Synthesis of Diene‐Functionalized Macromonomers via Functionalization with Hexa‐1,3,5‐triene

The reaction of poly(styryl)lithium (PSLi) with hexa‐1,3,5‐triene (HXT) was studied as a route to diene‐functionalized macromonomers. When PSLi was reacted with 1.5 molar equivalents of HXT for 2.5 h at ?10 °C in toluene, it was found that the diene‐functionalized macromonomer was obtained in high yield; however, oligomerization of the HXT was observed by matrix‐assisted laser absorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS). Oligomerization was eliminated by running the reaction with only 1.2 molar equivalents of HXT to PSLi and allowing the reaction to run for 15 min at ?10 °C in toluene. The resulting polymer exhibited high diene chain‐end functionality and no oligomerization was observed by MALDI‐TOF MS. ¹³C NMR spectroscopy and the attached‐proton test (APT), along with calculated chemical shifts, showed the presence of both the 1,2‐ and 1,4‐addition chain‐end structures. Further analysis by the reaction of the functional polymer with maleic anhydride indicated that 18 wt.‐% of the product was unreactive, either because of a 1,4‐addition chain‐end structure or a nonfunctional polymer. The structure of the maleic anhydride‐modified polymer was determined by MALDI‐TOF MS and ¹³C NMR spectroscopy. Preliminary work on the reactivity of the diene‐functionalized macromonomers was performed by the addition of a large excess of PSLi to a solution of macromonomer followed by characterization by size‐exclusion chromatography (SEC).

The formation of hexa‐1,3,5‐triene‐functionalized polystyrenes and their reaction with maleic anhydride.     

Modeling Deformation-Induced Fluid Flow in Cortical Bone’s Canalicular–Lacunar System

To explore the potential role that load-induced fluid flow plays as a mechano–transduction mechanism in bone adaptation, a lacunar–canalicular scale bone poroelasticity model is developed and implemented. The model uses micromechanics to homogenize the pericanalicular bone matrix, a system of straight circular cylinders in the bone matrix through which bone fluids can flow, as a locally anisotropic poroelastic medium. In this work, a simplified two-dimensional model of a periodic array of lacunae and their surrounding systems of canaliculi is used to quantify local fluid flow characteristics in the vicinity of a single lacuna. When the cortical bone model is loaded, microscale stress, and strain concentrations occur in the vicinity of individual lacunae and give rise to microscale spatial variations in the pore fluid pressure field. Furthermore, loading of the bone matrix containing canaliculi generates fluid pressures in the contained fluids. Consequently, loading of cortical bone induces fluid flow in the canaliculi and exchange of fluid between canaliculi and lacunae. For realistic bone morphology parameters, and a range of loading frequencies, fluid pressures and fluid–solid drag forces in the canalicular bone are computed and the associated energy dissipation in the models compared to that measured in physical in vitro experiments on human cortical bone. The proposed model indicates that deformation-induced fluid pressures in the lacunar–canalicular system have relaxation times on the order of milliseconds as opposed to the much shorter times (hundredths of milliseconds) associated with deformation-induced pressures in the Haversian system.     

Community Member and Faith Leader Perspectives on the Process of Building Trusting Relationships between Communities and Researchers

In the first phase of this research, we conducted, audio‐recorded, and transcribed seven focus groups with more than 50 English‐ or Spanish‐speaking women of childbearing age. Qualitative analysis revealed the following themes: (1) expectation that participation would involve relationships based on trust that is built over time and impacted by cultural factors; (2) perceived characteristics of research staff that would help facilitate the development of trusting relationships; (3) perceptions about the location of the visits that may affect trust; (4) perceptions of a research study and trust for the institution conducting the study may affect trust; (5) connecting the study to larger communities, including faith communities, could affect trust and willingness to participate. In the second phase of this research, we conducted, recorded, transcribed, and analyzed interviews with leaders from diverse faith communities to explore the potential for research partnerships between researchers and faith communities. In addition to confirming themes identified in focus groups, faith leaders described an openness to research partnerships between the university and faith communities and considerations for the formation of these partnerships. Faith leaders noted the importance of finding common ground with researchers, establishing and maintaining trusting relationships, and committing to open, bidirectional communication.     

