TLR2 Expression in Relation to IL-6 and IL-1β and their Natural Regulators Production by PMN and PBMC in Patients with Lyme Disease

Recently, it has been reported that TLR2 on macrophages plays a unique role in the inflammatory response and host defense to infection with Borrelia burgdorferi (Bb) which is an etiologic agent of Lyme disease. Experimental studies show that PMNs also play an essential role in infection control by Bb. However, there is no available data about TLR2 expression on PMN in the course of Lyme disease. In the present study, TLR2 expression and production of IL-1β and IL-6 as well as their natural regulators (sIL-1RII, IL-1Ra and sIL-6Rα, sgp130, resp) by PMN of peripheral blood in patients with Lyme disease were examined. For the purpose of comparison, the same activity of autologous peripheral blood mononuclear cells (PBMCs) was estimated. An effect of rhIL-15 on TLR2 and cytokine secretion was also studied. Increased TLR2 expression in unstimulated neutrophils suggests an important role of these cells in mechanism recognition of B burgdorferi in patients with Lyme disease. The relationship between IL-1β and IL-6 as well as their regulators by unstimulated PMN and PBMC, observed in the present study, may lead to enhanced IL-6- and to inhibition of IL-1β-mediated reactions in this patient group. Changes in the TLR2 expression after rhIL-15 stimulation appear to have a favorable effect on mechanism recognition of Bb. The relations between IL-6 and its regulators (sIL-6Rα and sgp130) as well as between IL-1β and its regulators (IL-1Ra and sIL-1RII) after rhIL-15 stimulation may lead to enhanced IL-1β- and IL-6-mediated inflammatory reactions in the course of Lyme disease

Role of interleukin-15 and interleukin-18 in the secretion of sIL-6R and sgp130 by human neutrophils.

BACKGROUND: Available data indicate that neutrophils (PMN) produce a wide range of cytokines with the potential to modulate immune response. Recent investigation have shown that interleukin (IL)-15 and IL-18 potentiated several functions of normal neutrophils. It has been reported that IL-18-induced cytokine production may be significantly enhanced by coincident addition of IL-15. AIMS: In the present study we compared the effect of recombinant human (rh)IL-15 and rhIL-18 as well as effect of a rhIL-15 and rhIL-18 combination on the induction secretion of sIL-6Ralpha and sgp130 by human neutrophils. METHODS: PMN were isolated from heparinized whole blood of healthy persons. The PMN were cultured for 18 h at 37 degrees C in a humidified incubator with 5% CO(2). rhIL-15 and/or rhIL-18 and lipopolysaccharide were tested to PMN stimulation. The culture supernatants of PMN were removed and examined for the presence of sIL-6R and sgp130 by human enzyme-linked immunosorbent assay kits. Cytoplasmic protein fractions of PMN were analysed for the presence of sIL-6R and sgp130 by western blotting using monoclonal antibodies capable of detecting these proteins. Cells were lysed and cytoplasmic proteins were electrophoresed on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The resolved proteins were transferred onto nitrocellulose and incubated with the primary monoclonal antibodies anti-sIL-6R and anti-sgp130. The membranes were incubated at room temperature with alkaline phosphatase anti-mouse immunoglobulin G. Immunoreactive protein bans were visualized by an AP Conjugate Substrate Kit. RESULTS AND CONCLUSIONS: The results of our investigation revealed that IL-15 alone, similarly to IL-18, has no significant ability for the regulation of both soluble IL-6 receptors, sIL-6R and sgp130, released by human neutrophils. It is interesting to note that the secretion of sgp130 was changed after PMN stimulation with rhIL-15 in the presence of rhIL-18. The combination of rhIL-15 and rhIL-18 was shown to induce PMN to secretion relatively higher amounts of sgp130 compared with the stimulation of PMN with rhIL-15 alone and rhIL-18 alone. The results obtained suggest that IL-15 and IL-18, belonging to the inflammatory cytokines, through the regulation of sgp130 secretion must be also considered as anti-inflammatory mediators that may influence the balance reactions mediated by the IL-6 cytokine family

A comparison of two approaches in teaching reading to low achieving adult women

This study was undertaken to determine which of two programs in reading - Words in Color or Traditional Basal - proved more effective as a short term course in reading for low achieving adult women. More specifically, answers to the following questions were sought. 1. Which of the two programs shows better total reading gains as measured by the California Reading Test? 2. Which of the two programs yields better results in comprehension as measured by the California Reading Test? 3. Which of the two programs shows better gains in word recognition as measured by the Wide Range (Reading Section) Test? 4. Were there any observations which might lead one to believe that one or the other program was more enthusiastically received? 5. Are these indications that these students will be favourably motivated to continue reading on their own

Role of soluble gp130 in the tumour necrosis factor-alpha expression and its production by peripheral blood mononuclear cells.

Background: In our previous study we found that rhsIL-6R, along with recombinant human interleukin-6, plays a regulatory role in the immune response by modulating the tumour necrosis factor-α (TNF-α) expression and its production by peripheral blood mononuclearcells (PBMC). We also suggested that sIL-6R with IL-6 secreted by human PMN (neutrophils) influenced the TNF-α expression and its production by autologous PBMC

The release of soluble forms of TRAIL and DR5 by neutrophils of oral cavity cancer patients.

