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Automated testing platforms facilitate the introduction of red cell genotyping of patients and blood donors. Fluidic microarray systems, such as Luminex XMAP (Austin, TX), are used in many clinical applications, including HLA and HPA typing. The Progenika ID CORE XT (Progenika Biopharma-Grifols, Bizkaia, Spain) uses this platform to analyze 29 polymorphisms determining 37 antigens in 10 blood group systems. Once DNA has been extracted, processing time is approximately 4 hours. The system is highly automated and includes integrated analysis software that produces a file and a report with genotype and predicted phenotype results. Immunohematology2015;31:62–68.

Correct donor D typing is critical to prevent recipient alloimmunization. No method can detect all variants, and the immunogenicity of many variants is unknown. Routine ABO and D serologic typings are performed in our laboratory by automated microplate testing. Until 2011, routine confirmation of D– status of first-time donors was performed by the manual tube indirect antiglobulin test (IAT); this was replaced by automated solid-phase testing including weak D testing by IAT. Selected donors are investigated by other methods. We describe four weak D type 67 (RHD*01W.67)donors whose samples tested as D– by automated microplate and manual methods but were later determined to be D+ by automated solid-phase and RHDgene analysis. Solid-phase serologic and molecular typing results of all four donors were identical. It was identified that the donors are of English-Irish descent; two are brothers and the others are cousins. Transfusion of blood from one of these donors likely resulted in alloimmunization to D in one of three recipients tested since no other documented exposures were identified. Lookback studies determined that two other D– recipients were not alloimmunized. Immunohematology2015;31:159–162.

Magnetospheric Multiscale observations are used to probe the structure and temperature profile of a guide field reconnection exhaust ~100 ion inertial lengths downstream from the X‐line in the Earth's magnetosheath. Asymmetric Hall electric and magnetic field signatures were detected, together with a density cavity confined near 1 edge of the exhaust and containing electron flow toward the X‐line. Electron holes were also detected both on the cavity edge and at the Hall magnetic field reversal. Predominantly parallel ion and electron heating was observed in the main exhaust, but within the cavity, electron cooling and enhanced parallel ion heating were found. This is explained in terms of the parallel electric field, which inhibits electron mixing within the cavity on newly reconnected field lines but accelerates ions. Consequently, guide field reconnection causes inhomogeneous changes in ion and electron temperature across the exhaust. Plasma heating and energization by magnetic reconnection is a fundamental process in space, solar, astrophysical, and planetary plasmas. Most reconnecting current sheets do not exhibit perfectly antialigned magnetic fields and a so‐called guide fieldis often present. Using new experimental data from NASA's Magnetospheric Multiscale mission, this article shows that far from the X‐line during guide field reconnection, the heating is substantially modified from the typically studied antiparallel case. More specifically, the new multipoint, high time resolution Magnetospheric Multiscale measurements of a guide field reconnection exhaust in the Earth's magnetosheath reveal inhomogenous ion and electron heating and cooling. This uncovers in new detail the structure of the exhaust, including predicted density cavity structure and electron holes, and indicates the importance of the parallel electric field. The results are important for the general understanding of reconnection heating and energization. The results will be of immediate and timely interest to the Geophysical Research Letters (GRL) community and beyond. A guide field reconnection exhaust was encountered by MMS in the magnetosheath ~100 ion inertial lengths downstream from the X‐lineA density cavity forms on one edge of the exhaust with embedded electron jetting toward the X‐line and electron holes on the cavity edgeThe parallel electric field causes electron cooling and ion heating in the cavity and inhomogeneous temperature profiles across the exhaust

We show observations of whistler mode waves in both the low‐latitude boundary layer (LLBL) and on closed magnetospheric field lines during a crossing of the dayside reconnecting magnetopause by the Magnetospheric Multiscale (MMS) mission on 11 October 2015. The whistlers in the LLBL were on the electron edge of the magnetospheric separatrix and exhibited high propagation angles with respect to the background field, approaching 40°, with bursty and nonlinear parallel electric field signatures. The whistlers in the closed magnetosphere had Poynting flux that was more field aligned. Comparing the reduced electron distributions for each event, the magnetospheric whistlers appear to be consistent with anisotropy‐driven waves, while the distribution in the LLBL case includes anisotropic backward resonant electrons and a forward resonant beam at near half the electron‐Alfvén speed. Results are compared with the previously published observations by MMS on 19 September 2015 of LLBL whistler waves. The observations suggest that whistlers in the LLBL can be both beam and anisotropy driven, and the relative contribution of each might depend on the distance from the X line. Whilstlers in both LLBL and magnetosphere observed during magnetopause crossingLLBL whistlers have large parallel electric field and more oblique propagation anglesThe presence of electron beam and anisotropy suggests two simultaneous generation mechanisms for whistlers in LLBL

