The limited proteolysis of tumor necrosis factor-α

The limited proteolysis of human recombinant TNF- by trypsin yields two stable products resulting from cleavage after Arg6 and Arg44. In solution these two products remain associated together in a trimer with a Stokes' radius slightly greater than the radius of intact TNF- and, therefore, could not be separated from each other under nondenaturing conditions. This limited digest retains at least 20% of the activity of the original TNF- sample, and has a tertiary structure that is similar to that of the native protein by circular dichroism. On the other hand, incorrectly folded, inactive TNF- undergoes extensive digestion following similar treatment with trypsin. These results indicate that the active form of TNF- has a tight core structure which is maintained afterN-terminal cleavage and removal.     

Is Atlantic salmon production limited by number of territories?

Thirty Atlantic salmon Salmo salar parr (mean 135 mm total length, L _T) in the River Alta, Norway, were radio‐tagged and tracked during a 11–13 day period. The parr stayed within defined home ranges with a 95% probability of localization within an average area of 1286 m²(241–3484 m²). On average, each parr had overlapping home ranges with 7·7 other parr, and the overlap between pairs of fish covered on average 24% of their home range areas. Mean length of the river stretch used was 90 m (22–383 m). On average, during 38%(16–77%) of the tracking surveys, the fish had moved >10 m since the previous survey. Mean total distance moved during the whole study was 402 m (208–862 m). The Atlantic salmon were most often recorded in riffles, but 27% of the parr alternated between riffles and pools. The extensive movements, flexible habitat utilization and relatively large home ranges, together with the fact that all the parr had home ranges partly overlapping with other radio‐tagged parr, indicate a flexible and variable behaviour and, thereby, a more complex social structure than a rigid defence of a fixed territory. Thus, Atlantic salmon production seems not to be limited by the highest potential number of territories for large parr in a given area.     

One cell–one immunoglobin. Origin of limited heterogeneity of myeloma proteins

Plasma-cell tumour 5563 forms a single molecular species of immunoglobulin IgG(2)a, i.e. one variant of heavy chain and one variant of light chain. The molecules formed are labile and undergo alterations in charge properties, which rapidly lead to heterogeneity of the myeloma protein after synthesis. The single immunoglobulin species originally formed is found only after the shortest time-intervals tested, i.e. 10min incubation. Two types of changes in charge properties take place: (1) The originally formed protein (component o) is converted via an intermediate o' into the most basic form of 5563 myeloma protein found in serum (component a). Charge differences between these components are suppressed at pH8.9, but can be studied by chromatography at pH6.5 or by analysis of isoelectric points by isoelectric focusing in polyacrylamide gel. The conversion of components o and o' into component a apparently commences soon after assembly of the molecules and proceeds to completion extracellularly. (2) The second type of charge difference that distinguishes components a, b, c and d is exhibited over the pH range 6.0-8.9, but not at acid pH, and has been studied by electrophoresis at pH8.9, by chromatography and by isoelectric focusing. Conversion of component a into components b, c, d and e is only partial so that all five components can be found at decreasing concentrations in serum. Both types of charge alteration can be effected in vitro in the presence of serum, with optimum pH8.0. None of the charge differences could be attributed to the secretion process, since a component with the same isoelectric point as component o was found in secreted myeloma protein (1h). We have found no evidence to support the idea that the first type of change from component o to component a is due to ring formation of N-terminal [(14)C]glutamine into pyrrolid-2-one-5-carboxylic acid; however, our findings do not exclude this process happening very rapidly to a precursor of component o, possibly the polypeptide chain during or immediately after synthesis. In studying this point we noted that not only the heavy chains but also the kappa-type light chain of mouse 5563 myeloma protein have a blocked N-terminus.     

