Baryon Electric Charge Correlation as a Magnetometer of QCD.

The correlation between net baryon number and electric charge, χ_{11}^{BQ}, can serve as a magnetometer of QCD. This is demonstrated by lattice QCD computations using the highly improved staggered quarks with physical pion mass of M_{π}=135 MeV on N_{τ}=8 and 12 lattices. We find that χ_{11}^{BQ} along the transition line starts to increase rapidly with magnetic field strength eB≳2M_{π}^{2} and by a factor 2 at eB≃8M_{π}^{2}. Furthermore, the ratio of electric charge chemical potential to baryon chemical potential, µ_{Q}/µ_{B}, shows significant dependence on the magnetic field strength and varies from the ratio of electric charge to baryon number in the colliding nuclei in heavy ion collisions. These results can provide baselines for effective theory and model studies, and both χ_{11}^{BQ} and µ_{Q}/µ_{B} could be useful probes for the detection of magnetic fields in relativistic heavy ion collision experiments as compared with corresponding results from the hadron resonance gas model.

Microscopic Encoding of Macroscopic Universality: Scaling Properties of Dirac Eigenspectra near QCD Chiral Phase Transition.

Macroscopic properties of the strong interaction near its chiral phase transition exhibit scaling behaviors, which are the same as those observed close to the magnetic transition in a three-dimensional classical spin system with O(4) symmetry. We show that the universal scaling properties of the chiral phase transition in quantum chromodynamics (QCD) at the macroscale are, in fact, encoded within the microscopic energy levels of its fundamental constituents, the quarks. We establish a connection between the cumulants of the chiral order parameter, i.e., the chiral condensate, and the correlations among the energy levels of quarks, i.e., the eigenspectra of the massless QCD Dirac operator. This relation elucidates how the fluctuations of the chiral condensate arise from the correlations within the infrared part of the energy spectra of quarks, and naturally leads to a generalization of the Banks-Casher relation for the cumulants of the chiral condensate. Then, through (2+1)-flavor lattice QCD calculations with varying light quark masses near the QCD chiral transition, we demonstrate that the correlations among the infrared part of the Dirac eigenvalue spectra exhibit same universal scaling behaviors as expected of the cumulants of the chiral condensate. We find that these universal scaling behaviors extend up to the physical values of the up and down quark masses. Our study reveals how the hidden scaling features at the microscale give rise to the macroscopic universal properties of QCD.

[Regulating the immune response to carbon tetrachloride-induced liver fibrosis in mice by blocking inducible co-stimulatory molecules and interleukin-33].

Objective: To investigate the effects of combined blockade of interleukin-33 (IL-33) and inducible co-stimulatory molecule (ICOS) on carbon tetrachloride-induced chronic liver fibrosis and imbalance of T helper lymphocyte subsets in mice. Methods: There were 40 BALB/c mice in each model and control group. Flow cytometry was used to determine the proportion of Th1/Th2/Th17 cells in the splenic lymphocyte suspension of mice, the expression levels of interferon γ, IL-4, and IL-17 in the splenic lymphocyte suspension of liver fibrosis mice after combined blockade of IL-33 and ICOS, and the pathological changes of liver histopathology in mice with liver fibrosis. Two independent sample t-test was used to compare data between groups. Results: Compared with the non-blocking group, the proportion of Th2 and Th17 cells in the IL-33/ICOS blocking group was significantly down-regulated (Th2: 65.96% ± 6.04% vs. 49.09% ± 7.03%; Th17: 19.17% ± 4.03% vs. 9.56% ± 2.03%), while the proportion of Th1 cells and Th1/Th2 ratio were up-regulated (Th1: 17.14% ± 3.02% vs. 31.93% ± 5.02%; Th1/Th2: 0.28 ± 0.06 vs. 0.62 ± 0.23), and the difference was statistically significant (t = 5.15, 6.03, 7.14, 4.28, respectively, with P < 0.05). After entering the chronic inflammation stage of liver fibrosis in mice (10 weeks), compared with the non-blocking group, the expression levels of IL-4 and IL-17 in the blockade group were significantly down-regulated [IL-4: (84.75 ± 14.35) pg/ ml vs. (77.88 ± 19.61) pg/ml; IL-17: (72.38 ± 15.13) pg/ml vs. (36.38 ± 8.65) pg/ml], while the expression of interferon γ was up-regulated [(37.25 ± 11.51) pg/ml vs. (77.88 ± 19.61) pg/ml], and the difference was statistically significant (t: IL-4: 4.71; IL-17: 5.84; interferon γ: 5.05, respectively, with P < 0.05). Liver histopathological results showed that hepatic necrosis, hepatic lobular structural disorder, and fibrous tissue hyperplasia were significantly lower in the blockade group than those in the non-blocking group at 13 weeks of liver fibrosis. Conclusion: Combined blockade of the ICOS signaling pathway and IL-33 can regulate Th2 and Th17 polarization, down-regulate the inflammatory response, and inhibit or prevent the occurrence and progression of fibrosis.

