Rhamnogalacturonan I with ß-(1,4)-Galactan Side Chains as an Ever-Present Component of Tertiary Cell Wall of Plant Fibers.

The cellulose-enriched tertiary cell walls present in many plant fibers have specific composition, architecture, machinery of formation, and function. To better understand the mechanisms underlying their mode of action and to reveal the peculiarities of fibers from different plant species, it is necessary to more deeply characterize the major components. Next to overwhelming cellulose, rhamnogalacturonan I (RG-I) is considered to be the key polymer of the tertiary cell wall; however, it has been isolated and biochemically characterized in very few plant species. Here, we add RG-I to the list from the phloem fibers of the Phaseolus vulgaris stem that was isolated and analyzed by nuclear magnetic resonance (NMR), dynamic light scattering, and immunolabeling, both within tissue and as an isolated polymer. Additionally, fibers with tertiary cell walls from nine species of dicotyledonous plants from the orders Malphigiales, Fabales, and Rosales were labeled with RG-I-related antibodies to check the presence of the polymer and compare the in situ presentation of its backbone and side chains. The obtained results confirm that RG-I is an obligatory polymer of the tertiary cell wall. However, there are differences in the structure of this polymer from various plant sources, and these peculiarities may be taxonomically related.

Dynamics of cell wall polysaccharides during the elongation growth of rye primary roots.

MAIN CONCLUSION: In cells of growing rye roots, xyloglucans and homogalacturonans demonstrate developmental stage specificity, while different xylans have tissue specificity. Mannans, arabinans and galactans are also detected within the protoplast. Mannans form films on sections of fresh material. The primary cell walls of plants represent supramolecular exocellular structures that are mainly composed of polysaccharides. Cell wall properties and architecture differ between species and across tissues within a species. We revised the distribution of cell wall polysaccharides and their dynamics during elongation growth and histogenesis in rye roots using nonfixed material and the spectrum of antibodies. Rye is a member of the Poaceae family and thus has so-called type II primary cell walls, which are supposed to be low in pectins and xyloglucans and instead have arabinoxylans and mixed-linkage glucans. However, rye cell walls at the earliest stages of cell development were enriched with the epitopes of xyloglucans and homogalacturonans. Mixed-linkage glucan, which is often considered an elongation growth-specific polysaccharide in plants with type II cell walls, did not display such dynamics in rye roots. The cessation of elongation growth and even the emergence of root hairs were not accompanied by the disappearance of mixed-linkage glucans from cell walls. The diversity of xylan motifs recognized by different antibodies was minimal in the meristem zone of rye roots, but this diversity increased and showed tissue specificity during root growth. Antibodies specific for xyloglucans, galactans, arabinans and mannans bound the cell content. When rye root cells were cut, the epitopes of xyloglucans, galactans and arabinans remained within the cell content, while mannans developed net-like or film-like structures on the surface of sections.

Direct antiglobulin test in the prediction of hyperbilirubinemia and predischarge bilirubin levels in infants with mother-infant blood type incompatibility.

BACKGROUND: This research evaluated the association between the mother-infant blood type or rhesus (ABO or Rh) incompatibility, the pattern of neonatal jaundice, and serum bilirubin (TSB) values obtained prior to discharge from hospital of healthy born neonates with gestational age >34 weeks and birth weight >2000 g. METHODS: We utilized a laboratory and neonatal database to identify the cord blood ABO/Rh and direct antiglobulin test (DAT) and TSB measured during hospitalization and re-admission with hyperbilirubinemia for phototherapy treatment. We used hour-specific TSB to analyze the TSB levels for ABO/Rh compatibility and isoimmunization using chi-square, analysis of variance, and regression models. RESULTS: Of the 901 infants studied, 158 (17.5%) had ABO/Rh incompatibility, including 27 with positive DAT. Hyperbilirubinemia was diagnosed in 33.3% DAT positive, 6.9% DAT negative, and 4.6% of infants with compatible blood types. Increased predischarge TSB was observed in DAT positive infants at 48-72 h of postnatal age (P < 0.001). After controlling for age at TSB testing and weight loss percentage, multiple regression analysis did not show any impact of ABO/Rh incompatibility and DAT results on the predischarge TSB levels. CONCLUSION: Blood type incompatibility increases the frequency of hyperbilirubinemia only in the DAT-positive infants. Irrespective of the isoimmunization status, it does not significantly affect the level of predischarge TSB.

