Arthritis Prediction of Advanced Hepatic Fibrosis in HFE Hemochromatosis.

OBJECTIVE: To evaluate whether arthritis predicts the likelihood of advanced hepatic fibrosis in HFE hemochromatosis. PATIENTS AND METHODS: We conducted a retrospective, cross-sectional analysis of 112 well-characterized patients with HFE hemochromatosis and liver biopsy-validated fibrosis staging recruited between January 1, 1983, and December 31, 2013. Complete clinical, biochemical, hematologic, and noninvasive serum biochemical indices (aspartate aminotransferase to platelet ratio index [APRI] and fibrosis 4 index [FIB4]) were available. Scheuer fibrosis stages 3 and 4, APRI greater than 0.44, or FIB4 greater than 1.1 were used to define advanced hepatic fibrosis. Comparisons between groups were performed using categorical analysis, unpaired or paired t test. RESULTS: Male (n=76) and female (n=36) patients were similar in age. Nineteen patients had advanced hepatic fibrosis, and 47 had hemochromatosis arthritis. Arthritis was significantly associated with the presence of advanced hepatic fibrosis as determined by liver biopsy (sensitivity, 84%, [95% CI, 62% to 95%]; negative predictive value, 95% [95% CI, 87% to 99%]; relative risk, 7.4 [95% CI, 2.5 to 23]; P<.001), APRI (sensitivity, 75% [95% CI, 55% to 88%]; negative predictive value, 91% [95% CI, 81% to 96%]; relative risk, 4.5 [95% CI, 2.0 to 10.2]; P<.001), or FIB4 (sensitivity, 61% [95% CI, 41% to 78%]; negative predictive value, 67% [95% CI, 68% to 90%]; relative risk, 2.2 [95% CI, 1.1 to 4.6]; P=.03). Mean cell volume values were significantly higher pretreatment in patients with F3-4 fibrosis (96.7±1.1 fL) compared with F0-2 fibrosis (93.4±0.5 fL; P=.004) and declined following treatment (F3-4, 93.2±0.9 fL, P=.01; F0-2, 91.7±0.6 fL, P=.01). CONCLUSION: Advanced hepatic fibrosis is strongly associated with arthritis in HFE hemochromatosis. The absence of arthritis predicts a low likelihood of advanced hepatic fibrosis, supporting its use as a clinical marker for advanced hepatic fibrosis in HFE hemochromatosis.

The incidence and outcome of clinically significant antibodies detected in Rhesus-D positive pregnant women of the Northern Territory.

BACKGROUND: Haemolytic disease of the fetus/newborn secondary to clinically significant non-Rhesus-D antibodies has risen in importance since the advent of immunoprophylactic anti-D administration to Rhesus-D negative women. Of interest is the incidence of these antibodies in Rhesus-D positive women, who receive less frequent antenatal alloantibody screening. This is of particular concern if the antibodies arise late in pregnancy and may go undetected. AIMS: To assess the proportion of Rhesus-D positive pregnant women with late developing clinically significant antibodies for haemolytic disease of the fetus/newborn, and whether these resulted in adverse fetal outcomes. MATERIALS AND METHODS: A retrospective analysis over a 12-month period at a tertiary hospital in the Northern Territory. Group and antibody screen results in addition to clinical data regarding pregnancy/newborn were collected. RESULTS: Sixty-four of 2612 women (2.5%) had red blood cell antibodies detected during their pregnancy. Of these, 21 clinically significant antibodies were detected in 19 women (0.7% of initial cohort). The most common antibody detected was anti-c (28.5%). In six of these women (0.23% of initial cohort), the antibodies were late developing. Mild jaundice was noted in three newborns with phototherapy required in one. CONCLUSIONS: Although clinically significant antibodies were detected during pregnancy, and in a small proportion of cases as a late developing antibody undetected in the first trimester screening, clinical outcomes for the newborn were mild. As such, the cost of retesting all Rhesus-D positive pregnant women in the third trimester would be considerable and unlikely to result in any meaningful clinical benefit.

CCN2 promotes keratinocyte adhesion and migration via integrin α5ß1.

BACKGROUND: CCN2, (a.k.a. connective tissue growth factor and CTGF) has emerged as a regulator of cell migration. While the importance of CCN2 for the fibrotic process in wound healing has been well studied, the effect of CCN2 on keratinocyte function is not well understood. In this study, we investigated the mechanism behind CCN2-driven keratinocyte adhesion and migration. MATERIALS AND METHODS: Adhesion assays were performed by coating wells with 10 µg/ml fibronectin (FN) or phosphate-buffered saline (PBS). Keratinocytes were seeded in the presence or absence of 200 ng/ml CCN2, 5 mmol/l ethylenediaminetetraacetic acid, 10 mmol/l cations, 500 µl arginine-glycine-aspartic acid (RGD), 500 µM arginine-glycine-glutamate-serine (RGES), and 10 µg/ml anti-integrin blocking antibodies. Migration studies were performed using a modified Boyden chamber assay. Quantitative PCR was used to study the effect of CCN2 on integrin subunit mRNA expression. To block intracellular pathways, keratinocytes were pretreated with 20 µM PD98059 (MEK-1 inhibitor) or 20 µM PF573228 (FAK inhibitor) for 60 min prior the addition of CCN2. Western blot was used to measure CCN2, p-ERK1/2, and ERK1/2. RESULTS: CCN2 enhanced keratinocyte adhesion to fibronectin via integrin α5ß1. The addition of anti-integrin α5ß1 antibodies reduced CCN2-mediated keratinocyte migration. In addition, CCN2 regulated mRNA and protein expression of integrin subunits α5 and ß1. CCN2 activated the FAK-MAPK signaling pathway, and pretreatment with MEK1-specific inhibitor PD98059 markedly reduced CCN2-induced keratinocyte migration. CONCLUSIONS: Our results demonstrate that CCN2 enhances keratinocyte adhesion and migration through integrin α5ß1 and activation of the FAK-MAPK signaling cascade.