Peptidoglycan recognition protein 3 and Nod2 synergistically protect mice from dextran sodium sulfate-induced colitis

Aberrant immune response and changes in the gut microflora are the main causes of inflammatory bowel disease (IBD). Peptidoglycan recognition proteins (Pglyrp1, Pglyrp2, Pglyrp3, and Pglyrp4) are bactericidal innate immunity proteins that maintain normal gut microbiome, protect against experimental colitis, and are associated with IBD in humans. Nucleotide-binding oligomerization domain 2 (Nod2) is an intracellular bacterial sensor and may be required for maintaining normal gut microbiome. Mutations in Nod2 are strongly associated with Crohn's disease, but the causative mechanism is not understood, and the role of Nod2 in ulcerative colitis is not known. Because IBD is likely caused by variable multiple mutations in different individuals, in this study, we examined the combined role of Pglyrp3 and Nod2 in the development of experimental colitis in mice. We demonstrate that a combined deficiency of Pglyrp3 and Nod2 results in higher sensitivity to dextran sodium sulfate-induced colitis compared with a single deficiency. Pglyrp3(-/-)Nod2(-/-) mice had decreased survival and higher loss of body weight, increased intestinal bleeding, higher apoptosis of colonic mucosa, elevated expression of cytokines and chemokines, altered gut microbiome, and increased levels of ATP in the colon. Increased sensitivity to dextran sodium sulfate-induced colitis in Pglyrp3(-/-)Nod2(-/-) mice depended on increased apoptosis of intestinal epithelium, changed gut microflora, and elevated ATP. Pglyrp3 deficiency contributed colitis-predisposing intestinal microflora and increased intestinal ATP, whereas Nod2 deficiency contributed higher apoptosis and responsiveness to increased level of ATP. In summary, Pglyrp3 and Nod2 are both required for maintaining gut homeostasis and protection against colitis, but their protective mechanisms differ

Infant and Young Child Feeding Practices among Mothers in Lahore, Pakistan: A Cross-Sectional Study

Background: Inadequate child feeding practices lead to malnutrition, higher under-five mortality rates and adverse effects on quality of life. This study aimed to assess the breastfeeding and complementary feeding practices of mothers as well as the influence of various sociodemographic factors on them in local families of Lahore. Methods: This is a cross-sectional, descriptive study. It was conducted in CMH (Combined Military Hospital), Lahore in 2018. It comprises a sample of 203 mothers with children of at least two years of age, from various urban areas of Lahore. The subjects were selected on the basis of the inclusion criteria. Mothers with psychiatric illnesses and children with congenital anomalies were excluded from the study. Mothers were approached in the paediatric outpatient departments of four tertiary care hospitals of Lahore. Responses were recorded using a modified version of the Action Contre La Faim (ACF) questionnaire. Independent sample t-test and chi-square test were applied for analysis of the data. Results: Early initiation of breastfeeding within one hour from birth was observed in 83.3% children. Most children were administered colostrum (69.5%). The rate of exclusive breastfeeding for the first six months was 45.3%.  A child was being breastfed 8.21 ± 6.67 (mean ± SD) times a day. Maternal educational status, total number of adults in a household, and access to free healthcare were identified as important factors influencing the practice of breastfeeding. Porridge, khichdi, eggs, fruit and yoghurt were the most frequently used complementary foods. Conclusions: A high rate of an early start of breastfeeding and a low rate of exclusive breastfeeding for at least six months were predominant in our population. Administration of colostrum was observed in approximately two-thirds of the study participants. Education of the mother, type of the family system (nuclear or combined), and access to free healthcare strongly influence the breastfeeding practices

Cervicovaginal mucus barrier properties during pregnancy are impacted by the vaginal microbiome

Introduction Mucus in the female reproductive tract acts as a barrier that traps and eliminates pathogens and foreign particles via steric and adhesive interactions. During pregnancy, mucus protects the uterine environment from ascension of pathogens and bacteria from the vagina into the uterus, a potential contributor to intrauterine inflammation and preterm birth. As recent work has demonstrated the benefit of vaginal drug delivery in treating women’s health indications, we sought to define the barrier properties of human cervicovaginal mucus (CVM) during pregnancy to inform the design of vaginally delivered therapeutics during pregnancy. Methods CVM samples were self-collected by pregnant participants over the course of pregnancy, and barrier properties were quantified using multiple particle tracking. 16S rRNA gene sequencing was performed to analyze the composition of the vaginal microbiome. Results Participant demographics differed between term delivery and preterm delivery cohorts, with Black or African American participants being significantly more likely to delivery prematurely. We observed that vaginal microbiota is most predictive of CVM barrier properties and of timing of parturition. Lactobacillus crispatus dominated CVM samples showed increased barrier properties compared to polymicrobial CVM samples. Discussion This work informs our understanding of how infections occur during pregnancy, and directs the engineering of targeted drug treatments for indications during pregnancy

Cervicovaginal mucus barrier properties during pregnancy are impacted by the vaginal microbiome

