Morphological variability of the human chromosomes in two Indian populations - Rajputs and Punjabis.

Karotypic evaluation of 100 Rajputs and 100 Punjabis revealed different frequencies of Y chromosome polymorphism and minor chromosome variants. Long Y chromosome were observed 5% of the Rajputs and 3% of the Punjabis. The Y indices of Rajputs were consistently higher than those of Punjabis. Significant differences were noted between Rajputs and Punjabis with respect to the 5 Y indices. Significant differences were also found when those 2 populations were compared with different populations of the world. Minor chromosome variants were observed in 18% of the Rajputs and 19% of the Punjabis.     

The characterisation and function of the polysaccharidases of human synovial fluid in rheumatoid and osteoarthritis.

A potential enzymic mechanism for the degradation of glycosaminogly cans was characterised using enzymes found in rheumatoid synovial fluid from the knee joint. This mechanism involves a true hyluronidase together with the concerted action of beta-glucuronidase and beta-N-acetylhexosaminidase. The contribution of the exopolysaccharidases to hyaluronate degradation was demonstrated by the use of specific inhibitors, while the distinct identity of a true hyaluronidase was shown by ammonium sulphate and agarose gel column fractionations. Only the hyluronidase fraction was capable of degrading high molecular weight hyaluronate. The exopolysaccharidase activities were shown to be markedly elevated in rheumatoid as compared to osteoarthritic synovial fluid and also normal serum. On the other hand, hyluronidase was similarly active in rheumatoid and osteoarthritic synovial fluids; both these levels were lower than that of normal human serum. Hyaluronidase in synovial fluid may thus be derived by diffusion from serum, since it is of relatively low molecular weight (60 000). The pH requirements of this enzyme system and the strong inhibition of hyaluronidase by synovial fluid make it unlikely that the mechanism operates extracellularly. It is proposed that as a lysosomal mechanism, however, it is an important contributing factor in the chronic erosion process characteristic of rheumatoid arthritis.     

Studies on the binding of 1-anilino-8-naphthalene sulfonate to very low density and high density himan serum lipoproteins.

Very low density and high density lipoproteins have been isolated from human plasma and their interaction with 1-anilin0-8-naphthalene sulfonate has been studied under different conditions of pH and added salt. Intrinsic fluorescence of bound 1-anilino-8-naphthalene sulfonate was higher for high density lipoproteins then for very low density lipoproteins, but was unaffected by salt in both systems. Binding of 1-anilino-8-naphthalene sulfonate by both these lipoproteins was saturable and was higher in the presence of added NaCl or CaCl2, Ca2+ having a greater effect than Na+ in enhancing fluorescence. The binding data were analyzed by Scatchard plots; the number of binding sites and the affinity of 1-anilino-8-naphthalene sulfonate for the site increased with increasing salt concentration. Fluorescence pH curves were similar to those published for phospholipids. From these and previous observations it is suggested that the phospholipids probably represent the major binding sites for 1-anilino-8-naphthalene sulfonate.     

Effect of 3-(3,4-dihydro-6-methoxy-2-naphthyl)2,2-dimethyl pentanoic acid--an antifertility agent--on in vitro uptake of estradiol 17beta-6, 7(3)-H in rat uterus.

The effect of 3-(3,4-dihydro-6-methoxy-2-naphthyl)-2,2-dimethyl-pentanoic acid, a potent nonsteroidal antifertility compound, on the uptake of estradiol-17beta-6, 7-tritiated in vitro in the rat uterus was studied. The estradiol uptake of estrogen-primed and compound-treated groups were the same. When estradiol and the compound were present in medium at the same time, estradiol uptake was significantly (p less than .01) increased. The results indicate that the compound synergizes the effect of estradiol.     

