The augmentation of tumor-specific immunity using haptenic muramyl dipeptide (MDP) derivatives

Summary A new haptenic compound, a muramyl dipeptide (MDP) derivative (designated as L4-MDP-ONB) cross-reactive with Bacillus Calmette Guerin (BCG) was synthesized. The cross-reactivity of L4-MDP hapten to BCG was demonstrated from the following evidence; (a) lymph node cells from BCG-primed C3H/HeN mice exhibited appreciable L4-MDP-specific proliferative responses to the in vitro stimulation of L4-MDP-modified syngeneic cells (L4-MDP-self); (b) inoculation of L4-MDP-self into footpads of BCG-primed C3H/HeN mice elicited ample delayed type-hypersensitivity (DTH) responses in vivo as measured by footpad swelling; and (c) BCG-primed mice contained L4-MDP-reactive helper T cell activity which functions to augment the generation of effector T cell responses to cell surface antigens. This crossreactivity between L4-MDP hapten and BCG as measured by the helper T cell activity was applied to enhanced induction of tumor-specific immunity. When BCG-primed C3H/HeN mice were immunized with L4-MDP-modified syngeneic X5563 tumor cells, these mice could generate augmented tumor-specific in vivo protective (tumor neutralizing) immunity as well as in vitro cytotoxic T cell responses. These results indicate the effectiveness of L4-MDP hapten in augmenting tumor-specific immunity. The present approach is discussed in the context of potential advantages of this new hapten for its future application to clinical tumor systems.     

Anterograde Axonal Transport of Peptidylglycine α-Amidating Monooxygenase in Rat Sciatic Nerves

Axonal transport of peptidylglycine alpha-amidating monooxygenase (PAM) activity was studied in rat sciatic nerves from 12 to 120 h after double ligations. The anterograde axonal transport increased and reached a plateau between 48 and 72 h and then decreased. The flow rate was 100 mm/day, and the molecular mass of the active entity was 70 kDa, which was determined by gel filtration. In contrast, there was no evidence for significant retrograde axonal transport. Anterograde axonal transport of immunoreactive cholecystokinin, a carboxy-terminal-amidated putative neuropeptide, was also found. These results suggest that PAM is transported by a rapid axonal flow and may play a role as a processing enzyme during transport or in the terminals of rat sciatic nerves.     

Mechanism of How Salt-Gradient-Induced Charges Affect the Translocation of DNA Molecules through a Nanopore

Experiments using nanopores demonstrated that a salt gradient enhances the capture rate of DNA and reduces its translocation speed. These two effects can help to enable electrical DNA sequencing with nanopores. Here, we provide a quantitative theoretical evaluation that shows the positive net charges, which accumulate around the pore entrance due to the salt gradient, are responsible for the two observed effects: they reinforce the electric capture field, resulting in promoted molecule capture rate; and they induce cationic electroosmotic flow through the nanopore, thus significantly retarding the motion of the anionic DNA through the nanopore. Our multiphysical simulation results show that, during the polymer trapping stage, the former effect plays the major role, thus resulting in promoted DNA capture rate, while during the nanopore-penetrating stage the latter effect dominates and consequently reduces the DNA translocation speed significantly. Quantitative agreement with experimental results has been reached by further taking nanopore wall surface charges into account.     

Physiological evaluation of temperature effect on the growth processes of the mysid, Neomysis intermedia Czerniawsky

Ingestion, respiration, and molting loss rates were measuredover the 3 – 29°C range in Neomysis intermedia. Weightspecific rates of these physiological processes ranged from2 to 140% body C day^–1 for ingestion, from 2 to 15% bodyC day^–1 for respiration, and from 0.1 to 5% body C day^–1for molting loss. All weight-specific rates showed a logarithmicdecrease with a logarithmic increase in body weight, and a logarithmicincrease with a linear increase in temperature below 20 or 25°C.The effect of temperature, however, was different between thephysiological rates, with a large temperature dependency foringestion (Q₁₀ = 2.6 –3.9) and molting loss (Q¹⁰ = 2.9– 3.6) and a moderate temperature dependency for respiration(Q₁₀ = 1.9 – 2.1). Calculated assimilation efficiencychanged with body size, but was constant over the temperaturerange examined. Allocation of assimilated materials varied witha change in temperature, reflecting the different temperaturedependence between physiological processes. It was deduced thatthe strong temperature dependency of the growth rate in N. intermediaobserved in the previous studies resulted from the large temperatureeffect on ingestion and assimilation rates, superimposed bythe different allocation of assimilated materials. ¹Present address: Department of Botany, University of Tokyo,Hongo, Tokyo 113, Japan     

