Double immunocytochemical staining for the in situ study of allotype distribution during an anti-trinitrophenyl immune response in chimeric rabbits

After incubation of tissue sections with anti-allotype-enzyme conjugates, the localization of immunoglobulin-allotype-bearing cells in the lymphoid tissues of conventional and chimeric rabbits could be established. The use of anti-allotype sera bearing distinct enzyme labels allowed simultaneous recognition of B cells producing immunoglobulin of one or the other parental types in heterozygous rabbits, or of B cells from the donor and recipient in chimeras. After immunization of chimeric rabbits with trinitrophenyl-keyhole limpet hemocyanin, anti-trinitrophenyl antibody-forming cells could be demonstrated through the use of a trinitrophenyl-alkaline phosphatase conjugate. Simultaneous incubation of sections with this reagent and with horseradish peroxidase coupled to (donor or recipient) anti-allotype sera made possible the determination of the origin (donor or recipient) of the antibody-forming cells. In agreement with the results of plaque assays and analyses of serum antibodies, all the anti-TNP producing cells were of donor origin when the chimeras had been created through injection of spleen or lymph node cells from trinitrophenyl primed donors. With this study we introduce a simple, direct method for the simultaneous identification of cells that produce antibody of a given allotype and a given specificity, applicable to appropriate studies in heterozygous or chimeric rabbits. The procedure has various advantages over previously reported methods.     

Physiological profile of world-class high-altitude climbers

The functional characteristics of six world-class high-altitude mountaineers were assessed 2-12 mo after the last high-altitude climb. Each climber on one or several occasions had reached altitudes of 8,500 m or above without supplementary O2. Static and dynamic lung volumes and right and left echocardiographic measurements were found to be within normal limits of sedentary controls (SC). Muscle fiber distribution was 70% type I, 22% type IIa, and 7% type IIb. Mean muscle fiber cross-sectional area was significantly smaller than that of SC (-15%) and of long-distance runners (LDR, -51%). The number of capillaries per unit cross-sectional area was significantly greater than that of SC (+ 40%). Total mitochondrial volume was not significantly different from that of SC, but its subsarcolemmal component was equal to that of LDR. Average maximal O2 consumption was 60 +/- 6 ml X kg-1 X min-1, which is between the values of SC and LDR. Average maximal anaerobic power was 28 +/- 2.5 W X kg-1, which is equal to that of SC and 40% lower that that of competitive high jumpers. All subjects were characterized by resting hyperventilation both in normoxia and in moderate (inspired O2 partial pressure = 77 Torr) hypoxia resulting in higher oxyhemoglobin saturation levels in hypoxia. The ventilatory response to four tidal volumes of pure O2 was similar to that of SC. It is concluded that elite high-altitude climbers do not have physiological adaptations to high altitude that justify their unique performance.     

Endurance training in humans: aerobic capacity and structure of skeletal muscle

The adaptation of muscle structure, power output, and mass-specific rate of maximal O2 consumption (VO2max/Mb) with endurance training on bicycle ergometers was studied for five male and five female subjects. Biopsies of vastus lateralis muscle and VO2max determinations were made at the start and end of 6 wk of training. The power output maintained on the ergometer daily for 30 min was adjusted to achieve a heart rate exceeding 85% of the maximum for two-thirds of the training session. It is proposed that the observed preferential proliferation of subsarcolemmal vs. interfibrillar mitochondria and the increase in intracellular lipid deposits are two possible mechanisms by which muscle cells adapt to an increased use of fat as a fuel. The relative increase of VO2max/Mb (14%) with training was found to be smaller by more than twofold than the relative increase in maximal maintained power (33%) and the relative change in the volume density of total mitochondria (+40%). However, the calculated VO2 required at an efficiency of 0.25 to produce the observed mass-specific increase in maximal maintained power matched the actual increase in VO2max/Mb (8.0 and 6.5 ml O2 X min-1 X kg-1, respectively). These results indicate that despite disparate relative changes the absolute change in aerobic capacity at the local level (maintained power) can account for the increase in aerobic capacity observed at the general level (VO2max).     

Binding of different antigen-enzyme and antibody-enzyme conjugates by intracellular antibodies in cytoplasm and Golgi complex of plasma cells

Summary Intracellular immunoglobulins in plasma cells were characterized by antigen-enzyme conjugates and anti-immunoglobulin antibody-enzyme conjugates applied in a double immunocytochemical approach. After their assemblage, immunoglobulins in the cytoplasm of anti-TNP antibody producing plasma cells can be demonstrated both by TNP-enzyme conjugates and by anti-immunoglobulin ( or chain specific) antibody-enzyme conjugates. Once arrived in the Golgi complex (GC) detection with TNP-enzyme conjugates remains possible, but anti-immunoglobulin antibody-enzyme conjugates did not bind to a detectable degree. Similar results were obtained in experiments where immunoglobulin-enzyme conjugates were used both as an antigen-enzyme conjugate and as an antibody-enzyme conjugate.     

