Subcommissural organ-Reissner's fiber complex of the teleost Clarias batrachus responds to GABA treatment

Subcommissural organ (SCO) is a highly specialized ependymal gland located in the roof of the third ventricle. The secretory products of the SCO, which condense to form Reissner's fiber (RF), were recently found to cross-react with the anti-calcitonin antibody. To understand the mechanisms regulating the formation of the RF and the possible function of these discrete structures, we studied the response of the SCO-RF complex to intracranially administered GABA, using immunocytochemical labeling with anti-calcitonin antibody. Although the SCO-RF complex of control fish was intensely immunostained, 1 h after GABA treatment, the ependymal cells revealed partial loss of immunoreactivity; the RF showed occasional loss of immunoreactivity with its diameter increased by about 56% of the control value. Following 2 h of GABA treatment, the SCO revealed dramatic loss of calcitonin-like immunoreactivity from the ependymal cells. The RF showed a dual response in this group, while in some segments the RF appeared conspicuously thick, elsewhere it appeared thin. The mean diameter was, however, not significantly different from the normal. Following 4 h of GABA treatment, while calcitonin-like immunoreactive material made its reappearance in the SCO, the RF diameter was uniformly reduced to about 35% of the control value. The responses by the RF as well as the SCO to intracranially administered GABA were blocked by pretreatment with bicuculline, a GABA(A) receptor antagonist. The results suggest that GABA, acting via GABA(A) receptors, may trigger the release of secretory material from the SCO and induce histomorphological changes in the RF indicative of discharge of stored material.     

Induction of neuronal death by alpha-synuclein

The molecular and cellular mechanisms underlying neuronal loss in neurodegenerative diseases are unclear. It is generally thought that aggregation of mutated, abnormally modified or abnormally folded proteins leads to the accumulation of extracellular, intracellular or intranuclear deposits that severely compromise cell physiology, leading to the death of the affected neurons. However, there is growing evidence that neuronal apoptosis in the absence of obvious pathological deposits could have a serious impact on the pathogenesis of neurodegenerative diseases. alpha-Synuclein has been implicated in aetiology and pathogenesis of certain neurodegenerative diseases, although the precise role of this protein in neurodegeneration is uncertain. The normal functions of alpha-synuclein and other members of the synuclein family in the development and function of the nervous system also remain elusive. Here we show that overexpression of wild-type and mutant forms of alpha-synuclein in cultured neurons, but not the closely related persyn (gamma-synuclein), causes apoptosis. These findings suggest that abnormalities of alpha-synuclein metabolism could lead to the neuronal loss occurring in certain forms of neurodegeneration before the formation of characteristic pathological lesions.     

Growth of asthmatic children is slower during than before treatment with inhaled glucocorticoids

Reports on the influence of inhaled glucocorticoids on growth have been controversial. We studied the growth of prepubertal asthmatic children prior to and during glucocorticoid therapy. We collected retrospectively the notes of 201 asthmatic children aged 1–11 years receiving inhaled beclomethasone dipropionate or budesonide. We calculated their height and height velocity standard deviation scores (HSDS and HVSDS, respectively) before the treatment and up to 5 years during the treatment and compared those with the growth of healthy peers. The dose of the medication was calculated and the severity of asthma was assessed. The asthmatic children grew similarly to their healthy peers before treatment with inhaled glucocorticoids: the mean HSDS was +0.02 and the mean HVSDS +0.01 for boys and -0.16 and +0.13 for girls, respectively. Growth retardation took place soon after the start of the treatment, the most profound decrease in the growth velocity (the change in the mean HVSDS from +0.05 to -0.88) occurring during the first year of treatment. The growth-retarding effect of inhaled glucocorticoids was not dose dependent. In the covariance analysis the increasing severity of asthma had a significant interaction with repeated measurements, showing more growth retardation along with more severe asthma, especially during long-term treatment. Asthma per se does not impair growth, but inhaled glucocorticoids may do so. Careful monitoring of the growth of all asthmatic children receiving inhaled glucocorticoids is necessary because the growth-retarding effect of the medication is not dose dependent. Individual sensitivity might explain the differences seen in the growth patterns of children receiving inhaled glucocorticoids.     

Pleomorphic adenoma of nasal septum

The pleomorphic adenoma is the most common benign tumour of the major and minor salivary glands. We report a 40-year-old female patient, who presented with progressive swelling and deformity of the left side of her nose for last 2 years. On a lateral rhinotomy a mass (4 cm X3 cm in size) was found to be arising from the cartilaginous part of the nasal septum. The mass was emicleated from its capsule and the walls excised. The histopathological examination revealed it to be pleomorphic adenoma.     

