Temporal variation and characterization of grunt sounds produced by Atlantic cod Gadus morhua and pollack Pollachius pollachius during the spawning season.

Fine-scale temporal patterning in grunt production and variation in grunt attributes in Atlantic cod Gadus morhua and pollack Pollachius pollachius was examined. Pollachius pollachius produced only a single sound type, the grunt, similar to that previously described for G. morhua. Sound production and egg production were correlated in P. pollachius but not in G. morhua. Only G. morhua displayed a strongly cyclical pattern, producing more grunts at night. Finer-scale temporal patterning in grunt production was observed in both species which produced significantly fewer grunts following a period of high grunt production. These quieter periods lasted up to 45 min for P. pollachius and up to 1 h in G. morhua. Grunts were not always produced in isolation but organized into bouts in both species. Longer bouts were more frequent during periods of increased sound activity and were linked with changes in grunt characteristics including increased grunt duration, pulse duration and repetition period of each pulse combined with decreased dominant frequency. This study provides the first evidence of acoustic signalling being used by spawning P. pollachius and presents the most detailed analysis of the complexity of gadoid sound production.

Investigating the prevalence of low energy availability, disordered eating and eating disorders in competitive and recreational female endurance runners.

Eating disorders (ED), disordered eating (DE) and low energy availability (LEA) can be detrimental to health and performance. Previous studies have independently investigated the prevalence of ED, DE or LEA; however, limited studies have combined methods identifying risk within female runners. The aim of this study was to identify the prevalence of ED, DE and LEA in United Kingdom-based female runners and associations between age, competition level and running distance. The Female Athlete Screening Tool (FAST) and Low Energy Availability in Females Questionnaire (LEAF-Q) were used in a cross-sectional study design. A total of n = 524 responses eligible for analysis were received. A total of n = 248 (47.3%), n = 209 (40%) and n = 49 (9.4%) athletes were at risk of LEA, DE and ED, respectively. LEAF-Q scores differed based upon age (Age: H(3) = 23.998, p ≤ .05) and competitive level (Comp: H(1) = 7.682, p ≤ .05) whereas FAST scores differed based on age (Age: F(3,523) = 4.753, p ≤ .05). Tukey's post-hoc tests showed significantly higher FAST scores in 18-24 years compared to all other age categories (p ≤ .05). Stepwise multiple regression demonstrated age and competitive level modestly predicted LEAF-Q scores (R2adj = 0.047, F(2,523) = 13.993, p ≤ .05, VIF = 1.0) whereas age modestly predicted FAST scores (R2adj = 0.022, F(1,523) = 12.711, p ≤ .05, VIF = 1.0). These findings suggest early identification, suitable screening methods and educational intervention programmes should be aimed at all levels of female endurance runners.Highlights A total of 524 female endurance completed a self-administered, online questionnaire screening for low energy availability, disordered eating and eating disorders risk.Age and competitive level modestly predicted low energy availability and age modestly predicted disordered eating and eating disorders in female endurance runners.A higher percentage of 18- to 24-year-old female endurance runners were at greater risk of low energy availability, disordered eating and eating disorders compared to other age categories.These findings highlight the need for regular screening in order to aid early interventions to prevent potential decrements in performance and health as endurance runners mature.

Undamaged cotton plants yield more if their neighbour is damaged: implications for pest management.

Understanding the compensatory responses of crops to pest damage is important in developing pest thresholds. Compensation for pest damage in crops can occur at the plant level, where the architecture, growth dynamics and allocation patterns of damaged plants are altered, allowing them to recover or, at the crop level, where differential damage between plants may alter plant-to-plant interactions. We investigated growth and yield of cotton (Gossypium hirsutum L.) following non-uniform manual defoliation of seedlings. This partially replicates real pest damage and is valuable in understanding crop-level responses to damage because it can be inflicted precisely. Damage distributions included damaging 0, 25, 50, 75 or 100% of the plants. Damage intensity for the damaged plants was varied by removing 100 or 75% of each true leaf when plants had two, four and six true leaves. At the crop level, yield loss increased as the proportion of plants damaged and intensity of damage per damaged plant increased. Neighbour interactions occurred; undamaged plants with damaged neighbours grew larger and yielded better than undamaged plants with undamaged neighbours, while the converse applied for damaged plants with undamaged neighbours. Neighbour interactions were influenced by the intensity of damage and were stronger when 100% of the leaf area was removed than when 75% was removed. At the crop level, when compared with yield estimates based on yield of plants from uniformly damaged or undamaged plots, these interactions resulted in higher yield than expected (+8%). This suggests that damage distribution may have to be considered in studies where artificial or real pest damage is inflicted uniformly on plants.

