Rac1 plays a crucial role in MCP-1-induced monocyte adhesion and migration.

Monocyte migration is an important process in inflammation and atherogenesis. Identification of the key signalling pathways that regulate monocyte migration can provide prospective targets for prophylactic treatments in inflammatory diseases. Previous research showed that the focal adhesion kinase Pyk2, Src kinase and MAP kinases play an important role in MCP-1-induced monocyte migration. In this study, we demonstrate that MCP-1 induces iPLA2 activity, which is regulated by PKCß and affects downstream activation of Rac1 and Pyk2. Rac1 interacts directly with iPLA2 and Pyk2, and plays a crucial role in MCP-1-mediated monocyte migration by modulating downstream Pyk2 and p38 MAPK activation. Furthermore, Rac1 is necessary for cell spreading and F-actin polymerization during monocyte adhesion to fibronectin. Finally, we provide evidence that Rac1 controls the secretion of inflammatory mediator vimentin from MCP-1-stimulated monocytes. Altogether, this study demonstrates that the PKCß/iPLA2/Rac1/Pyk2/p38 MAPK signalling cascade is essential for MCP-1-induced monocyte adhesion and migration.

Growth, haemato-biochemical, hormonal and disease characteristics in Black Bengal goats: a review.

The goats have been considered one of the noteworthy animals to provide food security and could promote socio-economic upliftment under challenging climatic scenarios in the coming decades, particularly in the tropics. Black Bengal goat (BBG) is one of the recognised native meat-type breeds of hot-humid tropics with distinguished characteristics, including superior-quality meat, excellent skin and high prolificacy. Smaller body mass, lower metabolic rate and efficient utilisation of high-fibre forages enable BBG to adapt to a wide range of harsh climates in the tropics. The BBG can maintain physiological homeostasis efficiently in terms of electrolyte profile, endocrine functions and haemato-biochemical traits in different life phases, including the gestation period, even in high-saline coastal areas of hot-humid tropics. Crossbreeding to improve its growth rate has been attempted, but the prolificacy has been decayed. This review is intended to attract global attention to the adaptive potentialities of Black Bengal goats in terms of growth and production, haemato-biochemical, endocrinological, salt tolerance and disease characteristics that could be an asset of climate-resilient agricultural farming.

A PKCß-LYN-PYK2 Signaling Axis Is Critical for MCP-1-Dependent Migration and Adhesion of Monocytes.

MCP-1-induced monocyte chemotaxis is a crucial event in inflammation and atherogenesis. Identifying the important signal transduction pathways that control monocyte chemotaxis can unravel potential targets for preventive therapies in inflammatory disease conditions. Previous studies have shown that the focal adhesion kinase Pyk2 plays a critical role in monocyte motility. In this study, we investigated the MCP-1-mediated activation of Pyk2 (particularly by the phosphorylation of Tyr402) in primary human peripheral blood monocytes. We showed that MCP-1 induces Src phosphorylation in a similar time frame and that the MCP-1-induced Pyk2 tyrosine phosphorylation is controlled by the Src family kinase. We also report, in this study, that PKCß, an isoform of PKC, is required for both Src and Pyk2 activation/phosphorylation in response to MCP-1 stimulation. We identified Lyn as the specific Src kinase isoform that is activated by MCP-1 and acts upstream of Pyk2 in primary monocytes. Furthermore, Lyn is found to be indispensable for monocyte migration in response to MCP-1 stimulation. Moreover, our coimmunoprecipitation studies in monocytes revealed that PKCß, Pyk2, and Lyn exist constitutively in a molecular complex. To our knowledge, our study has uncovered a novel PKCß-Lyn-Pyk2 signaling cascade in primary monocytes that regulates MCP-1-induced monocyte adhesion and migration.

Integrating 3Rs approaches in WHO guidelines for the batch release testing of biologicals: Responses from a survey of vaccines and biological therapeutics manufacturers.

