Extended Water-Level Drawdowns in Dammed Rivers Enhance Fish Habitat: Environmental Pool Management in the Upper Mississippi River.

Environmental Pool Management (EPM) can improve ecosystem function in rivers by restoring aspects of the natural flow regime lost to dam construction. EPM recreates summer baseflow conditions and promotes the growth of terrestrial vegetation which is inundated in the fall, thereby improving habitat heterogeneity for many aquatic taxa. A three-year experiment was conducted wherein terrestrial floodplain areas were dewatered through EPM water-level reductions and the resulting terrestrial vegetation was (1) allowed to remain or (2) removed in paired plots in Mississippi River pool 25. Fish assemblage and abundance were quantified in paired plots after inundation. Abundances of many fish species were greater in vegetated plots, especially for species that utilize vegetation during portions of their life history. Fish assemblages varied more between plot types when the magnitude of EPM water-level drawdowns was greater, which produced greater vegetation growth. Young-of-year individuals, especially from small, early maturing species and/or species reliant on vegetation for refuge, feeding, or life history, utilized vegetated plots more than devegetated plots. Vegetation growth produced under EPM was heavily used by river fishes, including young-of-year individuals, which may ultimately positively influence recruitment. Increased habitat heterogeneity may mitigate some of the negative impacts of dam construction and water-level regulation on river fishes. Annual variability in vegetation responses that occurs under EPM enhances natural environmental variability which could ultimately contribute to increased fish diversity. Low-cost programs like EPM can be implemented as a part of adaptive management plans to help maintain biodiversity and ecosystem health in anthropogenically altered rivers.

Precision shock tuning on the national ignition facility.

Ignition implosions on the National Ignition Facility [J. D. Lindl et al., Phys. Plasmas 11, 339 (2004)] are underway with the goal of compressing deuterium-tritium fuel to a sufficiently high areal density (ρR) to sustain a self-propagating burn wave required for fusion power gain greater than unity. These implosions are driven with a very carefully tailored sequence of four shock waves that must be timed to very high precision to keep the fuel entropy and adiabat low and ρR high. The first series of precision tuning experiments on the National Ignition Facility, which use optical diagnostics to directly measure the strength and timing of all four shocks inside a hohlraum-driven, cryogenic liquid-deuterium-filled capsule interior have now been performed. The results of these experiments are presented demonstrating a significant decrease in adiabat over previously untuned implosions. The impact of the improved shock timing is confirmed in related deuterium-tritium layered capsule implosions, which show the highest fuel compression (ρR~1.0 g/cm(2)) measured to date, exceeding the previous record [V. Goncharov et al., Phys. Rev. Lett. 104, 165001 (2010)] by more than a factor of 3. The experiments also clearly reveal an issue with the 4th shock velocity, which is observed to be 20% slower than predictions from numerical simulation.

In vitro uptake and processing of prezein and other maize preproteins by maize membranes.

A cell-free, mRNA-dependent system has been developed for the translation and processing of zein preproteins. A rough endoplasmic reticulum (RER)-enriched fraction, isolated by sucrose density gradients, can be treated with micrococcal nuclease to destroy endogenous messages. When these membranes are added to a wheat germ protein-synthesizing system together with zein mRNA, synthesis and processing of the polypeptides to the mature products takes place. The RER fraction from the endosperm has a different protein composition than that prepared from either the shoot or nucellar tissue and processes prezein more efficiently. The cleavage of the preproteins appears to be a cotranslational step as the completed preprotein chains cannot be processed, although they can be taken up to a limited extent. This small uptake, or absorption, or unprocessed zein seems to be an artifact and may be related to the unusual solubility properties of zein. Finally a sodium dodecyl sulfate (SDS)-urea polyacrylamide gel system has been developed which is particularly suited for the separation of low molecular weight proteins (less than 10,000 daltons). Using this method, we examined the products of in vitro zein processing and detected no presequence polypeptides. This suggests that the zein cleavage proteinase is probably an exopeptidase.

Three mutations in Zea mays affecting zein accumulation: a comparison of zein polypeptides, in vitro synthesis and processing, mRNA levels, and genomic organization.

