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Magnetic fields have an important role in the evolution of interstellar medium and star formation1,2. As the only direct probe of interstellar field strength, credible Zeeman measurements remain sparse owing to the lack of suitable Zeeman probes, particularly for cold, molecular gas3. Here we report the detection of a magnetic field of +3.8 ± 0.3 microgauss through the H I narrow self-absorption (HINSA)4,5 towards L15446,7-a well-studied prototypical prestellar core in an early transition between starless and protostellar phases8-10 characterized by a high central number density11 and a low central temperature12. A combined analysis of the Zeeman measurements of quasar H I absorption, H I emission, OH emission and HINSA reveals a coherent magnetic field from the atomic cold neutral medium (CNM) to the molecular envelope. The molecular envelope traced by the HINSA is found to be magnetically supercritical, with a field strength comparable to that of the surrounding diffuse, magnetically subcritical CNM despite a large increase in density. The reduction of the magnetic flux relative to the mass, which is necessary for star formation, thus seems to have already happened during the transition from the diffuse CNM to the molecular gas traced by the HINSA. This is earlier than envisioned in the classical picture where magnetically supercritical cores capable of collapsing into stars form out of magnetically subcritical envelopes13,14.
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Regulation of the first committed step of peptidoglycan precursor synthesis by MurA-enzyme homologs has recently taken center stage in many different bacteria. In different low-GC Gram-positive bacteria, regulation of this step has been shown to be regulated by phosphorylation of homologs of the IreB/ReoM regulatory protein by PASTA-domain Ser/Thr-protein kinases. In this issue, Mascari, Little, and Kristich determine this regulatory pathway and its links to resistance to cephalosporin ß-lactam antibiotics in the major human pathogen, Enterococcus faecalis (Efa). Unbiased genetic selections identified MurAA (MurA-family homolog) as the downstream target of IreB regulation in the absence of the IreK Ser/Thr-protein kinase. Physiological and biochemical approaches, including determination of MICs to ceftriaxone, Western blotting of MurAA cellular amounts, isotope incorporation into peptidoglycan sacculi, and thermal-shift binding assays of purified proteins, demonstrated that unphosphorylated IreB, together with proteins MurAB (MurZ-family homolog), and ReoY(Efa) negatively regulate MurAA stability and cellular amount by the ClpCP protease. Importantly, this paper supports the idea that ceftriaxone stimulates phosphorylation of IreB, which leads to increased cellular MurAA amount and precursor pathway flux required for E. faecalis cephalosporin resistance. Overall, findings in this paper significantly contribute to understanding variations of this central regulatory pathway in other low-GC Gram-positive bacteria.
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Ceftriaxona , Enterococcus , Humanos , Fosforilação , Enterococcus/metabolismo , Peptidoglicano/metabolismo , Enterococcus faecalis/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismoRESUMO
RodZ of rod-shaped bacteria functions to link MreB filaments to the Rod peptidoglycan (PG) synthase complex that moves circumferentially perpendicular to the long cell axis, creating hoop-like sidewall PG. Ovoid-shaped bacteria, such as Streptococcus pneumoniae (pneumococcus; Spn) that lack MreB, use a different modality for peripheral PG elongation that emanates from the midcell of dividing cells. Yet, S. pneumoniae encodes a RodZ homolog similar to RodZ in rod-shaped bacteria. We show here that the helix-turn-helix and transmembrane domains of RodZ(Spn) are essential for growth at 37°C. ΔrodZ mutations are suppressed by Δpbp1a, mpgA(Y488D), and ΔkhpA mutations that suppress ΔmreC, but not ΔcozE. Consistent with a role in PG elongation, RodZ(Spn) co-localizes with MreC and aPBP1a throughout the cell cycle and forms complexes and interacts with PG elongasome proteins and regulators. Depletion of RodZ(Spn) results in aberrantly shaped, non-growing cells and mislocalization of elongasome proteins MreC, PBP2b, and RodA. Moreover, Tn-seq reveals that RodZ(Spn), but not MreCD(Spn), displays a specific synthetic-viable genetic relationship with aPBP1b, whose function is unknown. We conclude that RodZ(Spn) acts as a scaffolding protein required for elongasome assembly and function and that aPBP1b, like aPBP1a, plays a role in elongasome regulation and possibly peripheral PG synthesis.