The UK National Centre for the Replacement, Refinement, and Reduction of Animals in Research (NC3Rs) has been tasked by the World Health Organization (WHO) to review the extent to which animal-based testing methods are described in their manuals, guidelines and recommendations for vaccines and biotherapeutics. The aim is to identify and recommend where updates to these documents can lead to an increased and more harmonised adoption of 3Rs principles (i.e. Replacement, Reduction and Refinement of animal tests) in the quality control and batch release testing requirements for vaccines and biotherapeutics. Developing recommendations that are widely applicable by both the manufacturers and national regulatory authorities for vaccines and biologicals globally requires a detailed understanding of how different organisations view the opportunities and barriers to better integration of the 3Rs. To facilitate this, we developed and distributed a survey aimed at vaccine and biotherapeutics manufacturers in July 2021. In this paper, we present the key findings from this survey and how these will help inform the recommendations for wider integration of 3Rs approaches by WHO in their guidance documents applicable to the quality control and batch testing of vaccines and biotherapeutics.

Integrating 3Rs approaches in WHO guidelines for the batch release testing of biologicals: Responses from a survey of National Control Laboratories and National Regulatory Authorities.

The UK National Centre for the Replacement, Refinement, and Reduction of Animals in Research (NC3Rs) is reviewing World Health Organization (WHO) manuals, guidelines and recommendations for vaccines and biotherapeutics to identify the extent to which animal-based testing methods are described. The aim is to recommend where updates to these documents can lead to an increased and more harmonised adoption of 3Rs principles (i.e. Replacement, Reduction and Refinement of animal tests) in the quality control and batch release testing requirements for vaccines and biotherapeutics. Improved adoption of 3Rs principles and non-animal testing strategies will help to reduce the delays and costs associated with product release testing. Developing recommendations that are widely applicable by both the manufacturers and national regulatory authorities for vaccines and biological therapeutics globally requires a detailed understanding of how different organisations view the opportunities and barriers to better integration of the 3Rs. To facilitate this, we developed and distributed a survey aimed at individuals who work for national regulatory authorities (NRAs) and/or national control laboratories (NCLs). In this paper, we present the key findings from this survey and how these will help inform the recommendations for wider integration of 3Rs approaches by WHO in their guidance documents applicable to the quality control and batch release testing of vaccines and biotherapeutics.

Effect of dietary oregano essential oil and milk replacer on physiological status and immunological responses of pre- and post-weaned Ghoongroo piglets.

Ghoongroo pigs have good adaptability in a low input production system with high prolificacy. The present study was conducted on pre-and post-weaned Ghoongroo piglets from 2-3 days to 12 weeks of age to evaluate the effect of a milk replacer and oregano essential oil (EO) on growth, physiological and immunological responses. Thirty six piglets were randomly divided into three groups. The control group (n = 12) was allowed to suck mother's milk. Second group piglets were provided milk replacer (MR) and piglets of the third group were provided milk replacer along with oregano EO at 500 mg/kg diet. After weaning, piglets were provided standard concentrate diets. The results showed that the body weight in MR and MR + EO groups were significantly higher compared with the control. The MR + EO group had better intestinal microbiota, greater nonspecific innate immunity with the phagocytosis efficacy of neutrophils, lower cortisol concentration and more stable thyroid hormones than the other groups. The better haematological status supported the rapid organ development and improved intestinal health status in both the experimental groups. In conclusion, milk replacer, especially with the inclusion of oregano EO, can lower weaning stress, enhance nonspecific immunity and improve growth and health status of piglets.

Accelerating Global Deletion of the Abnormal Toxicity Test for vaccines and biologicals. Planning common next steps. A workshop Report.