[Application of cortical bone trajectory screw and sacral alar screw internal fixation for surgical treatment of lumbar adjacent segment degeneration].

Objective: To explore the application of cortical bone trajectory screw (CBTS) and sacral alar screw (SAS) internal fixation in the treatment of lumbar adjacent segment degeneration (ASD) and evaluate its clinical effect. Methods: Data of 24 patients who were diagnosed with ASD and treated by CBTS or SAS in Beijing Chaoyang Hospital were retrospectively reviewed. There were 14 males and 10 females with a mean age of (67.9±8.2) years. The patients were followed-up for (2.6±0.4) years. Perioperative parameters including operation time, intraoperative blood loss and postoperative time on the ground were counted. All patients were followed-up for at least 2 years. Visual analogue scale (VAS) and the Oswestry disability index (ODI) were compared between pre-operation and at the last follow-up. The internal fixation-related complications, pseudarthrosis and adjacent re-degeneration were evaluated in the follow-up. Results: There were 14 proximal ASD patients, 8 distal ASD patients, 1 both ends ASD patient and 1 ASD patient in between the fusion surgeries. Bone mineral density (BMD) T score of the adjacent vertebrae was -1.98±0.91 on average. The ASD patients were re-operated with CBTS and SAS internal fixation technique. A small incision was made in the revision surgery and the original fixation was not completely cut open and removed. The mean operation time was (125±36) min, mean blood loss was (85±33) ml. The postoperative ambulation time was (3.1±1.9) days, and the hospitalization time was (9.0±2.6) days. Before the operation, the average VAS (back pain) score was 5.2±1.0, the average of VAS (leg pain) score was 6.8±1.9 and ODI was 56.6%±12.8%. VAS score was reduced to 1.4±0.6 (waist pain) and 0.9±0.4 (leg pain). ODI was improved to 13.8%±6.3%. All the difference between preoperative and the last follow-up was statically significant (all P<0.01). No internal fixation failure, pseudarthrosis and adjacent re-degeneration were observed in the final follow-up. Conclusion: The application of CBTS and SAS internal fixation techniques in the surgical treatment of lumbar ASD has the advantages of less trauma, faster postoperative recovery, reliable internal fixation, and fewer complications, especially in patients with low bone mineral density.

Correlated Dirac Eigenvalues and Axial Anomaly in Chiral Symmetric QCD.

We introduce novel relations between the derivatives [∂^{n}ρ(λ,m_{l})/∂m_{l}^{n}] of the Dirac eigenvalue spectrum [ρ(λ,m_{l})] with respect to the light sea quark mass (m_{l}) and the (n+1)-point correlations among the eigenvalues (λ) of the massless Dirac operator. Using these relations we present lattice QCD results for ∂^{n}ρ(λ,m_{l})/∂m_{l}^{n} (n=1, 2, 3) for m_{l} corresponding to pion masses m_{π}=160-55 MeV and at a temperature of about 1.6 times the chiral phase transition temperature. Calculations were carried out using (2+1) flavors of highly improved staggered quarks with the physical value of strange quark mass, three lattice spacings a=0.12, 0.08, 0.06 fm, and lattices having aspect ratios 4-9. We find that ρ(λ→0,m_{l}) develops a peaked structure. This peaked structure arises due to non-Poisson correlations within the infrared part of the Dirac eigenvalue spectrum, becomes sharper as a→0, and its amplitude is proportional to m_{l}^{2}. We demonstrate that this ρ(λ→0,m_{l}) is responsible for the manifestations of axial anomaly in two-point correlation functions of light scalar and pseudoscalar mesons. After continuum and chiral extrapolations we find that axial anomaly remains manifested in two-point correlation functions of scalar and pseudoscalar mesons in the chiral limit.