Elongating maize root: zone-specific combinations of polysaccharides from type I and type II primary cell walls.

The dynamics of cell wall polysaccharides may modulate the cell wall mechanics and thus control the expansion growth of plant cells. The unique composition of type II primary cell wall characteristic of grasses suggests that they employ specific mechanisms for cell enlargement. We characterized the transcriptomes in five zones along maize root, clustered the expression of genes for numerous glycosyltransferases and performed extensive immunohistochemical analysis to relate the changes in cell wall polysaccharides to critical stages of cell development in Poaceae. Specific patterns of cell wall formation differentiate the initiation, realization and cessation of elongation growth. Cell walls of meristem and early elongation zone represent a mixture of type I and type II specific polysaccharides. Xyloglucans and homogalacturonans are synthesized there actively together with mixed-linkage glucans and glucuronoarabinoxylans. Rhamnogalacturonans-I with the side-chains of branched 1,4-galactan and arabinan persisted in cell walls throughout the development. Thus, the machinery to generate the type I primary cell wall constituents is completely established and operates. The expression of glycosyltransferases responsible for mixed-linkage glucan and glucuronoarabinoxylan synthesis peaks at active or late elongation. These findings widen the number of jigsaw pieces which should be put together to solve the puzzle of grass cell growth.

AFM analysis reveals polymorphism of purified flax rhamnogalacturonans I of distinct functional types.

Functionally distinct polymers organized on the basis of rhamnogalacturonan I (RG-I) backbone with more than a half of rhamnose residues substituted by the side chains containing mostly galactose were purified from flaxseed mucilage, the primary cell wall of young hypocotyls and tertiary cell walls of bast fibers and characterized by atomic force microscopy. Seed mucilage RG-I with short side chains and unusual O3 substitution showed loose coils or star-like conformations. Primary cell wall RG-I, which included polygalacturonan (PGA) fragments, represented micellar objects and rare long chains. Pure RG-I with long galactan side chains, which was isolated as nascent polysaccharide before its incorporation into the tertiary cell wall of bast fibers was observed as long unbranched objects. RG-I entrapped by cellulose microfibrils in tertiary cell wall was visualized as compact micellar complexes. All types of flax RGs-I tended to aggregate. Relationships between RG-I structure and morphology are discussed.

Gelation of rhamnogalacturonan I is based on galactan side chain interaction and does not involve chemical modifications.

The article presents the structural principles of microwave-induced formation of new gel type from pectic rhamnogalacturonan I (RG-I). The backbone of gel-forming RG-I does not contain consecutive galacturonic residues and modifying groups that can be the cause of junction zone formation as it occurs in course of classical ways of pectin gelation. Microwave irradiation does not cause destruction and chemical modifications of RG-I. Removal of half of galactan chains from RG-I leads to loss of gelling capability pointing out on their leading role in this process. Rising of intensity of the bands attributed to galactose and glycosidic linkages in RG-I gel comparing to solution where this polymer exists as molecule associate indicates that the spatial organization of galactans in gel is changed. A model of the RG-I gelation is proposed: being destabilized at volumetric microwave heating RG-I associates are repacked forming network where RG-I molecules are entangled by galactan chains.

Prevalence and risk factors for vitamin D insufficiency and deficiency at birth and associated outcome.