This online workshop Accelerating Global Deletion of the Abnormal Toxicity Test for vaccines and biologicals. Planning common next steps was organized on October 14th, 2021, by the Animal Free Safety Assessment Collaboration (AFSA), the Humane Society International (HSI), the European Federation of Pharmaceutical Industries and Associations (EFPIA), in collaboration with the International Alliance of Biological Standardization (IABS). The workshop saw a participation of over a hundred representatives from international organizations, pharmaceutical industries and associations, and regulatory authorities of 28 countries. Participants reported on country- and region-specific regulatory requirements and, where present, on the perspectives on the waiving and elimination of the Abnormal Toxicity Test. With AFSA, HSI, EFPIA and IABS representatives as facilitators, the participants also discussed specific country/global actions to further secure the deletion of ATT from all regulatory requirements worldwide.

Functional Myoglobin Model Composed of a Strapped Porphyrin/Cyclodextrin Supramolecular Complex with an Overhanging COOH That Increases O2/CO Binding Selectivity in Aqueous Solution.

A water-soluble strapped iron(III)tetraarylporphyrin (FeIIIPor-1) bearing two propylpyridinium groups at the side chains and a carboxylic acid group at the overhanging position of the strap was synthesized to mimic the function of myoglobin with the distal polar functionality in aqueous solution. FeIIIPor-1 forms a stable 1:1 inclusion complex with a per-O-methylated ß-cyclodextrin dimer having a pyridine linker (Py3OCD), providing a hydrophobic environment and a proximal fifth ligand to stabilize the O2-complex. The ferrous complex (FeIIPorCD-1) binds both O2 and CO in aqueous solution. The O2 and CO binding affinities (P1/2O2 and P1/2CO) and half-life time (t1/2) of the O2 complex of FeIIPorCD-1 are 6.3 and 0.021 Torr, and 7 h, respectively, at pH 7 and 25 °C. The control compound without the strap structure (FeIIPorCD-2) has similar oxygen binding characteristics (P1/2O2 = 8.0 Torr), but much higher CO binding affinity (P1/2CO = 3.8 × 10-4 Torr), and longer t1/2 (30 h). The O2 and CO kinetics indicate that the strapped structure in FeIIPorCD-1 inhibits the entrance of these gaseous ligands into the iron(II) center, as evidenced by lower konO2 and konCO values. Interestingly, the CO complex of FeIIPorCD-1 is significantly destabilized (relatively larger koffCO), while the koffO2 value is much smaller than that of FeIIPorCD-2, resulting in significantly increased O2/CO selectivity (reduced M value, where M = P1/2O2/P1/2CO = 320) in FeIIPorCD-1 compared to FeIIPorCD-2 (M = 21000).

Electrocatalytic Water Oxidation by a Phosphorus-Nitrogen O═PN3-Pincer Cobalt Complex.

Water oxidation is a primary step in natural as well as artificial photosynthesis to convert renewable solar energy into chemical energy/fuels. Electrocatalytic water oxidation to evolve O2, utilizing suitable low-cost catalysts and renewable electricity, is of fundamental importance considering contemporary energy and environmental issues, yet it is kinetically challenging owing to the complex multiproton/electron transfer processes. Herein, we report the first cobalt-based pincer catalyst for catalytic water oxidation at neutral pH with high efficiency under electrochemical conditions. Most importantly, ligand (pseudo)aromaticity is identified to play an important role during electrocatalysis. A significant potential jump (∼300 mV) was achieved toward a lower positive value when the aromatized cobalt complex was transformed into a (pseudo)dearomatized cobalt species. The dearomatized species catalyzes the water oxidation reaction to evolve oxygen at a much lower overpotential (∼340 mV) on the basis of the onset potential (at a current density of 0.5 mA/cm2) of catalysis at pH 10.5, outperforming other Co-based molecular catalysts reported to date. These observations may provide a new strategy for the judicious design of earth-abundant transition-metal-based water oxidation catalysts.

Cyanomethyl Ether as an Orthogonal Participating Group for Stereoselective Synthesis of 1,2-trans-ß-O-Glycosides.