[Clinical application and prospect of lumbar cortical bone trajectory screw].

For a long time, lumbar pedicle screw fixation and fusion has been regarded as the gold standard for the treatment of the lumbar spine degenerative diseases. However, in patients with osteoporosis, pedicle screw fixation often fails to obtain long-term satisfactory. In recent years, the emergence of cortical bone trajectory screw has become an alternative option for dealing with these problems. With the development of clinical and basic research, scholars have found that cortical bone screw internal fixation can help improve fixation strength, reduce surgical trauma, and accelerate postoperative recovery. Based on the current evidence-based research and clinical application experience, this article analyzes the research hotspots of cortical bone screws, introduces the experience and pitfalls in clinical practice, and provides references for colleagues.

[Clinical application of sacral alar screw fixation in surgical treatment of lumbosacral degenerative disease].

Objective: To verify the safety and feasibility of sacral alar screw fixation and fusion surgery in lumbosacral area. Methods: The clinical and radiological data of patients receiving L5/S1 single-level cortical bone trajectory screw combined with sacral alar screw decompression fixation and fusion in Beijing Chaoyang Hospital due to lumbar spinal stenosis from January 2019 to January 2020 were retrospectively analyzed. The clinical data included operation duration, intraoperative blood loss, postoperative drainage, pain and function scores during follow-up. The radiological data included preoperative and postoperative follow-up X-rays and three-dimensional reconstruction of lumbar spine CT. The fusion status and complication of internal fixation, such as screw loosening and fixation failure were evaluated by the radiological examination. Results: According to the inclusion and exclusion criteria, a total of 16 patients (7 males and 9 females) were included, with an average age of (69.6±5.2) years. The patients were followed-up for (16.2±1.7) months. The average operation duration was (144±21) min, the intraoperative blood loss was (103±20) ml, and the postoperative drainage was (80±34) ml. The patient's low back pain visual analogue scale (VAS) score was 6.8±1.0 before surgery, and it was improved to 0.9±0.7 at the last follow-up; leg pain VAS score was 6.1±0.9 before the operation and it improved to 0.9±0.7 at the last follow-up; Oswestry disability index (ODI) function score was 66.2%±8.0% before the surgery and it decreased to 26.6%±7.2% at the last follow-up. No neurological complications, surgical site infection, screw loosening, internal fixation failure or cage displacement were observed during the follow-up. The fusion rate was 68.8% (11/16) at 6 months post operation. Conclusion: It is safe, feasible and effective to apply cortical screws combined with sacral alar screws in short-segment decompression, fixation and fusion surgery on the lumbosacral region to treat lumbosacral spinal stenosis.

[A long-term follow up study of cervical spondylotic myelopathy using diffusion tensor imaging].

Objective: To analyze the correlation between the parameters of diffusion tensor imaging (DTI) and the clinical function scores before and 5 years after anterior cervical discectomy and fusion (ACDF) by measuring the DTI signal of the maximum compression level (MCL) of the cervical spinal cord in patients with cervical spondylotic myelopathy (CSM). Methods: A prospective study was conducted and clinical data of 37 patients with CSM treated by ACDF in Beijing Chaoyang Hospital from February 2014 to February 2020 were analyzed. The MCL was studied with diffusion tensor imaging (DTI) of MRI. The changes of diffusion indexes (fractional anisotropy (FA)), and clinical function score including Modified Japanese Orthopedic Association (mJOA), neck disability index (NDI) and visual analog scale (VAS) of pain were compared among each follow-up point. The FA value and clinical function score were collected and the data was analyzed to determine whether the high signal intensity could be observed in T2-weighted imaging (T2WI) at each follow-up point. The changes of preoperative and postoperative data were compared, and the correlation between FA value and other two clinical function score were analyzed at each follow-up point. Results: There were 14 males and 23 females with a mean age of (55±10) years in this study. All patients received the surgery and the medullary symptoms improved significantly postoperatively. After 3 months, the mJOA and FA value were improved by 2.62±1.41 and 0.14±0.11 on average, respectively (both P<0.05); and there were significant differences in these two indexes between each follow-up point in two years after the operation (all P<0.05). At the MCL, there were strong correlations between the FA value and mJOA score pre-and postoperatively (rs=0.770, 0.729, both P<0.01). There was no significant correlation between mJOA, NDI and VAS (both P>0.05). Conclusions: The DTI sensitively reflects the improvement of spinal cord function and can be used as an important tool to predict and evaluate the state of spinal cord dysfunction in patients with CSM.

