Discoid lupus erythematosus-like lesions and stomatitis in female carriers of X-linked chronic granulomatous disease

The skin and oral mucosa were studied in an unselected series of carriers of X-linked chronic granulomatous disease, a hereditary condition in which phagocytic cells display a pronounced functional defect. Three carriers had discoid lupus erythematosus (DLE)-like skin lesions which histopathologically were consistent with DLE of the hypertrophic and profundus type. Four patients had experienced photosensitivity in childhood. Seven patients had recurrent aphthous-like stomatitis which should be distinguished from the recurrent aphthous stomatitis seen in otherwise healthy individuals. The remarkably high incidence of DLE-like symptoms in heterozygous carriers might be related to the presence of mixed populations of defective and normal phagocytes. The variable expression of skin symptoms may be related to uneven distribution of abnormal to normal phagocytes. Female patients with these clinical symptoms, especially the combination of DLE-like skin lesions and aphthous-like stomatitis, should be suspected of being carriers of chronic granulomatous disease and studies of phagocyte function in vitro should be performed, since the diagnosis of the carrier state is of utmost importance for genetic counselling before pregnancy. In order to describe in greater detail the clinicopathological findings and their frequency of expression in the skin and oral mucosa, we have undertaken a prospective study of nine unselected known carriers of X-linked CGD.     

Neutrophils mediate lipid peroxidation in human red cells

Activated neutrophils (ANs) are known to release reactive oxygen species that may cause oxidative damage to surrounding tissues. We determined if ANs could induce lipid peroxidation (LP) in human red cells and investigated the mechanism involved in this interaction. We studied neonatal glucose-6-phosphate dehydrogenase (G6PD) deficient, and sickle red cells, since each of these are known to be susceptible to oxidant injury. Neutrophils were isolated from whole blood and activated by incubation with opsonized zymosan. Mixtures of such neutrophils and red cells at a ratio of 1:100 were incubated for two hours at 37 degrees C, after which the malonyldialdehyde content in red cells was measured as an index of LP. All red cells underwent LP after AN treatment, and the degree of LP was proportional to the amount of AN in the mixture. Superoxide dismutase and catalase partially inhibited LP. When compared to normal red cells, only sickle cells demonstrated a significant increase in AN-mediated LP. Conversion of hemoglobin to carboxy-hemoglobin increased AN-mediated LP, whereas conversion to met- hemoglobin decreased AN-mediated LP. The protective effect of met- hemoglobin on LP was less in sickle cells than in normal cells. We conclude that AN can induce LP in red cells in vitro and that sickle cells are more susceptible to this process than normal cells. Hemoglobin can serve as an electron trap and protect the cell against peroxidative damage, but this mechanism is impaired in sickle cells. We speculate that the pathogenesis of hemolysis associated with infectious disease may include AN-induced red cell LP.     

Multiplexed immunobead‐based assay for detection of oral cancer protein biomarkers in saliva

Objective: For clinical applications of biomarkers, there is a need for multiplex assays using high throughput platforms. The objective of this study was to determine the efficacy of Luminex Multianalyte Profiling (xMAP) technology for measurement of salivary proteins and to evaluate whether multiplex assays are as effective as single‐plex assays and enzyme‐linked immunosorbent assay (ELISA). Results: The average levels of interleukin‐8 (IL‐8) from the single‐plex assay were 3313.2 ± 3759.8 pg ml^?1 [oral squamous cell carcinoma (OSCC), n = 20] and 1061.7 ± 1978.8 pg ml^?1 (control, n = 20). The IL‐1β average levels from the single‐plex assay were 945.2 ± 1134.8 pg ml^?1 (OSCC, n = 20) and 314.2 ± 444.8 pg ml^?1 (control, n = 20). The average levels of IL‐8 from the multiplex assay were 2834.9 ± 3385.6 pg ml^?1 (OSCC, n = 20) and 947.3 ± 2036.8 pg ml^?1 (control, n = 20). The IL‐1β average levels from the multiplex assay were 1013.5 ± 1221.1 pg ml^?1 (OSCC, n = 20) and 376.3 ± 576.3 pg ml^?1 (control, n = 20). The correlation coefficient between Luminex and ELISA assay for IL‐8 (n = 19) and IL‐1β (n = 19) was 0.91 and 0.84, respectively. Conclusion: Luminex xMAP single‐plex and multiplex assays are as effective as ELISA assays for quantification of proteins in saliva. Both IL‐8 and IL‐1β were expressed at significantly higher levels in OSCC subjects than in the matched healthy control subjects.     

Esthetic Provisional Restorations for Porcelain Veneer Preparations

A method to provisionalize porcelain laminate veneers, similar to previously described methods for provi-sionalizing crowns, is presented. The advantages of this system include the ability to have a prototype restoration for evaluation of esthetics and phonetics, thermal protection, and protection from preparation fracture. Additional advantages include improved tissue health and ease of progressing from the provisionaliza-tion stage to the final bonding stage.     

