TCR sequences from patients with autoimmune disease. Methods to validate and characterize patient-derived TCR data are needed, as well as relevant model systems that can support the development of antigen-specific tolerance inducing drugs. We have generated a pipeline to allow streamlined generation of 'artificial' T cells in a robust and reasonably high throughput manner for in vitro and in vivo studies of antigen-specific and patient-derived immune responses. Hereby chimeric (mouse-human) TCR alpha and beta constructs are re-expressed in three different formats for further studies: (i) transiently in HEK cells for peptide-HLA tetramer validation experiments, (ii) stably in the TCR-negative 58 â€‹T cell line for functional readouts such as IL-2 production and NFAT-signaling, and lastly (iii) in human HLA-transgenic mice for studies of autoimmune disease and therapeutic interventions. As a proof of concept, we have used human HLA-DRB1∗04:01 restricted TCR sequences specific for a type I diabetes-associated GAD peptide, and an influenza-derived HA peptide. We show that the same chimeric TCR constructs can be used in each of the described assays facilitating sequential validation and prioritization steps leading to humanized animal models.

Corneal reconstruction in chemically damaged cornea using temperature responsive surface assisted mesenchymal stem cell transplantation in rabbits.

PURPOSE: Transplantation of autologous stem cells over damaged cornea seems to be a promising approach for corneal reconstruction. Use of a biocompatible carrier is still a challenge in bedside translation of transplantation. We investigated corneal reconstruction and tissue remodelling by transplantation of mesenchymal stem cells (MSCs) using temperature responsive membranes in chemically damaged rabbit cornea model. METHODS: MSCs were cultured from rabbit's bone marrow and transplanted over alkali injured cornea, using either temperature responsive membrane or fibrin glue method. Endogenous levels of MSCs were assessed to decide the optimal time point for transplanting cells. MSC transplanted corneas were harvested at different time points post-transplantation. Corneal repair markers were evaluated using histopathology, immunohistochemistry (IHC) and real time qPCR. The quality of cornea reconstructed was evaluated and compared using corneal opacity scoring and immunohistochemistry (IHC). RESULTS: Use of temperature responsive surface as carrier resulted in uniform and homogenous delivery of MSCs sheet over the damaged corneal surface. Corneal transparency improved day 7 onwards post-MSC transplantation in rabbit chemically injured cornea. Complete re-epithelialization of injured cornea was observed 15 days after MSC transplantation. Restoration of vimentin, α-smooth muscle actin and collagen levels in MSC transplanted cornea was observed post-transplantation. Further, differentiation of MSCs into mature corneal epithelial cells was also observed upon transplantation. CONCLUSIONS: The extent of corneal repair was apparently better using temperature responsive surfaces. The surface provides biocompatible niche for MSCs and can be a method of choice in clinics for cell transplantation over the damaged ocular surfaces.

Mycobacterium Tuberculosis Modulates Fibroblast Growth Factor and Vascular Endothelial Growth Factor in Ocular Tuberculosis.

Purpose: To evaluate the role of angiogenic growth factors in the pathogenesis of intraocular tuberculosis.Methods: Retinal Pigment Epithelium (RPE) cells were infected with varying dilution of Mycobacterium tuberculosis (MTB), ranging from several thousand to a few MTB bacilli to replicate paucibacillary conditions. Angiogenesis growth factors were evaluated using multiplex fluorescent bead based flow cytometry in the culture supernatant of RPE cells infected with MTB, vitreous fluids and tear samples of uveitis patients visiting retina clinic.Results: Vascular endothelial growth factor (VEGF) levels were elevated and fibroblast growth factors (FGFs) were down regulated in RPE-infected MTB cells. Similar pattern of VEGF and FGF was observed in the vitreous of IOTB patients. However, no changes were observed in tear samples.Conclusions: MTB exploits the angiogenesis growth factors for pathogenesis by decreasing FGF with concomitant surge of VEGF in MTB infected RPE as well in the vitreous of IOTB patients.

Contrasting responses to climate change at Himalayan treelines revealed by population demographics of two dominant species.