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Peptidoglicano , Streptococcus pneumoniae , Peptidoglicano/metabolismo , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Parede Celular/metabolismo , Divisão Celular/genéticaRESUMO
Bacterial peptidoglycan (PG) synthesis requires strict spatiotemporal organization to reproduce specific cell shapes. In ovoid-shaped Streptococcus pneumoniae (Spn), septal and peripheral (elongation) PG synthesis occur simultaneously at midcell. To uncover the organization of proteins and activities that carry out these two modes of PG synthesis, we examined Spn cells vertically oriented onto their poles to image the division plane at the high lateral resolution of 3D-SIM (structured-illumination microscopy). Labeling with fluorescent D-amino acids (FDAA) showed that areas of new transpeptidase (TP) activity catalyzed by penicillin-binding proteins (PBPs) separate into a pair of concentric rings early in division, representing peripheral PG (pPG) synthesis (outer ring) and the leading-edge (inner ring) of septal PG (sPG) synthesis. Fluorescently tagged PBP2x or FtsZ locate primarily to the inner FDAA-marked ring, whereas PBP2b and FtsX remain in the outer ring, suggesting roles in sPG or pPG synthesis, respectively. Pulses of FDAA labeling revealed an arrangement of separate regularly spaced "nodes" of TP activity around the division site of predivisional cells. Tagged PBP2x, PBP2b, and FtsX proteins also exhibited nodal patterns with spacing comparable to that of FDAA labeling. Together, these results reveal new aspects of spatially ordered PG synthesis in ovococcal bacteria during cell division.
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Divisão Celular/fisiologia , Peptidoglicano/biossíntese , Streptococcus pneumoniae/metabolismo , Aminoaciltransferases/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Ciclo Celular/metabolismo , Corantes Fluorescentes , Proteínas de Ligação às Penicilinas/metabolismo , Peptidil Transferases/metabolismo , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/crescimento & desenvolvimentoRESUMO
Bacterial cell division and peptidoglycan (PG) synthesis are orchestrated by the coordinated dynamic movement of essential protein complexes. Recent studies show that bidirectional treadmilling of FtsZ filaments/bundles is tightly coupled to and limiting for both septal PG synthesis and septum closure in some bacteria, but not in others. Here we report the dynamics of FtsZ movement leading to septal and equatorial ring formation in the ovoid-shaped pathogen, Streptococcus pneumoniae Conventional and single-molecule total internal reflection fluorescence microscopy (TIRFm) showed that nascent rings of FtsZ and its anchoring and stabilizing proteins FtsA and EzrA move out from mature septal rings coincident with MapZ rings early in cell division. This mode of continuous nascent ring movement contrasts with a failsafe streaming mechanism of FtsZ/FtsA/EzrA observed in a ΔmapZ mutant and another Streptococcus species. This analysis also provides several parameters of FtsZ treadmilling in nascent and mature rings, including treadmilling velocity in wild-type cells and ftsZ(GTPase) mutants, lifetimes of FtsZ subunits in filaments and of entire FtsZ filaments/bundles, and the processivity length of treadmilling of FtsZ filament/bundles. In addition, we delineated the motion of the septal PBP2x transpeptidase and its FtsW glycosyl transferase-binding partner relative to FtsZ treadmilling in S. pneumoniae cells. Five lines of evidence support the conclusion that movement of the bPBP2x:FtsW complex in septa depends on PG synthesis and not on FtsZ treadmilling. Together, these results support a model in which FtsZ dynamics and associations organize and distribute septal PG synthesis, but do not control its rate in S. pneumoniae.
