Activated alpha 2-macroglobulin promotes mitogenesis in rat vascular smooth muscle cells by a mechanism that is independent of growth-factor-carrier activity

Vascular smooth muscle cell (vSMC) proliferation is important in atherosclerosis. We previously demonstrated that methylamine-activated alpha 2-macroglobulin (alpha 2M) and transforming growth factor beta 1 (TGF-beta 1) cause a synergistic proliferative response in quiescent rat aortic vSMCs [Stouffer, G. A., La-Marre, J., Gonias, S. L. & Owens, G. K. (1993) J. Biol. Chem. 268, 18,340-18,344]. The first goal of this study was to determine whether the synergy is due to the ability of alpha 2M-methylamine (alpha 2M-MeNH2) to bind TGF-beta 1 and target the growth factor to vSMCs that express the alpha 2M receptor. Receptor-recognized alpha 2M derivatives without TGF-beta 1-binding activity, including ternary alpha 2M-trypsin, an 18-kDa proteolytic fragment of the alpha 2M subunit, and the corresponding recombinant receptor-binding fragment (rRBF) increased vSMC [3H]thymidine incorporation and cell number in a manner similar to alpha 2M-MeNH2. In combination with TGF-beta 1, each alpha 2M derivative caused a synergistic vSMC proliferative response. vSMCs responded comparably when treated with alpha 2M-MeNH2 and TGF-beta 1 simultaneously or in sequence. Furthermore, alpha 2M-MeNH2-TGF-beta 1 complexes increased [3H]thymidine incorporation no more than alpha 2M-MeNH2 alone. These results indicate that TGF-beta 1 binding to alpha 2M is not responsible for the synergistic mitogenic activity. Additional studies were undertaken to determine whether activated alpha 2M independently induces a signal-transduction response in vSMCs. alpha 2M-MeNH2 and rRBF caused a rapid, transient increase in vSMC inositol 1,4,5-trisphosphate. This response was pertussis-toxin insensitive. Receptor-associated protein (RAP; 170 nmol/L) inhibited 91-95% of the specific binding of 125I-alpha 2M-MeNH2 and 125I-rRBF to vSMC; however, RAP did not affect the inositol 1,4,5-trisphosphate response or the mitogenic response. These studies suggest that vSMCs express a receptor, other than low-density-lipoprotein-receptor-related protein, that transduces a signal in response to activated alpha 2M. This receptor may mediate the mitogenic activity of alpha 2M in vSMC culture.     

Mechanism of colloidal particle uptake into the lymphatic system: basic study with percutaneous lymphography

The distribution and evolution of (CT)n microsatellites were examined in GenBank mammalian DNA sequences because these microsatellites are known to play important roles in the regulation of some genes in Drosophila melanogaster. A total of 236 (CT)n microsatellite loci were found in GenBank mammalian gene sequences. To determine whether (CT)n microsatellite arrays were conserved at orthologous positions in distantly related mammalian sequences, we determined whether orthologous sequences existed in GenBank for each of the 236 loci. A total of 47 sequence alignments could be made. For rodent x rodent comparisons, 7 of 8 (CT)n arrays were conserved at identical positions in each pair of orthologous sequences. Comparisons of orthologous sequences between different orders of mammals indicated that 11 of 39 (CT)n arrays occurred at orthologous positions or within 1 kb of orthologous positions in each pair of sequences. It appears that there is some level of conservation of (CT)n repeats in distantly related mammals. However, this level of conservation may not be greater than what might be expected to occur by chance. In 13 cases where (CT)n arrays were not conserved at orthologous positions, the lack of a (CT)n array in one sequence resulted from either nucleotide substitution within an array or nonexpansion of a shorter (CT)n element. In these cases, significant sequence identity could be detected throughout the entire region even though the repeat array was not detected in one of the sequences. In contrast, there was a disruption of sequence identity in the (CT)n microsatellite region that ranged from 24 to 1600 bp in 21 cases.(ABSTRACT TRUNCATED AT 250 WORDS)     

Local regulation of collateral ventilation by oxygen and carbon dioxide

The effects of changes in CO2 and O2 on the mechanics of collateral ventilation were studied in anesthetized paralyzed dogs. A doublelumen catheter was wedged into a peripheral airway obstructing a segment of lung distal to the catheter. Through one lumen of the catheter, air, 5% CO2 in air, 10% CO2 in air, 5% O2 in N2, or 5% CO2 in N2 was infused at a constant flow (V). Pressure (Ps) was monitored through the other lumen. At functional residual capacity the resistance to collateral flor Rcoll = Ps/V. When V was interrrupted , the time for Ps to fall 63% was defined as the time constant for collateral ventilation, Tcoll. The effective compliance (Cs') = Tcoll/Rcoll. When air was replaced by 5% CO2, Rcoll fell 46.3% (+/- SE 2.8) and Tcoll fell 41.5% (+/- SE 3.0). When the CO2 concentration was increased from 5% to 10%, Rcoll fell an additional 9.2% (+/- SE 2.2) and Tcoll fell an additional 5.1% (+/- SE 4.4). When air was replaced by 5% O2 in N2, Rcoll rose 36.6% (+/- SE 6.0) and Tcoll rose 13.6% (+/- SE 10.5). No significant changes in Cs' were noted. We conclude that varying concentrations of CO2 and O2 provide potent mechanisms for the control of collateral ventillation which may be of importance in the regulation of ventillation perfusion relationships at the local level.     

The influence of acute diazepam pretreatment on the action and disposition of [14C]pentobarbital in rats

Diazepam (DZP) pretreatment (100mg/kg, ip) of rats 6 h before pentobarbital administration (45 mg/kg, ip) prolonged the barbiturate-induced narcosis. The concentrations of [14C]pentobarbital and total pentobarbital derivatives in blood or brain showed no differences between control and DZP-pretreated animals. The brain and blood concentrations of pentobarbital, when measured at a time corresponding to the respective arousal times from pentobarbital narcosis, were lower in the DZP-pretreated group. These results indicate that acute DZP pretreatment increases the sensitivity of the rat brain to pentobarbital rather than inducing changes in the disposition of the barbiturate.     

Electron microscopic study of the rat submaxillary gland during development

Cytological changes in the submaxillary glands of rat embryos of the 14th-21st days of the intrauterine development and of 7-day-old rats were studied electron microscopically. The cytoplasm of the salivary gland primordium cells of the 14-day embryo was found to possess multiple polysomes, the canals of the granular cytoplasmic network (GCN) being solitary. The latter elongated in the process of embryonic development, and later predominant development of GCN was observed in basal portions of the acinar cells. At early stages of the gland development the GCN was often observed to approximate mitochindria. The laminar complex in the course of embryonic development enlarges its size and at later stages it has a well differentiated appearance in the acinar cells. Secretory inclusions make their appearance in the forming cells of acinuses long before birth, the composition of the embryo gland secretion being somewhat different from that of adult animals. Differentiation of acinuses and striated parts of the salivary tubules is well pronounced in the embryonic period while no signs of appearance of granular portions of the salivary tubules are observed both in light and electron microscopy either in embryos or in 1-week-old rats.     

Asymmetry of the pelvic ureters in normal females

A total of 65 cases showing asymmetry of the pelvic ureters of greater than 1.4 cm for which no cause could be identified are presented. Only one was a male. The right ureter was situated more medially in 61 of the 64 females. Comparison of the characteristics of these patients with 51 females with symmetrical pelvic ureters revealed no differences except for a significantly more frequent left-sided position of the uterus in the asymmetrical group. Gross medial displacement of the right ureter in a female probably represents a normal variant in a large majority of patients.