[Effect of posterior lumbar fusion on the degeneration of adjacent segments using cortical bone trajectory screw fixation].

Objective: To compare the degenerative changes of adjacent segments after posterior lumbar fusion surgery using cortical bone trajectory (CBT) screws and pedicle screws (PS) in lumbar spinal degenerative disease. Methods: According to certain inclusion and exclusion criteria, prospective non-randomized study on cases of the single-segment lumbar spinal fusion surgery using cortical bone trajectory screws fixation and pedicle screws fixation in Beijing Chao-Yang Hospital from January 2015 to February 2016 was performed. The intervertebral space height (ISH), foramen height (FH), foramen width (FW), cephalad adjacent segment (CAS) and intervertebral disc degeneration grades at MRI scans were measured before surgery and during follow-up. Clinical symptoms were evaluated by Oswestry Dysfunction Index (ODI) score and pain visual analogue scale (VAS) before and during the follow-up. Radiologic adjacent segment degeneration (R-ASD) and symptomatic adjacent segment disease (S-ASD) patients were diagnosed during the follow-up. Paired sample t test was performed when data were compared before surgery and during follow-up. Results: A total of 69 patients were included in the study, 33 in the CBT group (male/female, 15/18), with an average age of (65±11) years, and 36 in the PS group (male/female, 17/19), with an average age of (64±10) years. The follow-up time was no less than 36 months. At the last follow-up, the ISH of the adjacent segments in the CBT group were not statistically different from that before surgery; the ISH of the adjacent segments in the PS group were significantly reduced compared with that before surgery (t=6.78, P<0.05). The loss of ISH, FH, and FW in the adjacent segments in the CBT group were smaller than those in the PS group, and the differences were statistically significant. During follow-up, 4 cases (12.1%) of R-ASD and no S-ASD were found in the CBT group, while 12 cases (33.3%) of R-ASD appeared in the PS group, which was significantly higher than that in the CBT group (χ(2)=4.35, P=0.04). According to MRI, the adjacent discs of PS group had significantly more severe degeneration at the last follow-up than the CBT group (Z=-2.14, P=0.03). Conclusions: Compared with the PS fixation fusion, the CBT screw fixation can effectively reduce the occurrence of adjacent segment degeneration.

Chiral Phase Transition Temperature in (2+1)-Flavor QCD.

We present a lattice-QCD-based determination of the chiral phase transition temperature in QCD with two degenerate, massless quarks and a physical strange quark mass using lattice QCD calculations with the highly improved staggered quarks action. We propose and calculate two novel estimators for the chiral transition temperature for several values of the light quark masses, corresponding to Goldstone pion masses in the range of 58 MeV≲m_{π}≲163 MeV. The chiral phase transition temperature is determined by extrapolating to vanishing pion mass using universal scaling analysis. Finite-volume effects are controlled by extrapolating to the thermodynamic limit using spatial lattice extents in the range of 2.8-4.5 times the inverse of the pion mass. Continuum extrapolations are carried out by using three different values of the lattice cutoff, corresponding to lattices with temporal extents N_{τ}=6, 8, and 12. After thermodynamic, continuum, and chiral extrapolations, we find the chiral phase transition temperature T_{c}^{0}=132_{-6}^{+3} MeV.

[Comparison of cortical bone trajectory screw fixation and pedicle screw fixation in posterior lumbar fusion].

