The allele-specific synthetic lethality of prlA-prlG double mutants predicts interactive domains of SecY and SecE.

The secretion of proteins from the cytoplasm of Escherichia coli requires the interaction of two integral inner membrane components, SecY and SecE. We have devised a genetic approach to probe the molecular nature of the SecY-SecE interaction. Suppressor alleles of secY and secE, termed prlA and prlG, respectively, were analyzed in pair-wise combinations for synthetic phenotypes. From a total of 115 combinations, we found only seven pairs of alleles that exhibit a synthetic defect when present in combination with one another. The phenotypes observed are not the result of additive defects caused by the prl alleles, nor are they the consequence of multiple suppressors functioning within the same strain. In all cases, the synthetic defect is recessive to wild-type secY or secE provided in trans. The recessive nature argues for a defective interaction between the Prl suppressors. The extreme allele specificity and topological coincidence of the mutations represented by these seven pairs of alleles identify domains of interaction between SecY/PrlA and SecE/PrlG.     

Association of apolipoprotein E but not B with Alzheimer's disease

In our studies apolipoprotein E4 (APOE4) is associated with both early- and late-onset Alzheimer's disease. Alzheimer's patients from West Texas were screened for the APOE4 allele, which was found at frequencies of 0.43 and 0.59 in familial late- and early-onset cases. Sporadic cases had lower frequencies, but they still were 2–4 times higher than control spouses. To determine whether the APOE association may be a risk factor for coronary disease as well, we examined two APOB gene restriction sites that have previously been found to be associated with coronary artery disease, especially myocardial infarctions. The APOB alleles were found at similar frequencies in Alzheimer's patients and control spouses.     

Lymphocyte cytotoxicity in x-irradiation-induced rat small bowel adenocarcinoma. III. Blocking by 3 M KCL extract

Hypertonic salt extracts (3 M KCl) of x-irradiation-induced Holtzman rat small bowel adenocarcinomas blocked the in vitro destruction of allogeneic cultured cells of this malignancy by sensitized lymphoid cells obtained from tumor-bearing animals. The protective effect were mediated by a blocking action at both the effector and the target cell level. The extracts were separated into 50% ammonium sulfate soluble and insoluble fractions with the soluble fraction being more effective in blocking the cytotoxic responses through interaction with the lymphoid cells whereas the insoluble one had a greater effect upon tumor target cells. Associated with both fractions was the oncofetal glycoprotein previously identified with the cellular membrane of this x-ray-induced malignancy. Immunoglobulins were identified with insoluble fraction; some were able to bind the oncofetal protein, thus clasifying it as a fetal antigen. The protective effects of the soluble fraction and this neoantigen were found to be citric acid labile, whereas the effects due to the insoluble fraction were unchanged.     

Evaluation of lung recovery assay for schistosomula in mice immunized with x-irradiated cercariae of Schistosoma mansoni.

The lung recovery assay for schistosomula has recently been used as a rapid method for measuring the state of immunity of a challenged animal. Its merit rests on the fact that the peak day of schistosomular recovery in the lungs of both normal and immune hosts falls on the same day and the degree of immunity can be measured by the percent of schistosomular recovery on this particular day. An evaluation of this method for mice immunized with highly X-irradiated cercariae of Schistosoma mansoni was made. The results indicate that in the immunized mice the peak day for the schistosomular recovery was delayed and the number of schistosomula recovered on the peak day was decreased. The delay of the peak day was increased by the degree of host immunity, so its identification for a host of an unknown immunologic status was not a simple matter. In addition, this method measured only the immune effect which took place in the skin and on prepeak days in the lungs. It was not as competent as the conventional liver perfusion method which measures the total result of the immune effect of the skin, lungs, and liver. Our conclusion is that for measuring the immunizing effect of irradiated cercariae, the lung recovery method is not suitable for the determination of the state of immunity induced.     

The positional differentiation of abscission zones during the development of leaves of Sambucus nigra and the response of the cells to auxin and ethylene

Summary Abscission in the leaf rachis of Sambucus nigra L. is preceded by a positional differentiation of zone cells that enlarge and separate in response to ethylene but not to auxin. These cells are absent from youngest leaves, and such leaves do not abscind even in ethylene; other cells of the immature rachii will enlarge in response to auxin. These two classes of target cells are always recognisable by their opposing responses to auxin and ethylene. Prior to separation zone cells exposed to ethylene show considerable activation of the cytoplasm, many polysomes, elongate endoplasmic reticulum and highly dilated dictyosomes with many associated vesicles. Treatment with auxin precludes these changes, and abscission is always retarded: high levels of ethylene must be added to overcome the auxin inhibition. The differentiation of zone cells and their ethylene-stimulated growth and activation are prerequisites for rachis abscission in Sambucus. Such cell development may be of general occurrence prior to organ abscission in plants.Abbreviation IAA indole-3yl-acetic acid     

PGE1 metabolism by the perfused rat liver

The hepatic and biliary metabolites of PGE₁ have been isolated and identified after infusions of PGE₁ into isolated rat liver preparations. The results demonstrate that in general PGE₁ undergoes metabolism similar to that of PGE₂ in the rat and reveals the possibility of a selective PG metabolite transport system across the biliary canalicular membrane.     

