Heart and ovarian allografts from C57BL/6 to BALB/c mice had a significantly longer survival time in recipients treated 6 hours/day with HBO starting 48 hours prior to transplantation and continuing until allograft rejection than did those in untreated mice. Exposure to normobaric oxygen beginning 48 hours prior to transplantation also prolonged ovarian allograft survival, but did not have a statistically significant effect on survival of heart allografts. Exposure to HBO 6 hours/day beginning 24 hours after transplantation and continuing until the rejection of the allografts did not prolong the survival time of these tissues. Treatment of the recipients with hyperbaric air starting 48 hours prior to transplantation did not increase the survival time of either heart or ovarian allografts. Control grafts in syngeneic recipients functioned for at least 30 days after transplantation indicating that the transplantation technique and post-operational animal care were satisfactory.

A group of interrelated studies from these laboratories have shown that various cell-mediated and humoral immune processes were altered in rodents exposed to prolonged hyperbaric oxygen. The study here reported is based upon these findings and was undertaken to determine whether hyperoxygenemia would suppress cell-mediated rejection of foreign tissues and acceptance of xenogeneic tumor transplants. A Fischer 344 rat lymphoma (NRL 1871) was implanted intraperitoneally into C57BL/6, DBA/2 and BALB/c mice. Implants produced fatal neoplasms in 13 of 43 C57BL/6 and 6 of 18 DBA/2 mice given daily HBO for 30· days starting one day after implantation. None of the 17 BALB/c recipients given the same regimen of oxygen developed tumors nor did 117 untreated, tumor recipients observed over a period of 60 to 100 days. The xenogeneic growths metastasized extensively even after cessation of HBO therapy. The histologic appearance of these growths was identical with that of the parent lymphoma and they could be transplanted back to Fischer 344 rats but not to normal or HBO treated mice syngeneic with the primary xenogeneic host.

C-reactive protein, a normally occurring plasma protein, may become elevated as much as 3,000-fold during disease states involving acute inflammation or tissue damage. Through its binding to phosphorylcholine, in the presence of calcium, C-reactive protein has been shown to potentiate the activation of the classical and alternative complement pathways and stimulate phagocytosis as well as inhibit certain platelet and T cell reactivities. Its interaction with the various substances that are an integral part of immunological surveillance implies an important role for C-reactive protein in the host reaction to defense, disease and inflammation. The possible significance of this role is discussed.

Abstract: Background: Dental hygienists can be utilized to provide care to adolescents and young adults in a school-based setting. These dental health care professionals work to improve upon public oral health through educational practices, preventative methods, and referrals. The purpose of this study was to explore the knowledge and perceptions of elementary and middle school staff with regard to the utilization of a dental hygienist in a school setting. Methods: The international review board at approved the following study. A total of eight faculty and staff members from Unicoi County Elementary School and Unicoi County Middle School completed a 14-question survey. Participants included each school’s principal, nurse, physical education/wellness teacher and science teacher or K-6. All surveys were distributed by hand to each school. Results: All eight participants saw a need for their students and would be in support of having a hygienist assigned to their school. Questions in the survey revealed that the facility believed a hygienist would be beneficial, but a full-time nurse was the major care provider in their institution. The schools that had interactions with a dental hygienist could not give the correct answer for how frequently they were coming, what services they were providing, and who was sending the dental hygienists. Discussion: The Northeast Regional Health Office supply dental hygienist for student dental health centered care and application of preventative services. However, there are only three hygienists working with the Northeast Regional Health Office to provide care for seven counties in this region.

Polymethyl methacry late-antibiotic implants for the local treatment of osteomyelitis have been used in hospitals in the U.S. since the 1970s. This treatment requires a second surgery for removal of t he depleted implant. The purpose of this study is to investigate a new biodegradable implant system based upon poly(ortho esters) (POE) which can release tobramycin sulfate (TS) over a 21-day period (the usual clinical treatment period) with simultaneous and subsequent degradation of polymer in the tissues to harmless constituents. The POE-TS disks without any excipients showed a similar drug release pattern regardless of TS loading ( 4%, 8%, 12%, w/ w) and thickness (148, 244, 433 pm): 20%-60% TS-release within the first 24 hours followed by less than 10% release during the following 20 days with less than 3% weight loss of the polymer. These formulations did not meet therapeutic requirements of either drug release or polymer degradation rate. POE d egradation is known to be acid sensitive. To achieve uniform and high TS release over 21 days, various concentrations of lactic acid (LA), sorbic acid (SA), oleic acid (OA) and palmitoleic acids (PA) were incorporated in the 244pm-disks of 8% TS. The disks with LA demonstrated a pseudo zero-order TS release. With SA, OA & PA, the TS release was multi-phasic and the rate of polymer degradation was found to be acid concentration dependent. The TS release from these disks could be attributed to burst effect, polymer erosion and diffusion. Among these formulations, disks with 0.2% LA and 0 .9% OA gave the best release rate of 2mg TS/ day/ g implant; they achieved desirable therapeutic level of TS release and POE degradation. No physical or chemical interactio ns were detected by DSC, NMR and JR. Comparisons of POE weight loss and molecular weight loss with different acid catalysts revealed extremely different kinetics of acid catalysis. Uniform distribution of the drug in the disks was concluded based on content uniformity studies. Aqueous and solid state stability of TS were found to be suitable for clinical usage.

