Anti-Inflammatory Effect of Lemon Mucilage: In Vivo and In Vitro Studies

The mucilage extracted from a lemon juice centrifugation pulp was studied for its anti-inflammatory effect in rat. In vivo the lemon mucilage significantly inhibited carrageenan-induced edema in rat paw from 59% to 73.5% showing the highest effect at the third hour. In vitro, at the doses of 10^?8, 10^?6, 10^?4 or 10^?2 mg/mL the lemon mucilage stimulated the superoxide anion production in rat testing neutrophils in whole blood but inhibited it in FMLP stimulated cells at the dose of 10^?2 mg/mL. The neutrophils of rats receiving p.o. the lemon mucilage for 21 days showed a significant decrease of 45.5% in O₂^? generation after FMLP stimulation, and a not-significant increase after phorbol-12-myristate-13-acetate (PMA) or zymosan stimulation. Since the activity on zymosan- and PMA-induced O₂^? production was not significant, the inhibition exerted by FMLP in rat neutrophils occurred mainly through the blockade of phospholipase D.     

阳极氧化活化处理纯钛经皮种植体的体内外实验研究

为解决经皮器械在负重条件下的长期稳定和经皮密封问题，本文对比研究了阳极氧化表面以及光滑表面的纯钛种植体经皮植入动物体内的情况．并同时将人皮肤表皮细胞接种于这两种表面上进行培养，初步探讨了阳极氧化活化处理后的纯钛金属作为经皮植入体的可能性。结果表明：经皮部分钛金属的光滑表面与阳极氧化的微观粗糙表面对于术后炎性反应的影响无明显差异，阳极氧化活化表面不仅能与骨形成牢固结合，而且也可以与皮下组织紧密贴附。同时，在体内种植体表面形成的钙磷层可能也是形成经皮密封的原因之一，阳极氧化活化钛的微孔粗糙表面对表皮也有一定的锁合固定作用。因此阳极氧化的表面活化方法有可能成为有效的解决经皮生物密封的活化方法之一。     

Qualitative and Quantitative Assessment of the Benefit-Risk Ratio of Medium Potency Topical Corticosteroids In Vitro and In Vivo

Corticosteroids are widely used for the treatment of inflammatory skin disorders. However, systemic and local adverse drug reactions, especially skin atrophy, are potential complications that limit their use. Several attempts have been made to increase the safety of topical corticosteroid treatment, including new application schedules, special vehicles and new agents. In particular, the group of hydrocortisone and prednisolone double esters, with prednicarbate as the first and most often prescribed representative, seem to be equipotent alternatives to the gold standard betamethasone 17-valerate with respect to anti-inflammatory activity. At the same time, these new agents induce less skin atrophy, which may result from a unique skin metabolism and a specific influence on the cytokine network in the epidermis and dermis. On the basis of these effects, a new approach to in vitro quantification of the benefit-risk ratio has been developed. As already suggested by investigations in human volunteers, the benefit-risk ratio of the new compounds appears to be increased. Therefore, recent research has focused on drugs that selectively modulate cytokine release.     

Cellular Immunity In Vitro: Migration Inhibition and Phagocytosis

The activity of peritoneal exudate cells from candidin-sensitive and normal guinea pigs with and without antigen was studied in vitro with the aid of time-lapse, phase-contrast cinemicrography. Macrophages from normal animals migrate well on an agar medium and readily phagocytose Merthiolate-killed Candida albicans cells. A reduction in the migration of peritoneal macrophages from guinea pigs sensitized to C. albicans during the first 24 hr after exposure to antigen was accompanied by a decrease in phagocytosis of C. albicans cells. The macrophages were viable but comparatively immotile. Since the sensitized macrophages came from resistant donors, it is possible that the initial stage of cellular immunity to C. albicans is associated with a reduced activity of the phagocytic macrophages, apparently to limit spread of the pathogen from the infected area.     