In Innocenti et al. (2015) we have observed and characterized for the first time Petschek‐like switch‐off slow shock/rotational discontinuity (SO‐SS/RD) compound structures in a 2‐D fully kinetic simulation of collisionless magnetic reconnection. Observing these structures in the solar wind or in the magnetotail would corroborate the possibility that Petschek exhausts develop in collisionless media as a result of single X point collisionless reconnection. Here we highlight their signatures in simulations with the aim of easing their identification in observations. The most notable signatures include a four‐peaked ion current profile in the out‐of‐plane direction, associated ion distribution functions, increased electron and ion anisotropy downstream the SS, and increased electron agyrotropy downstream the RDs. Petschek‐like exhausts can be generated by collisionless reconnection; the ion current is linked to in‐plane magnetic field switch‐offSignatures of the switch‐off discontinuity are a four‐peaked ion current, ion distribution functions, and increased electron agyrotropyThese signatures can help the identification of the structure, should it be encountered by spacecraft, such as MMS

We present an analysis of the properties of the electron velocity distribution during island coalescence in asymmetric reconnection with and without guide field. In a previous study, three main domains were identified, in the case without guide field, as X, D, and Mregions featuring different reconnection evolutions. These regions are also identified here in the case with guide field. We study the departure from isotropic and gyrotropic behavior by means of different robust detection algorithms proposed in the literature. While in the case without guide field these metrics show an overall agreement, when the guide field is present, a discrepancy in the agyrotropy within some relevant regions is observed, such as at the separatrices and inside magnetic islands. Moreover, in light of the new observations from the Multiscale MagnetoSpheric mission, an analysis of the electron velocity phase‐space in these domains is presented. Asymmetric magnetic island coalescence with and without guide fieldElectron anisotropy and agyrotropy in asymmetric magnetic reconnectionFully kinetic simulation of asymmetric magnetic reconnection at the dayside magnetopause

The restriction of a monotone operator Pto the cone Ω of nonnegative decreasing functions from a weighted Orlicz space Lφ,vwithout additional a priori assumptions on the properties of theOrlicz function φand the weight function vis considered. An order-sharp two-sided estimate of the norm of this restriction is established by using a specially constructed discretization procedure. Similar estimates are also obtained for monotone operators over the corresponding Orlicz–Lorentz spaces Λφ,v. As applications, descriptions of associated spaces for the cone Ω and the Orlicz–Lorentz space are obtained. These new results are of current interest in the theory of such spaces.

We show observations from the Magnetospheric Multiscale (MMS) mission of whistler mode waves in the Earth's low‐latitude boundary layer (LLBL) during a magnetic reconnection event. The waves propagated obliquely to the magnetic field toward the X line and were confined to the edge of a southward jet in the LLBL. Bipolar parallel electric fields interpreted as electrostatic solitary waves (ESW) are observed intermittently and appear to be in phase with the parallel component of the whistler oscillations. The polarity of the ESWs suggests that if they propagate with the waves, they are electron enhancements as opposed to electron holes. The reduced electron distribution shows a shoulder in the distribution for parallel velocities between 17,000 and 22,000 km/s, which persisted during the interval when ESWs were observed, and is near the phase velocity of the whistlers. This shoulder can drive Langmuir waves, which were observed in the high‐frequency parallel electric field data. Whistler mode waves are observed on the magnetic reconnection separatrixThese waves are propagating toward the X lineSolitary bipolar parallel electric fields appear to be in phase with the wave and may correspond with electron enhancements

We report observations from the Magnetospheric Multiscale satellites of large-amplitude, parallel, electrostatic waves associated with magnetic reconnection at the Earth's magnetopause. The observed waves have parallel electric fields (E||) with amplitudes on the order of 100?mV/m and display nonlinear characteristics that suggest a possible net E||. These waves are observed within the ion diffusion region and adjacent to (within several electron skin depths) the electron diffusion region. They are in or near the magnetosphere side current layer. Simulation results support that the strong electrostatic linear and nonlinear wave activities appear to be driven by a two stream instability, which is a consequence of mixing cold (<10?eV) plasma in the magnetosphere with warm (~100?eV) plasma from the magnetosheath on a freshly reconnected magnetic field line. The frequent observation of these waves suggests that cold plasma is often present near the magnetopause. Magnetospheric Multiscale observations of large-amplitude, parallel, electrostatic waves associated with magnetic reconnectionSimulations support that the strong electrostatic linear and nonlinear wave activities appear to be driven by a two-stream instabilityThe frequent observation of these waves suggests that cold plasma is often present near the magnetopause

The Koyna‐Warna area of India is one of the best worldwide examples of reservoir‐induced seismicity, with the distinction of having generated the largest known induced earthquake (M6.3 on 10 December 1967) and persistent moderate‐magnitude (>M5) events for nearly 50 years. Yet, the fault structure and tectonic setting that has accommodated the induced seismicity is poorly known, in part because the seismic events occur beneath a thick sequence of basalt layers. On the basis of the alignment of earthquake epicenters over an ~50 year period, lateral variations in focal mechanisms, upper‐crustal tomographic velocity images, geophysical data (aeromagnetic, gravity, and magnetotelluric), geomorphic data, and correlation with similar structures elsewhere, we suggest that the Koyna‐Warna area lies within a right step between northwest trending, right‐lateral faults. The sub‐basalt basement may form a local structural depression (pull‐apart basin) caused by extension within the step‐over zone between the right‐lateral faults. Our postulated model accounts for the observed pattern of normal faulting in a region that is dominated by north‐south directed compression. The right‐lateral faults extend well beyond the immediate Koyna‐Warna area, possibly suggesting a more extensive zone of seismic hazards for the central India area. Induced seismic events have been observed many places worldwide, but relatively large‐magnitude induced events are less common because critically stressed, preexisting structures are a necessary component. We suggest that releasing bends and fault step‐overs like those we postulate for the Koyna‐Warna area may serve as an ideal tectonic environment for generating moderate‐ to large‐ magnitude induced (reservoir, injection, etc.) earthquakes. Koyna‐Warna seismicity occurs in a step‐over between NW trending dextral faultsNW trending faults may pose a more hazard to much of west‐central IndiaRight stepping dextral faults may be ideal hosts for large induced earthquakes

