Efficacy and safety of fixed-dose combination of Bilastine-Montelukast in adult patients with allergic rhinitis: a phase III, randomized, multi-center, double-blind, active controlled clinical study.

BACKGROUND: Histamine and cysteinyl leukotrienes (CysLTs) are potent inflammatory mediators in allergic rhinitis (AR). Studies involving other combinations of antihistaminics (Levocetirizine) and highly selective leukotriene receptor antagonist (LTA) (Montelukast) combination have shown additive benefits and are widely prescribed for AR. OBJECTIVE: Evaluate the efficacy and safety of Bilastine 20 mg and Montelukast 10 mg fixed-dose combination (FDC) therapy in patients with AR. METHODS: A randomized, double-blind, comparative, parallel, phase III study was conducted to evaluate efficacy and safety of Bilastine 20 mg and Montelukast 10 mg FDC at 16 tertiary care otolaryngology centres in India. Adult patients with AR for one year with IgE antibody positive and 12-h NSS score >36 in 3 days were randomized to receive either Bilastine 20 mg and Montelukast 10 mg or Montelukast 10 mg & Levocetirizine 5 mg tablets for 4 weeks. The change in total symptom score (nasal symptom scores (NSS) & non-nasal symptom scores (NNSS)) from baseline to week 4 was assessed as primary endpoint. Secondary endpoints included changes in TSS, NSS, NNSS, individual symptom scores (ISS), Rhinoconjunctivitis Quality of Life (RQLQ), discomfort due to rhinitis (VAS), and clinical global impression (CGI) scores. RESULTS: The change in mean TSS from baseline to week 4 in Test group (16.6 units) was comparable to reference group (17 units) (p= 0.8876). The difference in change in mean NSS, NNSS and ISS from baseline to day 7, 14, 28 were comparable. RQLQ improved from baseline to Day 28. Significant improvements were observed in discomfort due to AR measured by VAS and CGI scores from baseline to day 14 and 28. The safety and tolerability of patients were comparable between the groups. All adverse events (AEs) were mild to moderate in severity. No patient discontinued due to AEs. CONCLUSIONS: The FDC of Bilastine 20 mg and Montelukast 10 mg was efficacious and well tolerated in Indian patients with AR.

Endosidin1 defines a compartment involved in endocytosis of the brassinosteroid receptor BRI1 and the auxin transporters PIN2 and AUX1.

Although it is known that proteins are delivered to and recycled from the plasma membrane (PM) via endosomes, the nature of the compartments and pathways responsible for cargo and vesicle sorting and cellular signaling is poorly understood. To define and dissect specific recycling pathways, chemical effectors of proteins involved in vesicle trafficking, especially through endosomes, would be invaluable. Thus, we identified chemicals affecting essential steps in PM/endosome trafficking, using the intensely localized PM transport at the tips of germinating pollen tubes. The basic mechanisms of this localized growth are likely similar to those of non-tip growing cells in seedlings. The compound endosidin 1 (ES1) interfered selectively with endocytosis in seedlings, providing a unique tool to dissect recycling pathways. ES1 treatment induced the rapid agglomeration of the auxin translocators PIN2 and AUX1 and the brassinosteroid receptor BRI1 into distinct endomembrane compartments termed "endosidin bodies"; however, the markers PIN1, PIN7, and other PM proteins were unaffected. Endosidin bodies were defined by the syntaxin SYP61 and the V-ATPase subunit VHA-a1, two trans-Golgi network (TGN)/endosomal proteins. Interestingly, brassinosteroid (BR)-induced gene expression was inhibited by ES1 and treated seedlings displayed a brassinolide (BL)-insensitive phenotype similar to a bri1 loss-of-function mutant. No effect was detected in auxin signaling. Thus, PIN2, AUX1, and BRI1 use interactive pathways involving an early SYP61/VHA-a1 endosomal compartment.

Trehalose-6-phosphate synthase/phosphatase regulates cell shape and plant architecture in Arabidopsis.