Purification and characterization of β-structural domains of β-lactoglobulin liberated by limited proteolysis

Incubation of β-lactoglobulin with immobilized trypsin at 5–10°C results in a time-dependent release of several fragments of the core domain in yields approaching 15%. Digests were fractionated by ion-exchange chromatography with a Mono Q HR5/5 column and analyzed after disulfide reduction by polyacrylamide gel electrophoresis in sodium dodecylsulfate. Three fragments with approximate molecular weights of 13.8, 9.6, and 6.7 kD were identified. The fraction from ion-exchange chromatography yielding the 6.7 kD fraction after disulfide reduction was further characterized because it was most homogeneous and gave the highest yield. The C-terminal cleavage site of the 6.7 kD core fragment appeared to be Lys₁₀₀ or Lys₁₀₁ as determined by C-terminal amino acid analysis. The exact masses, after reduction with dithiothreitol, are 6195 and 6926 as determined by laser desorption mass spectrometry, corresponding to residues 48–101 and 41–100. Prior to reduction, β-lactoglobulin C-terminal residues 149–162 are connected to these core domain fragments as shown by C-terminal analysis and mass spectrometry. Structural studies indicate that these 7.9 and 8.6 kD core domain fragments released by immobilized trypsin retain much of their native structure. CD spectra indicate the presence of antiparallel β-sheet structure similar to the native protein but the α-helix is lost. Spectra in the aromatic region indicate the existence of tertiary structure. Moreover, structural transitions in urea are completely reversible as measured by CD spectra, although the extrapolated ΔG _D ^H20 and the urea concentration at the transition midpoint are lower than for the native protein. The core domain fragments also display apH-dependent binding to immobilizedtrans-retinal as does intact protein. A single endotherm is obtained for both core domain fragments and native protein upon differential scanning calorimetry, but again, the domain is less stable as indicated by a transition peak maxima of 56.9°C as compared with 81.1°C for native protein.     

Structure–function relationships of molluscan troponin T revealed by limited proteolysis

Molluscan troponin regulates muscle contraction through a novel Ca²⁺-dependent activating mechanism associated with Ca²⁺-binding to the C-terminal domain of troponin C. To elucidate the further details of this regulation, we performed limited chymotryptic digestion of the troponin complex from akazara scallop striated muscle. The results indicated that troponin T is very susceptible to the protease, compared to troponin C or troponin I. The cleavage occurred at the C-terminal extension, producing an N-terminal 33-kDa fragment and a C-terminal 6-kDa fragment. This extension is conserved in various invertebrate troponin T proteins, but not in vertebrate troponin T. A ternary complex composed of the 33-kDa fragment of troponin T, troponin I, and troponin C could be separated from the 6-kDa troponin T fragment by gel filtration. This complex did not show any Ca²⁺-dependent activation of the Mg-ATPase activity of rabbit-actomyosin–scallop-tropomyosin. In addition, the actin–tropomyosin-binding affinity of this complex was significantly decreased with increasing Ca²⁺ concentration. These results indicate that the C-terminal extension of molluscan troponin T plays a role in anchoring the troponin complex to actin–tropomyosin filaments and is essential for regulation.     

Muscular dystrophies,alterations in a limited number of cellular pathways?

Identification of new genes involved in muscle disorders has dramatically changed the traditional clinical classification of the large and heterogeneous group of the muscular dystrophies. Results obtained in recent years by positional candidate cloning have demonstrated the role of the sarcolemma and of the nuclear envelope in normal muscle function and have elucidated molecular pathways perturbed by mutations that lead to muscular dystrophy.     

Is reproduction of endemic plant species particularly pollen limited in biodiversity hotspots?

