Biomedical subjects

Robert D White

Publications and source records attributed to Robert D White.

7 recordsLinked to original sources

PubMed2005

Preparation of isotopically labeled spinach acyl-acyl carrier protein for NMR structural studies.

Acyl carrier proteins (ACPs) are important protein cofactors in fatty acid biosynthesis, but their acylated forms have not been well-studied. To permit detailed nuclear magnetic resonance studies of acylated spinach ACP isoform I, we have developed a new expression plasmid for recombinant production of the apo-protein and modified protocols for purifying the protein product and acylating it to form acyl-ACP. To solve plasmid stability problems associated with growth in minimal media, the ampicillin resistance gene from pSACP-2a was replaced with the tetA(C) gene from pBR322. The resulting plasmid, pSACP-2t, supported overexpression of apo-ACP in Escherichia coli BL21(DE3) cells in M9 medium containing 15NH4Cl as the sole nitrogen source. Apo-ACP was purified to homogeneity by means of polyethylene glycol precipitation and anion exchange. Two in vitro synthetic routes were used to produce acyl-ACPs. In one route, apo-ACP was converted to the holo form and the acyl form by a published protocol that employs a discrete enzymatic reaction for each step. As an alternative route to produce decanoyl-ACP, apo-ACP was directly converted to the acyl form by using holo-ACP synthase along with the non-natural substrate decanoyl-CoA. Two-dimensional 1H-15N NMR spectroscopy of decanoyl-ACP and stearoyl-ACP revealed that changes in the length of the covalently attached fatty acid do not affect the secondary structure of the protein but do influence the local conformation and dynamics.

Acyl Carrier Protein↗

PubMed2005

Microengineered hydromechanical cochlear model.

Micromachined fluid-filled variable impedance waveguides intended to mimic the mechanics of the passive mammalian cochlea have been fabricated and experimentally examined. The structures were microfabricated with dimensions similar to those of the biological system. Experimental tests demonstrate acoustically excited traveling fluid-structure waves with phase accumulations between 1.5 and 3 pi radians at the location of maximum response. The resulting measured frequency-position mapping function, with similarities to that observed in the cochlea, is presented. Results for both isotropic and orthotropic membranes are reported, demonstrating that the achieved orthotropy ratio of 8:1 in tension is insufficient to produce the sharp filtering observed in animal experiments and many computational models that use higher ratios. It is also shown experimentally that high viscosity fluids must be used to provide sufficient damping to avoid the formation of a nonphysiological standing wave pattern. A mathematical model incorporating a thin-layer viscous, compressible fluid approximation coupled to an orthotropic membrane model is validated against experimental results. The work presented herein is motivated by the possibility of producing microfabricated cochlear-like filters, thus the structure is designed for production in a scalable microfabrication process.

Animals↗

PubMed2005

Multicenter study to determine antibody concentrations and assess the safety of administration of INH-A21, a donor-selected human Staphylococcal immune globulin, in low-birth-weight infants.

Nosocomial or late-onset sepsis is a common complication among premature infants, with a frequency inversely correlated with birth weight. Increased susceptibility to infection is due in part to an immature humoral (antibody-mediated) immune response. This study investigated the pharmacokinetics (PKs) and safety of a donor-selected specific intravenous immune globulin (IVIG) preparation, INH-A21 (Veronate), for prevention of sepsis in premature infants. Thirty-six infants weighing between 500 and 1,250 g during the first postnatal week were eligible to begin a series of up to four intravenous infusions of 500 or 750 mg/kg of body weight INH-A21. Blood samples were analyzed for antibodies against the Ser-Asp dipeptide repeat G (SdrG) and clumping factor A (ClfA) surface proteins of staphylococci. Sparse sampling and population PK analyses were performed to derive PK parameters. Following administration of the 500- and 750-mg/kg doses, the estimated average steady-state levels of anti-ClfA were 6.1 U/ml and 9.2 U/ml, respectively, and those of anti-SdrG were 5.2 U/ml and 7.7 U/ml, respectively. The elimination half-lives for anti-ClfA and anti-SdrG were 719 h and 701 h, respectively, and the clearances were 0.18 ml/h and 0.21 ml/h, respectively. In the final model, the values of the PK parameters were independent of gestational age. Both doses of INH-A21 were well tolerated, and the safety profile was similar to those of other IVIG preparations. These results suggest that a shorter dosing interval should be utilized between the first and second doses to achieve and maintain higher titers of anti-ClfA and anti-SdrG antibodies. Further studies examining INH-A21 for the prevention of late-onset sepsis in infants within the weight range studied are warranted.