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Proteínas de Bactérias/genética , Proteínas de Membrana/genética , Proteínas de Ligação às Penicilinas/genética , Infecções Pneumocócicas/microbiologia , Streptococcus pneumoniae/genética , Divisão Celular/genética , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/ultraestrutura , Citoesqueleto/genética , Citoesqueleto/ultraestrutura , Escherichia coli/genética , GTP Fosfo-Hidrolases/genética , Humanos , Microscopia de Fluorescência , Peptidoglicano/biossíntese , Peptidoglicano/genética , Infecções Pneumocócicas/genética , Streptococcus pneumoniae/patogenicidade , Streptococcus pneumoniae/ultraestruturaRESUMO
Human milk is a complex and variable ecosystem fundamental to the development of newborns. This study aimed to investigate relationships between human milk oligosaccharides (HMO) and human milk bacterial profiles and infant body composition. Human milk samples (n = 60) were collected at two months postpartum. Infant and maternal body composition was measured with bioimpedance spectroscopy. Human milk bacterial profiles were assessed using full-length 16S rRNA gene sequencing and 19 HMOs were quantitated using high-performance liquid chromatography. Relative abundance of human milk bacterial taxa were significantly associated with concentrations of several fucosylated and sialylated HMOs. Individual human milk bacteria and HMO intakes and concentrations were also significantly associated with infant anthropometry, fat-free mass, and adiposity. Furthermore, when data were stratified based on maternal secretor status, some of these relationships differed significantly among infants born to secretor vs non-secretor mothers. In conclusion, in this pilot study the human milk bacterial profile and HMO intakes and concentrations were significantly associated with infant body composition, with associations modified by secretor status. Future research designed to increase the understanding of the mechanisms by which HMO and human milk bacteria modulate infant body composition should include intakes in addition to concentrations.
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Aleitamento Materno , Leite Humano , Bactérias/genética , Composição Corporal , Ecossistema , Feminino , Humanos , Lactente , Recém-Nascido , Lactação , Leite Humano/química , Oligossacarídeos/química , Projetos Piloto , RNA Ribossômico 16S/análise , RNA Ribossômico 16S/genéticaRESUMO
BACKGROUND: To investigate relationships between infant body composition (BC) and human milk (HM) immunomodulatory proteins (IMPs) during the first 12 months of lactation. METHODS: BC of breastfeeding dyads (n = 20) was measured with ultrasound skinfolds (infants) and bioimpedance spectroscopy (infants/mothers) at 2, 5, 9, and/or 12 months post partum. Breastfeeding frequency, 24-h milk intake, and IMP concentrations (lactoferrin, lysozyme, secretory immunoglobulin A (sIgA)) were measured, and calculated daily intakes (CDIs) were determined. We used linear regression/mixed-effects models and adjusted results for multiple comparisons. RESULTS: No associations were seen between maternal characteristics and IMP concentrations/CDIs or between IMP concentrations and infant BC. Lactoferrin CDI was negatively associated with infant fat-free mass index (P = 0.002); lysozyme CDI was positively associated with infant fat mass (P = 0.004) and fat mass index (P = 0.004) measured with ultrasound skinfolds. CONCLUSION: In this small cohort of infants breastfed on demand during first year of life, we report differential associations of HM IMPs with infant BC, showing that in addition to their critical role in shaping infant immunity, lactoferrin, and lysozyme also influence development of infant BC, highlighting the importance of breastfeeding for 12 months and beyond. IMPACT: HM IMPs (concentrations and, most importantly, daily intakes) time-dependently and differentially associate with development of infant lean mass and adiposity during first 12 months of lactation. There is no information on how intakes and concentrations of these components affect development of infant BC. HM contains IMPs-lactoferrin, lysozyme, and sIgA, which not only play a critical role in shaping infant's immunity, but also influence infant growth and development of BC, highlighting the importance of breastfeeding for 12 months and beyond and warranting careful consideration of the dose effects of supplemented formula.
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Composição Corporal , Lactação/imunologia , Leite Humano/química , Leite Humano/imunologia , Adiposidade , Antropometria , Austrália , Índice de Massa Corporal , Aleitamento Materno , Espectroscopia Dielétrica , Feminino , Humanos , Imunoglobulina A/sangue , Lactente , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido , Lactoferrina/sangue , Estudos Longitudinais , Muramidase/sangue , Projetos Piloto , Análise de Regressão , Fatores de Risco , UltrassonografiaRESUMO
PEACH is an important tool for evaluation of children's hearing development, used in age 2-7 years. It is also appropriate for amplification outcomes measurements. PEACH scale includes 13 questions. Parents fill the questionnaire after week observation of child's hearing behavior in different situations. The goal of the study was validation of Russian version of PEACH scale. Translation and cross-cultural adaptation were performed following international guidelines. 50 children with normal hearing and 50 hearing impaired children were involved in the validation process. All of the hearing-impaired children used hearing aids or cochlear implants. PEACH scores of the children with normal hearing have strong correlation with data of original version (ρ=0.998; p<0.05) and can be used as a normative data for Russian version. PEACH scores of the hearing-impaired children were worse in higher degrees of hearing loss, which shows sensitivity of the method. Test-retest reliability in children with normal hearing was ρ=1.0 (p<0.05), in hearing impaired children ρ=0.976 (p<0.05). Russian PEACH scale is free available at the official site of Center of Pediatric Audiology: https://dgsc.kzdrav.gov.spb.ru.
