51 research outputs found
Identification of a novel N-acetylmuramic acid (MurNAc) transporter in Tannerella forsythia.
Tannerella forsythia is a Gram-negative periodontal pathogen lacking the ability to undergo de novo synthesis of amino sugars N-acetylmuramic acid (MurNAc) and N-acetylglucosamine (GlcNAc) that form the disaccharide-repeating unit of the peptidoglycan backbone. T. forsythia relies on the uptake of these sugars from the environment, which is so far unexplored. Here, we identified a novel transporter system of T. forsythia involved in the uptake of MurNAc across the inner membrane and characterized a homolog of the Escherichia. coli MurQ etherase involved in the conversion of MurNAc-6P to GlcNAc-6P. The genes encoding these components were identified on a three gene cluster spanning Tanf_08375 to Tanf_08385 located downstream from a putative peptidoglycan recycling locus. We show that the three genes, Tanf_08375, Tanf_08380, and Tanf_08385, encoding a MurNAc transporter, a putative sugar kinase, and a MurQ etherase, respectively, are transcriptionally linked. Complementation of the Tanf_08375 and Tanf_08380 genes together in trans, but not individually rescued the inability of an E. coli mutant deficient in the PTS (phosphotransferase system)-dependent MurNAc transporter MurP as well as that of a double mutant deficient in MurP and components of the PTS system to grow on MurNAc. In addition, complementation with this two-gene construct in E. coli caused depletion of MurNAc in the medium, further confirming this observation. Our results show that the products of Tanf_08375 and Tanf_08380 constitute a novel non-PTS MurNAc transporter system that seems to be widespread among bacteria of the Bacteroidetes phylum. To the best of our knowledge, this is the first identification of a PTS-independent MurNAc transporter in bacteria. IMPORTANCE: In this study we report the identification of a novel transporter for peptidoglycan amino-sugar N-acetylmuramic acid (MurNAc) in the periodontal pathogen T. forsythia It has been known since the late 1980s that T. forsythia is a MurNAc auxotroph relying on environmental sources for this essential sugar. Most sugar transporters, and the MurNAc transporter MurP in particular require a PTS phosho-relay to drive the uptake and concurrent phosphorylation of the sugar through the inner membrane in Gram-negative bacteria. Our study uncovered a novel type of PTS-independent MurNAc transporter, and although so far unique to T. forsythia, may be present in a range of bacteria both of the oral cavity and gut especially of the phylum Bacteroidetes
Draft Genome Sequences of Three Clinical Isolates of Tannerella forsythia Isolated from Subgingival Plaque from Periodontitis Patients in the United States.
We report the genome sequences of three clinical isolates of Tannerella forsythia from the subgingival plaque of periodontitis patients attending clinics at the School of Dental Medicine, University at Buffalo. The availability of these genome sequences will aid the understanding of the pathogenesis of periodontitis
Cyclic vomiting syndrome in children: a nationwide survey of current practice on behalf of the Italian Society of Pediatric Gastroenterology, Hepatology and Nutrition (SIGENP) and Italian Society of Pediatric Neurology (SINP)
Background: Cyclic Vomiting Syndrome (CVS) is a rare functional gastrointestinal disorder, which has a considerable burden on quality of life of both children and their family. Aim of the study was to evaluate the diagnostic modalities and therapeutic approach to CVS among Italian tertiary care centers and the differences according to subspecialties, as well as to explore whether potential predictive factors associated with either a poor outcome or a response to a specific treatment. Methods: Cross-sectional multicenter web-based survey involving members of the Italian Society of Pediatric Gastroenterology, Hepatology and Nutrition (SIGENP) and Italian Society of Pediatric Neurology (SINP). Results: A total of 67 responses were received and analyzed. Most of the respondent units cared for less than 20 patients. More than half of the patients were referred after 3 to 5 episodes, and a quarter after 5 attacks. We report different diagnostic approaches among Italian clinicians, which was particularly evident when comparing gastroenterologists and neurologists. Moreover, our survey demonstrated a predilection of certain drugs during emetic phase according to specific clinic, which reflects the cultural background of physicians. Conclusion: In conclusion, our survey highlights poor consensus amongst clinicians in our country in the diagnosis and the management of children with CVS, raising the need for a national consensus guideline in order to standardize the practice
Extracellular matrix turnover and inflammation in chemically-induced TMJ arthritis mouse models.
The temporomandibular joint (TMJ) is a fibrocartilaginous tissue critical for chewing and speaking. In patients with temporomandibular disorders (TMDs), permanent tissue loss can occur. Recapitulating the complexity of TMDs in animal models is difficult, yet critical for the advent of new therapies. Synovial fluid from diseased human samples revealed elevated levels of tumor necrosis factor alpha (TNF-alpha). Here, we propose to recapitulate these findings in mice by subjecting murine TMJs with TNF-alpha or CFA (Complete Freund's Adjuvant) in mandibular condyle explant cultures and by local delivery in vivo using TMJ intra-articular injections. Both TNF-alpha and CFA delivery to whole mandibular explants and in vivo increased extracellular matrix deposition and increased cartilage thickness, while TNF-alpha treated explants had increased expression of inflammatory cytokines and degradative enzymes. Moreover, the application of TNF-alpha or CFA in both models reduced cell number. CFA delivery in vivo caused soft tissue inflammation, including pannus formation. Our work provides two methods of chemically induced TMJ inflammatory arthritis through a condyle explant model and intra-articular injection model that replicate findings seen in synovial fluid of human patients, which can be used for further studies delineating the mechanisms underlying TMJ pathology
[Immunological development in asymptomatic HIV infected children].
