Effect of malnutritional rehabilitation on tuberculin reactivity and complement level in rats

The purpose of this study was to clarify the effect of differing nutritional states on various components of the immune system, especially on the interplay of the complement system and cell-mediated immunity. Malnutrition was induced in Sprague—Dawley rats by feeding them diets containing 5% protein or 0.5% protein as compared with 18% protein in the diet of the controls. Nutritional rehabilitation was achieved in some experimental groups by transferring those fed 0.5% protein diet to the 18% protein diet. Malnutrition was confirmed by weight changes, biochemical findings in the sera, haematological observations and histological observation of the liver, and rehabilitation was confirmed by body weight increase and changes in other measurements. In rats suffering malnutrition, the tuberculin skin reactivity was suppressed. After feeding the 0.5% protein diet for 8 weeks, all the rats showed negative tuberculin skin reactions. In the malnourished rats, including those fed with 0.5% protein, the serum complement level decreased but did not show any significant differences as compared with the well nourished control group. After 1 week of nutritional rehabilitation, the tuberculin reactivity of six out of ten rats remained negative and after 2 weeks, all rats showed positive tuberculin reactions. After 1 week of nutritional rehabilitation, all the rats showed a normal or higher serum complement level. At this stage, two of the tuberculin-negative rats showed significantly higher titre of serum complement than even the controls.     

Activated microglia in a rat stroke model express NG2 proteoglycan in peri‐infarct tissue through the involvement of TGF‐β1

We investigated activated microglia in ischemic brain lesions from rats that had been subjected to transient middle cerebral artery occlusion. Activated microglia expressing NG2 chondroitin sulfate proteoglycan (NG2) were found only in the narrow zone (demarcation zone) that demarcated the peri‐infarct tissue and ischemic core. NG2^? activated microglia were abundantly distributed in the peri‐infarct tissue outside the demarcation zone. NG2⁺ microglia but not NG2^? microglia expressed both CD68 and a triggering receptor expressed on myeloid cells 2 (TREM‐2), suggesting that NG2⁺ microglia eliminated apoptotic neurons. In fact, NG2⁺ microglia often attached to degenerating neurons and sometimes internalized NeuN⁺ or neurofilament protein⁺ material. Kinetic studies using quantitative real‐time RT‐PCR revealed that expression of transforming growth factor‐β1 (TGF‐β1) was most evident in the ischemic core; with this marker produced mainly by macrophages located in this region. TGF‐β receptor mRNA expression peaked at 3 days post reperfusion (dpr) in the peri‐infarct tissue, including the demarcation zone. Primary cultured rat microglia also expressed the receptor mRNA. In response to TGF‐β1, primary microglia enhanced the expression of NG2 protein and TREM‐2 mRNA as well as migratory activity. A TGF‐β1 inhibitor, SB525334, abolished these effects. The present results suggest that TGF‐β1 produced in the ischemic core diffused toward the peri‐infarct tissue, driving activated microglial cells to eliminate degenerating neurons. Appropriate control of NG2⁺ microglia in the demarcation zone might be a novel target for the suppression of secondary neurodegeneration in the peri‐infarct tissue. GLIA 2014;62:185–198     

IDENTIFICATION OF A UNIVERSAL B CELL EPITOPE ON DNA TOPOISOMERASE I,AN AUTOANTIGEN ASSOCIATED WITH SCLERODERMA

Objective. To investigate the distribution of B cell autoepitopes of human DNA topoisomerase I (topo I), an autoantigen associated with scleroderma. Methods. A complementary DNA clone, T1B, was used to produce recombinant proteins of topo I as β-galactosidase fusion proteins. Immunoreactivity to these fusion proteins was then tested in 35 anti–topo I–positive sera from patients with scleroderma, by immunoblotting, enzyme-linked immunosorbent assay, and double immunodiffusion. Results. One epitope was found to be universally recognized by all sera tested. Thirty-two of the samples recognized multiple antigenic regions, but sera from the remaining 3 patients recognized only this universal epitope, and in longitudinal studies of 1 of these 3 patients, the serum recognized only this epitope for more than 2 years, even though multiple, potent, antigenic regions were found on topo I. Conclusion. Recognition of multiple epitopes in most patients suggests that the topo I molecule itself would drive the autoimmunity on topo I. However, antigen-driven autoimmunity could not explain the production of the monoreactive anti–topo I antibody seen in the 3 patients. We thus hypothesize that there is a process whereby recognition of the universal epitope by cross-reaction develops into antigen-driven autoimmunity.     