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Auxiliares de Audição , Prunus persica , Criança , Pré-Escolar , Humanos , Reprodutibilidade dos Testes , Federação Russa , Inquéritos e Questionários , TraduçõesRESUMO
Prolonged osteochondral tissue damage can result in osteoarthritis and decreased quality of life. Multiphasic scaffolds, where different layers model different microenvironments, are a promising treatment approach, yet stable joining between layers during fabrication remains challenging. Here, a bilayer scaffold for osteochondral tissue regeneration was fabricated using thermally-induced phase separation (TIPS). Two distinct polymer solutions were layered before TIPS, and the resulting porous, bilayer scaffold was characterized by seamless interfacial integration and a mechanical stiffness gradient reflecting the native osteochondral microenvironment. Chitosan is a critical component of both scaffold layers to facilitate cell attachment and the formation of polyelectrolyte complexes with other biologically relevant natural polymers. The articular cartilage region was optimized for hyaluronic acid content and stiffness, while the subchondral bone region was defined by higher stiffness and osteoconductive hydroxyapatite content. Following co-culture with chondrocyte-like (SW-1353 or mesenchymal stem cells) and osteoblast-like cells (MG63), cell proliferation and migration to the interface along with increased gene expression associated with relevant markers of osteogenesis and chondrogenesis indicates the potential of this bilayer scaffold for osteochondral tissue regeneration.
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Osso e Ossos/fisiologia , Cartilagem Articular/fisiologia , Quitosana/química , Quitosana/farmacologia , Regeneração/efeitos dos fármacos , Alicerces Teciduais/química , Alginatos/química , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Osso e Ossos/citologia , Osso e Ossos/efeitos dos fármacos , Cartilagem Articular/citologia , Cartilagem Articular/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Durapatita/química , Humanos , Fenômenos Mecânicos , Engenharia TecidualRESUMO
This study examined if plant sterols and walking reduce postprandial triacylglycerol (TAG) concentrations in Chinese men with elevated body mass index (≥ 23.5 kg/m2). Fifteen Chinese men (mean [SD]: age = 25 [3] years and body mass index = 26.2 [1.5] kg/m2] completed four 10-day trials in random order with a 7- to 10-day washout between trials: (a) daily consumption of a control margarine while sedentary (C-S), (b) daily consumption of margarine containing 2 g/day of plant sterols while sedentary (PS-S), (c) daily consumption of a control margarine with 30-min daily walking (C-W), and (d) daily consumption of margarine containing 2 g/day of plant sterols with 30-min daily walking (PS-W). On Day 11 of each trial, postprandial TAG was measured after a high-fat milkshake. The 5-hr total area under the TAG curve was 22%, 25%, and 12% lower on PS-W (mean [SD]: 8.9 [4.3] mmol·5 hr/L) than C-S (11.4 [4.5] mmol·5 hr/L; p = .005; d = 0.56), PS-S (11.9 [4.9] mmol·5 hr/L; p = .004; d = 0.67), and C-W (10.1 [4.4] mmol·5 hr/L; p = .044; d = 0.27) trials, respectively. Similarly, 5-hr incremental area for PS-W (4.5 [2.7] mmol·5 hr/L) was 31%, 32%, and 18% lower than C-S (6.6 [3.3] mmol·5 hr/L; p = .005; d = 0.62), PS-S (6.6 [3.4] mmol·5 hr/L; p = .004; d = 0.64), and C-W (5.5 [2.8] mmol·5 hr/L; p = .032; d = 0.29). Ten days of daily plant sterol intake combined with walking presents an intervention strategy to lower postprandial TAG in Chinese men with elevated body mass index.