The adsorptive characteristics of proteins for polystyrene and their significance in solid-phase immunoassays

The adsorptive characteristics of polystyrene tubes for seven proteins of different molecular weight and ionic charge have been studied. The stability of these adsorbed proteins was monitored throughout the various steps of the amplified-enzyme-linked immunosorbent assay (a-ELISA). Using ¹²⁵I-labeled proteins, it was observed that each had a characteristic adsorptive behavior not explainable by simple charge differences. The proportion of protein bound to a 6.5-cm² tube surface was independent of the amount added up to an input of about 100 ng. Binding increased proportionately with temperature and incubation time. Beyond the range of concentrations where the proportion bound was independent of input, i.e., the region of independence, the kinetics of adsorption changed. The amount of protein adsorbed at the upper limit of the region of independence was inversely proportional to molecular weight for all except α-lactalbumin. Quantitative measurement of bound protein using the a-ELISA suggests that beyond the region of independence, protein-to-protein rather than protein-to-polystyrene adsorption occurred. This was suggested by computation using the Stokes radii of the proteins which implied that the amount of protein bound at the upper limit of the region of independence was a layer of protein one molecule thick. The upper limit of the region of independence determines, therefore, the maximun amount of functional antigen for such assays. Once adsorbed, proteins remained stably bound throughout the assay steps. The presence of competing proteins had no effect on the adsorptive characteristics of a given protein provided that the total protein concentration of the mixture was within the range of independence. The results indicate that the adsorptive characteristics of proteins for polystyrene tubes routinely used in solid-phase immunoassays should be taken into consideration when designing and interpreting data obtained with such assays.     

Embryogenesis and germination in rye (Secale cereale L.)

Summary When rye embryos imbibe water they rapidly return to a condition of biochemical and structural complexity. Three stages of imbibition can be recognised: Phase I a short period (10 min) of physical wetting; Phase II a longer period (1 h) when little further imbibition occurs, followed by Phase III a continuous phase of active water uptake. The latter coincides with an increase in respiration rate and an increase both in the number of mitochondria and of cristae within them. Changes in fine structure become evident in all organelles in Phase III, after 2 h of imbibition. In the unimbibed embryo endoplasmic reticulum is present only as short crescents associated with electron lucent bodies, but in Phase III the endoplasmic reticulum proliferates to form many surrounding cirlets. After 6 h these circlets become fewer and instead the endoplasmic reticulum is seen in close association with the nuclear membrane. Concurrently incorporation of radioactive uridine and thymidine is first detectible. This suggests that the large increase in protein synthesis occurs on new ribosomes present on the reticulum associated with the nuclear membrane. For the first 6 h protein synthesis must occur either on polysomes within the dense packing of ribosomes or on these circlets of endoplasmic reticulum associated with electron lucent bodies.     

Soil Greenhouse Gas Emissions and Carbon Dynamics of a No-Till,Corn-Based Cellulosic Ethanol Production System

Crop residues like corn (Zea mays L.) stover perform important functions that promote soil health and provide ecosystem services that influence agricultural sustainability and global biogeochemical cycles. We evaluated the effect of corn stover removal from a no-till, corn-soybean (Glycine max (L.) Merr) rotation on soil greenhouse gas (GHG; CO₂, N₂O, CH₄) fluxes, crop yields, and soil organic carbon (SOC) dynamics. We conducted a 4-year study using replicated field plots managed with two levels of corn stover removal (none; 55 % stover removal) for four complete crop cycles prior to initiation of ground surface gas flux measurements. Corn and soybean yields were not affected by stover removal with yields averaging 7.28 Mg ha^?1 for corn and 2.64 Mg ha^?1 for soybean. Corn stover removal treatment did not affect soil GHG fluxes from the corn phase; however, the treatment did significantly increase (107 %, P?=?0.037) N₂O fluxes during the soybean phase. The plots were a net source of CH₄ (～0.5 kg CH₄-C ha^?1 year^?1 average of all treatments and crops) during the generally wet study duration. Soil organic carbon stocks increased in both treatments during the 4-year study (initiated following 8 years of stover removal), with significantly higher SOC accumulation in the control plots compared to plots with corn stover removal (0–15 cm, P?=?0.048). Non-CO₂ greenhouse gas emissions (945 kg CO₂-eq ha^?1 year^?1) were roughly half of SOC (0–30 cm) gains with corn stover removal (1.841 Mg CO₂-eq ha^?1 year^?1) indicating that no-till practices greatly improve the viability of biennial corn stover harvesting under local soil-climatic conditions. Our results also show that repeated corn stover harvesting may increase N loss (as N₂O) from fields and thereby contribute to GHG production and loss of potential plant nutrients.