Stereoselective formation of glycosidic linkages has been the prime focus for contemporary carbohydrate chemistry. Herein, we report cyanomethyl (CNMe) ether as an efficient and effective participating orthogonal protecting group for the stereoselective synthesis of 1,2-trans-ß-O-glycosides. The participating group facilitated good to high ß-selective glycosylation with a broad range of electron-rich and electron-deficient glycosyl acceptors. Detailed experimental and theoretical studies reveal the involvement of CNMe ether in the formation of a six-membered imine-type cyclic intermediate for the observed stereoselectivity. Rapid incorporation and selective removal of the CNMe ether group in the presence of benzyl ether and isopropylidene acetal protection have also been reported here. The nitrile group provided an opportunity for the glycodiversification through further derivatizations.

Regulation of monoamine oxidase A (MAO-A) expression, activity, and function in IL-13-stimulated monocytes and A549 lung carcinoma cells.

Monoamine oxidase A (MAO-A) is a mitochondrial flavoenzyme implicated in the pathogenesis of atherosclerosis and inflammation and also in many neurological disorders. MAO-A also has been reported as a potential therapeutic target in prostate cancer. However, the regulatory mechanisms controlling cytokine-induced MAO-A expression in immune or cancer cells remain to be identified. Here, we show that MAO-A expression is co-induced with 15-lipoxygenase (15-LO) in interleukin 13 (IL-13)-activated primary human monocytes and A549 non-small cell lung carcinoma cells. We present evidence that MAO-A gene expression and activity are regulated by signal transducer and activator of transcription 1, 3, and 6 (STAT1, STAT3, and STAT6), early growth response 1 (EGR1), and cAMP-responsive element-binding protein (CREB), the same transcription factors that control IL-13-dependent 15-LO expression. We further established that in both primary monocytes and in A549 cells, IL-13-stimulated MAO-A expression, activity, and function are directly governed by 15-LO. In contrast, IL-13-driven expression and activity of MAO-A was 15-LO-independent in U937 promonocytic cells. Furthermore, we demonstrate that the 15-LO-dependent transcriptional regulation of MAO-A in response to IL-13 stimulation in monocytes and in A549 cells is mediated by peroxisome proliferator-activated receptor γ (PPARγ) and that signal transducer and activator of transcription 6 (STAT6) plays a crucial role in facilitating the transcriptional activity of PPARγ. We further report that the IL-13-STAT6-15-LO-PPARγ axis is critical for MAO-A expression, activity, and function, including migration and reactive oxygen species generation. Altogether, these results have major implications for the resolution of inflammation and indicate that MAO-A may promote metastatic potential in lung cancer cells.

Formally Ferric Heme Carbon Monoxide Adduct.

Formally ferric carbonyl adducts are reported in a series of thiolate-bound iron porphyrins. Resonance Raman data indicate the presence of both Fe-S and Fe-CO bonds, and EPR data of this S = 1/2 species indicate a ligand-based electron hole, giving this complex an Fe(II)-thiyl radical electronic ground state. The FTIR data show that the C-O vibrations are substantially higher than in the corresponding ferrous-thiolate CO adducts. DFT calculations reproduce the spectroscopic features and indicate that backbonding to the low lying π* orbitals of the bound CO stabilizes the Fe 3d orbitals resulting in a stabilization of the ferrous-thiyl radical ground state compared to the five-coordinate ferric-thiolate precursor complexes. Access to stable thiyl radicals will help understand these elusive species that are mostly encountered as short-lived reactive reaction intermediates.

Resonance Raman Spectroscopy and Density Functional Theory Calculations on Ferrous Porphyrin Dioxygen Adducts with Different Axial Ligands: Correlation of Ground State Wave Function and Geometric Parameters with Experimental Vibrational Frequencies.