Assuntos
Fitosteróis/administração & dosagem , Período Pós-Prandial , Triglicerídeos/sangue , Caminhada , Adulto , Povo Asiático , Índice de Massa Corporal , China , Humanos , Masculino , Adulto JovemRESUMO
GpsB regulatory protein and StkP protein kinase have been proposed as molecular switches that balance septal and peripheral (side-wall like) peptidoglycan (PG) synthesis in Streptococcus pneumoniae (pneumococcus); yet, mechanisms of this switching remain unknown. We report that ΔdivIVA mutations are not epistatic to ΔgpsB division-protein mutations in progenitor D39 and related genetic backgrounds; nor is GpsB required for StkP localization or FDAA labeling at septal division rings. However, we confirm that reduction of GpsB amount leads to decreased protein phosphorylation by StkP and report that the essentiality of ΔgpsB mutations is suppressed by inactivation of PhpP protein phosphatase, which concomitantly restores protein phosphorylation levels. ΔgpsB mutations are also suppressed by other classes of mutations, including one that eliminates protein phosphorylation and may alter division. Moreover, ΔgpsB mutations are synthetically lethal with Δpbp1a, but not Δpbp2a or Δpbp1b mutations, suggesting GpsB activation of PBP2a activity. Consistent with this result, co-IP experiments showed that GpsB complexes with EzrA, StkP, PBP2a, PBP2b and MreC in pneumococcal cells. Furthermore, depletion of GpsB prevents PBP2x migration to septal centers. These results support a model in which GpsB negatively regulates peripheral PG synthesis by PBP2b and positively regulates septal ring closure through its interactions with StkP-PBP2x.
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Aminoaciltransferases/genética , Proteínas de Bactérias/genética , Divisão Celular/genética , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas de Ligação às Penicilinas/genética , Peptidoglicano/biossíntese , Streptococcus pneumoniae/genética , Fatores de Virulência/genética , Aminoaciltransferases/metabolismo , Proteínas de Bactérias/metabolismo , Composição de Bases/genética , Divisão Celular/fisiologia , Parede Celular/metabolismo , Mutação/genética , Proteínas de Ligação às Penicilinas/metabolismo , Fosforilação , Streptococcus pneumoniae/metabolismo , Fatores de Virulência/metabolismoRESUMO
BACKGROUND: This study examined language development in young children with hearing loss and different types of additional disabilities (ADs). METHOD: A population-based cohort of 67 children who were enrolled in the Longitudinal Outcomes of Children with Hearing Impairment study took part. Language ability was directly assessed at 3 and 5 years of age using the Preschool Language Scale, Fourth Edition and the Peabody Picture Vocabulary Test, Fourth Edition. Standard scores were used to enable comparison with age-based expectations for typically developing children. RESULTS: Analysis of variance showed that, across the total cohort, children's language scores remained stable over the 2-year period. However, this overall stability masked a significant difference between children with different types of ADs; in particular, children with autism, cerebral palsy and/or developmental delay showed a decline in standard scores, whereas children with other disabilities showed a relative improvement. In addition, larger improvements in receptive vocabulary were associated with use of oral communication only. CONCLUSIONS: The results suggest that type of AD can be used to gauge expected language development in the population of children with hearing loss and ADs when formal assessment of cognitive ability is not feasible.