A Highlights from MBoC Selection: Fast and parallel nanoscale three-dimensional tracking of heterogeneous mammalian chromatin dynamics

Chromatin organization and dynamics are critical for gene regulation. In this work we present a methodology for fast and parallel three-dimensional (3D) tracking of multiple chromosomal loci of choice over many thousands of frames on various timescales. We achieved this by developing and combining fluorogenic and replenishable nanobody arrays, engineered point spread functions, and light sheet illumination. The result is gentle live-cell 3D tracking with excellent spatiotemporal resolution throughout the mammalian cell nucleus. Correction for both sample drift and nuclear translation facilitated accurate long-term tracking of the chromatin dynamics. We demonstrate tracking both of fast dynamics (50 Hz) and over timescales extending to several hours, and we find both large heterogeneity between cells and apparent anisotropy in the dynamics in the axial direction. We further quantify the effect of inhibiting actin polymerization on the dynamics and find an overall increase in both the apparent diffusion coefficient D* and anomalous diffusion exponent α and a transition to more-isotropic dynamics in 3D after such treatment. We think that in the future our methodology will allow researchers to obtain a better fundamental understanding of chromatin dynamics and how it is altered during disease progression and after perturbations of cellular function.     

Cooperative engagement and subsequent selective displacement of SR proteins define the pre-mRNA 3D structural scaffold for early spliceosome assembly

We recently reported that serine–arginine-rich (SR) protein-mediated pre-mRNA structural remodeling generates a pre-mRNA 3D structural scaffold that is stably recognized by the early spliceosomal components. However, the intermediate steps between the free pre-mRNA and the assembled early spliceosome are not yet characterized. By probing the early spliceosomal complexes in vitro and RNA-protein interactions in vivo, we show that the SR proteins bind the pre-mRNAs cooperatively generating a substrate that recruits U1 snRNP and U2AF65 in a splice signal-independent manner. Excess U1 snRNP selectively displaces some of the SR protein molecules from the pre-mRNA generating the substrate for splice signal-specific, sequential recognition by U1 snRNP, U2AF65 and U2AF35. Our work thus identifies a novel function of U1 snRNP in mammalian splicing substrate definition, explains the need for excess U1 snRNP compared to other U snRNPs in vivo, demonstrates how excess SR proteins could inhibit splicing, and provides a conceptual basis to examine if this mechanism of splicing substrate definition is employed by other splicing regulatory proteins.     

Production of monoclonal antibodies to estriol and their application in the development of a sensitive nonisotopic immunoassay

The development of highly specific monoclonal antibodies to estriol and a nonisotopic immunoassay (EIA) for unconjugated estriol based on the use of these monoclonal antibodies have been described. The monoclonal antibodies show little cross reactivity with other steroids and steroid conjugates and can be used directly in immunoassays without any purification. The EIA described here can be performed in 96-well microtiter plates or polystyrene tubes that have been coated with estriol-bovine serum albumin conjugate. In this assay, estriol in the standard or clinical samples (serum or saliva) competes with the immobilized steroid on the plate or the tube for binding with the antibody. The assay shows good agreement with radioimmunoassay (RIA) and is highly sensitive and reliable. Since no prior processing or extraction of the clinical samples is necessary, the method is potentially applicable for routine use in fetal monitoring as well as in a steroid laboratory.     

Biotransformation of p-coumaric acid by Paecilomyces variotii

AIMS: To investigate the biotransformation of p-coumaric acid into p-hydroxybenzoic acid (p-HBA) by Paecilomyces variotii Bainier MTCC 6581. METHODS AND RESULTS: As a result of p-coumaric acid degradation by P. variotii, three phenolic metabolites, p-hydroxybenzaldehyde (p-HBAld), p-HBA and protocatechuic acid were formed. These phenolics were detected using TLC and HPLC. The identity of p-HBA and p-HBAld was further confirmed by mass spectrometry. Various analyses showed that 10.0 mmol l(-1) concentration of p-coumaric acid produced a maximum amount of p-hydroxybenzoic acid, 200 mg l(-1), into the medium at 37 degrees C with high-density cultures. CONCLUSIONS: A catabolic pathway of p-coumaric acid by the fungus P. variotii is suggested for the first time. During the process of p-coumaric acid degradation, p-HBA accumulated in the medium as the major degradation product. SIGNIFICANCE AND IMPACT OF THE STUDY: Microbial degradation of cinnamic acid and hydroxycinnamic acid has continued to be the focus of intensive study. The main goal was to identify the microbial species capable of converting these substances into commercially value-added products such as benzoic acid derivatives or aromatic aldehydes.