BACKGROUND: Occurrence and consequence of cord blood (CB) vitamin D insufficiency/deficiency has not been adequately explored despite rising concern regarding this topic in pediatrics. This study was designed to determine the rate, maternal risk factors, and clinical outcomes in infants in association with vitamin D insufficient/deficient status at birth. METHODS: American Academy of Pediatrics (AAP) defined levels (ng/mL) were utilized to categorize the vitamin D status in CB samples as deficient (5-15), insufficient (16-20), and sufficient (21-100). We used descriptive statistics and multiple regression models to identify the rate and factors associated with vitamin D deficiency/insufficiency and related outcomes in the enrolled mother-infant pairs. RESULTS: This prospective study was conducted at a single center on postpartum women and their infants. Vitamin D deficiency and insufficiency was recorded in 38.9 and 29.8% respectively of the 265 CB samples. Deficient CB vitamin D levels in infants were associated with maternal Black, Hispanic, or Asian race/ethnicity, younger age, and increased number of pregnancies. The likelihood for infants to be born with an insufficient vitamin D level increases with younger maternal age and the number of pregnancies as well as Asian ethnicity. We did not find an association between the vitamin D status at birth and pre-discharge clinical characteristics of the neonates. CONCLUSIONS: The likelihood for an infant to be born with vitamin D deficiency/insufficiency is relatively high and is related mainly to younger maternal age, gravidity, and non-White race/ethnicity. Our findings raise a question regarding the adequacy of the AAP recommended vitamin D supplementation requirements without knowing the infant's vitamin D status at birth.

Physicochemical properties of complex rhamnogalacturonan I from gelatinous cell walls of flax fibers.

The physicochemical properties of flax fiber cell wall rhamnogalacturonan I (RG-I) and its fragments, obtained after galactanase treatment (fraction G1), were characterized. RG-I retains its hydrodynamic volume after its molecular weight decreases by approximately half, as revealed by SEC. Two techniques, DLS and NMR, with different principles of diffusion experiment were used to establish the reasons for this property of RG-I. Three possible types of particles were revealed by DLS depending on the concentration of the RG-I and G1 solutions (2-2.5, 15-20, and 150-200 nm). It was determined by BPP-LED experiments that the backbone of the RG-I was 1.3-1.9-fold more mobile than the side chains. The obtained data suggest a novel type of pectin spatial organization-the formation of RG-I associates with the backbone at the periphery and the interaction between the side chains to form a core zone.

Vitamin D in the maternal-fetal-neonatal interface: clinical implications and requirements for supplementation.

Identification of the current evidence regarding the pathophysiological and clinical facets of vitamin D in the maternal-fetal-neonatal interface is of value because of the significance of the vitamin D endocrine system in human health and high prevalence of vitamin D deficiency in mothers and their infants. Although many questions have still not been answered by the existing literature, we found evidence that: (i) during pregnancy vitamin D participates in fetal skeletal mineralization and growth, (ii) neonatal vitamin D levels are dependent on the maternal vitamin D status at delivery, (iii) a vitamin D sufficient status at birth may decrease the risk for the development of asthma and type 1 diabetes mellitus in later life, (iv) recommendations for maintaining serum 25-hydroxyvitamin D [25(OH)D] levels ≥32 ng/mL to avoid secondary hyperparathyroidism in adults have not been applied to mothers and their infants, (v) American Academy of Pediatrics recommended supplementation of 400 IU of vitamin D per day is sufficient only for infants who are born with normal vitamin D levels and (vii) supplementation of lactating mothers with high doses of vitamin D (4000 IU/d) allows the achievement of optimal 25(OH)D concentrations (>32 ng/mL) in the maternal and infant serum without any risk of hypervitaminosis D in the mother. We believe that inconsistency in the recognition of sufficient levels of vitamin D in mothers and their infants affects the identification of adequate doses for vitamin D supplementation during pregnancy, lactation and infancy.

Management of neonatal hyperbilirubinemia: pediatricians' practices and educational needs.