Antibodies, Bacterial↗

PubMed2004

Lighting design in the neonatal intensive care unit: practical applications of scientific principles.

Meeting the varied lighting needs of infants, caregivers, and families has become more complex as our understanding of visual development and perception and the effect of light on circadian rhythms advances. Optimal lighting strategies are discussed for new unit construction, as well as modifications to consider for existing units. In either case, the key concept is that lighting should be provided for the individual needs of each person, rather than the full-room lighting schemes previously used. Ideas gleaned from nonhospital settings, re-introduction of natural light into the neonatal intensive care unit, and new devices such as light-emitting diodes will dramatically change the lighting and visual environment of future neonatal intensive care units.

Health Facility Environment↗

PubMed2004

Mothers' arms--the past and future locus of neonatal care?

With the advent of neonatal intensive care, medical professionals inserted themselves between the baby and its family. Even for healthy newborn, the mother could only get to her infant with permission from the medical staff and then only for limited periods and in a very restricted manner. Family-centered care restored the family's right to full access to their baby, but as generally practiced(and certainly as NICUs are currently designed) medical professionals still view the infant as a solitary individual who sleeps most of the time in a bed. Future NICU design should recognize that the baby must spend most of its time in its mother's arms to get the full benefit of her sensory environment as experienced throughout our evolution. NICUs must therefore be planned to facilitate this extended proximity as much as possible (ie, not just providing for parents at the bedside with the bed as the locus of care, but transforming the preferred and predominant locus of care from the bed to the parents' arms, with the design changes inherent to that concept). Designing our units to facilitate this interaction will not assure that it will always occur, but it will guarantee that we have not created permanent structural features that interfere with this crucial relationship.

Health Facility Environment↗

PubMed2003

Chain cleavage and sulfoxidation of thiastearoyl-ACP upon reaction with stearoyl-ACP desaturase.

The fatty acid analogues 9- and 10-thiastearate were converted to acyl-ACP derivatives by in vitro enzymatic synthesis and reacted with the reconstituted soluble stearoyl-ACP Delta9 desaturase complex. Electrospray ionization mass spectral analysis of the acyl chains purified from the reaction mixtures showed that 10-thiastearoyl-ACP was converted to the 10-sulfoxide as the sole product. In the presence of (18)O(2), the sulfoxide oxygen was found to be derived exclusively from O(2). This result confirms the ability of the soluble stearoyl-ACP desaturase to catalyze O atom transfer in the presence of the appropriate substrate analogue. Inhibition studies showed that 10-thiastearoyl-ACP was a mixed-type inhibitor of 18:0-ACP, with an apparent K(I) of approximately 10 microM. Comparable reactions of the stearoyl-ACP desaturase complex with 9-thiastearoyl-ACP gave the 9-sulfoxide as approximately 5% of the total products, with the O atom again exclusively derived from O(2). The remaining 95% of the total products arose from an acyl chain cleavage reaction between S-9 and C-10. Matrix-assisted laser desorption ionization time-of-flight mass spectral analysis showed that 9-thiastearoyl-ACP had a mass of 9443 amu while the acyl chain cleavage product had a mass of 9322 amu, corresponding to the calculated mass of 8-mercaptooctanoyl-ACP. Recovery of the acyl chain from the ACP product gave the disulfide of 8-mercaptooctanoate (mass of 349.1 amu), arising from the dimerization of 8-mercaptooctanoate during product workup. Gas chromatography-mass spectral analysis also showed the accumulation of nonanal in sealed reaction vials, accounting for the other product of the acyl chain cleavage reaction. The reactivity at both the 9 and 10 positions of the thia-substituted acyl-ACPs is consistent with the proximity of both positions to the diiron center oxidant in the enzyme-substrate complex. Moreover, the differential reactivity of the 9- and 10-thiastearoyl-ACPs also suggests position-dependent consequences of the reaction within the Delta9D active site. Mechanisms accounting for both sulfoxidation and acyl cleavage reactions by the stearoyl-ACP Delta9 desaturase are proposed.

Acyl Carrier Protein↗

PubMed2003

Individual rooms in the NICU - an evolving concept.

The provision of individual rooms for NICU patients has several attractive benefits, including the ability to provide the appropriate environmental stimuli for each child, and increased privacy and accessibility for families. This concept can put serious strain on caregivers, however, by isolating them from one another and from the multiple infants for whom they may have responsibility, and places additional financial and space demands on the institution. These problems are not insurmountable, however, and use of individual rooms for at least some patients is feasible and probably desirable if certain considerations are addressed in the planning and design process.

Family↗