Current evidence suggests that plants in biodiversity hotspots suffer more from pollen limitation of reproduction than those in lower diversity regions, primarily due to the response of self‐incompatible species. Species in biodiversity hotspots may thus be more at risk of limited reproduction and subsequent population decline. Should these species have restricted ranges (i.e. be endemics to a certain region), pollen limitation within highly diverse regions may pose an important threat to global plant biodiversity. We further dissect the global pattern by exploring whether pollen limitation of range‐restricted (endemic) species is distinctive and/or relates differently to species diversity than that of widespread (non‐endemic) species. To provide a preliminary test of this prediction we conducted both cross‐species and comparative phylogenetic meta‐analyses to determine the effect of endemism on the magnitude of pollen limitation and its relationship with regional species richness. Our data set included 287 plant species belonging to 78 families distributed world‐wide. Our results revealed that endemism and self‐compatibility contribute to the global association between pollen limitation and species richness. Self‐incompatible species were more pollen limited than self‐compatible ones, and the PICs analysis indicated that transitions to endemism were associated with transitions to self‐compatibility. The relationship between pollen limitation and species richness was significant only for the self‐incompatible species, and was monotonically increasing in non‐endemic species but accelerating in the endemic species. Thus, self‐incompatible endemic species from biodiversity hotspots are at the greatest risk of pollination failure, a previously unknown aspect suggesting this group of species as a top priority for future development of conservation strategies. In contrast, reproduction of self‐compatible species appears to be unrelated to plant diversity, although we caution that current data do not account for the reproductive limitation due to the quality of pollen received. Understanding the mechanisms underlying these patterns requires further investigation into plant–plant pollinator mediated interactions and the dynamics of pollen transfer in communities differing in species diversity.     

Are growth rates of Escherichia coli in batch cultures limited by respiration?

Batch cultures of Escherichia coli were grown in minimal media supplemented with various carbon sources which supported growth at specific growth rates from 0.2 to 1.3/h. The respiration rates of the cultures were measured continuously. With few exceptions, the specific rate of oxygen consumption was about 20 mmol of O2/h per g (dry weight), suggesting that the respiratory capacity was limited at this value. The adenosine triphosphate (ATP) required for the production of cell material from the different carbon sources was calculated on the basis of known ATP requirements in the biochemical pathways and routes of macromolecular synthesis. The calculated ATP requirements, together with the measured growth rates and growth yields on the different carbon sources, were used to calculate the rate of ATP synthesis by oxidative phosphorylation. This rate was closely related to the respiration rate. We suggest that aerobic growth of E. coli in batch cultures is limited by the rate of respiration and the concomitant rate of ATP generation through oxidative phosphorylation.     

Carbondioxide affects the growth rate of “oxygen limited”Escherichia coli at low concentrations of oxygen

Summary It is shown that addition of 1% CO₂ in Argon, containing traces of oxygen (<30 ppm O₂) to a described mineral salts medium leads to deblockage of the normal oxygen limited growth pattern ofEscherichia coli (ATCC 1524).     

Are invaders disturbance‐limited? Conservation of mountain grasslands in Central Argentina

Abstract. Extensive areas in the mountain grasslands of central Argentina are heavily invaded by alien species from Europe. A decrease in biodiversity and a loss of palatable species is also observed. The invasibility of the tall‐grass mountain grassland community was investigated in an experiment of factorial design. Six alien species which are widely distributed in the region were sown in plots where soil disturbance, above‐ground biomass removal by cutting and burning were used as treatments. Alien species did not establish in undisturbed plots. All three types of disturbances increased the number and cover of alien species; the effects of soil disturbance and biomass removal was cumulative. Cirsium vulgare and Oenothera erythrosepala were the most efficient alien colonizers. In conditions where disturbances did not continue the cover of aliens started to decrease in the second year, by the end of the third season, only a few adults were established. Consequently, disturbances are needed to maintain alien populations in tall‐grass mountain grasslands. Burning also increased the species richness of native species. We conclude that an efficient way to control the distribution of alien species is to decrease grazing pressure while burning as a traditional management tool may be continued.     

Is it legitimate to characterize muscle strength using a limited number of measures?