Avoiding calcineurin inhibitors in the early post-operative course in high-risk liver transplant recipients: The role of extracorporeal photopheresis

The aim of this work is to report on the results of a single-center, prospective study on the feasibility of calcineurin-inhibitor (CNI)-staggered immunosuppression by use of extracorporeal photopheresis (ECP) in liver transplant (LT) recipients at risk of renal and neurological complications. Patients were matched on a 1:1 basis with historical controls on standard CNI immunosuppression. ECP patients were treated with ECP plus antimetabolites and/or steroids, while CNIs were withheld until clinically indicated. Thirty-six patients were evaluated: 18 ECP patients and 18 controls. ECP was tolerated in 100% of cases. CNI were introduced at a median of 8 days (4-55) in 17 ECP patients, while one patient was on a fully CNI-sparing regimen 22 months after LT. Acute rejection occurred in 27.7% patients in ECP (5/18) versus 16.7% in controls (3/18) (P = ns) with a shorter time to rejection in ECP (36 +/- 31.3 days vs. 83.6 +/- 65.6 days; P = ns). All rejection episodes were amenable to medical treatment. Neurological and renal complications occurred in 22.2% (4/18) of patients in either group, but led to in-hospital mortality in 3 patients among controls versus 1 in ECP (P = ns). One-, 6-, and 12-month survival rates were 94.4, 88.1, and 88.1% in ECP versus 94.4, 77.7, and 72.2% among controls (P < 0.0001). ECP seems to allow for management of high-risk LT recipients in the early post-transplant course and reduction of CNI-related mortality. Continued data validation is favored to assess the impact of ECP on long-term graft and patient survival.     

Plasma levels of ionized and total calcium during storage of citrated platelet concentrate

Measurements of ionized and total calcium levels in supernatant plasma samples from citrated platelet concentrates (PCs) were made over 7 days of storage. Both ionized and total calcium increased significantly during the storage period: respectively, from 0.074 mM Ca2+ in fresh platelet-rich plasma to 0.084 mM in PCs stored for 7 days (p = 0.017), and from 1.94 mM total calcium to 2.06 mM (p = 0.014) over the same period. The increase in calcium was partially blocked by the addition of platelet activation inhibitors to the PCs. Platelet-poor plasma stored under similar conditions showed no significant change in ionized or total calcium, which indicated that the increases observed in PCs were due to the release of cellular calcium. Significant correlations (p less than 0.01) were found between ionized or total calcium levels and lactate concentration or pH, but not hypotonic shock recovery rate. The demonstration of non-zero levels of ionized calcium makes it likely that Ca2+-dependent enzyme systems such as calpain expression and thrombin generation are active in the plasma of citrated PCs and may contribute to the platelet storage lesion.     

Acute colonic pseudo-obstruction after elective total joint arthroplasty

A retrospective review of 31 patients who developed acute colonic pseudo-obstruction (ACPO) after total joint arthroplasty was undertaken to determine predisposing factors related to, and outcomes following, therapeutic intervention. Comparison with all patients who underwent total joint arthroplasty revealed an overall 1.2% incidence of ACPO. There was a higher incidence of ACPO in patients undergoing sequential bilateral total knee arthroplasty (3.4%) compared with unilateral total knee arthroplasty (0.3%) and a higher incidence in patients undergoing revision total hip arthroplasty (5.6%) compared with primary total hip arthroplasty (1.4%). Additional risk factors for developing ACPO included slow postoperative mobilization and male gender. No association was found with respect to body mass index, age, comorbidity, anesthetic type, international normalized ratio level, or postoperative analgesia. There were no deaths, and 2 patients required operative intervention. The remaining cases of ACPO resolved with nonsurgical treatment. In all cases, there was a prolonged length of hospitalization (mean, 13.2 days) compared with all other arthroplasties performed at our institution (mean, 7.5 days).     

NS6180, a new KCa3.1 channel inhibitor prevents T-cell activation and inflammation in a rat model of inflammatory bowel disease

Background and Purpose

The K_Ca3.1 channel is a potential target for therapy of immune disease. We identified a compound from a new chemical class of K_Ca3.1 inhibitors and assessed in vitro and in vivo inhibition of immune responses.

Experimental Approach

We characterized the benzothiazinone NS6180 (4-[[3-(trifluoromethyl)phenyl]methyl]-2H-1,4-benzothiazin-3(4H)-one) with respect to potency and molecular site of action on K_Ca3.1 channels, selectivity towards other targets, effects on T-cell activation as well as pharmacokinetics and inflammation control in colitis induced by 2,4-dinitrobenzene sulfonic acid, a rat model of inflammatory bowel disease (IBD).

Key Results

NS6180 inhibited cloned human K_Ca3.1 channels (IC₅₀ = 9 nM) via T250 and V275, the same amino acid residues conferring sensitivity to triarylmethanes such as like TRAM-34. NS6180 inhibited endogenously expressed K_Ca3.1 channels in human, mouse and rat erythrocytes, with similar potencies (15–20 nM). NS6180 suppressed rat and mouse splenocyte proliferation at submicrolar concentrations and potently inhibited IL-2 and IFN-γ production, while exerting smaller effects on IL-4 and TNF-α and no effect on IL-17 production. Antibody staining showed K_Ca3.1 channels in healthy colon and strong up-regulation in association with infiltrating immune cells after induction of colitis. Despite poor plasma exposure, NS6180 (3 and 10 mg·kg⁻¹ b.i.d.) dampened colon inflammation and improved body weight gain as effectively as the standard IBD drug sulfasalazine (300 mg·kg⁻¹ q.d.).

Conclusions and Implications

NS6180 represents a novel class of K_Ca3.1 channel inhibitors which inhibited experimental colitis, suggesting K_Ca3.1 channels as targets for pharmacological control of intestinal inflammation.