Deciphering the contribution of human meningothelial cells to the inflammatory and antimicrobial response at the meninges.

We have investigated the response of primary human meningothelial cells to Neisseria meningitidis. Through a transcriptome analysis, we provide a comprehensive examination of the response of meningothelial cells to bacterial infection. A wide range of chemokines are elicited which act to attract and activate the main players of innate and adaptive immunity. We showed that meningothelial cells expressed a high level of Toll-like receptor 4 (TLR4), and, using a gene silencing strategy, we demonstrated the contribution of this pathogen recognition receptor in meningothelial cell activation. Secretion of interleukin-6 (IL-6), CXCL10, and CCL5 was almost exclusively TLR4 dependent and relied on MyD88 and TRIF adaptor cooperation. In contrast, IL-8 induction was independent of the presence of TLR4, MyD88, and TRIF. Transcription factors NF-κB p65, p38 mitogen-activated protein kinase (MAPK), Jun N-terminal protein kinase (JNK1), IRF3, and IRF7 were activated after contact with bacteria. Interestingly, the protein kinase IRAK4 was found to play a minor role in the meningothelial cell response to Neisseria infection. Our work highlights the role of meningothelial cells in the development of an immune response and inflammation in the central nervous system (CNS) in response to meningococcal infection. It also sheds light on the complexity of intracellular signaling after TLR triggering.

Injuries to Ice Hockey Referees and Linesmen: A Survey of International Ice Hockey Federation Officials.

Background: Ice hockey referees and linesmen are at risk for musculoskeletal injuries because of the lack of protective equipment and contact with players, sticks, pucks, the ice surface and boards. Purpose: To quantify and analyze injuries reported by officials of the International Ice Hockey Federation (IIHF). Study Design: Descriptive epidemiology study. Methods: A 61-question survey tool was designed by an interdisciplinary team to evaluate musculoskeletal injuries experienced by ice hockey officials. This survey was administered to 600 active IIHF referees and linesmen. Only completed survey responses were included in the statistical analysis. Continuous variables were analyzed using unpaired t-tests, while categorical data were assessed utilizing chi-square tests. Results: Of the 600 surveys administered, 264 surveys were completed by officials from 45 countries (44% response rate). Of the respondents, 72% were male, and 28% were female, with a mean age of 31.1 ± 5.8 years. Officiating experience averaged 11.4 ± 6.0 years (6.3 ± 4.5 years with the IIHF). A total of 295 injuries were reported by 55% of the officials. Injuries occurred more frequently during games compared with training, and officials who worked year-round had more total injuries than those who took time off (P = .03). The most common injuries involved the wrist and hand (n = 64 [22%]), head and face (n = 58 [20%]), and the knee (n = 47 [16%]). Wrist and hand trauma included 23 fractures. Knee and shoulder injuries were most likely to require surgery compared with other body areas (P < .001); 30 officials underwent surgery because of an acute knee injury (10%). Injury prevention activities were effective at reducing injuries (P = .04). Conclusion: Most ice hockey officials experienced musculoskeletal injuries during their career. The risk of trauma to the wrist and hand can possibly be reduced via equipment modifications including protective gloves. A greater emphasis should be placed on injury prevention programs and time away from officiating competitions.

Rapid Rule Out of Culture-Negative Bloodstream Infections by Use of a Novel Approach to Universal Detection of Bacteria and Fungi.

BACKGROUND: Currently it can take up to 5 days to rule out bloodstream infection. With the low yield of blood cultures (approximately 10%), a significant number of patients are potentially exposed to inappropriate therapy that can lead to adverse events. More rapid rule out can accelerate deescalation or cessation of antimicrobial therapy, improving patient outcomes. METHODS: A method is described, termed enzymatic template generation and amplification (ETGA), that universally and sensitively detects DNA polymerase activity liberated from viable bacteria and fungi isolated from blood culture samples as a measure of bloodstream infection. ETGA was applied in a diagnostic test format to identify negative blood cultures after an overnight incubation. Performance data for a prototype (Cognitor) and automated (Magnitor) version of the test are presented. RESULTS: The Cognitor manual assay displayed analytical reactivity for a panel of the 20 most prevalent causes of bloodstream infection, with a detection range of 28-9050 CFU/mL. Validation with 1457 clinical blood cultures showed a negative predictive value of 99.0% compared to blood culture incubation for 5 days. Magnitor showed an improved detection range of 1-67 CFU/mL, allowing for detection of bacteria-supplemented blood cultures after 2-8 h incubation, and Candida albicans-supplemented blood cultures at 16-22 h, 5-15 h faster than blood culture. Removing an aliquot from a blood culture bottle and replacing the bottle into the incubator was shown not to result in contaminating organisms being introduced. CONCLUSIONS: The described method displays excellent breadth and detection for microbial cells and demonstrates the capability of confirming negative blood cultures after an overnight incubation in a blood culture instrument.