Massively parallel numerical simulations of magnetic reconnection are presented in this study. Electromagnetic full‐particle implicit code iPIC3D is used to study the dynamics and 3‐D evolution of reconnection outflows. Such features as Hall magnetic field, inflow and outflow, and diffusion region formation are very similar to 2‐D particle‐in‐cell (PIC) simulations. In addition, it is well known that instabilities develop in the current flow direction or oblique directions. These modes could provide for anomalous resistivity and diffusive drag and can serve as additional proxies for magnetic reconnection. In our work, the unstable evolution of reconnection transient front structures is studied. Reconnection configuration in the absence of guide field is considered, and it is initialized with a localized perturbation aligned in the cross‐tail direction. Our study suggests that the instabilities lead to the development of finger‐like density structures on ion‐electron hybrid scales. These structures are characterized by a rapid increase of the magnetic field, normal to the current sheet (Bz). A small decrease in the magnetic field component parallel to the reconnection Xline and the component perpendicular to the current sheet is observed in the region ahead of the front. The instabilities form due to fact that the density gradient inside the front region is opposite to the direction of the acceleration Lorentz force. Such density structures may possibly further develop into larger‐scale earthward flux transfer events during magnetotail reconnection. In addition, oscillations mainly in the magnetic and electric fields and the electron density are observed shortly before the arrival of the main front structure which is consistent with recent THEMIS observations. Three dimensional particle‐in‐cell simulation of reconnection in the magnetotailEvolution of dipolarization front at reconnection and associated plasma flowDevelopment of instabilities in the plasma population

Calcineurin‐inhibitor refractory bronchiolitis obliterans (BO) represents the leading cause of late graft failure after lung transplantation. T helper (Th)2 and Th17 lymphocytes have been associated with BO development. Taking advantage of a fully allogeneic trachea transplantation model in mice, we addressed the pathogenicity of Th cells in obliterative airway disease (OAD) occurring in cyclosporine A (CsA)‐treated recipients. We found that CsA prevented CD8+T cell infiltration into the graft and downregulated the Th1 response but affected neither Th2 nor Th17 responses in vivo. In secondary mixed lymphocyte cultures, CsA dramatically decreased donor‐specific IFN‐γ production, enhanced IL‐17 production and did not affect IL‐13. As CD4+depletion efficiently prevented OAD in CsA‐treated recipients, we further explored the role of Th2 and Th17 immunity in vivo. Although IL‐4 and IL‐17 deficient untreated mice developed an OAD comparable to wild‐type recipients, a single cytokine deficiency afforded significant protection in CsA‐treated recipients. In conclusion, CsA treatment unbalances T helper alloreactivity and favors Th2 and Th17 as coexisting pathways mediating chronic rejection of heterotopic tracheal allografts.

Calcineurin-inhibitor refractory bronchiolitis obliterans (BO) represents the leading cause of late graft failure after lung transplantation. T helper (Th)2 and Th17 lymphocytes have been associated with BO development. Taking advantage of a fully allogeneic trachea transplantation model in mice, we addressed the pathogenicity of Th cells in obliterative airway disease (OAD) occurring in cyclosporine A (CsA)-treated recipients. We found that CsA prevented CD8+T cell infiltration into the graft and downregulated the Th1 response but affected neither Th2 nor Th17 responses in vivo. In secondary mixed lymphocyte cultures, CsA dramatically decreased donor-specific IFN-γ production, enhanced IL-17 production and did not affect IL-13. As CD4+depletion efficiently prevented OAD in CsA-treated recipients, we further explored the role of Th2 and Th17 immunity in vivo. Although IL-4 and IL-17 deficient untreated mice developed an OAD comparable to wild-type recipients, a single cytokine deficiency afforded significant protection in CsA-treated recipients. In conclusion, CsA treatment unbalances T helper alloreactivity and favors Th2 and Th17 as coexisting pathways mediating chronic rejection of heterotopic tracheal allografts.

Because of the variability of D expression, one method may be inadequate to correctly classify donors with variant RHDalleles. We evaluated the use of a solid-phase automated platform (ImmucorGamma Galileo) to confirm D– test results obtained on first-time donors on the Beckman Coulter PK7300 automated microplate test system. Samples with discordant results were analyzed by serologic tube methods, RHDgenotyping using the BLOODchip platform (Progenika), and, if necessary, sequencing. We estimated the number of cases of alloimmunization in women younger than 50 years likely to be prevented by the addition of Galileo testing. From May 2011 to May 2012, 910,220 donor samples were tested; 15,441 were first-time donors with concordant D– results. Five donors tested D– on the PK7300 and weak D+ on the Galileo; one was found to be a false positive on further testing. On manual testing, the other four donors had positive indirect antiglobulin test results with one to three of the antisera used and were C+. On BLOODchip testing, two donors were classified as D+, and two were assigned a “no call.” D variants included weak D type 67, weak D type 9, and two novel variants. Approximately 10 percent of D– units are transfused to women younger than 50 years. Assuming an alloimmunization rate of 30 percent, use of the Galileo would prevent approximately one alloimmunization every 5 to 6 years in this patient group. We conclude that the yield of preventing alloimmunization in this population by adding a second automated serologic testing platform is very low. Immunohematology2013; 29:97–100.