Reevaluation of the Effect of Levamisole in Chronic Brucellosis: In Vitro and in Vivo Effect on Monocyte Phagocytosis

Abstract

The in vitro effect of levamisole on peripheral blood monocyte(P.B.M.) phagocytosis was studied in 32 patients with chronic brucellosis and 20 normal subjects. It was shown that levamisole enhances P.B.M. phagocytic capacity, not reaching however normal levels. A subgroup of 11 patients were treated with levamisole for 6 months and the drug effect on cellular and humoral immunity and monocyte phagocytosis was also studied. By the end of the 6 month treatment-study period, the following results were obtained: 1. six patients were symptom free while five had significantly improved. 2.T lymphocyte number and monocyte phagocytosis reached normal values. 3. Significant specific cellular immunity against both brucella antigens was noted. 4.B lymphocytes showed no significant changes. 5. Antiglobulin titers varied. These findings suggest that the good therapeutical effect of levamisole in patients with chronic brucellosis could probably be attributed to the enhancement of both T-cell function and monocyte phagocytosis.     

Reevaluation of the Effect of Levamisole in Chronic Brucellosis: In Vitro and in Vivo Effect on Monocyte Phagocytosis

The in vitro effect of levamisole on peripheral blood monocyte(P.B.M.) phagocytosis was studied in 32 patients with chronic brucellosis and 20 normal subjects. It was shown that levamisole enhances P.B.M. phagocytic capacity, not reaching however normal levels. A subgroup of 11 patients were treated with levamisole for 6 months and the drug effect on cellular and humoral immunity and monocyte phagocytosis was also studied. By the end of the 6 month treatment-study period, the following results were obtained: 1. six patients were symptom free while five had significantly improved. 2.T lymphocyte number and monocyte phagocytosis reached normal values. 3. Significant specific cellular immunity against both brucella antigens was noted. 4.B lymphocytes showed no significant changes. 5. Antiglobulin titers varied. These findings suggest that the good therapeutical effect of levamisole in patients with chronic brucellosis could probably be attributed to the enhancement of both T-cell function and monocyte phagocytosis.     

Apoptosis of Renal Cortical Cells in the Hemolytic-Uremic Syndrome: In Vivo and In Vitro Studies

This study examined apoptotic cell death associated with Shiga-like toxin (Stx)-producing Escherichia coli. Renal cortices from three children with postenteropathic hemolytic-uremic syndrome (HUS) and from mice infected with E. coli O157:H7 and pediatric renal tubular epithelial cells stimulated with Stx and E. coli O157:H7 extracts were examined for apoptotic changes. Apoptotic cells were detected by terminal dUTP nick end labeling of tubuli and glomeruli from HUS patients and from mice inoculated with Stx-2-positive and Stx-negative strains. Apoptosis was more extensive and severe ultramorphological nuclear and cytoplasmic changes were seen in the Stx-2-positive group. Stx caused DNA fragmentation and ultramorphological changes indicating apoptosis in cultured pediatric tubular cells. DNA fragmentation increased when cells were prestimulated with tumor necrosis factor alpha. Polymyxin extracts from Stx-2-positive and Stx-negative strains induced DNA fragmentation, but only extracts from Stx-2-positive strains caused ultramorphological changes and extensive DNA fragmentation. The results indicate that HUS is accompanied by increased apoptosis of kidney cells and that bacterial factors, possibly together with host cytokines in vivo, may activate apoptotic tissue injury.     

Transdermal Delivery of Amino Acids and Antioxidants Enhance Collagen Synthesis: In Vivo and In Vitro Studies