An evaluation of vancomycin pharmacokinetics was performed in 2 phases. In phase I, a vancomycin population pharmacokinetic model was developed based on data from 126 patients using a two-compartment model. Variables tested for inclusion in the model were creatinine clearance (Crcl), age, total body weight (wt) ICU status, gender, body surface area, ideal weight and height. Variables were included at the P < _ 0. 05 level. The final population pharmacokinetic model was as follows: Cl (L/hr) = (0.025 * Crcl) * (1 + 0.0165 *age), V1 (L) = 29.5, V2 (L) = 8.17 + 0.349 * Crcl and Q (L/hr.kg) = 0. 0639 * wt. For ICU patients, V2 was larger than the non ICU patients and it was V2 (L) = 16.258 + 0.694 * Crcl. In phase II, the performance of the derived model was evaluated and compared to the Moellering, Matzke, Birt & Chandler and Rodvold Methods. Predictability of .52 vancomycin serum concentrations was assessed in 3 0 new patients. In predicting all concentration types, mean prediction error (MPE) with 95% CI for Moellering,. Matzke, Birt & Chandler, Rodvold and current methods were -0.1 (-1.6, 1.4), -0.8 (-0.7, 2. 4) , o. o ( -1.5, 1. 6) , -2. 0 ( -3. 4, -o. 5) , and 2. 1 ( o. 9, 3. 4) mg/L respectively. When considering only troughs, MPE with 95% CI were 1.8 (0.2, 3.4), 2.7 (0.9, 4.6), -1.5 (-3.5, 0.5), -1.5 (-3.2, 0.1), and 0.8 (-0.8, 2.4) mg/L respectively. MPE with 95% for the peaks were -2.5 (-5.0, 0.0), -1.6 (-4.1, 1.0), 2.0 (-0.4, 4.4), -2.5 (-5.3, 0.3) and 3.8 (1.8, 5.8) mg/L respectively. Median absolute prediction error (MABPE) , 5% and 95% quantiles for all concentration types for Moellering, Matzke, Birt & Chandler, Rodvold and current methods were 3.4 (0.2, 10.1), 4.1 (0.3, 10.9), 3.9 (0.4, 11.7), 3.1 (0.2, 13.0) and 2.3 (0.1, 9.5) mg/L respectively. MABPE, 5% and 95% quantiles for the troughs were 2. 6 ( o. 2, 9. 4) , 4. o ( o. 4, 10. 3) , 4. 1 (0.7, 10.2), 2.9 (0.2, 9.4) and 2.0 (0.1, 9.2) mg/L respectively. MABPE, 5% and 95% quantiles for the peaks were 5.0 (0.7, 11.5·), 4.2 (0.3, 10.9), 3.8 (0.4, 11.7), 4.9 (0.6, 13.0) and 5.0 (1.1, 9.5) mg/L respectively. It is recommended that the current method be used while setting initial target peak at 30 mg/L with the initial target trough at 6 mgjL. This should frequently result in serum vancomycin concentration within the therapeutic window. Individualization of therapy should then be done when the measured concentrations are available.

No description available.