BACKGROUND: Early detection and treatment of neonatal hyperbilirubinemia is important in the prevention of bilirubin-induced encephalopathy. In this study, we evaluated the New Jersey pediatricians' practices and beliefs regarding the management of neonatal hyperbilirubinemia and their compliance with the recommendations made by the American Academy of Pediatrics (AAP) in 1994. METHODS: A survey questionnaire was mailed to a random sample of 800 pediatricians selected from a list of 1623 New Jersey Fellows of the AAP initially in October 2003 and then in February 2004 for the non-respondents. In addition to the physicians' demographic characteristics, the questionnaire addressed various aspects of neonatal hyperbilirubinemia management including the diagnosis, treatment, and follow up as well as the pediatricians' beliefs regarding the significance of risk factors in the development of severe hyperbilirubinemia. RESULTS: The adjusted response rate of 49.1% (n = 356) was calculated from the 725 eligible respondents. Overall, the practicing pediatricians reported high utilization (77.9%) of the cephalocaudal progression of jaundice and low utilization (16.1%) of transcutaneous bilirubinometry for the quantification of the severity of jaundice. Most of the respondents (87.4%) identified jaundice as an indicator for serum bilirubin (TSB) testing prior to the neonate's discharge from hospital, whereas post-discharge, only 57.7% felt that a TSB was indicated (P < 0.01). If the neonate's age was under 72 hours, less than one-third of the respondents reported initiation of phototherapy at TSB levels lower than the treatment parameters recommended by the AAP in 1994, whereas if the infant was more than 72 hours old, almost 60% were initiating phototherapy at TSB lower than the 1994 AAP guidelines. Most respondents did not regard neonatal jaundice noted after discharge and gestational ages 37-38 weeks as being significant in the development of severe hyperbilirubinemia. However, the majority did recognize the importance of jaundice presenting within the first 24 hours and Rh/ABO incompatibility. CONCLUSION: The pediatricians' practices regarding the low utilization of laboratory diagnosis for the quantification of jaundice after discharge and underestimation of risk factors that contribute to the development of severe hyperbilirubinemia are associated with initiation of phototherapy at lower than AAP recommended treatment parameters and recognition of neonatal hyperbilirubinemia as an important public health concern.

Transcutaneous bilirubinometry in preterm infants receiving phototherapy.

The accuracy of transcutaneous bilirubin (TcB) measurements obtained with a Bilicheck device in 70 preterm neonates before phototherapy and during phototherapy from the exposed and unexposed skin of the forehead (under the eye patch) was evaluated by comparing the TcB with the total serum bilirubin (TSB) that was performed within +/- 30 minutes of the TcB measurements. The correlation and proportion of similar, overestimated, and underestimated paired TcB/TSB measurements was calculated. A TSB point estimate of +/- 1.5 mg/dL was used for comparison of differences between the TcB and TSB. There was weaker correlation between TcB and TSB when the TcB was obtained from the exposed skin (r = 0.70; p > 0.05) as compared with an unexposed area (r = 0.77; p < 0.04) or before initiation of phototherapy (r = 0.86; p < 0.01). The TcB measurements from the area unexposed to phototherapy were predominantly (97.2%) in the similar (57%) or overestimation categories (40.2%). A significantly lower proportion of the paired TcB/TSB measurements obtained from exposed area were similar (42%) or overestimated (27%; p < 0.001). More importantly, in 31% of the measurements, the TSB levels were underestimated by the TcB. We conclude that the Bilicheck can be used for the evaluation of bilirubin levels in preterm neonates receiving phototherapy by using the unexposed skin of the forehead (under the eye patch) for TcB measurements; this may significantly reduce the number of blood draws.

The accuracy of transcutaneous bilirubin measurements in neonates: a correlation study.

A prospective observational study was conducted on 212 neonates born between 24 and 42 weeks of gestation who required blood sampling to determine total serum bilirubin (TSB) in the first week of life, prior to phototherapy. The transcutaneous bilirubin (TcB) measurements were performed on the infant's forehead using BiliCheck within +/-30 min of a blood sample being drawn. There was significant (r = 0.78) correlation between bilirubin levels obtained transcutaneously and those measured in the infant's blood. The correlation was not affected by birth weight and was dependent on the bilirubin levels. The negative nonsignificant correlation appears when TSB levels are greater than 11 mg/dl. Thus, TcB measurements can accurately predict TSB values lower than 11 mg/dl in a multiracial preterm and term neonatal population.