Metallohosts Derived from the Assembly of Sugars around Transition Metals: The Complexation of Alkali Metal Cations.

The diacetone glucose (DAGH, 1,2:5,6-di-O-isopropylidene-alpha-D-glucofuranose) monoanion DAG binds as a terminal alkoxo ligand to a variety of transition metals. When it is used in excess, with respect to the oxidation state of the metal, homoleptic anionic complexes [M'(DAG)(6)](3)(-) are formed. Such complexes contain oxygen-rich cavities between pairs of DAG ligands appropriate for binding alkali metal cations. The anionic complexes have been obtained by using [Li(DAG)], 1, and [Na(DAG)], 2, whose syntheses and characterization are reported here. The reaction of 1 and 2 with [V(DAG)(3)] gave [V(DAG)(6)Li(3)], 3, and [V(DAG)(6)Na(3)], 4, respectively. An alternative synthesis of 3 and 4 involves the metathesis reaction of 1 and 2 with [VCl(3)(thf)(3)]. This strategy also led to the synthesis of [Cr(DAG)(6)Li(3)], 5, and [Ti(DAG)(6)Li(3)], 6. Three pairs of DAG shape a cavity appropriate for three lithium cations in the case of complexes 3, 5, and 6; a cavity is formed for three sodium cations in the case of 4, where the alkali cation is in a tetrahedral O(4) environment. In the anionic manganese derivative [Mn(Cl)(DAG)(4)](3)(-), the four DAG units arrange in such a way as to bind four Li cations, which form a cationic cage [Mn(Cl)(DAG)(4)Li(4)](+), and Cl(-) is bound inside as [Mn(Cl)(DAG)(4)Li(4)(&mgr;(4)-Cl)], 7. Crystallographic details: 4, prism, P2(1), a = 14.735(10) Å, b = 15.033(9) Å, c= 21.021(10) Å, beta = 107.34(2) degrees, V= 4445(5) Å(3), Z = 2, and R = 7.60; 5, prismatic, C2, a = 22.671(9) Å, b = 18.785(5) Å, c = 13.886(4) Å, beta = 126.39(2) degrees, V= 4761(3) Å(3), Z = 2, and R = 7.32; 6, prismatic, P2(1), a= 13.888(5) Å, b = 18.750(5) Å, c= 17.933(5) Å, beta = 91.84(2) degrees, V = 4667(2) Å(3), Z = 2, and R = 8.75; 7, prismatic, P2(1), a = 13.306(7) Å, b= 21.311(11) Å, c = 13.376(6) Å, beta = 95.01(2) degrees, V = 3779(3) Å(3), Z = 2, and R = 9.33.

The Preparation, Stereochemistry and Reactions of Some [Co(cyclen)(NH(3))X](3+/2+) Complexes (X = OH(2), Cl, N(3), OH).

A number of new [Co(cyclen)(NH(3))X](3+/2+) complexes have been prepared (X = OH(2), Cl, N(3), OH), and their stereochemistries, interconversions, and substitution reactions have been explored. All have the syn,anti configuration about the two meridional amine centers of the cis-coordinated cyclen ligand (syn(NH(3)),anti(X)- and syn(X),anti(NH(3))-isomers for X = OH(2), Cl, N(3), OH; designated as 1-X and 2-X respectively) but a syn,syn intermediate is proposed for a number of the reactions. OH(-)-catalyzed hydrolysis (base hydrolysis) dominates the substitution chemistry in aqueous solution (k(OH)/M(-)(1) s(-)(1) values reported) and results from deprotonation at one of the relatively very acidic meridional NH centers (k(H)/M(-)(1) s(-)(1) values reported). A syn orientation of the NH proton in the reactant is considered important in these reactions, with the resulting lone pair assisting in the displacement of X, and with this proton being transferred to X = OH to facilitate an unusually fast and spontaneous solvent replacement reaction (k(ex) = 8 +/- 2 s(-)(1), I = 1.0 M (NaClO(4)), 25 degrees C). Anation by N(3)(-) has been studied, and this proceeds largely via 2-OH for both isomers. A crystal structure of syn(NH(3)),anti(N(3))-[Co(cyclen)(NH(3))N(3)]Cl(0.5)(ClO(4))(1.5). H(2)O is reported; monoclinic, P2(1)/n (No. 14), a = 9.008(6) Å, b = 28.690(15) Å, c = 14.528(7) Å, a = 90 degrees, beta = 104.12(7) degrees, gamma = 90 degrees, Z = 8, R = 0.0695.

Role of o-Methylene-Bridged Bis(aniline) as a Spectroscopic Probe for the Metal Coordination Environment.

