Magnetic isolation of particles suspended in synovial fluid for diagnostics of natural joint chondropathies

Millions of people are stricken with the degenerative joint disease known as osteoarthritis. Osteoarthritis is associated with biochemical and mechanical processes, and is characterized by loss of articular cartilage and hypertrophy of bone. As cartilage and bone particles are released into the synovial fluid, a variety of biomarkers have been suggested for the analysis of this fluid. Here we have developed a method for isolating bone and cartilage wear particles suspended in the synovial fluid of the hip, knee and ankle joints of humans, based on specific magnetization of collagens I and II. Bio-ferrography is used to capture the particles on glass slides, allowing microscopic, chemical and statistical analyses. The relations between the level of the disease and the number, dimensions, shape and chemical composition of the particles were established. The method, which was found to be sensitive and reliable, can easily be extended to other applications, such as diagnosis of cancer and infectious diseases, determination of the efficacy of drugs or optimization of implants.     

Ureteric sarcoidosis--a rare entity

Primary ureteric involvement in sarcoidosis is very rare; to our knowledge, only a few cases have been reported in the literature. We present here a rare case of ureteric sarcoidosis presenting with obstructive uropathy.     

Antioxidant activity of Potentilla fulgens: An alpine plant of western Himalaya

Antioxidant activities of the methanol extract, fractions and isolated compounds from the roots of Potentilla fulgens Lodd. were evaluated by three in vitro experiments, namely, ABTS, DPPH, and FRAP assays. PF-2 was characterized as a new biflavanoid and designated as Potifulgene on the basis of NMR and mass spectrum, whereas PF-1 was identified as epicatechin. The activities of aqueous methanolic extract and fractions could be correlated with their respective total phenolic content and compared with standard natural antioxidants such as quercetin, vitamin C and pyrogallol. The root powder of the plant was extracted with methanol/water (80:20) by cold extraction and its extract was further partitioned with ethyl acetate, butanol and water fractions. Among the three fractions (ethyl acetate, butanol and water fraction) and the total aqueous methanolic extract, the butanol fraction exhibited good scavenging response measured in terms of TEAC (mM Trolox equivalent/mg extract). The butanol fraction was found to possess strong antioxidant activity (2.54 ± 0.69, 2.41 ± 0.53, 3.57 ± 0.05 mM Trolox equivalent/mg extract) with ABTS, DPPH and FRAP assays, respectively. The chemical composition of extracts, studied in terms of total polyphenol content (TPC), was found in the range of 20.61 ± 0.38 to 33.28 ± 0.11 mg/g gallic acid equivalent. A significant correlation was observed between total polyphenolics and antioxidant activity, indicating participation of phenolics in antioxidant activities of extract and fractions. The antioxidant activity of new biflavanoid (Potifulgene) was found to be higher, i.e. 6.85 ± 0.38, 4.24 ± 0.41, 5.35 ± 0.53 than that of epicatechin, 2.13 ± 0.05, 1.50 ± 0.02, 1.57 ± 0.03 with ABTS, DPPH and FRAP assays.     

Factors and strategies for improving buccal absorption of peptides.

Peptides and polypeptides have important pharmacological properties but only a limited number (e.g. insulin, oxytocin, vasopressin) have been exploited as therapeutics because of problems related to their delivery. The buccal mucosa offers an alternative route to conventional, parenteral administration. Peptides are generally not well absorbed through mucosae because of their molecular size, hydrophilicity and the low permeability of the membrane. Peptide transport across buccal mucosa occurs via passive diffusion and is often accompanied by varying degrees of metabolism. This review describes various approaches to improve the buccal absorption of peptides including the use of penetration enhancers to increase membrane permeability and/or the addition of enzyme inhibitors to increase their stability. Other strategies including molecular modification with bioreversible chemical groups or specific formulations such as bioadhesive delivery systems are also discussed.     

Skin permeability enhancement by low frequency sonophoresis: lipid extraction and transport pathways

The objective of this study was to shed light on the mechanism(s) by which low-frequency ultrasound (20 KHz) enhances the permeability of the skin. The physical effects on the barrier and the transport pathway, in particular, were examined. The amount of lipid removed from the intercellular domains of the stratum corneum following sonophoresis was determined by infrared spectroscopy. Transport of the fluorescent probes nile red and calcein, under the influence of ultrasound, was evaluated by laser-scanning confocal microscopy. The results were compared with the appropriate passive control data and with data obtained from experiments in which the skin was exposed simply to the thermal effects induced by ultrasound treatment. A significant fraction ( approximately 30%) of the intercellular lipids of the stratum corneum, which are principally responsible for skin barrier function, were removed during the application of low-frequency sonophoresis. Although the confocal images from the nile red experiments were not particularly informative, ultrasound clearly and significantly (again, relative to the corresponding controls) facilitated transport of the hydrophilic calcein via discrete permeabilized regions, whereas other areas of the barrier were apparently unaffected. Lipid removal from the stratum corneum is implicated as a factor contributing the observed permeation enhancement effects of low-frequency ultrasound. However, microscopic observations imply that sonophoresis induces localized (aqueous?) permeation pathways at discrete sites.     

Cutaneous iontophoretic delivery of CGP69669A, a sialyl Lewis(x) mimetic, in vitro

The aim was to investigate the feasibility of using iontophoresis for the cutaneous delivery of the E-selectin antagonist CGP69669A, a sialyl Lewis(x) -glycomimetic with potential activity against inflammatory skin diseases. The effects of current density and formulation on iontophoretic transport were evaluated in porcine and human skin in vitro. Cumulative permeation of CGP69669A increased with current density (69.73±9.51, 113.97±26.80 and 160.44±13.79μg/cm(2) at 0.1, 0.3 and 0.5mA/cm(2) , respectively) and drug concentration (37.42±13.13, 78.96±23.13 and 160.44±13.79μg/cm(2) , at 1, 3 and 5mg/ml, respectively). In contrast, passive delivery was negligible. Although permeation from a 2% hydroxyethyl cellulose gel was lower than that from aqueous solution, skin deposition - more relevant for the local treatment of dermatological conditions - was 3-fold higher. The results demonstrated that although CGP69669A cannot be delivered passively into the skin it is an excellent candidate for transdermal iontophoresis, a technique that is ideally suited to the delivery of glycomimetics.