We studied three mutations, opaque-2 (o2), opaque-7 (o7), and floury-2(fI2), each of which causes a depression in zein synthesis. We examined the processing efficiencies of the rough endoplasmic reticulum membranes in vitro, the levels of RNA transcription using cloned zein probes, and the genomic organization of the zein sequences as possible sites for the genetic defects. The results obtained indicate that the steps in prezein translation and processing occurring on the protein body membranes are not accountable for the lowered zein content in any of the mutations. The o2 mutation that typically shows a paucity of 22.5-kdalton zein polypeptides was found to have a concomitant reduction in a particular subgroup of mRNAs coding for this size class. Southern analyses suggest that the o2 mutation is not the result of a large deletion of tandem-linked zein genes.

Transposon tagging and molecular analysis of the maize regulatory locus opaque-2.

Genetic analyses suggested that the opaque-2 (o2) locus in maize acts as a positive, transacting, transcriptional activator of the zein seed storage-protein genes. Because isolation of the gene is requisite to understanding the molecular details of this regulation, transposon mutagenesis with the transposable element suppressor-mutator (Spm) was carried out, and three mutable o2 alleles were obtained. One of these alleles contained an 8.3-kilobase autonomous Spm, another a 6.8-kilobase nonautonomous Spm, and the third an unidentified transposon that is unrelated to Spm. A DNA sequence flanking the autonomous Spm insertion was verified to be o2-specific and provided a probe to clone a wild-type allele. Northern blots indicated that the gene is expressed in wild-type endosperm but not in leaf tissues or in endosperms homozygous for a mutant allele of the O2 gene. A transcript was detected in endosperms homozygous for mutations at opaque-7 and floury-2, an indication that O2 expression is independent of these two other putative regulators of zein synthesis.

Recombinant inbreds for molecular mapping in maize: theoretical and practical considerations.

Molecular mapping is rapidly being extended to more species as one result of the current emphasis on genome initiatives. The choice of the population used for mapping can have important consequences on the efficiency and accessibility of the mapping information. Recombinant inbred lines offer certain advantages over other mapping populations for many species. These more or less permanent populations permit many geneticists to contribute to the mapping effort and to profit from each other's work. Recombinant inbred lines are used extensively in mouse genetics and have been used in maize to compile a detailed molecular map.

International Rice Genome Sequencing Project: the effort to completely sequence the rice genome.

The International Rice Genome Sequencing Project (IRGSP) involves researchers from ten countries who are working to completely and accurately sequence the rice genome within a short period. Sequencing uses a map-based clone-by-clone shotgun strategy; shared bacterial artificial chromosome/P1-derived artificial chromosome libraries have been constructed from Oryza sativa ssp. japonica variety 'Nipponbare'. End-sequencing, fingerprinting and marker-aided PCR screening are being used to make sequence-ready contigs. Annotated sequences are immediately released for public use and are made available with supplemental information at each IRGSP member's website. The IRGSP works to promote the development of rice and cereal genomics in addition to producing genome sequence data.

Controlling-element events at the shrunken locus in maize.

We have examined insertions of the controlling element Ds at the Shrunken locus of maize. A cDNA probe complementary to a portion of the Shrunken locus mRNA was prepared. This probe recognizes a unique sequence in maize DNA. Using lines carrying derivatives of the same short arm of chromosome 9, we have detected modifications at the nucleic acid level caused by Ds. The changes appear to be large insertions, one of which may be more than 20 kilobase pairs in length. These observations provide a basis for the isolation and molecular characterization of one of the maize controlling elements.

Changes in state of the Wx-m5 allele of maize are due to intragenic transposition of Ds.

The molecular basis for the unusual phenotype conditioned by the waxy(Wx)-m5 Ds allele has been elucidated. Unlike most Ds alleles, Wx-m5 is phenotypically wild-type in the absence of Ac. We find that the Wx-m5 gene contains a 2-kb Ds element at -470 relative to the start of Wx transcription, representing the most 5' insertion of any transposable element allele characterized to date in plants. Despite its wild type phenotype, Wx-m5 has reduced levels of Wx enzymatic activity indicating that Ds insertion influences Wx gene expression. In the presence of Ac, Wx-m5 kernels have sectors of null expression on a wild-type background and give rise to stable wx and unstable wx-m germinal derivatives. Seventeen of 20 derivatives examined are wx-m alleles and at least 15 of these appear to result from intragenic transposition of Ds from -470 to new sites within the Wx gene. Three wx-m alleles contain two Ds elements, one at -470 and a second in Wx coding sequences. Surprisingly, only 3 out of 20 derivatives are stable wx mutants and these have sustained gross rearrangements of Wx and flanking sequences. For most other maize transposable element alleles somatic sectors and germinal derivatives usually arise following element excision or deletions of element sequences. In contrast, element insertion following intragenic transposition is apparently responsible for most of the somatic sectors and germinal derivatives of Wx-m5.