One of the most visible changes associated with the aging process in humans relates to a progressive thinning of the skin. This results from a decline in both collagen and glycosaminoglycans, as well as from changes in their chemical structure and 3-dimentional organization. Transdermal administration of antioxidants, α -lipoic acid (LA) (0.5%) and proanthocyanidin PA) (0.3%) in a standard cosmetic vehicle base formulation supplemented with 2% benzyl alcohol as a penetration enhancer, a mixture of essential amino acids (0.2%), significantly enhanced collagen synthesis and deposition. The amino acid mixture was designed to mimic serum concentrations, with supplemental methionine added to provide additional sulfur. The histological appearance of the skin of mature female rats treated in this fashion reflected the increased deposition of collagen in the dermis as well as a thickened epidermal layer. The changes do not seem to be mediated by TGF- β or PDGF, two growth factors known to stimulate collagen synthesis. At lower concentrations, α -lipoic acid did not affect cell proliferation but at higher doses, while it had an inhibitory effect on ³H-thimidine uptake, it did enhance collagen production. Pronanthocyanidin did not affect cell proliferation but significantly increased collagen synthesis by cultured fibroblasts.     

In Vivo and In Vitro Studies of Delayed-Type Hypersensitivity to Toxoplasma gondii in Guinea Pigs

Delayed-type hypersensitivity develops late in the course of human toxoplasmosis, and a positive skin test is of some value for implicating chronic or eliminating acute forms of toxoplasmosis as a cause of disease. Toxoplasma-infected guinea pigs were studied to determine the onset and development of delayed-type hypersensitivity. Both the toxoplasmin skin test and the in vitro macrophage migration inhibition technique indicated that delayed hypersensitivity to toxoplasma antigen existed as early as 1 week after infection. The mechanism responsible for the observed inhibition of macrophage migration in vitro appeared to be an inhibitory factor(s) released from sensitized lymphoid cells in the presence of antigen.     

Menthol in Combination with Iontophoresis Promotes Natamycin Penetration through the Cornea: In Vitro and In Vivo Studies

Bulletin of Experimental Biology and Medicine - We studied whether menthol can promote penetration of natamycin, a representative antifungal macrolide agent, through the cornea. Natamycin...     

Anthelmintic Efficacy of Nauclea Latifolia Extract Against Gastrointestinal Nematodes of Sheep: In Vitro and In Vivo Studies

Direct effects of Nauclea latifolia extracts on different gastrointestinal nematodes of sheep is described. In vivo and in vitro studies were conducted to determine possible anthelmintic effect of leaf extracts of Nauclea latifolia toward different ovine gastro intestinal nematodes. A larval development assay was used to investigate in vitro, the effect of aqueous and ethanolic extracts of N. latifolia towards strongyles larvae. The development and survival of infective larvae (L₃) was assessed and best-fit LC₅₀ values were computed by global model of non-linear regression analysis curve-fitting (95% CI). Twenty sheep harbouring naturally acquired gastrointestinal nematodes were treated with oral administration of ethanolic extracts at a dose rate of 125 mg/kg, 250 mg/kg and 500mg/kg to evaluate therapeutic efficacy, in vivo.The presence of the extracts in the cultures decreased the survival of larvae. The LC₅₀ of aqueous and ethanolic extract were 0.704 and 0.650 mg/ml respectively and differ significantly (P<0.05, paired t test). Faecal egg counts (FEC) on day 12 after treatment showed that the extract is effective, relative to control (1-way ANOVA, Dunnett''s multiple comparison test), at 500mg/kg against Haemonchus spp, Trichostrongylus spp (p<0.05), Strongyloides spp (P < 0.01); at 250mg/kg against Trichuris spp (P < 0.01) and ineffective against Oesophagostomum spp (p>0.05). The effect of doses is extremely significant; the day after treatment is sometimes significant while interaction between dose and day after treatment is insignificant (2-way ANOVA).N. latifolia extract could therefore find application in the control of helminth in livestock, by the ethnoveterinary medicine approach.     