Cauliflower mosaic virus 35 S promoter directs S phase specific expression in plant cells

Summary An experimental system to study cell cycle specific gene expression in plant cells was developed using protoplasts from tobacco cells synchronized by aphidicolin treatment. Chimeric plasmids consisting either of the chloramphenicol acetyltransferase (CAT) gene downstream of the cauliflower mosaic virus (CaMV) 35 S promoter or the nopaline synthase (nos) promoter were introduced into synchronized protoplasts of four cell cycle stages by electroporation. In the case of the CaMV 35 S promoter cyclic oscillation of CAT activity was observed which paralleled the cell cycle of the recipient cells. The peak of CAT activity was found in the S phase, while no such cyclic change was observed in the case of the nos promoter. This system clearly shows that it is feasible to search for a cell cycle specific promoter. The significance of these observations is discussed in relation to the study of plant cells.     

Den distribution and den use by the red foxVulpes vulpes japonica in Kyushu

The distribution of dens and den use by the red fox was studied in Kumamoto Prefecture between 1968 and 1982. A total of 80 dens were classified into natal, residential, and temporarily-visited dens. Natal dens comprised 12.4% of all dens. Relative proportions were quite stable for a long period. This indicates a constant number of reproducing females in the area for a considerable length of time. Seasonally the utilization of den showed three peaks in February, May and October. The peaks coincided well with the peaks of reproductive activity of female foxes. The results of multiple regression analysis of distribution of dens with respect to selected environmental parameters around densites indicated that the fox selectively utilize open land, rather than densely vegetated areas. The persistence of dens, the selective utilization by foxes of open land and the sizes of dens were all related to the breeding activities of female foxes inhabiting the area. Reproductive females usually used plural dens (“den group”) in one reproductive season. The numbers of those females occuring in the area were maintained at a steady level for fairy long periods. These indicate that the existence of a stable social relationship between females mediated through occupancy of “den groups.”     

Determination of the source of immunoreactive relaxin in the cat

We have recently reported the secretory profile of relaxin throughout gestation in the cat. Because the appearance of relaxin begins at about Day 20 (Day O = ovulation) and because implantation begins shortly before this at Days 13-14, we hypothesized that relaxin was of feto-placental origin. To test this hypothesis, we used 4 experimental groups: 1) Control (laparotomy-only at Day 23 or 42, n = 4); 2) Early Ovariectomy (Ovx, bilateral ovariectomy between Days 23 and 26, n = 4); 3) Late Ovx (bilateral ovariectomy between Days 40 and 44, n = 4); 4) Tissue Removal (removal of feto-placental units, uterus, and one ovary on Days 16, 21, 28 and 35, n = 1 per day). Pregnancies were maintained in both Ovx groups by progesterone administration. Relaxin secretory patterns in Ovx groups were similar to the Control data. Relaxin was detectable in plasma beginning at about Day 20, with maximum concentrations reached by Day 30. Relaxin concentrations were highest (immunoactivity per mg tissue) in homogenates of placental tissues as compared to luteal, fetal, or uterine tissues. Altogether, these data indicate that the feto-placental unit is the source of relaxin in the cat.     

Properties of the binding of follicle-stimulating hormone to the fetal rat testis

Basic properties of the binding of [131I]-labeled rat FSH ([131I]rFSH) to the testicular homogenates of fetal rats were analyzed by micro-radioreceptor assay. Specific binding of FSH was detectable in the testicular preparations from 15.5-day fetuses, but it was very low. After 17.5 days of gestation, specific FSH binding was apparent in the testis and was effectively displaced by rat FSH but not by rat LH. The Scatchard plot analyses of the binding of FSH to the testicular preparations of fetuses showed straight lines similar to those of postnatal rats, suggesting the presence of a single class of binding sites. The mean dissociation constant (Kd) for FSH receptors in 17.5-day fetuses was 0.413 +/- 0.043 nM, which was significantly greater than that in postnatal rats at 50 days of age. However, the Kd in 19.5-day fetuses was not significantly different from those in 17.5-day fetuses and postnatal rats due to its considerable variance. The capacity of FSH binding sites was 0.51 +/- 0.01 fmol/testis in 17.5-day fetuses, which was significantly less than those of 19.5-day fetuses and postnatal rats.