A dioxygen adduct of ferrous porphyrin is an important chemical species in nature as it is a common intermediate in all oxygen transfer, storage, reducing, and activating heme enzymes. The ground state (GS) wave function of this complex has been investigated using several techniques like resonance Raman (rR), Mossbauer, and X-ray absorption spectroscopies. The Fe-O and O-O vibrations of these six-coordinated diamagnetic species show a positive correlation with each other in contrast to analogous ferrous carbonyl complexes where the Fe-CO vibration correlates negatively with the C-O vibration due to a synergistic effect. In this Article, three Fe-porphyrins with different axial ligands (imidazole, phenolate, and thiolate) are investigated using rR spectroscopy and density functional theory (DFT) calculations. The GS wave functions of these species are analyzed, and the contribution of the three primary bonding interactions in the Fe-O2 unit (a σ interaction from the in-plane π* of the superoxide to the vacant dz2 of Fe, a π donation from the out of plane (oop) π* to the dπ orbital of Fe, and a π back bonding interaction from the dπ orbital of Fe to the oop π* of the superoxide) to the calculated Fe-O and O-O vibrations and bond lengths are deconvoluted using a MO theory framework. The GS wave function provides a basis for the correlations observed between different vibrational and geometric parameters of these dioxygen adducts. Furthermore, the correlations obtained allows estimation of the GS wave function and geometry of these species (both natural and artificial) using their experimentally observed Fe-O/O-O vibrations. The wave functions thus extracted from the experimental vibrational data reported offer insight into the role of the axial ligand and hydrogen bonding on the geometric and electronic structures of these crucial chemical species in different protein active sites.

Vibrational spectra of Pb2Bi2Te3, PbBi2Te4, and PbBi4Te7 topological insulators: temperature-dependent Raman and theoretical insights from DFT simulations.

Herein, using Raman spectroscopy, we have presented the investigation of a temperature-dependent frequency shift and the line broadening of phonon modes by inserting the atomic layers of Pb and PbTe in the prototype 3D topological insulator Bi2Te3. Good quality single crystals of Pb2Bi2Te3, PbBi2Te4, and PbBi4Te7 were grown using the modified Bridgman technique. The Raman modes show progressive blue-shift with the decrease in temperature from 298 K to 93 K in Pb2Bi2Te3, PbBi2Te4, and PbBi4Te7 due to the anharmonic vibrations of the lattice as well as the increase in the strength of Bi-Te covalent interactions. The experimental results were complemented by extensive first principles calculations, where a reasonable matching between the experimental and computational data was found. Chemical pressure, induced by the insertion of Pb and PbTe layers in Bi2Te3, modified the interactions at the boundaries of the quintuple-layers, which was evident from the evolution of the A21u mode. The enhancement in the out-of-plane Bi-Te vibrations with respect to the in-plane Bi-Te vibrations was observed at low temperatures. The temperature coefficients of the Raman modes were useful in determining the thermal conductivity, which is a key design parameter for the fabrication of spintronic devices using topological insulators. The estimated first order temperature coefficient (χ') for Pb2Bi2Te3 signified the decrease in the thermal conductivity relative to Bi2Te3, which was caused by the insertion of the Pb layers in the van der Waals gaps of Bi2Te3.

Deregulation of LIMD1-VHL-HIF-1α-VEGF pathway is associated with different stages of cervical cancer.

To understand the mechanism of cellular stress in basal-parabasal layers of normal cervical epithelium and during different stages of cervical carcinoma, we analyzed the alterations (expression/methylation/copy number variation/mutation) of HIF-1α and its associated genes LIMD1, VHL and VEGF in disease-free normal cervix (n = 9), adjacent normal cervix of tumors (n = 70), cervical intraepithelial neoplasia (CIN; n = 32), cancer of uterine cervix (CACX; n = 174) samples and two CACX cell lines. In basal-parabasal layers of normal cervical epithelium, LIMD1 showed high protein expression, while low protein expression of VHL was concordant with high expression of HIF-1α and VEGF irrespective of HPV-16 (human papillomavirus 16) infection. This was in concordance with the low promoter methylation of LIMD1 and high in VHL in the basal-parabasal layers of normal cervix. LIMD1 expression was significantly reduced while VHL expression was unchanged during different stages of cervical carcinoma. This was in concordance with their frequent methylation during different stages of this tumor. In different stages of cervical carcinoma, the expression pattern of HIF-1α and VEGF was high as seen in basal-parabasal layers and inversely correlated with the expression of LIMD1 and VHL. This was validated by demethylation experiments using 5-aza-2'-deoxycytidine in CACX cell lines. Additional deletion of LIMD1 and VHL in CIN/CACX provided an additional growth advantage during cervical carcinogenesis through reduced expression of genes and associated with poor prognosis of patients. Our data showed that overexpression of HIF-1α and its target gene VEGF in the basal-parabasal layers of normal cervix was due to frequent inactivation of VHL by its promoter methylation. This profile was maintained during different stages of cervical carcinoma with additional methylation/deletion of VHL and LIMD1.