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Surdez/epidemiologia , Crianças com Deficiência/estatística & dados numéricos , Transtornos do Desenvolvimento da Linguagem/epidemiologia , Linguagem Infantil , Pré-Escolar , Estudos de Coortes , Comorbidade , Feminino , Seguimentos , Humanos , Desenvolvimento da Linguagem , Masculino , New South Wales/epidemiologiaRESUMO
Human milk (HM) contains a complex array of hormones, including members of the glucocorticoid family. The predominant glucocorticoids, cortisol and cortisone may influence the growth and behaviour of the breastfed infant. However, little is understood of the factors regulating the levels of these hormones within HM. The aim of the study was to examine HM cortisol and cortisone concentration, measured in samples collected at each feed during a 24 hour period. Twenty three exclusively breastfeeding mothers collected milk, prior to and after each breastfeeding session over 24 hour period at 3.2(1.60) months. HM cortisol and cortisone levels were measured using high pressure liquid chromatography mass spectroscopy. Cortisone was the predominant glucocorticoid (3.40 ng/ml), and cortisol was detected in all samples (1.62 ng/ml). A positive correlation was found between cortisone and cortisol (r = 0.61, y = 1.93 ± 0.24, p < 0.0001). Cortisol and cortisone concentrations were significantly higher in feeds in the morning (2.97 ng/ml and 4.88 ng/ml), compared to afternoon (1.20 ng/ml and 3.54 ng/ml), evening (0.69 ng/ml and 2.13 ng/ml) and night (1.59 and 3.27 ng/ml). No difference was found between glucocorticoids level of the milk expressed for collection either before or immediately after the breastfeed, or between milk collected from the left or right breast. This study shows that HM glucocorticoid concentrations exhibit a 24 hour pattern, with highest peak levels in the early morning, reflecting the circadian pattern as previously reported in plasma. Thus, HM glucocorticoid concentrations are likely to reflect those in the maternal circulation.
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Glucocorticoides/química , Glucocorticoides/metabolismo , Leite Humano/química , Adulto , Aleitamento Materno/métodos , Cortisona/química , Cortisona/metabolismo , Feminino , Humanos , Hidrocortisona/química , Hidrocortisona/metabolismoRESUMO
Streptococcus pneumoniae is an ovoid-shaped Gram-positive bacterium that grows by carrying out peripheral and septal peptidoglycan (PG) synthesis, analogous to model bacilli, such as Escherichia coli and Bacillus subtilis In the model bacilli, FtsZ and FtsA proteins assemble into a ring at midcell and are dedicated to septal PG synthesis but not peripheral PG synthesis; hence, inactivation of FtsZ or FtsA results in long filamentous cells unable to divide. Here, we demonstrate that FtsA and FtsZ colocalize at midcell in S. pneumoniae and that partial depletion of FtsA perturbs septum synthesis, resulting in elongated cells with multiple FtsZ rings that fail to complete septation. Unexpectedly, complete depletion of FtsA resulted in the delocalization of FtsZ rings and ultimately cell ballooning and lysis. In contrast, depletion or deletion of gpsB and sepF, which in B. subtilis are synthetically lethal with ftsA, resulted in enlarged and elongated cells with multiple FtsZ rings, with deletion of sepF mimicking partial depletion of FtsA. Notably, cell ballooning was not observed, consistent with later recruitment of these proteins to midcell after Z-ring assembly. The overproduction of FtsA stimulates septation and suppresses the cell division defects caused by the deletion of sepF and gpsB under some conditions, supporting the notion that FtsA shares overlapping functions with GpsB and SepF at later steps in the division process. Our results indicate that, in S. pneumoniae, both GpsB and SepF are involved in septal PG synthesis, whereas FtsA and FtsZ coordinate both peripheral and septal PG synthesis and are codependent for localization at midcell.IMPORTANCEStreptococcus pneumoniae (pneumococcus) is a clinically important human pathogen for which more therapies against unexploited essential targets, like cell growth and division proteins, are needed. Pneumococcus is an ovoid-shaped Gram-positive bacterium with cell growth and division properties that have important distinctions from those of rod-shaped bacteria. Gaining insights into these processes can thus provide valuable information to develop novel antimicrobials. Whereas rods use distinctly localized protein machines at different cellular locations to synthesize peripheral and septal peptidoglycans, we present evidence that S. pneumoniae organizes these two machines at a single location in the middle of dividing cells. Here, we focus on the properties of the actin-like protein FtsA as an essential orchestrator of peripheral and septal growth in this bacterium.
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BACKGROUND: Accurate, noninvasive, and inexpensive methods are required to measure infant body composition. Ultrasound (US) and bioimpedance spectroscopy (BIS) have been validated in adults and introduced in pediatric populations. The aim of this study was to evaluate the performance of both methods in determining percentage fat mass (%FM) in breastfed infants. METHODS: %FM of 2, 5, 9, and 12 mo-old healthy, breastfed term infants (n = 58) was calculated using BIS-derived total body water equations and skinfold equations then compared with reference models. Skinfolds were measured with US at two and four sites (biceps, suprailiac and/or triceps, and subscapular). RESULTS: %FM differed widely within and between methods, with the degree of variation affected by infant age/sex. Not a single method/equation was consistent with the distributions of appropriate reference values for all age/sex groups. Moderate number of matches with references values (13-24 out of 36) was seen for both types of equations. High number of matches (25-36) was seen for US skinfold-based equations. %FM values calculated from US and BIS were not significantly different (P = 0.35). CONCLUSION: Both BIS and US are practical for predicting %FM in infants. BIS calculations are highly dependent upon an appropriate set of validated age-matched equations.