New chiral reverse phase HPLC method for enantioselective analysis of ketorolac using chiral AGP column

A simple, specific, precise, sensitive and rapid reverse phase-HPLC method was developed for determination of ketorolac enantiomers, a potent nonnarcotic analgesic in pharmaceutical formulations. The method was developed on a chiral AGP column. Mobile phase was 0.1 M sodium phosphate buffer (pH 4.5): Isopropanol (98:2, v/v), at a flow rate of 1 mL/min with run time of 15 min. Ultraviolet detection was made at 322 nm. The linearity range was 0.02–10 μg/mL for each of the enantiomers. The mobile phase composition was systematically studied to find the optimum chromatographic conditions. Validation of the method under the conditions selected showed that it was selective and precise and that the detector response was linear function of ketorolac.     

Quantification of coronary arterial stenoses in CTA using fuzzy distance transform

Quantification of coronary arterial stenoses is useful for the diagnosis of several coronary heart diseases. Being noninvasive, economical and informative, computed tomographic angiography (CTA) has become a common modality for monitoring disease status and treatment effects. Here, we present a new method for detecting and quantifying coronary arterial stenosis in CTA using fuzzy distance transform (FDT) approach and a new coherence analysis of observed data for computing expected local diameter. FDT allows computing local depth at each image point in the presence of partial voluming and thus, eliminates the need for binarization, commonly, associated with inclusion of additional errors. In the current method, coronary arterial stenoses are detected and their severities are quantified by analyzing FDT values along the medial axis of an arterial tree obtained by its skeletonization. A new skeletal pruning algorithm has been developed toward improving the quality of medial axes and thereby, enhancing the accuracy of stenosis detection and quantification. Further, we have developed a new method to estimate “expected diameter” along a given arterial branch using a new coherence analysis of observed diameter values along the branch. The overall method is completed in the following steps - (1) fuzzy segmentation of coronary artery in CTA, (2) FDT computation of coronary arteries, (3) medial axis computation, (4) estimation of observed and expected diameters along arteries and (5) detection of stenoses and quantification of arterial blockage. The performance of this method has been quantitatively evaluated on a realistic coronary artery phantom dataset with randomly simulated stenoses and the results have been compared with a binary distance transform based and a conventional binary algorithm. The method has also been applied on a clinical CTA dataset from thirteen heart patients and the results have been compared with an expert's quantitative assessment of stenoses. Results of the phantom experiment indicate that the new method (error: 0.53%) is significantly more accurate as compared to both binary distance transform based (error 2.11%) and conventional binary (error 3.71%) methods. Also, the results of the clinical study indicate that the new FDT-based method (kappa coefficient = 87.9%) is highly in agreement with the expert's assessments and, in this respect, outperforms the other two methods (kappa coefficients = 75.2% and 69.5%).     

Tumor growth inhibition through targeting liposomally bound curcumin to tumor vasculature

Increasing number of Phase I/II clinical studies have demonstrated clinical potential of curcumin for treatment of various types of human cancers. Despite significant anti-tumor efficacies and bio-safety profiles of curcumin, poor systemic bioavailability is retarding its clinical success. Efforts are now being directed toward developing stable formulations of curcumin using various drug delivery systems. To this end, herein we report on the development of a new tumor vasculature targeting liposomal formulation of curcumin containing a lipopeptide with RGDK-head group and two stearyl tails, di-oleyolphosphatidylcholine (DOPC) and cholesterol. We show that essentially water insoluble curcumin can be solubilized in fairly high concentrations (~ 500 μg/mL) in such formulation. Findings in the Annexin V/Propidium iodide (PI) binding based flow cytometric assays showed significant apoptosis inducing properties of the present curcumin formulation in both endothelial (HUVEC) and tumor (B16F10) cells. Using syngeneic mouse tumor model, we show that growth of solid melanoma tumor can be inhibited by targeting such liposomal formulation of curcumin to tumor vasculature. Results in immunohistochemical staining of the tumor cryosections are consistent with tumor growth inhibition being mediated by apoptosis of tumor endothelial cells. Findings in both in vitro and in vivo mechanistic studies are consistent with the supposition that the presently described liposomal formulation of curcumin inhibits tumor growth by blocking VEGF-induced STAT3 phosphorylation in tumor endothelium. To the best of our knowledge, this is the first report on inhibiting tumor growth through targeting liposomal formulation of curcumin to tumor vasculatures.