Valence tautomerism in synthetic models of cytochrome P450.

CytP450s have a cysteine-bound heme cofactor that, in its as-isolated resting (oxidized) form, can be conclusively described as a ferric thiolate species. Unlike the native enzyme, most synthetic thiolate-bound ferric porphyrins are unstable in air unless the axial thiolate ligand is sterically protected. Spectroscopic investigations on a series of synthetic mimics of cytP450 indicate that a thiolate-bound ferric porphyrin coexists in organic solutions at room temperature (RT) with a thiyl-radical bound ferrous porphyrin, i.e., its valence tautomer. The ferric thiolate state is favored by greater enthalpy and is air stable. The ferrous thiyl state is favored by entropy, populates at RT, and degrades in air. These ground states can be reversibly interchanged at RT by the addition or removal of water to the apolar medium. It is concluded that hydrogen bonding and local electrostatics protect the resting oxidized cytP450 active site from degradation in air by stabilizing the ferric thiolate ground state in contrast to its synthetic analogs.

Activation of Wnt-ß-catenin pathway in basal-parabasal layers of normal cervical epithelium comparable during development of uterine cervical carcinoma.

In this study, importance of Wnt-ß-catenin pathway in the development of uterine cervical carcinoma was evaluated. For this purpose, the profiles (expression/methylation/deletion) of ß-catenin, p-ß-catenin (Y654), Wnt3a, and APC were studied in disease free normal cervical epithelium (n = 9), adjacent normal cervical epithelium of primary tumors (n = 70), CIN (n = 28), CACX (n = 102) samples, and two CACX cell lines (HeLa and SiHa). Immunohistochemical analysis revealed high/medium (74-95%) expression of ß-catenin/p-ß-catenin (Y654) and Wnt3a and low expression (23-26%) of APC in proliferating basal-parabasal layers contrary to differentiated spinous layer in normal cervix irrespective of HPV16 infection. The expression profile of the genes in the basal-parabasal layers did not change significantly during development of CACX. High (66%) promoter methylation of APC was seen in basal-parabasal layers and the cervical lesions (42-69%), unlike in spinous layers (25%). The promoter methylation status of APC was validated by in vitro demethylation experiments using 5-aza-dC in CACX cell lines. However, additional deletion of APC was significantly increased from CIN (12%) to stage I/II (40%) and became comparable in stage III/IV (48%) of the tumor. Patients with alterations (deletion/methylation) of APC and high/medium expression of Wnt3a/ß-catenin/p-ß-catenin (Y654) showed significantly poor survival. Thus our data indicate that cumulative effect of Wnt3a overexpression and APC inactivation are needed for overexpression of ß-catenin during the development of CACX.

Risk of high-grade precancerous lesions and invasive cancers in high-risk HPV-positive women with normal cervix or CIN 1 at baseline-A population-based cohort study.