Gene mapping with recombinant inbreds in maize.

Recombinant inbred lines of maize have been developed for the rapid mapping of molecular probes to chromosomal location. Two recombinant inbred families have been constructed from F2 populations of T232 X CM37 and CO159 X Tx303. A genetic map based largely on isozymes and restriction fragment length polymorphisms has been produced that covers virtually the entire maize genome. In order to map a new gene, an investigator has only to determine its allelic distribution among the recombinant inbred lines and then compare it by computer with the distributions of all previously mapped loci. The availability of the recombinant inbreds and the associated data base constitute an efficient means of mapping new molecular markers in maize.

Quantitation of intracellular Mac-1 (CD11b/CD18) pools in human neutrophils.

The adhesive glycoprotein Mac-1 (CD11b/CD18) of the CD11/CD18 complex contributes to multiple neutrophil inflammatory functions. Activation of neutrophils by chemotactic stimuli results in a rapid, protein synthesis-independent increase in surface Mac-1 derived from incompletely defined intracellular compartments. Therefore, we developed a novel quantitative lectin immunoblot technique to define intracellular pools of Mac-1 in subcellular neutrophil fractions resolved on discontinuous Percoll gradients. In cavitates of unstimulated neutrophils, 30% and 26% of total Mac-1 was identified in beta [1.10 gm/ml; vitamin B12 binding protein (vit B12 B.P.)-rich] or pre-gamma (1.07 gm/ml; vit B12 B.P.-poor) granular fractions, respectively, whereas 24% was associated with the plasma membrane-rich gamma (1.06 gm/ml) fractions. N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation (10(-8) M, 15 min, 37 degrees C) significantly diminished Mac-1 in pre-gamma (-18% of total, P less than 0.05) but not beta fractions (+6% of total). Under these conditions, the content of Mac-1 in gamma fractions increased 13% in association with four- to eightfold increase in surface Mac-1 expression (OKM-1 binding). These findings suggest that chemotactic stimuli increase plasma membrane and/or surface Mac-1 on human neutrophils by mobilizing a novel intracellular granule pool.

An evaluation of selected methods for the decontamination of cattle hides prior to skinning.

The effectiveness of different decontamination treatments in reducing microbial loads on cattle hides was assessed. The 10-s hide treatments were conducted using a wet-and-dry vacuum cleaner filled with one of the liquids (heated to 50 °C) indicated below, followed or not by 10-min drying in the air. Also, the hide was clipped, followed or not by 10-s singeing using a hand-held blowtorch. Before and after each decontamination treatment, the hide was sampled (100 cm(2) areas) by a sponge-swabbing method to compare the total viable counts of bacteria (TVC). The largest bacterial reduction (P<0.001; 2.31log(10) cfu/cm(2)) was achieved by singeing of previously clipped hide. Treatment of hide with a food industry sanitizer solution (10% Betane Plus) resulted in significant reductions of 1.80 (P<0.001) and 1.98log(10) cfu/cm(2) (P<0.001) without and with subsequent drying, respectively. Treatment of hide with a food industry disinfectant (P3-Topactive DES) significantly reduced TVC by 0.97 (P<0.001) and 1.18log(10) cfu/cm(2) (P<0.001) without and with subsequent drying, respectively. Treatments of hide with water alone or with a food-safe detergent solution (Formula 963B), or hide clipping alone, did not produce significant decontamination effects. Since hide contamination is associated with microbial contamination of the carcasses, the results indicate that post-killing/pre-skinning hide decontamination (used alone, or in combination with carcass decontamination) has a potential to improve microbial meat safety. Nevertheless, further research is required to optimise the efficacy of these treatments in the reduction of specific pathogens under commercial conditions.

Commonly used antibacterial and antifungal agents for hospitalised paediatric patients: implications for therapy with an emphasis on clinical pharmacokinetics.

