HUBUNGAN PENGETAHUAN, SIKAP, DAN DUKUNGAN KELUARGA DENGAN KEPATUHAN BEROBAT PADA PASIEN TUBERKULOSIS PARU DI RS PARU KOTA PALEMBANG

Tuberkulosis (TB) merupakan penyakit menular yang disebabkan oleh infeksi bakteri Mycobacterium tuberculosis. Indonesia menempati posisi ke-3 di dunia setelah India dan Cina sebagai negara dengan jumlah pasien TB terbanyak. Angka prevalensi TB di Indonesia sebesar 647/100.000 penduduk, angka insidensi sebesar 399/100.000 penduduk dan angka mortalitas sebesar 25/100.000 penduduk. Perilaku sesorang ditentukan oleh tiga faktor yaitu predisposing factors (pengetahuan dan sikap), enabling factors, dan reinforcing factors (dukungan keluarga) yang dapat mempengaruhi kepatuhan berobat. Mengetahui hubungan pengetahuan TB paru, sikap pasien dan dukungan keluarga dengan kepatuhan berobat pada pasien Tuberkulosis Paru di RS Paru Kota Palembang Tahun 2017. Penelitian ini merupakan penelitian analitik observasional dengan desain penelitian cross sectional. Sampel adalah pasien tuberkulosis paru dewasa usia ³ 15 tahun yang berobat di RS Paru Kota Palembang 01 Juni 2017 - 30 November 2017 yang memenuhi kriteria inklusi dan eksklusi dengan jumlah sampel 62 orang. Data yang digunakan dalam penelitian ini adalah data primer dan data sekunder. Data primer dilakukan dengan menggunakan kuesioner, pengisisan kuesioner dilakukan secara langsung oleh peneliti dengan menggunakan teknik wawancara. Data sekunder didapatkan dari buku register TB paru di RS Paru Kota Palembang. Pengolahan data menggunakan uji statistik Chi-square yang dibantu perangkat lunak IBM SPSS Statistics.Hasil penelitian menunjukkan bahwa variabel yang terdapat hubungan signifikan terhadap kepatuhan berobat adalah dukungan keluarga (p=0,000). Variabel yang tidak memiliki hubungan yang signifikan terhadap kepatuhan berobat adalah pengetahuan TB paru (p=0,059) dan sikap pasien terhadap TB paru (p=0,213). Pengetahuan TB paru dan sikap pasien terhadap TB paru tidak terdapat hubungan yang signifikan terhadap kepatuhan berobat di RS Paru Kota Palembang Tahun 2017. Terdapat hubungan yang signifikan antara dukungan keluarga dan kepatuhan berobat di RS Paru Kota Palembang Tahun 2017

A new software tool for carbohydrate microarray data storage, processing, presentation, and reporting

Publisher Copyright: © 2022 The Author(s) 2022. Published by Oxford University Press. This project is supported by Wellcome Trust Biomedical Resource grants (WT099197/Z/12/Z, 108430/Z/15/Z and 218304/Z/19/Z); March of Dimes European Prematurity Research Centre grant 22-FY18-82 and NIH Commons Fund 1U01GM125267-01Glycan microarrays are essential tools in glycobiology and are being widely used for assignment of glycan ligands in diverse glycan recognition systems. We have developed a new software, called Carbohydrate microArray Analysis and Reporting Tool (CarbArrayART), to address the need for a distributable application for glycan microarray data management. The main features of CarbArrayART include: (i) Storage of quantified array data from different array layouts with scan data and array-specific metadata, such as lists of arrayed glycans, array geometry, information on glycan-binding samples, and experimental protocols. (ii) Presentation of microarray data as charts, tables, and heatmaps derived from the average fluorescence intensity values that are calculated based on the imaging scan data and array geometry, as well as filtering and sorting functions according to monosaccharide content and glycan sequences. (iii) Data export for reporting in Word, PDF, and Excel formats, together with metadata that are compliant with the guidelines of MIRAGE (Minimum Information Required for A Glycomics Experiment). CarbArrayART is designed for routine use in recording, storage, and management of any slide-based glycan microarray experiment. In conjunction with the MIRAGE guidelines, CarbArrayART addresses issues that are critical for glycobiology, namely, clarity of data for evaluation of reproducibility and validity.publishersversionpublishe