Infection with high-risk human papillomavirus (HR-HPV) is transient and clears on its own in majority of the women. Only a few women who have persistent infection may finally develop cervical intraepithelial neoplasia (CIN) or cervical cancer in later years. The risk of progression in the HR-HPV-positive women with normal cervix or low-grade lesion on colposcopy and histopathology at baseline is less studied. We performed a longitudinal study on 650 HR-HPV-positive women with colposcopy and/or histopathology-proved normal or CIN1 diagnosis at baseline to assess the cumulative risk of development of high-grade CIN. After a mean follow-up of 2.1 person years of observation (PYO) (range 0.1-5.1), the cumulative incidence of CIN2+ (6.4%; 3.0/100 PYO) was significantly higher in women who had persistent HR-HPV infection compared to those who cleared the infection (adjusted HR 6.28; 95% CI 2.87-13.73). The risk of viral persistence in women aged 50-60 years was two times higher compared to women aged 40-49 years and three times higher compared to women aged 30-39 years. The probability of having persistent infection increased progressively with higher viral load at baseline (adjusted HR 3.29, 95% CI 2.21-4.90 for RLU ≥100; adjusted HR 2.69, 95% CI 1.71-4.22 for RLU 10-100). Women with increasing viral load at follow-up had four times higher risk of developing CIN2 or worse lesions as compared to those with decreasing load (20.9% vs 4.8%; p < 0.001). In the context of developing countries where cytology or genotyping triaging is not feasible, colposcopy referral of HR-HPV-positive women with advancing age, viral persistence, and increasing viral load may be considered.

Detection of a cancer biomarker protein on modified cellulose paper by fluorescence using aptamer-linked quantum dots.

The development of point-of-care bioassays for sensitive screening of protein-based cancer biomarkers would improve the opportunity for early stage diagnosis. A strategy for a fluorescence resonance energy transfer (FRET)-based bioassay has been investigated that makes use of modified cellulose paper for the detection of an epithelial cell adhesion molecule (EpCAM), which is a transmembrane glycoprotein that is overexpressed in several tumors of epithelial origin. The paper matrix was a substrate for immobilized aptamer-linked quantum dots (QDs-Apt) and Cy3 labeled complementary DNA (cDNA), which served as a donor and an acceptor, respectively. Competitive binding of EpCAM displaced the cDNA, resulting in the reduction of FRET. The paper-based bioassay was able to detect EpCAM in buffer solution as well as in 10% bovine serum solution using a reaction time of no more than 60 minutes. The dynamic range was 1-100 nM in buffer with a precision better than 4%, and the limit of detection was 250 pM in buffer and 600 pM in 10% serum.

HPV detection-based cervical cancer screening program in low-resource setting: lessons learnt from a community-based demonstration project in India.

PURPOSE: A demonstration project was conducted to assess feasibility of implementing HPV detection-based cervical cancer screening in primary care settings in India and to generate local evidence on feasibility and effectiveness of HPV detection in primary screening. METHODS: The project was implemented by setting up screening clinics at primary health centers. Eligible women were screened by HPV DNA test (Hybrid capture 2). All samples were processed and tested in a single laboratory. Colposcopy services were provided to screen-positive women at the same community clinics. Project utilized services of community health workers for community mobilization, recall of screen-positive and disease-positive women. Women with ≥CIN2 diagnosis were treated at tertiary hospital. RESULTS: Totally, 44,110 women were screened and HPV positivity was 4.7 %. Compliance to recall of HC2-positive women for colposcopy was 78 %. Detection rate of CIN3+ by HPV test was 3.9/1,000 women. Compliance of women to treatment was 80.1 %. However, compliance of HPV-positive women for follow-up at 1 year was poor (23.2 %). Concurrent use of VIA to screen the women did not have any advantage but increased number of unnecessary colposcopies and biopsies. CONCLUSIONS: Our project demonstrated that it was possible to implement HPV detection-based screening using existing primary healthcare infrastructure. Performing colposcopy at primary setting is feasible, improves compliance and reduces over-treatment. In settings with low to moderately high HPV prevalence, direct referral of HPV-positive women is advisable. Community health workers can be effectively used for recalling the positive women.