The purpose of this systematic review was to examine the legitimacy of using a single measure or a small set of measures of strength to characterize an individual's overall strength. Briefly, the methods involved: (a) a search of electronic databases, article reference lists, and personal files to identify relevant literature; and (b) a summarizing of that literature. As a result of the searches, 25 relevant articles were identified. The articles reported correlation coefficients, Cronbach's alpha, and factor analysis. Together, these statistics suggest a tendency for different strength measures to be related. A close examination of the relationships, however, suggests that caution should be exercised in characterizing overall strength using a single measure such as grip strength. In conclusion, it may be legitimate to use one or several measures obtained from a single limb to characterize the strength of that limb but not the entire body. What this means practically is that the practitioner interested in characterizing strength of a limb can reduce test burden by testing a limited number of muscle actions of that limb.     

Is the development of falciparum malaria in the human host limited by the availability of uninfected erythrocytes?

Background  

The development and propagation of malaria parasites in their vertebrate host is a complex process in which various host and parasite factors are involved. Sometimes the evolution of parasitaemia seems to be quelled by parasite load. In order to understand the typical dynamics of evolution of parasitaemia, various mathematical models have been developed. The basic premise ingrained in most models is that the availability of uninfected red blood cells (RBC) in which the parasite develops is a limiting factor in the propagation of the parasite population.     

Are plant populations seed limited? A critique and meta-analysis of seed addition experiments

We examine the relative importance of processes that underlie plant population abundance and distribution. Two opposing views dominate the field. One posits that the ability to establish at a site is determined by the availability of suitable microsites (establishment limitation), while the second asserts that recruitment is limited by the availability of seeds (seed limitation). An underlying problem is that establishment and seed limitation are typically viewed as mutually exclusive. We conducted a meta-analysis of seed addition experiments to assess the relative strength of establishment and seed limitation to seedling recruitment. We asked (1) To what degree are populations seed and establishment limited? (2) Under what conditions (e.g., habitats and life-history traits) are species more or less limited by each? (3) How can seed addition studies be better designed to enhance our understanding of plant recruitment? We found that, in keeping with previous studies, most species are seed limited. However, the effects of seed addition are typically small, and most added seeds fail to recruit to the seedling stage. As a result, establishment limitation is stronger than seed limitation. Seed limitation was greater for large-seeded species, species in disturbed microsites, and species with relatively short-lived seed banks. Most seed addition experiments cannot assess the relationship between number of seeds added and number of subsequent recruits. This shortcoming can be overcome by increasing the number and range of seed addition treatments.     

Are plant populations seed‐limited? A review of seed sowing experiments

We define seed limitation to be an increase in population size following seed addition. Here, we briefly consider how theoretical models deal with seed limitation and how seed sowing experiments can be used to unravel the extent of seed limitation in natural systems. We review two types of seed addition experiments: seed augmentation studies where seeds are added to existing populations; and seed introductions where seeds are sown in unoccupied sites. Overall, approximately 50% of seed augmentation experiments show evidence of seed limitation. These studies show that seed limitation tends to occur more commonly in early successional habitats and in early successional species. Most of the studies have concentrated on simply categorising populations as seed- or microsite-limited, but we believe that seed sowing experiments could be used to reveal much more about community structure, and we discuss possible future directions. In 53% of introduction studies (where seeds were sown at sites from which the species was known to be absent) the introduced species was recorded in at least one of the experimental sites following sowing. However, of the subset of studies where both seedlings and adult plants were recorded, 64% of sites contained seedlings while only 23% contained adults. This implies that, for many species, conditions for establishment are more stringent than conditions for germination. The successful establishment of plants in unoccupied patches indicates the potential for immigration to enhance local diversity (the spatial mass effect). Few studies continued monitoring for long enough to determine whether or not self-sustaining populations were successfully established, and no study attempted to link introduction sites to a putative natural source of propagules, or considered the dynamics of the metapopulation as a whole.     

Localization of pancreatic polypeptide cells in a limited lobe of the human neonate pancreas: Remnant of the ventral primordium?

The localization of pancreatic polypeptide (PP) cells was studied in the pancreas of four human neonates by specific immunocytochemical techniques. PP cells were detected in all parts of the pancreas. However, examination at low magnification showed that they were considerably more numerous in a small lobe, located at the posterior-inferior part of the head region. It is suggested that this lobe corresponds to the part of the pancreas that is derived from the ventral primordium. Both in the lobe rich in PP cells and in the remainder of the pancreas, approximately 75% of PP cells were present in the islets and 25% distributed among acini and ducts.     