Reaction of the bidentate ligand 2,2'-methylenebis(5-(dimethylamino)aniline) (2, L) with PdCl(2) and K(2)PtCl(4) and the bidentate ligand 2,2'-methylenebis(5-(acetylamino)aniline (5, L') with K(2)PtCl(4) in polar solvents gave the complexes [PdCl(2)(L)] (6), [PtCl(2)(L)] (8), and [(L')PtCl(2)] (7), respectively. Treatment of 2 with K(2)PtCl(4) in the presence of DMSO resulted in the solvolysis of a chloride ion to give [(L)Pt(Cl)(DMSO)]Cl (9). The resulting eight-membered metallocyclic rings have rigid elongated chair conformations forcing one of the two bridging methylene hydrogens (H(endo)) in close proximity to the metal centre. An (1)H NMR study revealed that they are now diastereomeric (H(endo), H(exo)); the higher field signal position does not vary irrespective of the ligand or metal, whereas the proton corresponding to the lower field signal is apparently more sensitive to the metal coordination environment. The ability of this spectroscopic probe to predict the bonding mode of the metal was also studied by reacting 2 with K[PtCl(3)(C(2)H(4))].H(2)O to give [Pt(2)(&mgr;-L)Cl(4)(C(2)H(4))(2)] (10), which occurred irrespective of the L/Pt ratio. The corresponding methylene hydrogens were not diastereomeric, showing only one singlet in the (1)H NMR spectrum. Crystal structures of 6 and 9 have been determined: 6 is triclinic, space group P&onemacr;, a = 10.179(2) Å, b = 12.136(2) Å, c = 9.686(2) Å, alpha = 97.79(1) degrees, beta = 95.98(2) degrees, gamma = 78.02(1) degrees, V = 1156.1(4), Z = 2, and R = 0.034; 9 is triclinic, space group P&onemacr;, a = 12.348(1) Å, b = 12.407(1) Å, c = 10.159(1) Å, alpha = 103.02(1) degrees, beta = 104.87(2) degrees, gamma = 117.09(1) degrees, V = 1313.0(3), Z = 2, and R = 0.035.

17O NMR Study of Solvent Exchange in Some Aqueous [Co(tren)(X)(OH(2)/OH)](n)()(+), [Co(cyclen)(X)(OH(2)/OH)](n)()(+), and [Co(N-Mecyclen)(X)(OH(2)/OH)](n)()(+) Systems (X = NH(3), OH(2)/OH; n = 3, 2, 1)(1).

A (17)O NMR study (I = 1.0 M, NaClO(4) or NaOSO(2)CF(3), 25.0 degrees C) of solvent exchange in labeled p- and t-[Co(tren)(NH(3))OH(2)](3+), [Co(tren)(OH(2))(2)](3+), [Co(cyclen)(OH(2))(2)](3+), and [Co(N-Mecyclen)(OH(2))(2)](3+) ions (ca. 30% (17)O) in aqueous solution has shown that loss of coordinated OH(2) is slow for all of the complexes (k(ex)/s(-)(1) = 1.1 x 10(-)(5), 1.2 x 10(-)(5), 3.7 x 10(-)(5) (p-site)/8.7 x 10(-)(6) (t-site), 2 x 10(-)(4), and 2 x 10(-)(4), respectively). Values of k(ex) for solvent exchange in [Co(tren)(OH)(2)](+) have been determined as 9.7 x 10(-)(5) s(-)(1) (p-site) and 2.2 x 10(-)(7) s(-)(1) (t-site) Coordinated OH(-) in both p- and t-[Co(tren)(NH(3))OH](2+) also exchanges only slowly with solvent (k(ex)/s(-)(1)= 1.7 x 10(-)(4), and <1 x 10(-)(6), respectively), whereas exchange of coordinated solvent in the aqua-hydroxo complexes [Co(tren)(t-OH(2))(p-OH)](2+), [Co(cyclen)(OH(2))OH](2+), and [Co(N-Mecyclen)(OH(2))OH](2+) is much more rapid (k(ex)/s(-)(1) = 0.03 (p-site)/0.01 (t-site), 12 and 15, respectively). Ligand-OH(-) exchange in these latter systems is interpreted as occurring via (indirect) S(N)1(CB)-type processes on the corresponding aqua complexes: [Co(amine)OH](2+) right harpoon over left harpoon [Co(amine-H)OH(2)](2+) --> exchange. This type of pathway is seen to be more efficient when leaving-group departure is synchronous with proton transfer, and this appears to be more important for exchange in the cyclen and N-Mecyclen complexes where a reasonably acidic, adjacent syn NH proton is involved.

Medial patellofemoral ligament reconstruction: indications and technique.

Lateral instability of the patella is a common problem that often requires reconstruction of the proximal soft tissue restraints. The medial patellofemoral ligament (MPFL) has been demonstrated to be the major soft tissue stabilizer to prevent abnormal lateral displacement of the patella. In this chapter we will discuss the anatomy, biomechanics, indications, and technique for surgical reconstruction of the medial patellofemoral ligament.