Plant antibodies for human antifungal therapy

There is an increasing interest in the development of therapeutic antibodies (Ab) to improve the control of fungal pathogens, but none of these reagents is available for clinical use. We previously described a murine monoclonal antibody (mAb 2G8) targeting β-glucan, a cell wall polysaccharide common to most pathogenic fungi, which conferred significant protection against Candida albicans, Aspergillus fumigatus and Cryptococcus neoformans in animal models. Transfer of this wide-spectrum, antifungal mAb into the clinical setting would allow the control of most frequent fungal infections in many different categories of patients. To this aim, two chimeric mouse-human Ab derivatives from mAb 2G8, in the format of complete IgG or scFv-Fc, were generated, transiently expressed in Nicotiana benthamiana plants and purified from leaves with high yields (approximately 50 mg Ab/kg of plant tissues). Both recombinant Abs fully retained the β-glucan-binding specificity and the antifungal activities of the cognate murine mAb against C. albicans. In fact, they recognized preferentially β1,3-linked glucan molecules present at the fungal cell surface and directly inhibited the growth of C. albicans and its adhesion to human epithelial cells in vitro. In addition, both the IgG and the scFv-Fc promoted C. albicans killing by isolated, human polymorphonuclear neutrophils in ex vivo assays and conferred significant antifungal protection in animal models of systemic or vulvovaginal C. albicans infection. These recombinant Abs represent valuable molecules for developing novel, plant-derived immunotherapeutics against candidiasis and, possibly, other fungal diseases

Generation and characterization of β1,2-gluco-oligosaccharide probes fromBrucella abortuscyclic β-glucan and their recognition by C-type lectins of the immune system

The β1,2-glucans produced by bacteria are important in invasion, survival and immunomodulation in infected hosts be they mammals or plants. However, there has been a lack of information on proteins which recognize these molecules. This is partly due to the extremely limited availability of the sequence-defined oligosaccharides and derived probes for use in the study of their interactions. Here we have used the cyclic β1,2-glucan (CβG) of the bacterial pathogen Brucella abortus, after removal of succinyl side chains, to prepare linearized oligosaccharides which were used to generate microarrays. We describe optimized conditions for partial depolymerization of the cyclic glucan by acid hydrolysis and conversion of the β1,2-gluco-oligosaccharides, with degrees of polymerization 2-13, to neoglycolipids for the purpose of generating microarrays. By microarray analyses we show that the C-type lectin receptor DC-SIGNR, like the closely related DC-SIGN we investigated earlier, binds to the β1,2-gluco-oligosaccharides, as does the soluble immune effector serum mannose-binding protein. Exploratory studies with DC-SIGN are suggestive of the recognition also of the intact CβG by this receptor. These findings open the way to unravelling mechanisms of immunomodulation mediated by β1,2-glucans in mammalian systems

Mannan detecting C-type lectin receptor probes recognise immune epitopes with diverse chemical, spatial and phylogenetic heterogeneity in fungal cell walls

Funding Information: This work was supported by the Wellcome Trust Investigator, Collaborative, Equipment, Strategic and Biomedical Resource awards (086827, 075470, 097377, 101873, 200208, 093378 and 099197), the Applied Molecular Biosciences Unit-UCIBIO (FCT/MCTES UID/Multi/04378/2019), Wellcome Trust Biomedical Resource grant (108430/Z/15/Z), March of Dimes (Arlington, Virginia, U.S.A.) Prematurity Research Center grant (22-FY18-821) and by the MRC Centre for Medical Mycology (N006364/1). The University of Aberdeen funded a studentship to IV as part of NARG?s Wellcome Senior Investigator Award. https://wellcome.ac.uk/ - Wellcome. https://mrc.ukri.org/ - MRC. https:// www.requimte.pt/ucibio/ - the Applied Molecular Biosciences Unit-UCIBIO. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: © 2020 Vendele et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Peer reviewedPublisher PD

Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus

This work was supported by Fundação para a Ciência e a Tecnologia (FCT-MCTES), Portugal, through project grant PTDC/BIA-MIB/31730/2017 (to A.S.P.), fellowships PD/BD/105727/2014 (to V.G.C.) and SFRH/BD/143494/2019 (to F.T.), and program contract DL-57/2016 (to B.A.P. and C.N.) and by Wellcome Trust Biomedical Resource grants number WT108430/Z/15/Z and WT218304/Z/19/Z, a March of Dimes (Arlington, VA, USA) Prematurity Research Center grant (number 22-FY18-821) for the funding to the Carbohydrate Microarray Facility, Associate Laboratory projects LAQV-REQUIMTE (UIDB/50006/2020) and CICECO-Aveiro Institute of Materials (UIDB/50011/2020 & UIDP/50011/2020), and by the Applied Molecular Biosciences Unit (UCIBIO), which is financed by Portuguese national funds from FCT-MCTES (UIDP/04378/2020 and UIDB/04378/2020).A multigene polysaccharide utilization locus (PUL) encoding enzymes and surface carbohydrate (glycan)-binding proteins (SGBPs) was recently identified in prominent members of Bacteroidetes in the human gut and characterized in Bacteroides ovatus. This PUL-encoded system specifically targets mixed-linkage β1,3-1,4-glucans, a group of diet-derived carbohydrates that promote a healthy microbiota and have potential as prebiotics. The BoSGBPMLG-A protein encoded by the BACOVA_2743 gene is a SusD-like protein that plays a key role in the PUL's specificity and functionality. Here, we perform a detailed analysis of the molecular determinants underlying carbohydrate binding by BoSGBPMLG-A, combining carbohydrate microarray technology with quantitative affinity studies and a high-resolution X-ray crystallography structure of the complex of BoSGBPMLG-A with a β1,3-1,4-nonasaccharide. We demonstrate its unique binding specificity toward β1,3-1,4-gluco-oligosaccharides, with increasing binding affinities up to the octasaccharide and dependency on the number and position of β1,3 linkages. The interaction is defined by a 41-Å-long extended binding site that accommodates the oligosaccharide in a mode distinct from that of previously described bacterial β1,3-1,4-glucan-binding proteins. In addition to the shape complementarity mediated by CH-π interactions, a complex hydrogen bonding network complemented by a high number of key ordered water molecules establishes additional specific interactions with the oligosaccharide. These support the twisted conformation of the β-glucan backbone imposed by the β1,3 linkages and explain the dependency on the oligosaccharide chain length. We propose that the specificity of the PUL conferred by BoSGBPMLG-A to import long β1,3-1,4-glucan oligosaccharides to the bacterial periplasm allows Bacteroidetes to outcompete bacteria that lack this PUL for utilization of β1,3-1,4-glucans. IMPORTANCE With the knowledge of bacterial gene systems encoding proteins that target dietary carbohydrates as a source of nutrients and their importance for human health, major efforts are being made to understand carbohydrate recognition by various commensal bacteria. Here, we describe an integrative strategy that combines carbohydrate microarray technology with structural studies to further elucidate the molecular determinants of carbohydrate recognition by BoSGBPMLG-A, a key protein expressed at the surface of Bacteroides ovatus for utilization of mixed-linkage β1,3-1,4-glucans. We have mapped at high resolution interactions that occur at the binding site of BoSGBPMLG-A and provide evidence for the role of key water-mediated interactions for fine specificity and affinity. Understanding at the molecular level how commensal bacteria, such as prominent members of Bacteroidetes, can differentially utilize dietary carbohydrates with potential prebiotic activities will shed light on possible ways to modulate the microbiome to promote human health.publishersversionpublishe

Club cell CREB regulates the goblet cell transcriptional network and pro-mucin effects of IL-1B