The alloimmunization potential of many RHDvariants is unknown, and it can be explored by lookback and traceback studies. Héma-Québec (HQ) investigated the RHDstatus of 3980 D– repeat blood donors. Thirteen were found to be RHDpositive: 4 RHD*ψ,and 1 RHD*487delACAG,which show a D– phenotype; and 1 RHD*885Tand 7 RHD*(93–94insT)causing a DEL phenotype when C antigen is present. Lookback studies were done to verify the alloimmunization potential of these eight DEL donors. Coincidentally, Canadian Blood Services (CBS) performed a traceback study by investigating the RHDstatus of donors after a D– recipient developed anti-D after transfusion of two D– red blood cell (RBC) units. Donor genotyping was done either manually (HQ) or using the Progenika Bloodchip platform (CBS). Donations were traced through computer records. Letters were sent to hospital blood bank physicians to verify the presence of anti-D in recipients and to donors to request repeat samples. A total of 118 RBC units were transfused, 82 to D– recipients. Anti-D was found in three patients transfused with RHD*(93– 94insT)DEL red blood cells. One donor presenting the same DEL variant was involved in the traceback study. Even without strong evidence clearly demonstrating the alloimmunization potential of DEL variants, whenever HQ or CBS identifies a donor harboring a DEL phenotype, his or her D status will be changed from D– to D+ to protect against the potential alloimmunization risk. Immunohematology2013;29:136–140.

Immune responses in newborn mice are known to be biased toward the helper type 2 phenotype. This may account for their propensity to develop tolerance. Herein, we evaluated the effects of IL-4 deprivation on CD4+T-cell activities elicited by neonatal exposure to allogeneic spleen cells. We showed that chimerism, Th2-type polarization and pathology, as well as skin allograft acceptance were inhibited in BALB/c mice immunized at birth with (A/J x BALB/c) F1spleen cells upon in vivoIL-4 neutralization. While IL-4 neutralization inhibited the development of Th2 cells in this model, it led to the accumulation of IL-17A, IL-17F, IL-22, IL-6 and RORγt mRNA in the spleen or graft tissues. Moreover, IL-4 deprivation led to the differentiation of donor-specific Th17 cells with a concomitant Th1 response characterized by IFN-γ production. The Th17-type response emerging in IL-4-deprived mice was found to mediate both intragraft neutrophil infiltration and the abrogation of B-cell chimerism. Neutralization of this Th17 response failed however to restore functional skin graft acceptance. Collectively, our observations indicate that the neonatal Th2 response opposes the development of Th17 cells, and that Th17 cells are responsible for controlling lymphoid chimerism in mice neonatally injected with semiallogeneic cells.

Immune responses in newborn mice are known to be biased toward the helper type 2 phenotype. This may account for their propensity to develop tolerance. Herein, we evaluated the effects of IL‐4 deprivation on CD4+T‐cell activities elicited by neonatal exposure to allogeneic spleen cells. We showed that chimerism, Th2‐type polarization and pathology, as well as skin allograft acceptance were inhibited in BALB/c mice immunized at birth with (A/J x BALB/c) F1spleen cells upon in vivoIL‐4 neutralization. While IL‐4 neutralization inhibited the development of Th2 cells in this model, it led to the accumulation of IL‐17A, IL‐17F, IL‐22, IL‐6 and RORγt mRNA in the spleen or graft tissues. Moreover, IL‐4 deprivation led to the differentiation of donor‐specific Th17 cells with a concomitant Th1 response characterized by IFN‐γ production. The Th17‐type response emerging in IL‐4‐deprived mice was found to mediate both intragraft neutrophil infiltration and the abrogation of B‐cell chimerism. Neutralization of this Th17 response failed however to restore functional skin graft acceptance. Collectively, our observations indicate that the neonatal Th2 response opposes the development of Th17 cells, and that Th17 cells are responsible for controlling lymphoid chimerism in mice neonatally injected with semiallogeneic cells.

Inflammation and cancer are associated with impairment of T-cell responses by a heterogeneous population of myeloid-derived suppressor cells (MDSCs) coexpressing CD11b and GR-1 antigens. MDSCs have been recently implicated in costimulation blockade-induced transplantation tolerance in rats, which was under the control of inducible NO synthase (iNOS). Herein, we describe CD11bGR-1MDSC-compatible cells appearing after repetitive injections of lipopolysaccharide (LPS) using a unique mechanism of suppression. These cells suppressed T-cell proliferation and Th1 and Th2 cytokine production in both mixed lymphocyte reaction and polyclonal stimulation assays. Transfer of CD11b cells from LPS-treated mice in untreated recipients significantly prolonged skin allograft survival. They produced large amounts of IL-10 and expressed heme oxygenase-1 (HO-1), a stress-responsive enzyme endowed with immunoregulatory and cytoprotective properties not previously associated with MDSC activity. HO-1 inhibition by the specific inhibitor, SnPP, completely abolished T-cell suppression and IL-10 production. In contrast, neither iNOS nor arginase 1 inhibition did affect suppression. Importantly, HO-1 inhibition before CD11b cell transfer prevented the delay of allograft rejection revealing a new MDSC-associated suppressor mechanism relevant for transplantation.