Objective: To evaluate the efficacy and safety of cortical screw fixation in posterior lumbar spine fixation with cortical bone trajectory (CBT) fixation or pedicle screw (PS) fixation in posterior lumbar fusion. Methods: It was a prospective study and CBT or PS for the treatment of lumbar disease from August 2015 to August 2017 in Beijing Chaoyang Hospital were analyzed. There were 53 males and 51 females, aged 56-78 years (mean age, (67±5) years). The patient's gender, age, operative time, intraoperative blood loss, postoperative bleeding, postoperative hospital stay, and postoperative patient satisfaction were collected. The Oswestry disability index (ODI) and visual analogue scale (VAS) of back pain were used to evaluate preoperative and half-year, one-year and two-year postoperative function and quality of life. Complications such as wound infection, cage displacement, screw extraction and fixation fracture were compared between the two groups. The data of normal distribution were compared with Student t test between the two groups. Results: There was no significant difference in the operation time between the CBT group (n=50) and the PS group (n=54) ((223±17) min vs (221±16) min, t=0.74, P=0.46). The intraoperative blood loss and wound drainage volume were significantly lower in the CBT group than those in the PS screw group (t=-24.20, -57.62, both P<0.05). The average length of hospital stay in the CBT group was significantly shorter than that in the PS group (t=-7.65, P=0.00). Patients with CBT screws had better postoperative satisfaction than patients in PS group (91±6 vs 86±7, t=3.89, P=0.00). The ODI score in the CBT group was significantly lower than that in the PD screw group half a year after the operation (3.9±1.9 vs 5.8±3.1, t=-3.66, P=0.00). The VAS and ODI scores in the CBT group were significantly lower than those in the PS group 1 year after the surgery (t=-3.03, -4.09, both P<0.05). There was no significant differences in wound infection, cage displacement, screw extraction and fixation fracture between the two groups. Conclusion: CBT screw technique is minimally invasive, with short operation time, less intraoperative blood loss, rapid postoperative recovery and low complications. It can be effectively used in posterior lumbar cone fusion.

Additional strange hadrons from QCD thermodynamics and strangeness freezeout in heavy ion collisions.

We compare lattice QCD results for appropriate combinations of net strangeness fluctuations and their correlations with net baryon number fluctuations with predictions from two hadron resonance gas (HRG) models having different strange hadron content. The conventionally used HRG model based on experimentally established strange hadrons fails to describe the lattice QCD results in the hadronic phase close to the QCD crossover. Supplementing the conventional HRG with additional, experimentally uncharted strange hadrons predicted by quark model calculations and observed in lattice QCD spectrum calculations leads to good descriptions of strange hadron thermodynamics below the QCD crossover. We show that the thermodynamic presence of these additional states gets imprinted in the yields of the ground-state strange hadrons leading to a systematic 5-8 MeV decrease of the chemical freeze-out temperatures of ground-state strange baryons.

QCD phase transition with chiral quarks and physical quark masses.

We report on the first lattice calculation of the QCD phase transition using chiral fermions with physical quark masses. This calculation uses 2+1 quark flavors, spatial volumes between (4 fm)(3) and (11 fm)(3) and temperatures between 139 and 196 MeV. Each temperature is calculated at a single lattice spacing corresponding to a temporal Euclidean extent of N(t) = 8. The disconnected chiral susceptibility, χ(disc) shows a pronounced peak whose position and height depend sensitively on the quark mass. We find no metastability near the peak and a peak height which does not change when a 5 fm spatial extent is increased to 10 fm. Each result is strong evidence that the QCD "phase transition" is not first order but a continuous crossover for m(π) = 135 MeV. The peak location determines a pseudocritical temperature T(c) = 155(1)(8) MeV, in agreement with earlier staggered fermion results. However, the peak height is 50% greater than that suggested by previous staggered results. Chiral SU(2)(L) × SU(2)(R) symmetry is fully restored above 164 MeV, but anomalous U(1)(A) symmetry breaking is nonzero above T(c) and vanishes as T is increased to 196 MeV.

Strangeness at high temperatures: from hadrons to quarks.

Appropriate combinations of up to fourth order cumulants of net strangeness fluctuations and their correlations with net baryon number and electric charge fluctuations, obtained from lattice QCD calculations, have been used to probe the strangeness carrying degrees of freedom at high temperatures. For temperatures up to the chiral crossover, separate contributions of strange mesons and baryons can be well described by an uncorrelated gas of hadrons. Such a description breaks down in the chiral crossover region, suggesting that the deconfinement of strangeness takes place at the chiral crossover. On the other hand, the strangeness carrying degrees of freedom inside the quark gluon plasma can be described by a weakly interacting gas of quarks only for temperatures larger than twice the chiral crossover temperature. In the intermediate temperature window, these observables show considerably richer structures, indicative of the strongly interacting nature of the quark gluon plasma.

Freeze-out conditions in heavy ion collisions from QCD thermodynamics.