Introduction: Club cells are precursors for mucus-producing goblet cells. Interleukin 1β (IL-1B) is an inflammatory mediator with pro-mucin activities that increases the number of mucus-producing goblet cells. IL-1B-mediated mucin production in alveolar adenocarcinoma cells requires activation of the cAMP response element-binding protein (CREB). Whether the pro-mucin activities of IL-1B require club cell CREB is unknown.Methods: We challenged male mice with conditional loss of club cell Creb1 and wild type littermates with intra-airway IL-1B or vehicle. Secondarily, we studied human “club cell-like” H322 cells.Results: IL-1B increased whole lung mRNA of secreted (Mucin 5ac, Mucin 5b) and tethered (Mucin 1, Mucin 4) mucins independent of genotype. However, loss of club cell Creb1 increased whole lung mRNA of member RAS oncogene family (Rab3D), decreased mRNA of the muscarinic receptor 3 (M3R) and prevented IL-1B mediated increases in purinergic receptor P2Y, (P2ry2) mRNA. IL-1B increased the density of goblet cells containing neutral mucins in wildtype mice but not in mice with loss of club cell Creb1. These findings suggested that club cell Creb1 regulated mucin secretion. Loss of club cell Creb1 also prevented IL-1B-mediated impairments in airway mechanics. Four days of pharmacologic CREB inhibition in H322 cells increased mRNA abundance of forkhead box A2 (FOXA2), a repressor of goblet cell expansion, and decreased mRNA expression of SAM pointed domain containing ETS transcription factor (SPDEF), a driver of goblet cell expansion. Chromatin immunoprecipitation demonstrated that CREB directly bound to the promoter region of FOXA2, but not to the promoter region of SPDEF. Treatment of H322 cells with IL-1B increased cAMP levels, providing a direct link between IL-1B and CREB signaling.Conclusion: Our findings suggest that club cell Creb1 regulates the pro-mucin properties of IL-1B through pathways likely involving FOXA2

The Role of Sialyl Glycan Recognition in Host Tissue Tropism of the Avian Parasite Eimeria tenella

Eimeria spp. are a highly successful group of intracellular protozoan parasites that develop within intestinal epithelial cells of poultry, causing coccidiosis. As a result of resistance against anticoccidial drugs and the expense of manufacturing live vaccines, it is necessary to understand the relationship between Eimeria and its host more deeply, with a view to developing recombinant vaccines. Eimeria possesses a family of microneme lectins (MICs) that contain microneme adhesive repeat regions (MARR). We show that the major MARR protein from Eimeria tenella, EtMIC3, is deployed at the parasite-host interface during the early stages of invasion. EtMIC3 consists of seven tandem MAR1-type domains, which possess a high specificity for sialylated glycans as shown by cell-based assays and carbohydrate microarray analyses. The restricted tissue staining pattern observed for EtMIC3 in the chicken caecal epithelium indicates that EtMIC3 contributes to guiding the parasite to the site of invasion in the chicken gut. The microarray analyses also reveal a lack of recognition of glycan sequences terminating in the N-glycolyl form of sialic acid by EtMIC3. Thus the parasite is well adapted to the avian host which lacks N-glycolyl neuraminic acid. We provide new structural insight into the MAR1 family of domains and reveal the atomic resolution basis for the sialic acid-based carbohydrate recognition. Finally, a preliminary chicken immunization trial provides evidence that recombinant EtMIC3 protein and EtMIC3 DNA are effective vaccine candidates

Model uncertainties in climate change impacts on Sahel precipitation in ensembles of CMIP5 and CMIP6 simulations

The impact of climate change on Sahel precipitation suffers from large uncertainties and is strongly model-dependent. In this study, we analyse sources of inter-model spread in Sahel precipitation change by decomposing precipitation into its dynamic and thermodynamic terms, using a large set of climate model simulations. Results highlight that model uncertainty is mostly related to the response of the atmospheric circulation to climate change (dynamic changes), while thermodynamic changes are less uncertain among climate models. Uncertainties arise mainly because the models simulate different shifts in atmospheric circulation over West Africa in a warmer climate. We linked the changes in atmospheric circulation to the changes in Sea Surface Temperature, emphasising that the Northern hemispheric temperature gradient is primary to explain uncertainties in Sahel precipitation change. Sources of Sahel precipitation uncertainties are shown to be the same in the new generation of climate models (CMIP6) as in the previous generation of models (CMIP5)