Inflammation and cancer are associated with impairment of T-cell responses by a heterogeneous population of myeloid-derived suppressor cells (MDSCs) coexpressing CD11b and GR-1 antigens. MDSCs have been recently implicated in costimulation blockade-induced transplantation tolerance in rats, which was under the control of inducible NO synthase (iNOS). Herein, we describe CD11b+GR-1+MDSC-compatible cells appearing after repetitive injections of lipopolysaccharide (LPS) using a unique mechanism of suppression. These cells suppressed T-cell proliferation and Th1 and Th2 cytokine production in both mixed lymphocyte reaction and polyclonal stimulation assays. Transfer of CD11b+ cells from LPS-treated mice in untreated recipients significantly prolonged skin allograft survival. They produced large amounts of IL-10 and expressed heme oxygenase-1 (HO-1), a stress-responsive enzyme endowed with immunoregulatory and cytoprotective properties not previously associated with MDSC activity. HO-1 inhibition by the specific inhibitor, SnPP, completely abolished T-cell suppression and IL-10 production. In contrast, neither iNOS nor arginase 1 inhibition did affect suppression. Importantly, HO-1 inhibition before CD11b+ cell transfer prevented the delay of allograft rejection revealing a new MDSC-associated suppressor mechanism relevant for transplantation.

The growing development of composite tissue allografts (CTA) highlights the need for tolerance induction protocols. Herein, we developed a mouse model of heterotopic limb allograft in a stringent strain combination in which potentially tolerogenic strategies were tested taking advantage of donor stem cells in the grafted limb. BALB/c allografts were transplanted into C57BL/6 mice treated with anti-CD154 mAb, nondepleting anti-CD4 combined to either depleting or nondepleting anti-CD8 mAbs. Some groups received additional rapamycin. Both depleting and nondepleting mAb combinations without rapamycin only delayed limb allograft rejection, whereas the addition of rapamycin induced long-term allograft survival in both combinations. Nevertheless, robust donor-specific tolerance, defined by the acceptance of a fresh donor-type skin allograft and simultaneous rejection of third-party grafts, required initial CD8+T-cell depletion. Mixed donor-recipient chimerism was observed in lymphoid organs and recipient bone marrow of tolerant but not rejecting animals. Tolerance specificity was confirmed by the inability to produce IL-2, IFN-γ and TNF-α in MLC with donor antigen while significant alloreactivity persisted against third- party alloantigens. Collectively, these results show that robust CTA tolerance and mixed donor-recipient chimerism can be achieved in response to the synergizing combination of rapamycin, transient CD8+T-cell depletion and costimulation/coreceptor blockade.

The growing development of composite tissue allografts (CTA) highlights the need for tolerance induction protocols. Herein, we developed a mouse model of heterotopic limb allograft in a stringent strain combination in which potentially tolerogenic strategies were tested taking advantage of donor stem cells in the grafted limb. BALBc allografts were transplanted into C57BL6 mice treated with anti-CD154 mAb, nondepleting anti-CD4 combined to either depleting or nondepleting anti-CD8 mAbs. Some groups received additional rapamycin. Both depleting and nondepleting mAb combinations without rapamycin only delayed limb allograft rejection, whereas the addition of rapamycin induced long-term allograft survival in both combinations. Nevertheless, robust donor-specific tolerance, defined by the acceptance of a fresh donor-type skin allograft and simultaneous rejection of third-party grafts, required initial CD8T-cell depletion. Mixed donor-recipient chimerism was observed in lymphoid organs and recipient bone marrow of tolerant but not rejecting animals. Tolerance specificity was confirmed by the inability to produce IL-2, IFN-? and TNF-? in MLC with donor antigen while significant alloreactivity persisted against third- party alloantigens. Collectively, these results show that robust CTA tolerance and mixed donor-recipient chimerism can be achieved in response to the synergizing combination of rapamycin, transient CD8T-cell depletion and costimulationcoreceptor blockade.