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Tecido Adiposo/diagnóstico por imagem , Antropometria/métodos , Composição Corporal , Aleitamento Materno , Espectroscopia Dielétrica , Índice de Massa Corporal , Feminino , Humanos , Lactente , Masculino , Obesidade Infantil/diagnóstico por imagem , Valores de Referência , Reprodutibilidade dos Testes , Dobras Cutâneas , UltrassonografiaRESUMO
BackgroundInfants breastfed on demand exhibit a variety of feeding patterns and self-regulate their nutrient intake, but factors influencing their gastric emptying (GE) are poorly understood. Despite research into appetite regulation properties of leptin, there is limited information about relationships between human milk leptin and infant GE.MethodsGastric volumes were calculated from ultrasound scans of infants' stomachs (n=20) taken before and after breastfeeding, and then every 12.5 min (median; range: 3-45 min) until the next feed. Skim milk leptin and macronutrient concentrations were measured and doses were calculated.ResultsThe leptin concentration was (mean±SD) 0.51±0.16 ng/ml; the leptin dose was 45.5±20.5 ng per feed. No relationships between both concentration and dose of leptin and time between the feeds (P=0.57; P=1, respectively) or residual stomach volumes before the subsequent feed (P=0.20; P=0.050) were found. Post-feed stomach volumes (GE rate) were not associated with leptin concentration (P=0.77) or dose (P=0.85).ConclusionGE in term breastfed infants was not associated with either skim milk leptin concentration or dose. Further investigation with inclusion of whole-milk leptin and other hormones that affect gastrointestinal activity is warranted.
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Aleitamento Materno , Alimentos , Esvaziamento Gástrico , Leptina/química , Leite Humano/química , Ingestão de Energia , Comportamento Alimentar , Feminino , Humanos , Lactente , Recém-Nascido , Mães , Estômago/fisiologia , Fatores de TempoRESUMO
The emergence of methicillin-resistant Staphylococcus aureus (MRSA) infections with multi-drug resistance needs effective and alternative control strategies. In this study we investigated the adjuvant effect of a novel furan fatty acid, 7,10-epoxyoctadeca-7,9-dienoic acid (7,10-EODA) against multidrug-resistant S. aureus (MDRSA) strain 01ST001 by disc diffusion, checker board and time kill assays. Further the membrane targeting action of 7,10-EODA was investigated by spectroscopic and confocal microscopic studies. 7,10-EODA exerted synergistic activity along with ß-lactam antibiotics against all clinical MRSA strains, with a mean fractional inhibitory concentration index below 0.5. In time-kill kinetic study, combination of 7,10-EODA with oxacillin, ampicillin, and penicillin resulted in 3.8-4.2 log10 reduction in the viable counts of MDRSA 01ST001. Further, 7,10-EODA dose dependently altered the membrane integrity (p < 0.001) and increased the binding of fluorescent analog of penicillin, Bocillin-FL to the MDRSA cells. The membrane action of 7,10-EODA further facilitated the uptake of several other antibiotics in MDRSA. The results of the present study suggested that 7,10-EODA could be a novel antibiotic adjuvant, especially useful in repurposing ß-lactam antibiotics against multidrug-resistant MRSA.
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The Phr peptides of the Bacillus species mediate quorum sensing, but their identification and function in other species of bacteria have not been determined. We have identified a Phr peptide quorum-sensing system (TprA/PhrA) that controls the expression of a lantibiotic gene cluster in the Gram-positive human pathogen, Streptococcus pneumoniae. Lantibiotics are highly modified peptides that are part of the bacteriocin family of antimicrobial peptides. We have characterized the basic mechanism for a Phr-peptide signaling system in S. pneumoniae and found that it induces the expression of the lantibiotic genes when pneumococcal cells are at high density in the presence of galactose, a main sugar of the human nasopharynx, a highly competitive microbial environment. Activity of the Phr peptide system is not seen when pneumococcal cells are grown with glucose, the preferred carbon source and the most prevalent sugar encountered by S. pneumoniae during invasive disease. Thus, the lantibiotic genes are expressed under the control of both cell density signals via the Phr peptide system and nutritional signals from the carbon source present, suggesting that quorum sensing and the lantibiotic machinery may help pneumococcal cells compete for space and resources during colonization of the nasopharynx.