We present a determination of freeze-out conditions in heavy ion collisions based on ratios of cumulants of net electric charge fluctuations. These ratios can reliably be calculated in lattice QCD for a wide range of chemical potential values by using a next-to-leading order Taylor series expansion around the limit of vanishing baryon, electric charge and strangeness chemical potentials. From a computation of up to fourth order cumulants and charge correlations we first determine the strangeness and electric charge chemical potentials that characterize freeze-out conditions in a heavy ion collision and confirm that in the temperature range 150 MeV ≤ T ≤ 170 MeV the hadron resonance gas model provides good approximations for these parameters that agree with QCD calculations on the 5%-15% level. We then show that a comparison of lattice QCD results for ratios of up to third order cumulants of electric charge fluctuations with experimental results allows us to extract the freeze-out baryon chemical potential and the freeze-out temperature.

Optimization of a bacterial consortium for nitrobenzene degradation.

Three bacterial strains, Arthrobacter sp. NB1, Serratia sp. NB2 and Stenotrophomonas sp. NB3, were isolated from contaminated sludge by using nitrobenzene as a sole source of carbon and nitrogen. It was observed that all three strains could degrade nitrobenzene at 400 mg/L initial concentration and mixed-cultivation of these strains could enhance the degradation of nitrobenzene compared with mono-cultivation. Mixture design was used for adjusting the proportions of each strain and the optimal ratio of inoculation size was NB1:NB2:NB3 = 4:4:5, where the nitrobenzene degradation percentage was two times higher than for by the single strain. The results of Plackett-Burman design indicated that Mg(2+), Ca(2+), Fe(2+), Zn(2+) and Mn(2+) had a positive effect on the degradation of nitrobenzene, while Cu(2+) and Co(2+) had a negative effect on it.

Characterization of a thermally stable and organic solvent-adaptative NAD+ -dependent formate dehydrogenase from Bacillus sp. F1.

AIMS: To characterize a robust NAD(+) -dependent formate dehydrogenase firstly obtained from a nonmethylotroph, Bacillus sp. F1. METHODS AND RESULTS: The Bacillus sp. F1 NAD(+) -dependent formate dehydrogenase (BacFDH) gene was cloned by TAIL-PCR and heterologous expressed in Escherichia coli. BacFDH was stable at temperatures below 55°C, and the half-life at 60°C was determined as 52·9 min. This enzyme also showed a broad pH stability and retained more than 80% of the activities after incubating in buffers with different pH ranging from 4·5 to 10·5 for 1 h. The activity of BacFDH was significantly enhanced by some metal ions. Moreover, BacFDH exhibited high tolerance to 20% dimethyl sulfoxide, 60% acetone, 10% methanol, 20% ethanol, 60% isopropanol and 20% n-hexane. Like other FDHs, BacFDH displayed strict substrate specificity for formate. CONCLUSION: We isolated a robust formate dehydrogenase, designated as BacFDH, which showed excellent thermal stability, organic solvent stability and a broad pH stability. SIGNIFICANCE AND IMPACT OF THE STUDY: The multi-aspect stability makes BacFDH a competitive candidate for coenzyme regeneration in practical applications of chiral chemicals and pharmaceuticals synthesis with a relatively low cost, especially for the catalysis performed in extreme pH conditions and organic solvents.

L-selectin shedding does not regulate human neutrophil attachment, rolling, or transmigration across human vascular endothelium in vitro.

Current models of the multistep adhesion cascade for leukocyte-endothelial interactions predict loss of L-selectin from the leukocyte surface before transendothelial migration. We have tested this hypothesis using in vitro adhesion and transendothelial migration assays and a zinc-dependent metalloproteinase inhibitor, Ro 31-9790 (N-2-((2s)-[(hydroxycarbamoyl)methyl)-4-methylvaleryl]-N-1,3 -dimethyl-L-valinamide), which prevents chemoattractant-induced (e.g., IL-8, FMLP, C5a, platelet-activating factor) L-selectin endoproteolytic cleavage from isolated human neutrophils. Inhibitor and vehicle-treated neutrophils exhibited identical behavior during both adhesive interactions with 4- and 24-h TNF-alpha-activated HUVEC monolayers under flow, (including rate of initial attachment, rolling velocities, stable adhesion, and transmigration) and in static adhesion assays. Flow cytometric analysis of transmigrated neutrophils with mAb to L-selectin revealed that vehicle treated neutrophils had minimal detectable surface L-selectin, whereas inhibitor-treated neutrophils retained comparable levels of L-selectin on their surface as neutrophils maintained at 37 degrees C. In addition, mAb to L-selectin that induce rapid shape change and homotypic adhesion (LAM1-116) did not enhance the rate or extent of neutrophil transmigration under flow or static conditions. Neutrophils preincubated with LAM 1-116 displayed similar behavior to neutrophils preincubated with the control anti-L-selectin mAb, LAM1-101. In summary, these results demonstrate that there is no requirement for L-selectin to be shed from the surface of neutrophils before, or during, their migration across endothelial monolayers, and that prevention of surface L-selectin proteolytic cleavage does not enhance or inhibit neutrophil-endothelial cell adhesive interactions.