In the present study we examined the release of the soluble form of TRAIL by neutrophils (PMN) derived from patients with oral cavity cancer. Simultaneously, we estimated the ability of PMNs of these patients to release the soluble form of DR5 receptor, a natural regulatory protein of TRAIL. The obtained results were confronted with the serum levels of sTRAIL and sDR5. The cells were isolated from 21 patients with squamous cell carcinoma of oral cavity at diagnosis and three weeks after surgery treatment. For comparative purposes we performed similar examinations in autologous peripheral blood mononuclear cells (PBMC). Cytoplasmic protein fractions of the cells were analyzed for the presence of TRAIL and DR5 by western blotting. Soluble TRAIL and soluble DR5 concentrations in the culture supernatants of cells were confronted with their serum levels using ELISA kit. PMN and PBMC of the whole cancer patient group expressed decreased TRAIL protein and unchanged expression of DR5 receptor in comparison with the control group. Unchanged release of sTRAIL by PMNs of patients in Stage II was accompanying the decrease of the ability of PBMC to secrete this protein. In patients in Stage IV the secretion of sTRAIL by PMNs and PBMC was impaired. In contrast to changes in sTRAIL secretion by PMN and PBMC of oral cavity cancer patients, the secretion of sDR5 by these cells was unchanged. The serum levels of sTRAIL were increased in patients in Stage II before treatment and decreased in the same patients after treatment. The altered ability of PMN of PBMC to secrete sTRAIL may have different implications for the immune response of patients with oral cavity cancer cells at different stages of disease

Role of Stem Cells and Extracellular Matrix in the Regeneration of Skeletal Muscle

Adult skeletal muscle has a remarkable capacity to initiate a rapid and extensive repair process after damage due to injury or degenerative disease. Although satellite cells are the primary skeletal muscle stem cells, there are many reports of non-satellite cell populations with myogenic capacity resident within skeletal muscle. The activity of muscle-resident stem cells during the regeneration process is tightly controlled through the dynamic interactions between intrinsic factors within the cells and extrinsic factors constituting the muscle stem cell niche. The extracellular matrix (ECM) in skeletal muscle plays an integral role in force transmission, structural maintenance, and regulation of stem cell niche. ECM interacts with stem cells either directly by binding cell surface receptors or indirectly through growth factor presentation, and maintains a balance between their quiescence, self-renewal, and differentiation. These interactions are reciprocal since the stem cells can remodel the niche and secrete or degrade ECM components. Natural ECM scaffolds, derived from decellularized tissues can influence stem cell activity both in vitro and in vivo and are widely being investigated for skeletal muscle repair. In this chapter, we discuss the regenerative potential of stem cell populations and ECM bioscaffolds in the treatment of skeletal muscle injury and disease

Short-term treatment with nitrate is not sufficient to induce in vivo antithrombotic effects in rats and mice

In humans, short-term supplementation with nitrate is hypotensive and inhibits platelet aggregation via an nitric oxide (NO)-dependent mechanism. In the present work, we analyzed whether short-term treatment with nitrate induces antithrombotic effects in rats and mice. Arterial thrombosis was evoked electrically in a rat model in which renovascular hypertension was induced by partial ligation of the left renal artery. In mice expressing green fluorescent protein, laser-induced thrombosis was analyzed intravitally by using confocal microscope. Sodium nitrate (NaNO(3)) or sodium nitrite (NaNO(2)) was administered orally at a dose of 0.17 mmol/kg, twice per day for 3 days. Short-term nitrate treatment did not modify thrombus formation in either rats or mice, while nitrite administration led to pronounced antithrombotic activity. In hypertensive rats, nitrite treatment resulted in a significant decrease in thrombus weight (0.50 ± 0.08 mg vs. VEH 0.96 ± 0.09 mg; p < 0.01). In addition, nitrite inhibited ex vivo platelet aggregation and thromboxane B(2) (TxB(2)) generation and prolonged prothrombin time. These effects were accompanied by significant increases in blood NOHb concentration and plasma nitrite concentration. In contrast, nitrate did not affect ex vivo platelet aggregation or prothrombin time and led to only slightly elevated nitrite plasma concentration. In mice, nitrate was also ineffective, while nitrite led to decreased platelet accumulation in the area of laser-induced endothelial injury. In conclusion, although nitrite induced profound NO-dependent antithrombotic effects in vivo, conversion of nitrates to nitrite in rats and mice over short-term 3-day treatment was not sufficient to elicit NO-dependent antiplatelet or antithrombotic effects

Methemoglobinemia Secondary to Benzocaine Topical Anesthetic.

An 80-year-old white woman with a history of hypertension presented to the Emergency Department with bright red bleeding from the rectum. She was treated with 5 mg of midazolam and benzocaine topical anesthetic spray prior to undergoing colonoscopy and esophageal gastroduodenoscopy. Thirty minutes after endoscopy, the patient became cyanotic and dyspneic; she was suffering from methemoglobinemia, a rare complication secondary to the use of benzocaine topical anesthetic spray. After i.v. administration of methylene blue, 120 mg (2 mg/kg) in 100 cc of normal saline solution, the cyanosis and dyspnea resolved