Response to sirolimus in capillary lymphatic venous malformations and associated syndromes: Impact on symptomatology,quality of life,and radiographic response

Background

Capillary lymphatic venous malformations (CLVM) and associated syndromes, including Klippel–Trenaunay syndrome (KTS) and congenital lipomatous overgrowth, vascular malformation, epidermal nevi, skeletal, and spinal syndrome (CLOVES), are underrecognized disorders associated with high morbidity from chronic pain, recurrent infections, bleeding, and clotting complications. The rarity of these disorders and heterogeneity of clinical presentations make large-scale randomized clinical drug trials challenging. Identification of PIK3CA (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha [gene]) mutations in CLVM has made targeted medications, such as sirolimus, attractive treatment options. The aim of this study was to investigate the safety and efficacy of sirolimus therapy in CLVM.

Procedure

A combined prospective and retrospective cohort of pediatric and young adult patients with CLVM treated with sirolimus was evaluated for disease response, including symptom improvement, quality of life (QOL), and radiologic response. Sirolimus dosing regimens and toxicities were also assessed.

Results

Twenty-nine patients with CLVM, including KTS and CLOVES, were included. Ninety-three percent of patients reported improved QOL, and 86% had improvement in at least one symptom. Most significantly, improvement was noted in 100% of patients with bleeding and 89% with thrombotic complications with corresponding decreases in mean D-dimer (p = .008) and increases in mean fibrinogen (p = .016). No patients had progressive disease on sirolimus. Most common side effects included neutropenia, lymphopenia, infection, and aphthous ulcers/stomatitis. No toxicities were life-threatening, and none required long-term discontinuation of sirolimus.

Conclusion

Sirolimus appears to be effective at reducing complications and improving QOL in patients with CLVM and associated syndromes. In this patient cohort, sirolimus was well tolerated and resulted in few treatment-related toxicities.     

Fish consumption and advisory awareness in the Great Lakes Basin

More than 61 million adults live in the eight U.S. states bordering the Great Lakes. Between June 2001 and June 2002, a population-based, random-digit-dial telephone survey of adults residing in Great Lakes (GL) states was conducted to assess consumption of commercial and sport-caught fish and awareness of state-issued consumption advisories for GL fish. On the basis of the weighted survey data, approximately 84% of the adults living in these states included fish in their diets. Seven percent (an estimated 4.2 million adults) consumed fish caught from the Great Lakes. The percentage of residents who had consumed sport-caught fish (from any water source) varied regionally and was highest among those who lived in Minnesota (44%) and Wisconsin (39%). Consumption of GL sport fish was highest among residents of Michigan (16%) and Ohio (12%). Among residents who had eaten GL fish, awareness of consumption advisories varied by gender and race and was lowest among women (30%) and black residents (15%). However, 70% of those who consumed GL sport-caught fish twice a month or more (an estimated 509,000 adults across all eight states) were aware of the advisories. Findings from this survey indicate that exposure to persistent contaminants found in GL fish is likely limited to a relatively small subpopulation of avid sport-fish consumers. Results also underscore the public health importance of advisories for commercial fish because an estimated 2.9 million adults living in these states consume more than 104 fish meals per year and may be at risk of exceeding the reference doses for methylmercury, polychlorinated biphenyls, and other bioaccumulative contaminants.     

Prophylaxis of acute respiratory virus infections using nucleic acid-based drugs

Acute respiratory virus infections such as SARS and pandemic influenza are highly contagious diseases that cause global crisis, and inflict severe human mortality and morbidity. Vaccines against these viruses are either unavailable or do not provide adequate protection. In the absence of effective vaccines, nucleic acid-based immunomodulators have the potential to offer effective, broad-spectrum protection against these deadly pathogens. Poly ICLC and CpG oligonucleotides are promising gene-based drugs which have been shown in animal studies to protect against acute respiratory virus infections. Poly ICLC is a synthetic double-stranded RNA (dsRNA), and an effective interferon-inducer and natural killer cell activator. When encapsulated in liposomes, poly ICLC offers complete protection (100% survival rate in pretreated group versus 0% survival in control group) against a lethal respiratory challenge of influenza A virus in mice. This antiviral effect has been shown to persist for up to 3 weeks post-drug treatment. Poly ICLC pretreatment also protects mice against a respiratory challenge of western equine encephalitis (WEE) virus, at a level comparable to inactivated WEE vaccine. CpG oligos in liposomes also provided high level of protection against the lethal influenza challenge. Together, these studies suggest nucleic acid-based immunomodulators are promising antiviral agents which can offer effective and non-specific protection against acute respiratory virus infections.     