Assuntos
Bacteriocinas/biossíntese , Bacteriocinas/genética , Regulação Bacteriana da Expressão Gênica , Família Multigênica , Percepção de Quorum/fisiologia , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/metabolismo , Bacillus/genética , Bacillus/metabolismo , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/genética , Percepção de Quorum/genéticaRESUMO
The relative localization patterns of class B penicillin-binding proteins Pbp2x and Pbp2b were used as positional indicators of septal and peripheral (side-wall-like) peptidoglycan (PG) synthesis, respectively, in the mid-cell regions of Streptococcus pneumoniae cells at different stages of division. We confirm that Pbp2x and Pbp2b are essential in the strain D39 genetic background, which differs from that of laboratory strains. We show that Pbp2b, like Pbp2x and class A Pbp1a, follows a different localization pattern than FtsZ and remains at division septa after FtsZ reappears at the equators of daughter cells. Pulse-experiments with fluorescent D-amino acids (FDAAs) were performed in wild-type cells and in cells in which Pbp2x activity was preferentially inhibited by methicillin or Pbp2x amount was depleted. These experiments show that Pbp2x activity separates from that of other PBPs to the centres of constricting septa in mid-to-late divisional cells resolved by high-resolution 3D-SIM microscopy. Dual-protein and protein-fluorescent vancomycin 2D and 3D-SIM immunofluorescence microscopy (IFM) of cells at different division stages corroborate that Pbp2x separates to the centres of septa surrounded by an adjacent constricting ring containing Pbp2b, Pbp1a and regulators, StkP and MreC. The separate localization of Pbp2x suggests distinctive roles in completing septal PG synthesis and remodelling.
Assuntos
Divisão Celular , Proteínas de Ligação às Penicilinas/metabolismo , Peptidoglicano/biossíntese , Streptococcus pneumoniae/citologia , Proteínas de Ligação às Penicilinas/genética , Transporte Proteico , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/metabolismoRESUMO
Selective fluorescent ß-lactam chemical probes enable the visualization of the transpeptidase activity of penicillin-binding proteins (PBPs) at different stages of bacterial cell division. To facilitate the development of new fluorescent probes for PBP imaging, we evaluated 20 commercially available ß-lactams for selective PBP inhibition in an unencapsulated derivative of the D39 strain of Streptococcus pneumoniae. Live cells were treated with ß-lactam antibiotics at different concentrations and subsequently incubated with Bocillin FL (Boc-FL; fluorescent penicillin) to saturate uninhibited PBPs. Fluorophore-labeled PBPs were visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorescence scanning. Among 20 compounds tested, carbapenems (doripenem and meropenem) were coselective for PBP1a, PBP2x, and PBP3, while six of the nine penicillin compounds were coselective for PBP2x and PBP3. In contrast, the seven cephalosporin compounds tested display variability in their PBP-binding profiles. Three cephalosporin compounds (cefoxitin, cephalexin, and cefsulodin) and the monobactam aztreonam exhibited selectivity for PBP3, while only cefuroxime (a cephalosporin) was selective for PBP2x. Treatment of S. pneumoniae cultures with a sublethal concentration of cefuroxime that inhibited 60% of PBP2x activity and less than 20% of the activity of other PBPs resulted in formation of elongated cells. In contrast, treatment of S. pneumoniae cultures with concentrations of aztreonam and cefoxitin that inhibited up to 70% of PBP3 activity and less than 30% of other PBPs resulted in no discernible morphological changes. Additionally, correlation of the MIC and IC50s for each PBP, with the exception of faropenem, amdinocillin (mecillinam), and 6-APA, suggests that pneumococcal growth inhibition is primarily due to the inhibition of PBP2x.