In HIV infected children, CD45+CD4+ T lymph. reconstitution has been related to efficient thymopoiesis. Because human thymus undergoes spontaneous involution at a relatively young age, institution of antiretroviral therapy early in the course of infection has been recommended. 12 HIV vertically infected children aged 4-8 years were investigated T-cell subsets for four years. 7 children were naive for therapy (group NT); 5 experienced nucleoside analogues only (group T). CD45RA+ and CD45RO+ CD4+ values were compared to predicted values of healthy children. The two groups showed similar clinical and immunological baseline characteristics (CDC class N-A). Mean VL at t0 was 4.26 log10 (SD 0.71) in gr. NT and 4.01 log10 (SD 0.57) in gr. T; median CD4 T lymph values were 27% in gr. NT and 23.5% in gr. T. Median CD45RA+ values were 62.8% in gr. NT and 71.3% in gr. T. No differences in VL, CD4+ T lymph., CD45RA+, CD45RO+ were found in between groups or within each group at each time evaluation. Median CD45RA+ values were not different from predicted values of healthy children. None of the children changed CDC class during the study period. Although the number of subjects was small, our study evidenced the possibility of a normal immunological development in HIV-1 vertically infected asymptomatic children naive for HAART during the first decade, even in the presence of significant viremia
Follow-up immunologico di bambini con infezione asintomatica da HIV
In HIV infected children, CD45+CD4+ T lymph. reconstitution has been related to efficient thymopoiesis. Because human thymus undergoes spontaneous involution at a relatively young age, institution of antiretroviral therapy early in the course of infection has been recommended. 12 HIV vertically infected children aged 4-8 years were investigated T-cell subsets for four years. 7 children were naive for therapy (group NT); 5 experienced nucleoside analogues only (group T). CD45RA+ and CD45RO+ CD4+ values were compared to predicted values of healthy children. The two groups showed similar clinical and immunological baseline characteristics (CDC class N-A). Mean VL at t0 was 4.26 log10 (SD 0.71) in gr. NT and 4.01 log10 (SD 0.57) in gr. T; median CD4 T lymph values were 27% in gr. NT and 23.5% in gr. T. Median CD45RA+ values were 62.8% in gr. NT and 71.3% in gr. T. No differences in VL, CD4+ T lymph., CD45RA+, CD45RO+ were found in between groups or within each group at each time evaluation. Median CD45RA+ values were not different from predicted values of healthy children. None of the children changed CDC class during the study period. Although the number of subjects was small, our study evidenced the possibility of a normal immunological development in HIV-1 vertically infected asymptomatic children naive for HAART during the first decade, even in the presence of significant viremia
Clinical and immunological effects of the long term dietary supplementation with a probiotic fermented milk in pre-school children allergic to inhalants
Background and Aims: Probiotics may have
immunomodulatory functions, exerting beneficial
effects in allergic disease. We investigated whether
prolonged daily consumption of Lactobacillus casei DN-
114 001 fermented milk improve clinical and
immunological condition of children aged 3-6 years
allergic to inhalants.
Methods: In a randomised-controlled multicentric trial
patients were randomly assigned to receive for one year
100 ml per day of L. casei DN-114 001 fermented milk
(109 cells/ml), or no probiotics (controls). Clinical
evaluations occurred every 3 months. Faecal flora
composition was assessed every 6 months (30 treated,
15 controls). Outcome measures were allergy-related
signs incidence, infections incidence/severity, IgE-G-A
serological levels (baseline vs 12 months).
Results: Participants (187; 92 treated) were similar
regarding gestational age, breastfeeding, family
smokers, pets, siblings, day-care admission.
Supplemented children exibited less rhinitis episodes
than controls up to T6 [mean-SD: 0,6 (1) vs 0,9 (1,1) at
T3, p= 0,031; 1 (2,6) vs 1,6 (2) at T6, p=0,027]. Asthma
and infections incidence/severity did not differ between
groups. IgE mean % variation increased in controls at
T12 vs baseline (+ 25%; p< 0,01) but not in the treated
group (+8%; p=0,59). IgG and IgA mean % variation
significantly increased in treated group (+6,3% and
+11,5% respectively; p< 0,01) but not in controls (2,4%
and 3,7% respectively; p=0,34 and 0,19). Faecal
analysis showed Lactobacillus casei DN-114 001 in the
gut flora of 78% of supplemented children through the
study period.
Conclusions: Long-term daily consumption of
Lactobacillus Casei DN-114 001 fermented milk may
positively influence on the clinical and immunological
status of allergic childre
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