The macroinvertebrate fauna of littoral and bottom substrata as a tool for biotypology of Frisian waters

During a biotypological research of surface waters in the province of Friesland, The Netherlands, the macroinvertebrate fauna was sampled, both from the littoral and bottom substrata. Results of this investigation in different types of water on 60 and 55 stations in 1981 and 1982, respectively, are presented. The species richness in the littoral samples exceeded that of the bottom samples, and the samples per station had relatively few species in common. Even in many cases the most abundant taxon was a different one in both samples of the same spot. With multivariate analysis of the data sets from bottom, littoral and total samples a fairly good resemblance was found between these data sets. It is concluded that the semi-quantitative processing of macroinvertebrate fauna data leads to reliable results. One can use one sample per station, wherein different substrata are combined. Distinct clusters were recognized in more extreme environmental conditions such as brackish and acid waters. With the help of the macroinvertebrate fauna the other types of water could not easily be differentiated.     

Estimation of the influx and the radius of the depletion zone developing around a root during nutrient uptake

An approximation to the analytical solution of the one-dimensional diffusion equation in cylindrical coordinates is derived. The solution applies to a semi-infinite system with a prescribed concentration at the root surface. From this approximate solution, simple formulas for calculating the radius of the depletion zone and the nutrient flux at the root surface as a function of time are obtained.     

Conjugate formation in urea: coupling of insoluble peptides to alkaline phosphatase for ELISA and in situ detection of antibody-forming cells

We report here a new method to produce synthetic peptide/alkaline phosphatase (AP) conjugates in the presence of urea. The method allows the use of peptides that are not soluble to a sufficient degree in aqueous buffers. The presence of 8 M urea during the construction of the synthetic peptide/AP conjugates does not influence enzyme activity nor the affinity of the anti-peptide antibodies for the conjugated peptide. We demonstrate that these synthetic peptide/AP conjugates can be used for detection of specific antipeptide antibody-forming cells (AFC) in vivo. This method for constructing enzyme conjugates with insoluble proteins or peptides suggest not only new possibilities for detection of specific AFC in vivo but also for applications in receptor-ligand studies, ELISA (enzyme-linked immunosorbent assay), and spot ELISA for detection of antibody-secreting cells in vitro.     

Immunological discrimination between the human apolipoprotein E2(Arg158----Cys) and E3 isoforms.

A specific anti-apoE2(Arg158----Cys) monoclonal antibody was raised by means of immunization of mice with a variant specific synthetic peptide. The peptide sequences used were homologous to apolipoprotein E of human and mouse. Consequently, the mouse immune system was tolerant to most of the selected sequences. Immunization with only one of selected peptides (amino acids 154-172) evoked an anti-peptide and anti-native protein response. Surprisingly, this peptide was predicted to have a low antigenicity index, in contrast to the other used peptides. The variant specific anti-peptide MAb that was generated with this sequence, recognizes apoE2(Arg158----Cys) and not apoE3. We here describe a sensitive, time saving, and simple immunoblot assay to detect apoE2(Arg158----Cys) in human sera without prior isoelectric focusing of serum proteins.     

In situ analysis of antibody-forming cells from the BALB/c CRIc idiotype family: idiotopic heterogeneity among clustered cells

Immunocytochemical staining methods were applied toward defining the in situ expansion of the BALB/c mouse antibody-forming cells (AFC) that express the CRIc idiotype (id) family associated with the antibody response against the p-azophenylarsonate (Ar) hapten. CRIc+ AFC dominated the early primary anti-Ar response but represented a decreased fraction of the anti-Ar AFC during secondary responses. Two subsets of the CRIc family, CRIc1 and CRIc2, differed in their relative expression, with CRIc1 more pronounced during primary responses and CRIc2 better expanded in secondary responses. Idiotopic differences among AFC in very close proximity suggested convergent migration among B cells having similar but not identical V regions or possibly V region mutations among clonally derived cells. The approach also allowed in situ idiotypic analysis of polyclonally activated B AFC.     

Mineralogical controls on the bacterial oxidation of refractory Barberton gold ores

Abstract: The effect of mineralogical characteristics of gold ore minerals on the nature of sulphide oxidation during a bacterial leaching process was investigated. Three different ore types from the South African goldmines were used, i.e. an arsenopyritic-pyritic ore (Sheba goldmine), a pyritic ore (Agnes goldmine) and a loellingitic-arsenopyritic ore (New Consort goldmine). Detailed mineralogical characterization of each ore was performed. Thereafter, polished sections of the sulphides were suspended in a bacterial leach pulp in an air-stirred vessel for various periods of time. The effect of bacterial oxidation on the sulphides was monitored. Different types of gold-bearing arsenopyrite exist, each type having its own characteristic behaviour during the bacterial oxidation process. The rate of oxidation is controlled by the amount of defects in the crystal structure, and the amount of defects is again controlled by the composition of the arsenopyrite crystal. The distribution of refractory gold in the sulphide minerals can be correlated with the presence of compositional zones and structural deviations. These same mineralogical features also control the sites and rates of bacterial oxidation. Thus. refractory gold occurs at sites which are preferentially leached by the bacteria. The rate of gold liberation from sulphides is therefore being enhanced during the early stages of bacterial oxidation. Defects in a crystal structure influence the rate of bio-oxidation, and can be related directly to the crystal structure of the sulphide mineral, the crystallographic orientation of the exposed surfaces, and differences in chemical compositional and mechanical deviations in the crytals. A combination of all of these mineralogical factors influences the bacterial oxidation process. To optimize and to understand the leaching of an individual ore it is important to establish its controlling factors.