Background: Prior studies have established that botulinum toxin type A (BTX-A) is a safe and effective treatment of coarse dynamic rhytids. However, BTX-A has little known effect on the mottled hyperpigmentation, tactile roughness, skin thinning, and fine wrinkles that develop on periorbital skin as a result of cumulative photodamage. In contrast, topical retinoids have demonstrated the capacity to reverse clinical signs of cumulative photodamage in a number of studies. We hypothesize that the combination of BTX-A plus a topical retinoid (tazarotene 0.1% cream) for the treatment of lateral canthal rhytids (crow's feet) may be synergistic.Objective: To evaluate the effect of tazarotene 0.1% cream on crow's feet and mottled hyperpigmentation after a single treatment with BTX-A.Methods: This is a 16-week, single-center, double-blind, randomized, placebo-controlled study comparing the safety and efficacy of a single 12-U bilateral injection of BTX-A (Botox, Allergan Inc, Irvine, Calif) into the lateral third of the orbicularis oculi muscle at day 0 followed by nightly application of tazarotene 0.1% cream (Avage, Allergan) to one side and placebo cream to the contralateral side. Lateral canthal rhytids and mottled hyperpigmentation were scored by blinded physician evaluators at baseline and monthly intervals based on physical examination of the subjects and high-quality digital photographs. Overall improvement and adverse side effects were rated by patients at each visit via a patient questionnaire.Results: Sixteen of 20 subjects (80%) completed the trial. The mean baseline wrinkle scores for the lateral canthal areas randomized to tazarotene or placebo cream were 3.53 and 3.41, respectively (3 = moderate wrinkling). There were no statistically significant differences in the wrinkle scores between the two groups at baseline (P = .54). According to the blinded physician evaluators, the decreases from baseline of the mean wrinkle scores at the end of the trial (day 112) were 1.37 and 0.75 for the tazarotene-treated and the placebo-treated groups, respectively. There was 1.8-fold greater improvement from baseline of the mean wrinkle score of the tazarotene-treated group compared with placebo at the completion of the trial (P < .001). Treatment satisfaction scores assigned by the subjects were consistent with the physicians' evaluations. At the end of the trial (day 112), the subjects saw 2-fold greater improvement from baseline on the sides treated with tazarotene 0.1% cream versus those treated with placebo cream (P < .001). The mean mottled hyperpigmentation score for the sides randomized to tazarotene treatment improved 40%, from 4.07 (severe) at baseline to 2.4 (2 = mild, 3 = moderate) at the end of the trial (P < .001). Adverse side effects were few and transient and did not cause any subject to discontinue the study.Conclusion: Our results suggest that the therapeutic combination of BTX-A plus topical tazarotene 0.1% cream is a safe and more efficacious treatment of crow's feet and mottled hyperpigmentation than BTX-A treatment alone.

Background: Laparoscopic cholecystectomy (LC) is safe in acute cholecystitis, but the exact timing remains ill-defined. This study evaluated the effect of timing of LC in patients with acute cholecystitis. Methods: Prospective data from the hospital registry were reviewed. All patients admitted with acute cholecystitis from June 1994 to January 2004 were included in the cohort. Results: Laparoscopic cholecystectomy was attempted in 1,967 patients during the study period; 80% were women, mean patient age was 44 years (range, 20–73 years). Of the 1,967 LC procedures, 1,675 were successful, and 292 were converted to an open procedure (14%). Mean operating time for LC was 1 h 44 min (SD ± 50 min), versus 3 h 5 min (SD ± 79 min) when converted to an open procedure. Average postoperative length of stay was 1.89 days (± 2.47 days) for the laparoscopic group and 4.3 days (± 2.2 days) for the conversion group. No clinically relevant differences regarding conversion rates, operative times, or postoperative length of stay were found between patients who were operated on within 48 h compared to those patients who were operated on post-admission days 3–7. Conclusions: The timing of laparoscopic cholecystectomy in patients with acute cholecystitis has no clinically relevant effect on conversion rates, operative times, or length of stay.

Since several years ago, interleukin(IL)-12 is known to be responsible for the differentiation of naive CD4T cells into type 1 helper T cells producing interferon-?. Recently, two other cytokines of the IL-12 family, IL-23 and IL-27, were shown to play key roles in experimental autoimmune disorders mediated by Th17 cells, a novel pro-inflammatory CD4T-cell subset secreting IL-17. As our knowledge of IL-12 family members is rapidly growing and changing, it will be important to specify their involvement in the induction and regulation of allograft rejection in animal models as well as in clinical settings. Herein, we review key features of cytokines belonging to the IL-12 family and discuss their potential relevance to transplantation immunity.

Since several years ago, interleukin(IL)-12 is known to be responsible for the differentiation of naive CD4+T cells into type 1 helper T cells producing interferon-γ. Recently, two other cytokines of the IL-12 family, IL-23 and IL-27, were shown to play key roles in experimental autoimmune disorders mediated by Th17 cells, a novel pro-inflammatory CD4+T-cell subset secreting IL-17. As our knowledge of IL-12 family members is rapidly growing and changing, it will be important to specify their involvement in the induction and regulation of allograft rejection in animal models as well as in clinical settings. Herein, we review key features of cytokines belonging to the IL-12 family and discuss their potential relevance to transplantation immunity.

Gas temperature evolution is investigated in the corona gap of a steady state dielectric barrier discharge under conditions optimal for many chemical applications, i.e. with relatively high energy injection. Under these conditions, quite high temperatures may be reached and these temperatures may be favorable or not for specific applications. Emission spectroscopy measurements have been used and an acoustic method developed, to thermally characterize the discharge channel. The discharge is produced in air at atmospheric pressure, and ozone used as a thermal indicator. Through experimental results supported by kinetic calculations, it is shown that, depending upon the applied voltage frequency, ozone formation can, at least partially, take place not in the discharge channel itself, but in its immediate vicinity.

Hydrogen peroxide production was studied in an atmospheric pressure dielectric barrier discharge device. The discharge atmosphere was an argon/oxygen gas mixture of variable ratio with water vapor adjunction; H2O2measurements were performed in liquid water droplets or films submitted to the post discharge effluent. With a coaxial reactor allowing an easy scaling, a parametric study has been performed with an argon and water vapor discharge plasma, taking in account the gas flow rate, the water content of this gas flow and the input power. Our best results on H2O2production (800 ppm, 1.7 g.kWh-1) were obtained with 10 min runs in this reactor.

