The final destiny of acantholytic cells in pemphigus is Fas mediated

Background  Pemphigus is an autoimmune disease characterized by the formation of intra-epidermal blisters. Patients develop auto-antibodies against desmoglein 1 and 3 proteins and induce acantholysis.
Objective  This work addresses the issue of whether the Fas pathway mediates acantholysis. Furthermore, the possible suppliers of the Fas pathway were investigated.
Methods  Seventeen biopsies of pemphigus patients were studied by haematoxylin and eosin staining, and apoptosis was defined by TUNEL. The expression of Fas, FasL and caspase 3 was studied by in situ hybridization and immunohistochemistry. Cell infiltrates were studied by immunofluorescence with monoclonal anti-CD3, CD4, CD8, CD19 and CD69.
Results  All of the biopsies showed intra-epidermal blisters, acantholytic cells and inflammatory infiltrates. The blisters expressed Fas, FasL and caspase 3. Cell infiltrates were composed of CD8 and a few CD4⁺CD69⁺ cells. Additionally, CD19⁺ cells were detected. Interestingly, the Fas expression was increased in acantholytic cells and perilesional keratinocytes. Incidentally, these cells exhibited apoptotic features. Interestingly, the CD8 cells expressed FasL.
Conclusion  This paper presents the morphological evidence that apoptosis and acantholysis are linked. Therefore, the Fas pathway is associated with CD8 cells in pemphigus lesions.

Conflicts of interest

None declared.     

Doppler studies evaluating the effect of a physical therapy screening protocol on vertebral artery blood flow

General and isolated cervical positional tests are used to screen for potential vertebro-basilar insufficiency (VBI). There is limited research evaluating vertebral artery blood flow in these positions to justify the rationale of progressive mechanical stress occurring to the arteries. The purpose of the study was to determine vertebral artery blood flow in six cervical positions used in clinical practice. A comprehensive cervical assessment was conducted on 22 men and women (mean age 35) with no known vascular pathology. Vertebral artery peak systolic (PS), end diastolic (ED) flow rates and resistive index (RI) were measured using duplex colour Doppler sonography (sampling at C3-C5) in neutral, rotation, extension, combined rotation-extension, combined rotation-extension-traction, deKelyn's position and a C1-C2 pre-manipulative hold. Results showed there was a significant decrease in PS and ED in the contra-lateral artery during the pre-manipulative hold, and a decrease in ED in the contra-lateral artery during rotation. There was no effect of age, gender or mobility restriction on these blood flow changes. The pre-manipulative hold had the greatest response with 34% of the arteries demonstrating a complete cessation of ED flow. In conclusion the pre-manipulative hold and rotation created the greatest mechanical stress to the contra-lateral vertebral artery. These two positions may be useful screening positions to identify individuals at risk for VBI due to inadequate collateral blood flow.     

Nuclear RNA sequences coding for alpha and beta globins in erythroid cells: evidence for multiple intermediate molecules

The poly (A)-containing nuclear RNA from dimethylsulfoxide-induced Friend leukemia cells was fractionated by acrylamide gel electrophoresis in denaturing conditions and analyzed for alpha and beta globin RNA sequences. The results indicate that nuclear RNA contains one species of large-size RNA (0.6 X 10(6) daltons), which is the putative precursor for beta globin mRNA only. In addition, it was shown by electrophoretic analysis that the complex of RNA molecules not resolved by sucrose gradient centrifugation (11S) comprises sequences of decreasing size (0.34, 0.28, and 0.26 X 10(6) daltons), which might be the precursors of alpha and beta globin mRNA.     

Myocardial lactate metabolism at rest and during exercise in ischemic heart disease

Cardiac dynamics and myocardial extraction of oxygen and lactate were investigated at rest in subjects with normal coronary arteries and in patients with coronary heart disease. Coronary artery disease was considered significant when there was obstruction or more than 50 per cent narrowing of one or more coronary arteries. Half of the patients were also studied during moderate exercise. The hemodynamic findings were almost comparable in all cases. On exercise, in patients with significant coronary disease, there was evidence of abnormal left ventricular function, and increases in cardiac index were mediated through a much higher left ventricular end-diastolic pressure.

Coronary arteriovenous difference and the myocardial extraction of oxygen were normal at rest and unchanged on exercise in all cases. Marked rise in the arterial level and a significant increase in the arteriovenous difference of lactate were present in all cases. The extraction ratio of lactate by the myocardium remained normal and increased only slightly in control subjects. However, this ratio increased to normal levels during exercise in all patients with significant coronary artery disease. Possible explanations for the increased coronary arteriovenous difference and myocardial extraction ratio of lactate on exercise in these patients are discussed.     

The FLK2/FLT3 ligand synergizes with interleukin-7 in promoting stromal- cell-independent expansion and differentiation of human fetal pro-B cells in vitro

The effects of a novel cytokine FLK2/FLT3 ligand (FL) on human fetal bone marrow-derived CD34+CD19+ pro-B cells were analyzed in a stromal- cell-independent, serum-deprived culture system. FL, like interleukin-3 (IL-3), synergized with IL-7 in promoting pro-B cell growth, and differentiation of these cells into CD34-CD19+clgM+slgM- pre-B cells, whereas a small proportion of these cells even differentiate into more mature slgM+ B cells. In contrast, KIT ligand (KL) and granulocyte- macrophage colony-stimulating factor (GM-CSF) were ineffective in promoting IL-7-dependent pro-B cell growth and differentiation. Maximal levels of pro-B cell expansion, generally resulting in 15- to 30-fold increases in cellularity, were obtained in cultures supplemented with optimal doses of FL + IL-7 + IL-3. The addition of mouse bone marrow stromal cells further enhanced the proliferation and differentiation of pro-B cells obtained in the presence of these three cytokines. Under these conditions, cultures could be maintained for more than 4 weeks, and in general 40- to 50-fold increases in cell numbers were observed by 3 weeks of culture. The percentages of clgM+ and slgM+ B cells increased 1.5- to 3-fold and 2-fold, respectively, suggesting that stromal cells may provide additional costimulatory signals for human B- cell growth and differentiation that are different from IL-7, IL-3, and FL. Collectively, our results indicate that FL, in contrast to KL, strongly promotes long-term expansion and differentiation of human pro- B cells in the presence of IL-7 or in combination of IL-7 and IL-3, which is a novel property of this hematopoietic growth factor.