Cloning and expression of a novel, tissue specifically expressed member of the UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase family

We report the cloning and expression of the fifth member of the mammalian UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase (ppGaNTase) family. Degenerate polymerase chain reaction amplification and hybridization screening of a rat sublingual gland (RSLG) cDNA library were used to identify a novel isoform termed ppGaNTase-T5. Conceptual translation of the cDNA reveals a uniquely long stem region not observed for other members of this enzyme family. Recombinant proteins expressed transiently in COS7 cells displayed transferase activity in vitro. Relative activity and substrate preferences of ppGaNTase-T5 were compared with previously identified isoforms (ppGaNTase-T1, -T3, and -T4); ppGaNTase-T5 and -T4 glycosylated a restricted subset of peptides whereas ppGaNTase-T1 and -T3 glycosylated a broader range of substrates. Northern blot analysis revealed that ppGaNTase-T5 is expressed in a highly tissue-specific manner; abundant expression was seen in the RSLG, with lesser amounts of message in the stomach, small intestine, and colon. Therefore, the pattern of expression of ppGaNTase-T5 is the most restricted of all isoforms examined thus far. The identification of this novel isoform underscores the diversity and complexity of the family of genes controlling O-linked glycosylation.     

Estrogen, the ovary, and neutotransmitters: factors associated with aging

Our studies in the C57BL/6J mouse have been designed to examine the interactions of aging and the ovary, and their mutual effects on neuroendocrine function. In the pituitary, ovarian status and not age determines responsiveness to gonadotropin hormone releasing hormone (GnRH), but estrogen (E2) is an important mediator in CNS changes, and removal of the ovary (OVX) is deleterious to the neuroendocrine hypothalamus. OVX for just six days in young animals results in synaptic loss between noradrenergic terminals and gonadotropin hormone releasing hormone (GnRH) neurons. Long-term OVX, hypothesized to protect against neuroendocrine aging, fails to guard against any studied age-related changes. Some age-related changes occur as early as midlife. Although neuron number remains constant at middle age, opiatergic neurons undergo significant functional changes by producing opiate antagonist peptides. This change appears to be caused by alterations in the prohormone convertases, which cleave propeptide to peptide. Altered peptides may trigger the loss of reproductive capacity. The midlife shift in opiate peptide production is a component of natural developmental processes that begin in the neonate and continue through old age. In the cholinergic system, E2 mediates numbers of cholinergic receptors, cholinergic neurons, and cholinergic-modulated memory systems in both young and old animals. Regardless of age, ovarian steroids, if present at physiologic levels, are beneficial to the neuroendocrine CNS, and long-term deprivation from ovarian-produced factors is deleterious in the systems we have examined. Our studies have shown that deprivation from ovarian steroid hormones in the female appears to be a major factor in the health of the CNS and in events associated with aging.     

Ordering of polymorphic markers in the chromosome region 3p21

Starting from five markers, with a well-defined order from distal to proximal 3p21, nine other markers could be inserted in this 3p21 map. Five were precisely mapped genetically. The other markers were ordered by FISH and/or deletion hybrid mapping. The overall 3p21 order from distal to proximal is as follows: D3S1298-D3S1260-(D3S966, D3S1448 (= D3S1449)-D3S1029-D3S32-D3S643-D3F15S2 -D3S2968-D3S1235-D3S1289-D3S1447-D3S1295.     

Evaporation and pervaporation of a binary mixture from an inert carrier liquid

Bench-Scale experiments were carried out with the ternary mixture 2-propanol/water/glycerol. In a cylindrical vessel, the stirred and thermostated liquid was exposed to a preheated and either dry or humidified air stream. The mixture was evaporated either from a free surface or from the surface of a porous plate. On evaporating the mixture from a porous plate into a dry air stream, the less volatile component water escapes preferentially; in the case of evaporation from a free surface, water is retained in the mixture. In the former case, selectivity is liquid diffusion controlled, in the latter, the gas-side mass transfer and the thermodynamic equilibrium are the controlling mechanisms. On reducing the gas flow rate, the more volatile alcohol evaporates preferentially in both cases; the selectivity is controlled only by thermodynamic equilibrium. Humidification of the air stream with water vapour causes preferential evaporation of the alcohol at an increased rate. Furthermore, instabilities in the liquid boundary layer due to density gradients and surface tension effects may affect the selectivity of evaporation. Experimental results show that the selectivity of evaporation can be manipulated by choosing appropriate evaporation conditions. The theory, which applies the generalized Stefan-Maxwell equations to diffusion in the liquid, can describe these effects.     

Antibody-dependent cell-mediated cytotoxicity in rheumatoid arthritis. Effect of rheumatoid serum fractions on normal lymphocytes

Reduced antibody-dependent cell-mediated cytotoxicity (ADCC) was demonstrated in lymphocytes of patients with rheumatoid arthritis (RA). Rheumatoid factor (RF) positive sera inhibited ADCC of normal lymphocytes by reacting both with effector and target cells (sensitized chicken red blood cells). These sera were fractionated by specific adsorption or gradient ultracentrifugation, and isolated RF or RF negative fractions were tested for their ability to inhibit ADCC by reacting with normal human lymphocytes or target cells. RF was ineffective on normal lymphocytes but it strongly inhibited the reaction by interaction with target cells. IgG RF negative fractions of certain sera were inhibitory by direct interaction with effector cells.     

Brown fat retention in sudden infant death syndrome

This study was undertaken to confirm the findings of Naeye with regard to the pathological retention of the so-called brown fat cells in the periadrenal adipose tissue of infants who die of the sudden infant death syndrome. The percent of multilocular fat cells was determined in histological sections of periadrenal adipose tissue obtained from a series of 289 autopsies. In general, our morphological observations confirm his. However, our results indicate that the percent of periadrenal multilocular cells cannot be used as a diagnostic criterion in any single case. Futhermore, we consider that the mechanism for this delay in transformation remains unclear.     

Die analytische Beurteilung von Tribromsalicylanilid

Analytical Assay of Tribromosalicyl Anilide Methods published till now for the investigation of TBS, especially those involving ion-exchange, thin-layer chromatography, a combination of the two and gas-chromatography are reviewed. Using these methods it is possible, by simple means, to separate, identify and quantitatively determine the major components of the mixture.