Low cost polymer intramedullary nails for fracture fixation: a biomechanical study in a porcine femur model.

INTRODUCTION: Whilst intramedullary nailing is a commonly accepted technique for lower limb fracture fixation, the cost of nails can be prohibitive in hospitals in developing nations. In these institutions bone cement has found many off label applications, that whilst are effective do not meet manufacturers guidelines. The aim of this study was to examine the biomechanics of one such application, fracture fixation using a bone cement intramedullary nail. MATERIALS AND METHODS: Five porcine femurs underwent a mid-shaft osteotomy and were fixed using a nail made from antibiotic simplex bone cement. The torsional and flexural stiffness and shear modulus of these constructs were compared to five intact porcine femurs. RESULTS: The bone cement intramedullary nail was able to achieve relative stability in both torsion, with a mean shear modulus of 0.17 GPa and in flexion with a mean flexural stiffness of 358 N/mm. This corresponds to 47 and 22% of the respective measurements in the intact femurs. The mean ultimate flexural strength of fracture/nail constructs was 936 +/- 350 N, which is 20% of the ultimate flexural strength of an intact porcine femur (4,820 +/- 698 N). CONCLUSION: Intramedullary nails made from bone cement were able to provide sufficient promise in this situation to warrant further investigation for their applicability as a low cost alternative for use in developing countries.

Superparamagnetic gadonanotubes are high-performance MRI contrast agents.

We report the nanoscale loading and confinement of aquated Gd3+n-ion clusters within ultra-short single-walled carbon nanotubes (US-tubes); these Gd3+n@US-tube species are linear superparamagnetic molecular magnets with Magnetic Resonance Imaging (MRI) efficacies 40 to 90 times larger than any Gd3+-based contrast agent (CA) in current clinical use.

Transformation of Aspergillus nidulans with the hygromycin-resistance gene, hph.

Aspergillus nidulans strain G191 was transformed to hygromycin resistance using plasmid pDH25, which contains the bacterial hygromycin B phosphotransferase gene (hph) fused to promoter elements of the A. nidulans trpC gene. Southern hybridizations of transformants revealed multiple, integrated copies of the vector. A pleiotropic effect conferring increased hygromycin B sensitivity was found to be associated with the A. nidulans pyrG89 allele. Plasmid pDH25 features a ClaI site immediately preceding the hph start codon thus permitting convenient replacement of the trpC sequences with other eukaryotic promoters.

Primary structure of Mucor miehei aspartyl protease: evidence for a zymogen intermediate.

The gene encoding the aspartyl protease of the filamentous fungus Mucor miehei has been cloned in Escherichia coli and the DNA sequenced. The deduced primary translation product contains an N-terminal region of 69 amino acid (aa) residues not present in the mature protein. By analogy to the evolutionarily related mammalian gastric aspartyl proteases it is inferred that the primary secreted product is a zymogen containing a 47-aa propeptide. This propeptide is presumably removed in the later steps of the secretion process or upon secretion into the medium. To study the effects of modifications of the protease structure on its maturation by enzyme-engineering methods, an efficient expression system was sought. In E. coli, transcription of the preproenzyme coding sequence from a bacterial promoter results primarily in the accumulation of unsecreted, enzymatically inactive polypeptides, immunologically related to the authentic protease. In Aspergillus nidulans expression of the cloned gene, probably from its own promoter, results in the secretion into the culture medium of polypeptides which, compared to the authentic protease, are similar in specific activity, but differ in the character of their asparagine-linked oligosaccharides.

Molecular cloning and deletion of the gene encoding aspergillopepsin A from Aspergillus awamori.