Alpine treelines are expected to shift upward due to recent climate change. However, interpretation of changes in montane systems has been problematic because effects of climate change are frequently confounded with those of land use changes. The eastern Himalaya, particularly Langtang National Park, Central Nepal, has been relatively undisturbed for centuries and thus presents an opportunity for studying climate change impacts on alpine treeline uncontaminated by potential confounding factors.We studied two dominant species, Abies spectabilis (AS) and Rhododendron campanulatum (RC), above and below the treeline on two mountains. We constructed 13 transects, each spanning up to 400 m in elevation, in which we recorded height and state (dead or alive) of all trees, as well as slope, aspect, canopy density, and measures of anthropogenic and animal disturbance.All size classes of RC plants had lower mortality above treeline than below it, and young RC plants (<2 m tall) were at higher density above treeline than below. AS shows little evidence of a position change from the historic treeline, with a sudden extreme drop in density above treeline compared to below. Recruitment, as measured by size-class distribution, was greater above treeline than below for both species but AS is confined to ~25 m above treeline whereas RC is luxuriantly growing up to 200 m above treeline. Synthesis. Evidence suggests that the elevational limits of RC have shifted upward both because (a) young plants above treeline benefited from facilitation of recruitment by surrounding vegetation, allowing upward expansion of recruitment, and (b) temperature amelioration to mature plants increased adult survival. We predict that the current pure stand of RC growing above treeline will be colonized by AS that will, in turn, outshade and eventually relegate RC to be a minor component of the community, as is the current situation below the treeline.

Natural course of asymptomatic walled off pancreatic necrosis.

INTRODUCTION: There is paucity of data on natural course of asymptomatic walled off necrosis (WON). OBJECTIVE: To study the natural course as well as outcome of conservative management in patients with asymptomatic WON. METHODS: Retrospective analysis of prospectively maintained data base of patients with asymptomatic WON presenting to us 4-6 weeks after an episode of acute necrotising pancreatitis (ANP). RESULTS: Forty three patients (37 M; mean age: 38.2 ±â€¯10.4 years) with asymptomatic WON were studied. The size of WON ranged from 5 to 16 cm (mean 8.2 ±â€¯2.2 cm). The site of WON was head, body and tail in 5 (11%), 34 (79%) and 4 (10%) patients respectively. Thirty of 43 patients (70%) patients did not have any complications during the expectant management period of 3 weeks-32 months with 13 (30%) patients having spontaneous resolution within 6.2 ±â€¯3.4 months. Thirteen (30%) patients became symptomatic or developed complication within 3.2 ±â€¯1.3 months. These were refractory pain (n = 7), infection (n = 4), spontaneous rupture into gastrointestinal tract (n = 5; stomach in 3, duodenum in 1 and colon in 1 patient respectively) and bleeding from splenic artery pseudoaneursym in 1 patient. CONCLUSIONS: Majority of patients with asymptomatic WON have an uneventful clinical course. However, one third patients will develop symptoms/complications requiring interventional treatment.

Direct Engagement of TLR9 Ligand with T Helper Cells Leads to Cell Proliferation & Up-regulation of Cytokines.

PURPOSE: Toll like receptor (TLR) engagement is primarily a function of the innate immune cells. The purpose of the study was to assess direct uptake of ODN 2216 in T helper cells and effects on cell proliferation and cytokine expression. METHODS: We isolated CD4+ CD25- T helper cells by magnetic sorting and studied the uptake of ODN 2216 using flow cytometry and confocal microscopy. We then studied the effect of ODN 2216 engagement on cell proliferation and cytokine expression using flow cytometry and gene expression of TLR9 signaling genes using real time RT-PCR. RESULTS: We made a chance observation that purified T helper cells from healthy individuals consistently bind to the TLR9 ligand ODN 2216. In PBMCs, on the other hand, 98% of monocytes preferentially bound to ODN 2216 FITC, indicating that they competed with the lymphocytes. We confirmed intracellular localization of ODN 2216 FITC as well as intracellular expression of TLR9 in Thelper cells. Furthermore, ODN 2216 FITC was also co-localized with the lysosomal membrane associated protein 1. The uptake of TLR9 ligand culminated in cellular proliferation, up-regulation of cytokines and increased mRNA expression of TLR9 and IRF7 in T helper cells, in the absence of antigen presenting cells. ODN 2216 uptake was inhibited by promethazine as well as by TLR9 antagonist. CONCLUSIONS: Our results show a direct engagement of TLR9 ligand in T helper cells and suggest involvement of TLR9 signalling in CD4+T cells, which may envisage novel targets for TLR inhibitors.