Paracoccidioides brasiliensis, a thermodimorphic fungus, is the causative agent of paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in Latin America. Pathogenicity appears to be intimately related to the dimorphic transition from the hyphal to the yeast form, which is induced by a shift from environmental temperature to the temperature of the mammalian host. Little information is available on the P. brasiliensis genes that are necessary during the pathogenic phase. We have therefore undertaken Suppression Subtraction Hybridization (SSH) and macroarray analyses with the aim of identifying genes that are preferentially expressed in the yeast phase. Genes identified by both procedures as being more highly expressed in the yeast phase are involved in basic metabolism, signal transduction, growth and morphogenesis, and sulfur metabolism. In order to test whether the observed changes in gene expression reflect the differences between the growth conditions used to obtain the two morphological forms rather than differences intrinsic to the cell types, we performed real-time RT-PCR experiments using RNAs derived from both yeast cells and mycelia that had been cultured at 37°C and 26°C in either complete medium (YPD or Sabouraud) or minimal medium. Twenty genes, including AGS1 (α-1,3-glucan synthase) and TSA1 (thiol-specific antioxidant), were shown to be more highly expressed in the yeast cells than in the hyphae. Although their levels of expression could be different in rich and minimal media, there was a general tendency for these genes to be more highly expressed in the yeast cells.

Open abdominal aortic aneurysm repair has been reported to be associated with impairment of sexual function in men, most likely because of autonomic nerve injury and pelvic blood flow changes. Endovascular aneurysm repair does not involve dissection in the area of the iliac bifurcation and therefore may be associated with lower incidence of sexual dysfunction as compared to open repair. We conducted a retrospective study of males after open and endovascular abdominal aortic aneurysm repair to determine if there is a significant difference in the incidence of sexual dysfunction between the two procedures. A modified International Index of Erectile Function Questionnaire was used to access sexual function before and after aneurysm repair. The questionnaire was mailed to all male patients who underwent abdominal aortic aneurysm repair from January 1, 1999 to July 15, 2002. The questionnaire asked patients questions regarding their sexual function before and 3 months after the repair. Questionnaire scores for domains of sexual function (erectile function, orgasmic function, intercourse satisfaction, and overall satisfaction) as well as the total questionnaire score were analyzed. The chi-square and Wilcoxon’s signed ranks test were used for statistical comparisons, with p< 0.05 considered significant. Logistic regression was used to examine association. Two hundred ninety-three questionnaires were mailed and 90 were returned completed. There was no difference for the total questionnaire score or the erectile function score before the procedure. Based on the questionnaire score, erectile function worsened after open AAA repair (p= 0.002). Orgasmic function also deteriorated after open AAA repair (p= 0.001). Endovascular repair was not accompanied by decreased erectile or orgasmic function (p= 0.057 and p= 0.068, respectively). Impairment of erectile function was not associated with age, diabetes, or the number of patent hypogastric arteries after aneurysm repair, but there was a significant association between impaired erectile function and open aneurysm repair (p= 0.036). Endovascular repair of abdominal aortic aneurysms is associated with significantly less impairment of erectile and orgasmic function than that with open repair. Preservation of sexual function after endovascular as compared to open repair should be among the factors considered when weighing treatment options for an abdominal aortic aneurysm in a sexually active male.Open abdominal aortic aneurysm repair has been reported to be associated with impairment of sexual function in men, most likely because of autonomic nerve injury and pelvic blood flow changes. Endovascular aneurysm repair does not involve dissection in the area of the iliac bifurcation and therefore may be associated with lower incidence of sexual dysfunction as compared to open repair. We conducted a retrospective study of males after open and endovascular abdominal aortic aneurysm repair to determine if there is a significant difference in the incidence of sexual dysfunction between the two procedures. A modified International Index of Erectile Function Questionnaire was used to access sexual function before and after aneurysm repair. The questionnaire was mailed to all male patients who underwent abdominal aortic aneurysm repair from January 1, 1999 to July 15, 2002. The questionnaire asked patients questions regarding their sexual function before and 3 months after the repair. Questionnaire scores for domains of sexual function (erectile function, orgasmic function, intercourse satisfaction, and overall satisfaction) as well as the total questionnaire score were analyzed. The chi-square and Wilcoxon’s signed ranks test were used for statistical comparisons, with p< 0.05 considered significant. Logistic regression was used to examine association. Two hundred ninety-three questionnaires were mailed and 90 were returned completed. There was no difference for the total questionnaire score or the erectile function score before the procedure. Based on the questionnaire score, erectile function worsened after open AAA repair (p= 0.002). Orgasmic function also deteriorated after open AAA repair (p= 0.001). Endovascular repair was not accompanied by decreased erectile or orgasmic function (p= 0.057 and p= 0.068, respectively). Impairment of erectile function was not associated with age, diabetes, or the number of patent hypogastric arteries after aneurysm repair, but there was a significant association between impaired erectile function and open aneurysm repair (p= 0.036). Endovascular repair of abdominal aortic aneurysms is associated with significantly less impairment of erectile and orgasmic function than that with open repair. Preservation of sexual function after endovascular as compared to open repair should be among the factors considered when weighing treatment options for an abdominal aortic aneurysm in a sexually active male.