Due to normal growth and development, hospitalised paediatric patients with infection require unique consideration of immune function and drug disposition. Specifically, antibacterial and antifungal pharmacokinetics are influenced by volume of distribution, drug binding and elimination, which are a reflection of changing extracellular fluid volume, quantity and quality of plasma proteins, and renal and hepatic function. However, there is a paucity of data in paediatric patients addressing these issues and many empiric treatment practices are based on adult data. The penicillins and cephalosporins continue to be a mainstay of therapy because of their broad spectrum of activity, clinical efficacy and favourable tolerability profile. These antibacterials rapidly reach peak serum concentrations and readily diffuse into body tissues. Good penetration into the cerebrospinal fluid (CSF) has made the third-generation cephalosporins the agents of choice for the treatment of bacterial meningitis. These drugs are excreted primarily by the kidney. The carbapenems are broad-spectrum beta-lactam antibacterials which can potentially replace combination regimens. Vancomycin is a glycopeptide antibacterial with gram-positive activity useful for the treatment of resistant infections, or for those patients allergic to penicillins and cephalosporins. Volume of distribution is affected by age, gender, and bodyweight. It diffuses well across serous membranes and inflamed meninges. Vancomycin is excreted by the kidneys and is not removed by dialysis. The aminoglycosides continue to serve a useful role in the treatment of gram-negative, enterococcal and mycobacterial infections. Their volume of distribution approximates extracellular space. These drugs are also excreted renally and are removed by haemodialysis. Passage across the blood-brain barrier is poor, even in the face of meningeal inflammation. Low pH found in abscess conditions impairs function. Toxicity needs to be considered. Macrolide antibacterials are frequently used in the treatment of respiratory infections. Parenteral erythromycin can cause phlebitis, which limits its use. Parenteral azithromycin is better tolerated but paediatric pharmacokinetic data are lacking. Clindamycin is frequently used when anaerobic infections are suspected. Good oral absorption makes it a good choice for step-down therapy in intra-abdominal and skeletal infections. The use of quinolones in paediatrics has been restricted and most information available is in cystic fibrosis patients. High oral bioavailability is also important for step-down therapy. Amphotericin B has been the cornerstone of antifungal treatment in hospitalised patients. Its metabolism is poorly understood. The half-life increases with time and can be as long as 15 days after prolonged therapy. Oral absorption is poor. The azole antifungals are being used increasingly. Fluconazole is well tolerated, with high bioavailability and good penetration into the CSF. Itraconazole has greater activity against aspergillus, blastomycosis, histoplasmosis and sporotrichosis, although it's pharmacological and toxicity profiles are not as favourable.

Lime pretreatment of switchgrass.

Lime (calcium hydroxide) was used as a pretreatment agent to enhance the enzymatic digestibility of switchgrass. After studying many conditions, the recommended pretreatment conditions are: time = 2 h, temperature = 100 degrees C and 120 degrees C, lime loading = 0.1 g Ca(OH)(2)/g dry biomass, water loading = 9 mL/g dry biomass. Studies on the effect of particle size indicate that there was little benefit of grinding below 20 mesh; even coarse particles (4-10 mesh) digested well. Using the recommended pretreatment conditions, the 3-d reducing sugar yield was five times that of untreated switchgrass, the 3-d total sugar (glucose + xylose) yield was seven times, the 3-d glucose yield was five times, and the 3-d xylose yield was 21 times. A material balance study showed that little glucan (approx 10%) was solubilized as a result of the lime pretreatment, whereas about 26% of xylan and 29% of lignin became solubilized.

The first-year family practice resident. An identity crisis.

A definite identity crisis was recognized among first-year family practice residents prior to this year. A specific solution was found through a carefully planned one month family practice rotation that enabled the residents to become fully acquainted with the Family Practice Clinic operation while firmly establishing their identity as unique family practice residents. There is good evidence that this approach has been highly successful in dealing with the problem, and the alleviation of identity crises in first-year residents has, in addition, strengthened other areas of the residency program.

The tired patient.

One of the most challenging diagnostic problems that comes into the physician's office is the patient who complains of "being tired." It can prove to be a frustrating experience for the physician (especially if the physician is also tired) and to the patient. This article will propose and describe a two-pronged systematic approach to the patient that can increase the probability of a satisfying outcome for both physician and patient. This approach assumes the "complaint" of every patient has both informational value ("I think there's something wrong.") and transactional meaning (Please take care of me."). Tiredness, fatigue, "exhaustion" need to be seen simultaneously as information and need. Both aspects require simultaneous work-up.