Secretion of human atrial natriuretic peptide in response to atrial pacing and its function in patients with ventricular septal defect

The capacity to secrete human atrial natriuretic peptide (hANP) in response to atrial pacing and the resulting changes in diuresis and urinary electrolyte excretion were compared in children with and without a ventricular septal defect (VSD). The subjects examined were 9 children with a history of Kawasaki disease (as controls) and 11 patients with a VSD, including 5 patients with congestive heart failure (CHF). Their ages ranged from 6 to 40 months old. Atrial pacing resulted in a significant increase in the plasma hANP level from 40 +/- 19 to 140 +/- 37 pg/ml in the controls and from 757 +/- 762 to 1540 +/- 1160 pg/ml in the VSD patients. In control children, the urinary flow rate increased 2.3-fold, urinary sodium excretion increased 6.2-fold and urinary chloride excretion increased 7.6-fold, but these values increased only slightly in VSD patients, especially in those patients with CHF, in spite of the marked increase in their plasma hANP level. These results indicate that the capacity for hANP secretion was increased in VSD patients who had chronic volume overloading of the left atrium, but that their diuresis and urinary electrolyte excretion in response to hANP were attenuated.     

Reactive oxygen species-producing site in hydrogen peroxide-induced apoptosis of human peripheral T cells: involvement of lysosomal membrane destabilization

In our previous study, we examined the effect of exogenous hydrogen peroxide, which causes a potent oxidative stress and has been demonstrated to be a potent apoptosis-inducer in many kinds of cells. We found that the addition of 1 or 10 mM hydrogen peroxide induced reactive oxygen species (ROS) formation, oxidative DNA damage, dysfunction of the mitochondrial membrane potential, and early apoptotic changes in the human osteosarcoma cell line HS-Os-1. We therefore concluded that intracellular ROS formation was involved in the hydrogen peroxide-induced apoptosis of HS-Os-1 cells. In contrast to the osteosarcoma cell line HS-Os-1, human peripheral T cells are considered to be easily susceptible to oxidative stress, because these cells lack peroxidase activity. Therefore, in this study, we investigated the site of ROS formation by utilizing MitoCapture, H2DCFDA (succinimidyl ester of dichloro-dihydrofluorescein diacetate), DAPI (4',6-diamidino-2-phenylindole), and LysoSensor. Our results showed that ROS formation was apparently diffusely distributed in T cells oxidatively stressed with 0.1 mM hydrogen peroxide. Moreover, lysosomal swelling and deformity, possibly revealing lysosomal membrane destabilization, were observed in these cells. Based on the above results, there exists an apoptotic cascade involving early lysosomal membrane destabilization in the hydrogen peroxide-induced apoptosis of human peripheral T cells. Therefore, the possible involvement of lysosomal protease leakage caused by hydroxyl radical formation in lysosomes (possibly resulting in mitochondrial membrane dysfunction) is considered to play an important role in hydrogen peroxide-induced T cell apoptosis.     

Increased prenatal blood manganese may induce gestational blood pressure

Objective: Pregnancy hypertension is the most common gestational complication and poses a critical risk for mother and fetus. Whether environmental factors may play an important role in disease occurrence is not fully determined. Methods: To investigate the effects of prenatal manganese (Mn) exposure on gestational blood pressure, 386 women were examined. Results: Early pregnancy blood Mn was significantly (p < 0.05) correlated with blood pressure through gestation. A significant association between odds of pre-hypertension with blood Mn was shown (OR:1.150, 95% CI:1.052–1.258). Conclusion: The current study results might suggest the blood Mn level during early stage of pregnancy as a potential risk factor for increasing the risk of gestational blood pressure.