We have cloned genomic pepA sequences encoding the aspartic proteinase aspergillopepsin A (PEPA) from Aspergillus awamori using a synthetic oligodeoxyribonucleotide probe. Nucleotide sequence data from the pepA gene revealed that it is composed of four exons of 320, 278, 249, and 338 bp. Three introns which interrupt the coding sequence are 51, 52, and 59 bp in length. Directly downstream from the putative start codon lies a sequence encoding 69 amino acids (aa) which are not present in mature PEPA. Based on similarities to other aspartic proteinases, this region may represent a 20-aa signal peptide followed by a 49-aa propeptide that is rich in basic aa residues. Northern blots of total cellular RNA extracted from A. awamori cells indicate that pepA is transcribed as a single 1.4-kb mRNA. Mutants of A. awamori lacking the pepA structural gene were derived by the following gene replacement strategy. First, we constructed a plasmid in which a 2.4-kb SalI fragment containing the entire pepA coding region was deleted from a 9-kb Eco RI genomic DNA clone and replaced by a synthetic DNA polylinker. Second, a selectable argB gene was inserted into the polylinker. Third, the EcoRI fragment which contained the argB marker flanked by pepA sequences was excised from the plasmid and used to transform an argB auxotroph of A. awamori. From 16-40% of the resulting prototrophic transformants were found to have a PEPA-deficient phenotype when screened with an immunoassay using antibodies specific for PEPA. Southern hybridization experiments confirmed that these mutants resulted from a gene replacement event at the pepA locus.

Replacement of an inherited stretch receptor by a newly evolved stretch receptor in hippid sand crabs.

Primary sensory neurons that are motoneuron-like in morphology and often nonspiking (transmit afferent signals as graded depolarizations) characterize an unusual type of stretch receptor in decapod crustaceans. Nonspiking and spiking receptors occur in similar positions at homologous joints in different species and have been presumed to be homologous, the spiking one considered "primitive". To better understand the evolutionary origin of these stretch receptors and why some are nonspiking, we examined the spiking telson-uropod stretch receptors in the spiny sand crab Blepharipoda occidentalis (Albuneidae) and the squat lobster Munida quadrispina (Galatheidae) and compared them with the nonspking telson-uropod stretch receptor of the mole sand crab Emerita analoga (Hippidae). The position, morphology and responses to stretch of the sensory neurons, and the ultrastructure of the elastic strand portion of the receptor are similar in M. quadrispina and B. occidentalis, except that in B. occidentalis the receptor muscles are substantially smaller and the extracellular matrix of the elastic receptor strand is both more extensive and more organized, reminiscent of the ultrastructure of E. analoga's nonspiking receptor. We conclude that the spiking telson-uropod stretch receptors of albuneids and galatheids are homologous. The differences in the ultrastructure of their receptor strands imply that the efficiency of coupling receptor length change to deformation of the dendritic termini increases in the order M. quadrispina < B. occidentalis < E. analoga. The spiking and nonspiking telson-uropod stretch receptors differ anatomically in three major respects that appear to preclude their homology. (1) The receptor strands are on opposite sides of a conserved muscle. (2) The sensory somata are in different regions of the sixth abdominal ganglion: a lateral cluster of somata for the spiking sensory neurons and two medial clusters, one anterior, one posterior, for the nonspiking sensory neurons. (3) The neuropil projections of the sensory neurons are different. We conclude that the hippid's nonspiking telson-uropod stretch receptor evolved de novo and not by modification of the ancestral anomuran telson-uropod stretch receptor (which Hippidae have lost).

Functional morphology of the telson-uropod stretch receptor in the sand crab Emerita analoga.