The vacuole occupies most of the volume of plant cells; thus, the tonoplast marker delta-tonoplast intrinsic protein-green fluorescent protein delineates cell shape, for example, in epidermis. This permits rapid identification of mutants. Using this strategy, we identified the cell shape phenotype-1 (csp-1) mutant in Arabidopsis thaliana. Beyond an absence of lobes in pavement cells, phenotypes included reduced trichome branching, altered leaf serration and stem branching, and increased stomatal density. This result from a point mutation in AtTPS6 encoding a conserved amino-terminal domain, thought to catalyze trehalose-6-phosphate synthesis and a carboxy-terminal phosphatase domain, is catalyzing a two-step conversion to trehalose. Expression of AtTPS6 in the Saccharomyces cerevisiae mutants tps1 (encoding a synthase domain) and tps2 (encoding synthase and phosphatase domains) indicates that AtTPS6 is an active trehalose synthase. AtTPS6 fully complemented defects in csp-1. Mutations in class I genes (AtTPS1-AtTPS4) indicate a role in regulating starch storage, resistance to drought, and inflorescence architecture. Class II genes (AtTPS5-AtTPS11) encode multifunctional enzymes having synthase and phosphatase activity. We show that class II AtTPS6 regulates plant architecture, shape of epidermal pavement cells, and branching of trichomes. Thus, beyond a role in development, we demonstrate that the class II gene AtTPS6 is important for controlling cellular morphogenesis.

Protein geranylgeranyltransferase I is involved in specific aspects of abscisic acid and auxin signaling in Arabidopsis.

Arabidopsis (Arabidopsis thaliana) mutants lacking a functional ERA1 gene, which encodes the beta-subunit of protein farnesyltransferase (PFT), exhibit pleiotropic effects that establish roles for protein prenylation in abscisic acid (ABA) signaling and meristem development. Here, we report the effects of T-DNA insertion mutations in the Arabidopsis GGB gene, which encodes the beta-subunit of protein geranylgeranyltransferase type I (PGGT I). Stomatal apertures of ggb plants were smaller than those of wild-type plants at all concentrations of ABA tested, suggesting that PGGT I negatively regulates ABA signaling in guard cells. However, germination of ggb seeds in response to ABA was similar to the wild type. Lateral root formation in response to exogenous auxin was increased in ggb seedlings compared to the wild type, but no change in auxin inhibition of primary root growth was observed, suggesting that PGGT I is specifically involved in negative regulation of auxin-induced lateral root initiation. Unlike era1 mutants, ggb mutants exhibited no obvious developmental phenotypes. However, era1 ggb double mutants exhibited more severe developmental phenotypes than era1 mutants and were indistinguishable from plp mutants lacking the shared alpha-subunit of PFT and PGGT I. Furthermore, overexpression of GGB in transgenic era1 plants partially suppressed the era1 phenotype, suggesting that the relatively weak phenotype of era1 plants is due to partial redundancy between PFT and PGGT I. These results are discussed in the context of Arabidopsis proteins that are putative substrates of PGGT I.

High-throughput fluorescent tagging of full-length Arabidopsis gene products in planta.

We developed a high-throughput methodology, termed fluorescent tagging of full-length proteins (FTFLP), to analyze expression patterns and subcellular localization of Arabidopsis gene products in planta. Determination of these parameters is a logical first step in functional characterization of the approximately one-third of all known Arabidopsis genes that encode novel proteins of unknown function. Our FTFLP-based approach offers two significant advantages: first, it produces internally-tagged full-length proteins that are likely to exhibit native intracellular localization, and second, it yields information about the tissue specificity of gene expression by the use of native promoters. To demonstrate how FTFLP may be used for characterization of the Arabidopsis proteome, we tagged a series of known proteins with diverse subcellular targeting patterns as well as several proteins with unknown function and unassigned subcellular localization.

Identification of an allele of CLA1 associated with variegation in Arabidopsis thaliana.

