Tetra substituted thiophenes as anti-inflammatory agents: exploitation of analogue-based drug design

A series of 17 novel tetra substituted thiophenes was designed, synthesized, and screened for anti-inflammatory activity in carrageenin induced rat paw edema model, an acute in vivo model. The lead molecule selected was Tenidap, a dual COX/LOX inhibitor. Compounds I (43%), III (60%), IV (60%), and VIII (64%) showed moderate to good anti-inflammatory activity. The best candidate among the whole series was VIII, which gave 64% protection to the inflamed paw. The side chain of candidate VIII had resemblance to that of Romazarit, a DMARD, which was withdrawn due to its toxicity profile. A probable reason for the metabolic stability of the proposed side chain not having the possibility of generating peroxy type radicals or acrylic acid moieties, unlike Romazarit, is discussed. The biological activity exhibited by the three designed series was in the order of methyl amino series > ethyl amino series > carbethoxy amino series. The –(CO)–CH₂–COOR side chain at the fifth position as in candidate VIII, the methyl amino group at the second position, and the ester at the third position of the thiophene can be considered as a three-point pharmacophore for designing better anti-inflammatory agents. The present study is a classical example of the exploitation of an analogue based drug design, which culminated in the development of good anti-inflammatory agents that have the potential of becoming dual inhibitors.     

Validation of the LC-MS/MS method for the quantification of mevalonic acid in human plasma and determination of the matrix effect

A simple, specific, and sufficiently sensitive liquid chromatography-tandem mass spectrometry (negative-ion electrospray ionization) methodology to determine mevalonic acid (MVA) in human plasma is described, and its application to the analysis of rat plasma MVA levels after rosuvastatin administration is demonstrated. The method was validated over the linearity range of 0.5-50.0 ng/ml (r(2) > 0.99) using deuterated MVA as an internal standard. The lower limit of quantification was 0.5 ng/ml. The assay procedure involved the isolation of MVA from plasma samples using solid-phase extraction. Chromatographic separation was achieved on a HyPurity Advance column with a mobile phase consisting of ammonium formate buffer (10 mM, pH 8.0) and acetonitrile (70:30, v/v). Excellent precision and accuracy were observed. MVA and deuterated mevalonolactone were stable in water and plasma under different storage and processing conditions. The recovery observed was low, which was attributable to a significant matrix effect. A significant decrease (30-40%; P < 0.05) was observed in rat plasma MVA levels after rosuvastatin administration.     

Cytotoxicity of two proteins isolated from Bacillus thuringiensis var. israelensis crystals to insect and mammalian cell lines

Abstract Two toxic proteins (proteins A and B) were isolated from Bacillus thuringiensis var. israelensis crystals using ion-exchange chromatography. A cell cytotoxicity assay based on the uptake of [³H]uridine by tissue culture cells following exposure to the toxic proteins, was used to measure the viability of the cells. Proteins A and B were cytotoxic to Aedes aegypti and MA104 cells. Protein B was also cytotoxic to Spodoptera frugiperda cells.     

Tolypocladium cylindrosporum (Deuteromycotina: Moniliales), a fungal pathogen of the mosquito Aedes australis

A blastoconidial preparation of the New Zealand isolate of T. cylindrosporum was tested in two field trials against larvae of the freshwater mosquito Aedes subalbirostris and the brackish water species Ae. australis. There was some reduction in the population of the Ae. subalbirostris larvae and larvae collected from the test pools exhibited signs of fungal infection when reared in the laboratory. In the brackish pools there was no observable decrease in the larval populations even though Ae. australis is the natural host of T. cylindrosporum [NZ]. Water volume fluctuations in the test pools were a major difficulty experienced when monitoring the results.Tests against field-collected non-target fauna indicated that T. cylindrosporum is non-specific in action. Species of daphnid, dixid larvae and copepods were all susceptible to infection at varying degrees.     