The telson-uropod stretch receptor in Emerita analoga belongs to a segmental array of axial-coxal receptors having sensory neurons with central somata. It consists of an elastic strand innervated by four giant, nonspiking, mechanoreceptive neurons. We examined the elastic (sensory) strand by light and electron microscopy to discover whether 1) functional differences between the four sensory neurons could be surmised from their peripheral terminations and 2) the process of mechano-electric transduction in this type of receptor might be revealed by the ultrastructure of the dendritic terminals. The elastic strand is divided into nonoverlapping domains by the mechanoreceptive neurons. We discerned no differences other than their serial order along the strand. Each neuron has three morphological zones in the periphery. In the zone of dendrite entry, the dendrite bifurcates into 40-60 microns primary branches that enter the strand at acute to right angles and turn rostrally and caudally along the strand's axis. The zone of branching consists of 4-35 microns diameter dendritic branches within the strand. Approximately 21,000 dendritic tips (diameters, 0.1-0.7 micron; lengths, 3-20 microns) per sensory neuron arise via short stubs from all levels of the zone of branching and constitute the zone of dendrite termination. The repeated bifurcations of dendrites within the zone of branching take place by radial intrusion of longitudinally oriented, partitions of extracellular matrix each of which extends the length of the pair of dendritic branches it divides. Tips project into these partitions. Since they change shape when the elastic strand is stretched, which compresses the extracellular matrix of the partitions, they are probably the site of mechanosensory transduction. Cross-sectional profiles of dendritic tips are significantly larger but less numerous in stretched than in relaxed receptors. We propose a model in which stretch compresses distal portions of tips until they are too small to be recognized. The large profiles are the proximal portions of tips that have been expanded by hydrostatic pressure. The very large number of dendritic tips/nonspiking mechanoreceptor may be what endows each neuron with 1) high sensitivity to stretch of the elastic strand and 2) ability to respond to a broader range of tensions than is usual for single sensory cells (the entire range experienced by the stretch receptor).

Medically safe rate of weight loss for the treatment of obesity: a guideline based on risk of gallstone formation.

OBJECTIVE: Currently recommended rates of weight loss for obese persons are not based on demonstrated health risks. The authors attempt to determine a medically safe rate of weight loss based on currently available data on the risk of gallstone formation during active weight loss. METHODS: All prospective studies published in the English language on the formation of gallstones in obese persons during active weight loss were reviewed and evaluated. A statistical analysis was performed using studies of at least 4 weeks' duration that included information on diet composition, rates of weight loss, length of follow-up, and incidence of gallstone formation. A statistical model was fit to the data from the studies that met the criteria. RESULTS: Nine subject groups derived from five published reports met the criteria for inclusion in the analysis. When the results of these groups were evaluated statistically, a curvilinear relationship between incidence of gallstone formation and rate of weight loss was demonstrated. The relationship was best described by an exponential regression curve, with an adjusted r2 of 0.98 and a dramatically increasing risk of gallstone formation at rates of weight loss above 1.5 kg per week. CONCLUSION: Risk of gallstone formation in obese persons during active weight loss seems to increase in an exponential fashion. The data suggest that rates of weight loss should not exceed an average of 1.5 kg per week.

Traceless solid-phase synthesis of 2,4-diaminoquinazolines.

[reaction: see text] The solid-phase synthesis of 2,4-diaminoquinazolines is presented. The chemistry involves the sequential condensation of 2-aminobenzonitriles and amines starting from an acyl isothiocyanate resin via a traceless cleavage and cyclization. The alpha-1 antagonist prazosin was synthesized, as well as several other examples, in good yields and purity.

Identification of donor melanoma in a renal transplant recipient.

A patient with chronic renal failure received a closely matched cadaveric kidney. Approximately 3 months after transplantation, the patient developed a metastatic malignant melanoma. A large retroperitoneal mass consisting of large pleomorphic polygonal neoplastic cells was found close to the donated kidney. This tumor was diagnosed as a malignant melanoma. DNA analysis of this tumor, the donated kidney, and the recipient indicated that the melanoma originated from the donor. Although this is not the first report of a donated melanoma, it is the first report of definitive DNA analysis of the origin of the malignant cells.

An epidemiologic study of fatigue with relevance for the chronic fatigue syndrome.

We surveyed households in four rural Michigan communities to confirm a reported cluster of cases resembling chronic fatigue syndrome (CFS) and to study the epidemiology of fatigue in a rural area. Data were collected from 1698 households. We did not confirm the reported cluster. The prevalence of households containing at least one fatigued person was similar between communities thought to harbor the cluster and communities selected for comparison. Symptoms and features of generic forms of fatigue were very similar to those often attributed to CFS.

Acrylamide and iodide fluorescence quenching as a structural probe of tryptophan microenvironment in bovine lens crystallins.