We have identified a mutant of Arabidopsis thaliana (lvr111) that exhibits a variegated phenotype, reduced isoprenoid pigmentation, and dwarfism in comparison with wild-type plants. Segregation analysis indicated that this phenotype was caused by a single, semi-dominant mutation and PCR-based marker mapping placed the mutation near position 56 on the RI map of chromosome IV. The lvr111 lesion was identified by genomic PCR and sequence analysis as a missense mutation (D306N) in the CLA1 gene (AT4g15560) and complementation analysis confirmed the allelic relationship between lvr111 and CLA1. CLA1 encodes 1-deoxy-d-xylulose 5-phosphate synthase, which catalyses the first step of the non-mevalonate isoprenoid biosynthetic pathway. These observations demonstrate that, unlike the albinism caused by severe alleles of CLA1, weaker alleles are associated with leaf variegation.

Prenylcysteine alpha-carboxyl methyltransferase expression and function in Arabidopsis thaliana.

Farnesylated proteins undergo a series of post-translational modifications, including carboxyl terminal isoprenylation, proteolysis, and methylation. In Arabidopsis thaliana, protein farnesylation has been shown to be necessary for negative regulation of ABA signaling. However, the role of post-isoprenylation protein processing in ABA signal transduction has not been described. Here, we show that the A. thaliana genome contains two distinct genes on chromosome V, AtSTE14A and AtSTE14B, which encode functional prenylcysteine alpha-carboxyl methyltransferases. AtSTE14B encodes a methyltransferase with lower apparent Kms for prenylcysteine substrates and higher specific activities than the previously described AtSTE14A-encoded methyltransferase. Furthermore, whereas AtSTE14A transcription is restricted to root and shoot tips, young leaves, and vascular tissue, AtSTE14B transcription is observed in all organs except hypocotyls and petioles. Pharmacological inhibitors of prenylcysteine alpha-carboxyl methyltransferase activity cause increased ABA sensitivity, seed dormancy, and stomatal closure, consistent with the hypothesis that prenylcysteine alpha-carboxyl methylation is necessary for negative regulation of ABA signaling. These results suggest that carboxyl methylation, which is a reversible and potentially regulated step in the processing, targeting, and function of isoprenylated plant proteins, may be an important biochemical target for introducing altered ABA sensitivity and drought tolerance into plants.

Plasma membrane-associated ROP10 small GTPase is a specific negative regulator of abscisic acid responses in Arabidopsis.

Abscisic acid (ABA) is an important plant hormone that modulates seed germination and plant growth and stress responses, but its signaling remains poorly understood. We investigated the role of ROP10, a member of the Arabidopsis Rop subfamily of Rho GTPases, in ABA signaling. A null rop10 mutant exhibits enhanced responses to ABA in seed germination, root elongation, and stomatal closure assays and in the induction of expression of the transcription factor MYB2, but it shows wild-type levels of ABA and normal responses to other hormones. Consistently, transgenic expression of a constitutively active form of ROP10 reduces ABA inhibition of seed germination, whereas dominant-negative mutants of ROP10 enhance ABA response and partially suppress abi2. Furthermore, ABA specifically downregulates ROP10 transcription in root tips. ROP10 is localized to the plasma membrane (PM), and PM localization is crucial for its function. These results suggest that ROP10 is a PM-localized signaling molecule that is involved specifically in the negative regulation of ABA signaling.

The dose-response relationship of controlled-release codeine (Codeine Contin) in chronic cancer pain.