INDUCED DIFFERENTIATION OF GERANIUM PLANTS FROM UNDIFFERENTIATED CALLUS IN VITRO

Geranium callus produced from explants of stem tip, internode pith with vascular tissue on synthetic media with or without coconut milk and 2,4-dichlorophenoxyacetic acid grew well for many generations, but only tracheids were induced in them. If callus produced on these media was subcultured immediately on Murashige and Skoog medium with 0.1 mg/liter/α-naphthalene acetic acid (NAA) and 10.0 mg/liter kinetin (K) and incubated at 16/8-hr light/dark cycle, shoots were induced in 8-10 weeks and roots in another 8-10 weeks. Callus produced on the MS medium with the same supplements of NAA and K, subcultured on the same medium and incubated in 16/8-hr light/dark cycle, produced shoots in 6-8 weeks. However, on callus subcultured more than three times, shoots differentiated with greater difficulty and none differentiated after six subcultures. Some abnormal shoot-like structures were also produced, the cells of which showed virus-like inclusion bodies. Requirements of the different varieties for differentiating organs differed. Among 12/12-, 15/9-, 16/8-, and 20/4-hr light/dark photoperiods that induced differentiation, 15/9- and 16/8-hr were more effective than the others. Continuous illumination did not induce differentiation. Differentiated shoots formed roots more readily on a medium with reversed proportions of auxin and kinetin. On agar roots were devoid of root hairs. Root hairs were formed when the shoots and plantlets were cultured on filter-paper bridges. Many “mother” stock plants were produced. These are being studied for their growth qualities and for possible viruses and other pathogens.     

Effect of recent antigen exposure on the functional expression of B cell subpopulations

We investigated whether the definition of functional B cell subpopulations changes after the exposure of B cells to specific antigen. Recent in vivo priming with fluorescein- (FL) coupled T-independent (TI) antigens leads to an augmentation of the subsequent in vitro responses of B cells to FL-coupled TI antigens, including FL-polymerized flagellin, FL-lipopolysaccharide, and FL-Brucella abortus, as well as a FL-coupled T-dependent (TD) antigen, FL-keyhole limpet hemocyanin (KLH). This effect, which is evident 7 days after priming, is of short duration in that B cells from FL-Ficoll injected mice display normal responsiveness 3 wk after priming. When mice are primed with FL-KLH, the TD antigen, B cells responsive to a subsequent FL-KLH challenge are expanded, but not short-term cross-priming for any of the TI antigen challenges is observed. Limiting dilution precursor analysis shows that B cell populations responding to different FL-coupled TD and TI antigens overlap in unimmunized animals. FL-TI antigen priming induces not only quantitative changes in the responding B cells (increased precursor frequencies) but also results in functional changes in FL-specific B cells. The primed B cells now respond to FL-hapten in a carrier-restricted manner and behave as independent (non-overlapping) subpopulations. We suggest that B cell responses to different forms of the same hapten are influenced in part by their recent "history" of antigenic exposure.     

Interaction between RNA polymerase and a ribosomal RNA promoter of E. coli.

The interaction between RNA polymerase and the E. coli ribosomal (r) RNA promoter(s) of the rrnE operon has been studied by the filter-binding method. The extent of complex formation between RNA polymerase and rrnE promoter(s) is salt-dependent; ppGpp specifically inhibits interaction of RNA polymerase with the rrnE promoter(s). A tentative model is proposed for the molecular events in the early steps of rRNA initiation: a transition of the primarily formed, labile RNA polymerase-rRNA promoter complex to a more stable form is the determining step. This step is salt-sensitive; ppGpp acts on this "isomerization".     

Root apical organization in gymnosperms

Summary The roots of Ephedra foliata have a long cap. The root apical meristem can be distinguished into (1) a discrete layer of stelar initials, (2) the columella and its initials, and (3) a common initiating region for the cortex and the peripheral region of the cap. The study of the cell lineages brings out that the first zone and its derivatives show a Körper complex of cells, the second and its derivatives divide in a transverse plane, and the third zone exhibits the Kappe complex.This organization resembles the gymnospermous type mentioned in the older literature in the discreteness of the plerome.A possible line of evolution of the root apical structures of gymnosperms is suggested.Now the Birla Institute of Technology & Science.