Is growth of soil microorganisms in boreal forests limited by carbon or nitrogen availability?

To study whether the biomass of soil microorganisms in a boreal Pinus sylvestris-Vaccinium vitis-idaea forest was limited by the availability of carbon or nitrogen, we applied sucrose from sugar cane, a C₄ plant, to the organic mor-layer of the C₃–C dominated soil. We can distinguish between microbial mineralization of the added sucrose and respiration of endogenous carbon (root and microbial) by using the C₄-sucrose as a tracer, exploiting the difference in natural abundance of ¹³C between the added C₄-sucrose (¹³C –10.8) and the endogenous C₃–carbon (¹³C –26.6 ). In addition to sucrose, NH₄Cl (340 kg N ha^–1) was added factorially to the mor-layer. We followed the microbial activity for nine days after the treatments, by in situ sampling of CO₂ evolved from the soil and mass spectrometric analyses of ¹³C in the CO₂. We found that microbial biomass was limited by the availability of carbon, rather than nitrogen availability, since there was a 50% increase in soil respiration in situ between 1 h and 5 days after adding the sucrose. However, no further increase was observed unless nitrogen was also added. Analyses of the ¹³C ratios of the evolved CO₂ showed that increases in respiration observed between 1 h and 9 days after the additions could be accounted for by an increase in mineralization of the added C₄–C.     

Effect of uniconazole and limited water on growth,water relations,and mineral nutrition of “Lalandei” Pyracantha

Pyracantha (Pyracantha coccinea M. J. Roem. Lalandei) plants were treated with uniconazole at 0.5 mg ai container^–1 as a medium drench, 150 mg ai L^–1 as a foliar spray, or left untreated. Plants from all treatments were placed under three water regimes: drought acclimated, nonacclimated and later exposed to drought, or nonstressed. Acclimated plants were conditioned by seven 4-day stress cycles (water withheld), while nonacclimated were well watered prior to a single 4-day stress cycle at the same time as the seventh drought cycle of acclimated plants. Nonstressed plants were well watered throughout the study. Nonstressed plants had higher leaf water potentials and leaf conductances than acclimated and nonacclimated plants, and transpiration rates were higher in nonacclimated than acclimated plants. Uniconazole did not affect leaf water potential, leaf conductance, or transpiration rate. Acclimated plants had smaller leaf areas and leaf, stem, and root dry weights than nonacclimated or nonstressed plants. Plants drenched with uniconazole had the lowest stem and root dry weights. Acclimated plants also contained higher N concentrations than nonacclimated or nonstressed plants, and higher P concentrations than nonacclimated plants. Uniconazole medium drench treatments increased levels of Mn and P. Calcium concentration was increased in plants receiving either medium drench or foliar applications.     

Is plant performance limited by abundance of arbuscular mycorrhizal fungi? A meta-analysis of studies published between 1988 and 2003

We conducted meta-analyses of 290 published field and glasshouse trials to determine the effects of various agricultural practices on mycorrhizal colonization in nonsterile soils, and the consequence of those effects on yield, biomass, and phosphorus (P) concentration. Mycorrhizal colonization was increased most by inoculation (29% increase), followed by shortened fallow (20%) and reduced soil disturbance (7%). The effect of crop rotation depended on whether the crop was mycorrhizal. Increased colonization resulted in a yield increase in the field of 23% across all management practices. Biomass at harvest and shoot P concentration in early season were increased by inoculation (57 and 33%, respectively) and shortened fallow (55 and 24%). Reduced disturbance increased shoot P concentration by 27%, but biomass was not significantly affected. Biomass was significantly reduced in 2% of all trials in which there was a significant increase in colonization. Irrespective of management practice, an increased mycorrhizal colonization was less likely to increase biomass if either soil P or indigenous inoculum potential was high.