Bordetella avium Virulence Measured In Vivo and In Vitro

Bordetella avium causes an upper-respiratory-tract disease called bordetellosis in birds. Bordetellosis shares many of the clinical and histopathological features of disease caused in mammals by Bordetella pertussis and Bordetella bronchiseptica. In this study we determined several parameters of infection in the domestic turkey, Meleagris galapavo, and compared these in vivo findings with an in vitro measure of adherence using turkey tracheal rings. In the in vivo experiments, we determined the effects of age, group size, infection duration, and interindividual spread of B. avium. Also, the effect of host genetic background on susceptibility was tested in the five major commercial turkey lines by infecting each with the parental B. avium strain and three B. avium insertion mutants. The mutant strains lacked either motility, the ability to agglutinate guinea pig erythrocytes, or the ability to produce dermonecrotic toxin. The susceptibilities of 1-day-old and 1-week-old turkeys to B. avium were the same, and challenge group size (5, 8, or 10 birds) had no effect upon the 50% infectious dose. Two weeks between inoculation and tracheal culture was optimal, since an avirulent mutant (unable to produce dermonecrotic toxin) persisted for a shorter time. Communicability of the B. avium parental strain between confined birds was modest, but a nonmotile mutant was less able to spread between birds. There were no host-associated differences in susceptibility to the parental strain and the three B. avium mutant strains just mentioned: in all turkey lines tested, the dermonecrotic toxin- and hemagglutination-negative mutants were avirulent whereas the nonmotile mutants showed no loss of virulence. Interestingly, the ability of a strain to cause disease in vivo correlated completely with its ability to adhere to ciliated tracheal cells in vitro.     

Anti-septic Effects of Fisetin In Vitro and In Vivo

Sepsis is a state of disrupted inflammatory homeostasis that is initiated by infection. High mobility group box 1 (HMGB1) protein acting as a late mediator of severe vascular inflammatory conditions, such as sepsis and endothelial cell protein C receptor (EPCR), is involved in vascular inflammation. Fisetin, an active compound from the family Fabaceae, was reported to have antiviral, neuroprotective, and anti-inflammatory activities. Here, we determined the anti-septic effects of fisetin on HMGB1-mediated inflammatory responses and on the shedding of EPCR in vitro and in vivo, for the first time. First, we monitored the effects of post-treatment fisetin on lipopolysaccharide (LPS) and cecal ligation and puncture (CLP)-mediated release of HMGB1 and HMGB1-mediated regulation of pro-inflammatory responses in human umbilical vein endothelial cells (HUVECs) and septic mice. Post-treatment fisetin was found to suppress LPS-mediated release of HMGB1 and HMGB1-mediated cytoskeletal rearrangements. Fisetin also inhibited HMGB1-mediated hyperpermeability and leukocyte migration in septic mice. Fisetin induced potent inhibition of phorbol-12-myristate 13-acetate (PMA) and CLP-induced EPCR. Fisetin also inhibited the expression and activity of tumor necrosis factor-α converting enzyme, induced by PMA in endothelial cells. In addition, fisetin inhibited the production of tumor necrosis factor-α and the activation of AKT, nuclear factor-κB, and extracellular regulated kinases 1/2 by HMGB1 in HUVECs. Fisetin also down-regulated CLP-induced release of HMGB1, production of interleukin 1β, and reduced septic mortality. Collectively, these results suggest that fisetin may be a candidate therapeutic agent for the treatment of vascular inflammatory diseases via inhibition of the HMGB1 signaling pathway.     

In Vitro and In Vivo Characterization of MEMS Microneedles

Transdermal drug delivery TDD systems have many advantages but are conventionally limited by the low permeability of skin. The idea of using microneedles to painlessly penetrate the topmost impermeable stratum corneum has previously been put forward. In this paper, the fabrication of solid and hollow silicon microneedles with straight side-walls and with the following dimensions: 20–100 m in diameter and 100–150 m in length is described. In vitro tests demonstrate that with prior solid microneedle application, transdermal drug transport is significantly increased by 10–20 times, with the degree of enhancement being related to needle diameter. In vivo tests in diabetic animals, however, were unable to demonstrate any delivery of insulin through the hollow microneedles. It is proposed that two factors, microneedle length and tip sharpness, have to be improved for systemic drug delivery to be seen in vivo.     