Weak polar interactions confer albumin binding site selectivity for haloether anesthetics

BACKGROUND: Enflurane and isoflurane are structural isomers with different anesthetic potencies and side effects. It is not clear whether these differences are produced by differing occupancy of common protein binding sites or by occupancy of different sites, but the very similar molecular properties make the latter possibility unlikely. In this study, the authors examined binding site selectivity of these anesthetics in human serum albumin (HSA). METHODS: Binding of isoflurane and enflurane with HSA was determined with isothermal titration calorimetry. Competition with known ligands (propofol) allowed localization of binding sites within the HSA molecule. Molecular properties of isoflurane and enflurane were calculated. RESULTS: Isoflurane binds HSA with higher affinity but smaller total enthalpy than enflurane. Enthalpogram analysis suggested that isoflurane bound a single site, whereas enflurane bound two. Competition experiments indicated that enflurane and isoflurane share one binding site, which also binds propofol. The additional enflurane site binds propofol but not isoflurane. Increased salt concentration decreased the affinity for isoflurane but not for enflurane. The dipole moment of isoflurane is higher than that of enflurane, and the isoflurane binding site is more polar. CONCLUSION: These data indicate two binding sites of different character for the haloether anesthetics on HSA. One site is more polar and prefers isoflurane, presumably because of its larger dipole. The second site prefers the less polar enflurane. Therefore, weak polar interactions confer considerable selectivity, and differences in drug action may arise from occupancy of different protein sites.     

Comparison of splice sites in mammals and chicken

We have carried out an initial analysis of the dynamics of the recent evolution of the splice-sites sequences on a large collection of human, rodent (mouse and rat), and chicken introns. Our results indicate that the sequences of splice sites are largely homogeneous within tetrapoda. We have also found that orthologous splice signals between human and rodents and within rodents are more conserved than unrelated splice sites, but the additional conservation can be explained mostly by background intron conservation. In contrast, additional conservation over background is detectable in orthologous mammalian and chicken splice sites. Our results also indicate that the U2 and U12 intron classes seem to have evolved independently since the split of mammals and birds; we have not been able to find a convincing case of interconversion between these two classes in our collections of orthologous introns. Similarly, we have not found a single case of switching between AT-AC and GT-AG subtypes within U12 introns, suggesting that this event has been a rare occurrence in recent evolutionary times. Switching between GT-AG and the noncanonical GC-AG U2 subtypes, on the contrary, does not appear to be unusual; in particular, T to C mutations appear to be relatively well tolerated in GT-AG introns with very strong donor sites.     

Noquist: reduced field-of-view imaging by direct Fourier inversion.

A novel technique called "Noquist" is introduced for the acceleration of dynamic cardiac magnetic resonance imaging (CMRI). With the use of this technique, a more sparsely sampled dynamic image sequence is reconstructed correctly, without Nyquist foldover artifact. Unlike most other reduced field-of-view (rFOV) methods, Noquist does not rely on data substitution or temporal interpolation to reconstruct the dynamic image sequence. The proposed method reduces acquisition time in dynamic MRI scans by eliminating the data redundancy associated with static regions in the dynamic scene. A reduction of imaging time is achieved by a fraction asymptotically equal to the static fraction of the FOV, by omitting acquisition of an appropriate subset of phase-encoding views from a conventional equidistant Cartesian acquisition grid. The theory behind this method is presented along with sample reconstructions from real and simulated data. Noquist is compared with conventional cine imaging by retrospective selection of a reduced data set from a full-grid conventional image sequence. In addition, a comparison is presented, using real and simulated data, of our technique with an existing rFOV technique that uses temporal interpolation. The experimental results confirm the theory, and demonstrate that Noquist reduces scan time for cine MRI while fully preserving both spatial and temporal resolution, but at the cost of a reduced signal-to-noise ratio (SNR).