Background Krüppel-like Factor 2 (KLF2) plays an important role in vessel maturation during embryonic development. In adult mice, KLF2 regulates expression of the tight junction protein occludin, which may allow KLF2 to maintain vascular integrity. Adult tamoxifen-inducible Krüppel-like Factor 4 (KLF4) knockout mice have thickened arterial intima following vascular injury. The role of KLF4, and the possible overlapping functions of KLF2 and KLF4, in the developing vasculature are not well-studied. Results Endothelial breaks are observed in a major vessel, the primary head vein (PHV), in KLF2-/-KLF4-/- embryos at E9.5. KLF2-/-KLF4-/- embryos die by E10.5, which is earlier than either single knockout. Gross hemorrhaging of multiple vessels may be the cause of death. E9.5 KLF2-/-KLF4+/- embryos do not exhibit gross hemorrhaging, but cross-sections display disruptions of the endothelial cell layer of the PHV, and these embryos generally also die by E10.5. Electron micrographs confirm that there are gaps in the PHV endothelial layer in E9.5 KLF2-/-KLF4-/- embryos, and show that the endothelial cells are abnormally bulbous compared to KLF2-/- and wild-type (WT). The amount of endothelial Nitric Oxide Synthase (eNOS) mRNA, which encodes an endothelial regulator, is reduced by 10-fold in E9.5 KLF2-/-KLF4-/- compared to KLF2-/- and WT embryos. VEGFR2, an eNOS inducer, and occludin, a tight junction protein, gene expression are also reduced in E9.5 KLF2-/-KLF4-/- compared to KLF2-/- and WT embryos. Conclusions This study begins to define the roles of KLF2 and KLF4 in the embryonic development of blood vessels. It indicates that the two genes interact to maintain an intact endothelial layer. KLF2 and KLF4 positively regulate the eNOS, VEGFR2 and occludin genes. Down-regulation of these genes in KLF2-/-KLF4-/- embryos may result in the observed loss of vascular integrity.

Polarized cell growth is a fundamental biological process that is tightly regulated spatially and temporally. In plants the key systems to study polar cell growth are pollen tubes and root hairs. In recent years a lot of work has focused on elucidating the mechanisms that mediate this process. The actin cytoskeleton plays a key role in polarized cell growth. Different studies in plant and animal models show that signaling mediated through small GTP-binding proteins is a common theme in actin signaling. In recent years many groups have shown that small GTP binding proteins regulate actin dynamics through the activity of Actin Binding Proteins (ABP). In this study I explored the function of two ABPs from Arabidopsis Thaliana: Actin depolymerizing factor (ADF) and a novel actin-binding guanine exchange factor (GEF). I used Arabidopsis protoplasts as a system to study the function of these proteins. We showed through over-expression of the GFP labeled GEF8 under the constitutively active 35 S promoter, that GEF8 labels the prominent cable like structures inside the cell. Using actin and tubulin binding drugs such as Latrunculin and Oryzalin we showed that GEF8 labels actin cables. Using the Yeast 2 Hybrid system we determined that GEF8 binds actin filaments directly. We established that GEF8 interacts with actin through the unique N terminus of the protein. Finally, using the Basic Local Alignment Search tool we showed that the N terminus of GEF8 is homologous to the Actin Binding Protein 140, a well-established protein marker in Yeast. ADF is an established key regulator of the actin cytoskeleton. Much is known about ADF regulation in animal systems. In plants it has been shown that the small Rho type GTP binding proteins, called RAC/ROPS, regulate ADF activity and that overexpression of RAC/ROPs causes the inactivation of ADF through the phosphoryaltion on Serine 6. However, little is known about the proteins that transduce the signal from small GTP binding proteins to the ADF. Here we show some evidence that upon overexpression of Ric 4 (a RAC/ROP effector known to play a role in actin polymerization), ADF gets displaced from the filament. Moreover, ADF is known to be inactivated by phosphorylation at Ser6; the kinase responsible for this phosphorylation has not been identified in plant. We observed that over-expression of Calcium Dependent Protein Kinase 16 (CDPK16) in protoplasts also induced dissociation of ADF from actin cables. These results suggest that both of RIC4 and CDPK16 may play a role in the pathways that regulate ADF activity.

Protein phosphatases regulate a wide array of proteins through post-translational modification and are required for a plethora of intracellular events in eukaryotes. While some core components of the protein phosphatase complexes are well characterized, many subunits of these large complexes remain unstudied. Here we characterize a murine loss-of-function allele of the protein phosphatase 1 regulatory subunit 35 (Ppp1r35) gene. Homozygous embryos lacking Ppp1r35 initiate developmental delay beginning at E7.5 and have obvious morphological defects at slightly later stages. Mutant embryos do not initiate turning and fail to progress beyond the size and relative development of an E8.5 embryo. Consistent with recent in vitro studies linking PPP1R35 with the microcephaly protein Rotatin and a role in centrosome elongation, we find that the Ppp1r35 mutant embryos lack primary cilia. Histological and molecular analysis of Ppp1r35 mutants revealed that notochord development is irregular and discontinuous and that the floor plate of the neural tube is not specified, consistent with defects in primary cilia. Similar to other mutant embryos with defects in centriole function, Ppp1r35 mutants display increased cell death that is prevalent in the neural tube and an increased number of proliferative cells in prometaphase. We hypothesize that loss of Ppp1r35 function abrogates centriole homeostasis, resulting in both a failure to produce functional primary cilia and cell death and/or cell cycle delay that leads to embryonic lethality. Taken together, these results highlight the essential function of Ppp1r35 during early mammalian development and implicate this gene as a candidate for human microcephaly.