Investigations were carried out on point-to-plane dielectric barrier discharges with two different gas gap lengths (d = 0 and 2 mm), energized with two different high voltage power supplies providing one an ac signal at 15 kHz and the other a pulsed signal with a same repetition rate. Correlations between the electrical properties and the behavior of the plasma in these different situations were established through ozone generation, and by the way O radicals net production, checking for this purpose that in our operating conditions gas­heating influence remained minor. Using the representation ozone concentration vs. charge amount transferred through current pulses, the specific electrical and thus physical properties of pulsed surface discharges are put in light; in these discharges, compared to ac and pulsed volume discharges, current pulses exhibit, for a given charge per pulse, the highest amplitudes and the shortest durations.

Most severe cases of neonatal alloimmune thrombocytopenia (NAIT) are due to anti-HPA-la (anti-PlA1) antibodies. We report a case of NAIT due to anti-HPA-2b that resulted in in utero intracranial hemorrhage. A 33-year-old G2P1A0 Caucasian woman had a routine ultrasound at 34 weeks. The fetus appeared to have a left hemispheric hematoma. ΓVIG, lg/kg, was started immediately and administered weekly until delivery. One day after receiving the first dose of ΓVIG, fetal platelet count was 18 × 109/L, and Hb was 116 g/L. Eleven mL of matched platelets compatible by monoclonal antibody immobilization of platelet antigens (MAIPA) assay were transfused in utero, raising the platelet count to 62 × 109/L. Repeat transfusions were done later that week and 1 week later, with pretransfusion counts of 19 × 109/L and 16 × 109/L, respectively. Delivery by C section was done at 35.5 weeks, after the third platelet transfusion. Platelet count at birth was 77 × 109/L. Drainage of the hematoma was performed after transfusion. Testing with a solid phase ELISA revealed reactivity against GPlb/IX. MAIPA testing after platelet treatment with the protease inhibitor leupeptin demonstrated the presence of anti-HPA-2b. On PCR-SSP the mother was HPA-2a homozygous, the father was HPA-2a/2b. Antibodies against the HPA-2b antigen located on the GPlb/IX complex have been reported in rare cases of NAIT Testing is complicated by proteolytic degradation of the antigen-bearing fragment. Compatible platelets are easily found since approximately 85 percent of donors are HPA-2a/2a. Immunohematotogy2003;19:43-46.

This paper reports on the identification of the glow current components also called pseudo-continuous components associated to non-impulsive currents, in a point-to-plane dielectric barrier discharge (DBD) energized by a sinusoidal alternating high voltage. Thanks to a detailed analysis of both photographic and discharge current recordings, coupled to a close comparison with dc corona behaviours, the existence of both positive and negative glow discharges in DBDs is evidenced. The negative glow current can represent the major part of the current in the negative alternation. The relatively low amplitudes of the glow currents (~0.1–1 mA), compared to streamers pulses amplitudes, often make them quite difficult to detect although they often represent more than 50&percent; of the power delivered to the discharge cell. Discharge photographs taken with a fast camera demonstrate that the negative glow (in the negative alternation of the voltage) can, for high voltage levels, extend across the whole gaseous gap with higher current amplitudes than in dc coronas and without any spark breakdown risks. Thus, the ionization and thermal phenomena identified under negative dc corona may play a more important role in the DBD chemical reactivity, besides the streamer type discharges, in particular through their influence on the gas temperature and chemical species profiles in and out of the discharge volume; this last point is evoked in the discussion closing the paper.

Trypanosoma cruzi, the etiological agent of Chagas' disease, may persist for many years in its mammalian host. This suggests escape from the immune response and particularly a suboptimal CD8⁺ T cell response, since these cells are involved in infection control. In this report, we show that T. cruzi inhibits the lipopolysaccharide (LPS)-induced up-regulation of MHC class I molecules at the surface of human dendritic cells (DC). To further investigate the functional consequences of this inhibition, a trypomastigote surface antigen-derived peptide (TSA-1_514-522 peptide) was selected for its stable binding to HLA-A*0201 molecules and used to generate a primary T. cruzi-specific human CD8⁺ T cell line in vitro. We observed that DC infected with T. cruzi or treated with T. cruzi-conditioned medium (TCM) had a weaker capacity to present this peptide to the specific CD8⁺ T cell line as shown in an IFN-γ ELISPOT assay. Interestingly, T. cruzi or TCM also reduced the antigen presentation capacity of DC to CD8⁺ T cell lines specific for the influenza virus M_58-66 or HIV RT_476-484 epitopes. This dysfunction appears to be linked essentially to reduced MHC class I molecule expression since the stimulation of the RT_476-484 peptide-specific CD8⁺ T cell line was shown to depend mainly on the MHC class I-TCR interaction and not on the co-stimulatory signals which, however, were also inhibited by T. cruzi. This impairment of DC function may represent a novel mechanism reducing in vivo the host's ability to combat efficiently T. cruzi infection.

To examine the reliability of HCG as a biomarker for early pregnancy loss, five experienced researchers independently assessed data from 153 menstrual cycles, determining whether each cycle represented 'no conception,' a 'continuing conception' or a 'conception lost.'