Adhesion of memory lymphocytes to vascular cell adhesion molecule-1-transduced human vascular endothelial cells under simulated physiological flow conditions in vitro.

The accumulation of mononuclear leukocytes is an early and persistent finding in atherosclerotic plaques. These mononuclear leukocytes are mostly monocyte-derived, but up to 20% are lymphocytes, predominantly CD4+ CD45RO+ (memory) T cells. To evaluate the potential of adenovirus vectors for studies of mononuclear leukocyte recruitment in vitro, we studied the effects of adenovirus vectors per se on human umbilical vein endothelial cells (HUVECs), a well-characterized in vitro model of vascular endothelium. A recombinant adenovirus containing the seven-domain isoform of rabbit vascular cell adhesion molecule-1 (rVCAM-1) was constructed and used to study lymphocyte adhesion under defined laminar flow conditions in transduced HUVEC monolayers. No increase in basal HUVEC surface expression of the inducible endothelial adhesion molecules and markers of activation, E-selectin and VCAM-1, was noted across a broad range of multiplicity of infection. A modest dose-dependent increase in surface intercellular adhesion molecule-1 expression was detectable by flow cytometry at an MOI of > 30 plaque-forming units per cell. Under defined laminar flow from 1.5 to 0.5 dyne/cm2, the adenovirus vector carrying rVCAM-1 mediated stable adhesion of both a Jurkat T-cell line and primary human CD4+ CD45RO+ (memory) T cells. Monoclonal antibodies to alpha 4-integrin or rVCAM-1 abolished adhesion, whereas monoclonal antibodies to CD18 or P-selectin had no effect. We conclude that adenoviral gene transfer in useful for studies of VCAM-1-dependent leukocyte adhesion in vitro and that endothelial expression of VCAM-1 alone, in the absence of over endothelial cell activation, is sufficient under simulated physiological flow conditions to support adhesion of memory T cells, the predominant lymphocyte subset in atherosclerotic plaque.

Adhesion of monocytes to vascular cell adhesion molecule-1-transduced human endothelial cells: implications for atherogenesis.

To study the role of vascular cell adhesion molecule-1 (VCAM-1) in monocyte recruitment and atherogenesis, we constructed a recombinant adenovirus, AdRSVrVCAM-1, carrying the rabbit VCAM-1 cDNA. We have previously shown that AdRSVrVCAM-1-transduced human umbilical vein endothelial cells (HUVECs) support the adhesion of CD4+ CD45RO+ memory T lymphocytes under laminar flow conditions. We now demonstrate that AdRSVrVCAM-1-transduced HUVECs support the adhesion of peripheral blood monocytes at a shear stress of < or = 1.5 dyne/cm2. Although VCAM-1 supported only firm adhesion of lymphocytes, it was able to mediate monocyte rolling, firm adhesion, and transmigration when expressed in the context of otherwise unactivated vascular endothelium. VCAM-1-transduced HUVECs supported the adhesion of as many as 4-fold more monocytes than T cells under laminar flow. The greater monocyte adhesion was explained at least in part by leukocyte-leukocyte interactions (secondary adhesions), which were not seen with T cells. These secondary monocyte interactions were specifically blocked by monoclonal antibodies to L-selectin and P-selectin glycoprotein ligand-1. These data demonstrate that VCAM-1 expressed in the context of unactivated vascular endothelium supports the adhesion of the leukocyte populations present in atherosclerotic plaque and may contribute to the predominance of monocytes over lymphocytes.