Cedrus atlantica Extract Suppress Glioblastoma Growth through Promotion of Genotoxicity and Apoptosis: In Vitro and In Vivo Studies

Glioblastoma (GBM) is the most common malignant primary brain tumor in humans, exhibiting highly infiltrative growth and drug resistance to conventional chemotherapy. Cedrus atlantica (CAt) extract has been shown to decrease postoperative pain and inhibit the growth of K562 leukemia cells. The aim of this study was to assess the anti-GBM activity and molecular mechanism of CAt extract in vitro and in vivo. The results showed that CAt extract greatly suppressed GBM cells both in vitro and in vivo and enhanced the survival rate in subcutaneous and orthotopic animal models. Moreover, CAt extract increased the level of ROS and induced DNA damage, resulting in cell cycle arrest at the G₀/G₁ phase and cell apoptosis. Western blotting results indicated that CAt extract regulates p53/p21 and CDK4/cyclin D1 protein expression and activates extrinsic and intrinsic apoptosis. Furthermore, CAt extract enhanced the cytotoxicity of Temozolomide and decreased AKT/mTOR signaling by combination treatment. In toxicity assays, CAt extract exhibited low cytotoxicity toward normal cells or organs in vitro and in vivo. CAt extract suppresses the growth of GBM by induction of genotoxicity and activation of apoptosis. The results of this study suggest that CAt extract can be developed as a therapeutic agent or adjuvant for GBM treatment in the future.     

Cocaine Inhibits Human Neutrophil Phagocytosis and Phagolysosomal Acidification In Vitro

Abstract

Cocaine, used intravenously, increases the risk of infections, but its effects on neutrophil phagocytosis have not been examined in vitro. Human neutrophils were suspended in cocaine hydrochloride 0, 1, 10, 50, 100 or 200 μg/ml in Hank's balanced salt solution to which was added a phagocytic meal of killed Saccharomyces cerevisiae stained with the pH indicator dye bromcresol purple. Yeast per phagocytosing neutrophil and the percent neutrophils phagocytosing yeast were reduced in neutrophils treated with cocaine 100 and 200 μg/ml (P < 0.05). When examined for percent of yeast phagocytosed after 10 minutes, neutrophils treated with cocaine 1-200 μg/ml demonstrated a decrease (P < 0.05). However, at 60 minutes only neutrophils treated with cocaine 50 and 100 μg/ml still showed a decrease in percent of yeast phagocytosed. Phagolysosomal acidification was impaired in neutrophils treated with 50, 100 and 200 μg/ml cocaine. Thus, cocaine inhibits neutrophil phagocytosis and phagolysosomal acidification in vitro, offering a reason for cocaine users/abusers to have impaired host defense and to be potentially at higher risk for infections.     

Neuroprotective Effects of Methylene Blue In Vivo and In Vitro

Bulletin of Experimental Biology and Medicine - Focal unilateral traumatic brain injury in the sensorimotor cortical region disturbed the functions of contralateral limbs controlled by the damaged...     

Hyperoside Inhibits High-Glucose-Induced Vascular Inflammation In Vitro and In Vivo

Hyperoside, an active compound from the genera of Hypericum and Crataegus, was reported to have antioxidant, antihyperglycemic, anticancer, anti-inflammatory, and anticoagulant activities. Vascular inflammatory process has been suggested to play a key role in initiation and progression of atherosclerosis, a major complication of diabetes mellitus. Thus, in this study, we attempted to determine whether hyperoside can suppress vascular inflammatory processes induced by high glucose (HG) in human umbilical vein endothelial cells (HUVECs) and mice. Data showed that HG induced markedly increased vascular permeability, monocyte adhesion, expressions of cell adhesion molecules (CAMs), formation of reactive oxygen species (ROS), and activation of nuclear factor (NF)-κB. Remarkably, all of the above-mentioned vascular inflammatory effects of HG were attenuated by pretreatment with hyperoside. Vascular inflammatory responses induced by HG are critical events underlying development of various diabetic complications; therefore, our results suggest that hyperoside may have significant therapeutic benefits against diabetic complications and atherosclerosis.