The improved pain control provided by regular dosing of opioid analgesics in patients with severe cancer pain has been well established. However, the treatment of mild-to-moderate cancer pain is often limited to "as needed" dosing with fixed combinations of codeine or oxycodone plus a nonopioid analgesic, which do not allow optimal titration of the individual components. This randomized double-blind study was designed to evaluate the efficacy of controlled-release codeine (Codeine Contin) in patients with cancer pain, and to estimate its dose equivalence to a standard combination of acetaminophen plus codeine. Twenty-four patients with at least moderate cancer pain were randomized to Codeine Contin 100, 200, or 300 mg every 12 hr or acetaminophen plus codeine (600 mg/60 mg) every 6 hr. On days 1 and 4 of dosing, pain intensity and pain relief were assessed hourly for 12 hr. The sum of pain intensity differences (SPID) from baseline and the total pain relief (TOTPAR) scores demonstrated a dose-response relationship for Codeine Contin on days 1 and 4 that was statistically significant on day 1 and suggested greater analgesic efficacy on day 4, compared with day 1. Codeine Contin 150 mg every 12 hr was estimated to be equianalgesic to acetaminophen plus codeine (600 mg/60 mg) given every 6 hr. Because a similar equivalence was also demonstrated from analysis of adverse event data, it is concluded that Codeine Contin 150 mg produces analgesia and a side-effect profile similar to a 40% lower dose of codeine provided by the combination.(ABSTRACT TRUNCATED AT 250 WORDS)

Direct measurement of interstitial convection and diffusion of albumin in normal and neoplastic tissues by fluorescence photobleaching.

Macromolecular transport through the interstitial space of a tissue occurs by convection and diffusion. The convective component of transport results from interstitial fluid flow. There have been no direct measurements of the magnitude or direction of interstitial fluid flow in tissues to date. Using fluorescence recovery after photobleaching, we have measured interstitial fluid velocities and the diffusion coefficient of bovine serum albumin in normal and neoplastic tissues grown in a thin, transparent window in the ear of a rabbit. A well-defined laser beam was focused on a region within the interstitium of the fluorescence-bathed tissue. A short pulse of laser irradiation extinguished the fluorescence emanating from this selected region. The recovery of fluorescence due to diffusion and convection within the medium was monitored and analyzed to yield values of the diffusion coefficient and the fluid velocity. The average fluid velocity was about 0.6 microns/s, and albumin diffusion coefficients were 5.8 +/- 1.3 x 10(-7) cm2/s and 6.3 +/- 1.9 x 10(-7) cm2/s in normal and neoplastic tissues, respectively. The interstitial fluid flow, in general, was directed into postcapillary venules. The results obtained in this study should provide the impetus for further investigation into the diffusion and convection in various tissues under normal and pathological conditions.

A clinical trial evaluating cholestyramine to prevent diarrhea in patients maintained on low-fat diets during pelvic radiation therapy.

A prospective randomized trial to determine the value of a low fat diet with or without cholestyramine in the treatment of acute intestinal complications of pelvic irradiation is presented. A total of 35 patients receiving pelvic irradiation were entered in the study and all patients had received a 40 gm fat diet. The group was then randomized to receive either placebo (17 patients) or cholestyramine (18 patients). Diarrhea occurred in six out of 16 evaluable patients in the control group and only one of the 17 evaluable patients in the cholestyramine group. The frequency of diarrhea and the diarrhea scale remained high in the placebo group in the entire observation period. Statistical analysis had revealed better diarrhea control in the cholestyramine group (p = less than 0.05). In this report mechanism by which diarrhea occurs following pelvic irradiation is discussed. The adverse effects associated with the use of cholestyramine have been presented. It was concluded that cholestyramine is effective in preventing acute diarrhea induced by pelvic irradiation in patients receiving a low fat diet but is associated with side effects.

Dissolution of retained bile duct stones using heparin.

Five cases of retained bile duct stones treated with heparin in normal saline are reported. In 2 cases the stones were retained in the left hepatic duct, proximal to the T tube. Infusion of heparin in normal saline was found to be simple and effective in treating retained bile duct stones without obvious side-effects. If stones are inadvertently left in the common bile duct following surgery, attempts should be made to encourage them to pass by heparin infusion through the T tube before a second operation is contemplated. Sufficient time must be allowed before the treatment is considered to have failed.