IntroductionMucus in the female reproductive tract acts as a barrier that traps and eliminates pathogens and foreign particles via steric and adhesive interactions. During pregnancy, mucus protects the uterine environment from ascension of pathogens and bacteria from the vagina into the uterus, a potential contributor to intrauterine inflammation and preterm birth. As recent work has demonstrated the benefit of vaginal drug delivery in treating women’s health indications, we sought to define the barrier properties of human cervicovaginal mucus (CVM) during pregnancy to inform the design of vaginally delivered therapeutics during pregnancy.MethodsCVM samples were self-collected by pregnant participants over the course of pregnancy, and barrier properties were quantified using multiple particle tracking. 16S rRNA gene sequencing was performed to analyze the composition of the vaginal microbiome.ResultsParticipant demographics differed between term delivery and preterm delivery cohorts, with Black or African American participants being significantly more likely to delivery prematurely. We observed that vaginal microbiota is most predictive of CVM barrier properties and of timing of parturition. Lactobacillus crispatus dominated CVM samples showed increased barrier properties compared to polymicrobial CVM samples.DiscussionThis work informs our understanding of how infections occur during pregnancy, and directs the engineering of targeted drug treatments for indications during pregnancy

Severe autosomal recessive retinitis pigmentosa maps to chromosome 1p13.3–p21.2 between D1S2896 and D1S457 but outside ABCA4

A severe form of autosomal recessive retinitis pigmentosa (arRP) was identified in a large Pakistani family ascertained in the Punjab province of Pakistan. All affected individuals in the family had night blindness in early childhood, early complete loss of useful vision, and typical RP fundus changes plus macular degeneration. After exclusion of known arRP loci, a genome-wide scan was performed using microsatellite markers at about 10 cM intervals and calculating two-point lod scores. PCR cycle dideoxynucleotide sequencing was used to sequence candidate genes inside the linked region for mutations. RP in this family shows linkage to markers in a 10.5 cM (8.9 Mbp) region of chromosome 1p13.3–p21.2 between D1S2896 and D1S457. D1S485 yields the highest lod score of 6.54 at θ=0. Sequencing the exons and intron–exon boundaries of five candidate genes and six ESTs in this region, OLFM3, GNAI3, LOC126987, FLJ25070, DKFZp586G0123, AV729694, BU662869, BU656110, BU171991, BQ953690, and CA397743, did not identify any causative mutations. This novel locus lies approximately 4.9 cM (7.1 Mbp) from ABCA4, which is excluded from the linked region. Identification and study of this gene may help to elucidate the phenotypic diversity of arRP mapping to this region.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/47597/1/439_2005_Article_54.pd

Genetic Association of Peptidoglycan Recognition Protein Variants with Inflammatory Bowel Disease

<div><p>Inflammatory bowel disease (IBD) is a common disease, includes Crohn's disease (CD) and ulcerative colitis (UC), and is determined by altered gut bacterial populations and aberrant host immune response. Peptidoglycan recognition proteins (PGLYRP) are innate immunity bactericidal proteins expressed in the intestine. In mice, PGLYRPs modulate bacterial populations in the gut and sensitivity to experimentally induced UC. The role of PGLYRPs in humans with CD and/or UC has not been previously investigated. Here we tested the hypothesis that genetic variants in <i>PGLYRP1</i>, <i>PGLYRP2</i>, <i>PGLYRP3</i> and <i>PGLYRP4</i> genes associate with CD and/or UC and with gender and/or age of onset of disease in the patient population. We sequenced all <i>PGLYRP</i> exons in 372 CD patients, 77 UC patients, 265 population controls, 210 familial CD controls, and 24 familial UC controls, identified all polymorphisms in these populations, and analyzed the variants for significant association with CD and UC. We identified 16 polymorphisms in the four <i>PGLYRP</i> genes that significantly associated with CD, UC, and/or subgroups of patient populations. Of the 16, 5 significantly associated with both CD and UC, 6 with CD, and 5 with UC. 12 significant variants result in amino acid substitutions and based on structural modeling several of these missense variants may have structural and/or functional consequences for PGLYRP proteins. Our data demonstrate that genetic variants in <i>PGLYRP</i> genes associate with CD and UC and may provide a novel insight into the mechanism of pathogenesis of IBD.</p></div

Chromosomal locations of <i>PGLYRPs</i> and schematic gene, cDNA, and protein structures, with positions of polymorphisms significantly associated with CD and/or UC.

<p>Genes show exons and introns, cDNAs show exons and start and stop codons, and proteins show domains and locations of significant polymorphisms.</p

<i>PGLYRP-4</i> variant associates with CD patients.

<p>Complete data are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0067393#pone.0067393.s002" target="_blank">Tables S2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0067393#pone.0067393.s014" target="_blank">S14</a>.</p><p>*CA, Cochran Armitage Trend test data for the recessive model.</p

Amino acid sequence homology in mammalian PGLYRPs in the regions of non-synonymous polymorphisms significantly associated with CD and/or UC.

<p>Amino acids identical to human Ensembl reference sequences are shaded gray, conservation of the reference polymorphism is highlighted in yellow, and the predicted structure of the region is shown underneath the sequence. No sequence indicates truncated sequence, “–” indicates a gap in alignment, and “X” indicates incomplete sequence.</p

Demographic characteristics of the populations in this IBD study.

<p>Demographic characteristics of the populations in this IBD study.</p