Fluorescence quenching using acrylamide and iodide quenchers has been used to investigate the microenvironments of tryptophan residues in bovine alpha-, beta-, and gamma-crystallin fractions. Acrylamide quenching is very sensitive to the degree of tryptophan accessibility to the solvent containing the acrylamide. Since acrylamide is able to diffuse into the interior of the protein, accessibility to acrylamide may result from Trp residues lying at the surface of the protein or from the existence of channels leading to the interior of the protein. Iodide ion is hydrated and is limited by its large size and charge to quenching of tryptophan residues lying at or near the surface of proteins. Tryptophan residues in the lens crystallin fractions were found to be highly accessible to acrylamide, yet the rate of quenching by acrylamide was very low, indicating that the tryptophan residues of the lens crystallin fractions occupy predominately hydrophobic environments. The high accessibility to acrylamide likely results from diffusion of acrylamide into the interior of the protein. Accessibility to iodide was much lower, as was the rate of quenching by iodide, adding support to the conclusions from acrylamide quenching.

The hagfish oocyte at late stages of oogenesis: structural and metabolic events at the micropylar region.

The hagfishes (Class Agnatha), primitive vertebrates and massive secretors of mucus, occupy a unique niche in a marine benthic environment and exists in abundance despite an infrequent ovulation of a low number (12-45) of notably large, prolate ellipsoidal (approximately 8 mm x 28 mm, width x length, respectively) eggs. To establish factors that might contribute to survivability of one species (Eptatretus stouti), we examined three groups of mature females and document structural events characteristic of oocytes at late developmental stages: attached to the gonad, after ovulation into the body cavity, and after deposition on aquaria substrata. Quantitative autoradiographic analyses following administration in vivo of [(3)H]-leucine was useful to confirm post-vitellogenic stages and late metabolic events at the micropylar region located at one end of each egg. After egg disassembly, a combination of light microscopy and scanning electron microscopy was used to delineate structural features and 3-D relationships in juxtaposition to the micropylar cup and canal. A thick (approximately 200 microm) complex tripartite chorion surrounds a large micropylar cup (approximately 300 microm, width at base) consisting of fused polygonal (mainly hexagonal) substructural units (3-4 microm, width) and collectively serves to stabilize the micropylar canal (of size to prevent polyspermy) that penetrates the ooplasm near the germinal vesicle. A tuft composed of numerous chorionic appendages ('anchor filaments', approximately 2-3 mm length) is enmeshed in a gel matrix and surrounds the cup. When the eggs are deposited, intermediate filament aggregates (IFA) from slime gland mucus become interspersed in the tuft/gel complex. A similar IFA/tuft/gel region found on the vegetal pole-end facilitates firm egg-end attachments and possibly assists localization of egg clutches to a favorable substratum. Although the mode of fertilization is as yet unknown, the collective structural characteristics of the hagfish egg arc indicative of physical strength and are markedly different from eggs of the other group of cyclostomes (lampreys) and from teleosts. The hagfish produces fewer but larger eggs which are prolate ellipsoidal rather than spherical and have a thicker chorion, a greater chorion to egg width ratio, a much larger micropylar cup with a unique substructure, larger and more localized chorionic appendages, and appendage tufts interspersed with adhesive substances derived not only from the follicular epithelium but also from the slime glands.

Modelling the spatial distribution of ammonia emissions from seabirds in the UK.

Knowledge of the sources and distribution of ammonia (NH3) emissions underpins our understanding of the nitrogen budget. Research has focused on quantifying NH3 emissions from anthropogenic sources, whilst those from natural sources have received little attention internationally. Seabirds excrete large quantities of nitrogen, making seabird colonies a major natural source of NH3. Ammonia emissions from each UK seabird species were estimated and combined with population distribution data to model their spatial distribution. Total NH3 emissions from UK seabirds were estimated at 2.7 kt per year. Seabird emissions are concentrated in remote parts of the UK where anthropogenic emissions are small, so that seabirds often represent the main source of NH3 emissions in these areas. Seabird NH3 emissions were found to have increased by 34% since the 1970s. This corresponds to population changes which may be influenced by human activities, showing that even this natural source can be anthropogenically modified.

Faraday magnetic balance capable of high resolution measurements on metalloprotein solutions from 6.5 to 300 K.

A Faraday balance, constructed from commercially available components, is reported which is convenient to operate and capable of high resolution measurements on metalloprotein solutions over a 6.5-300 K temperature range. The measurements are as accurately determined as for other, more expensive devices which usually require superconducting magnetic equipment. As an example, the temperature dependence of the magnetic susceptibility of metmyoglobin fluoride is reported and compared to other available data.

Ten central elements of family practice.

Previous studies of the content of family practice have analyzed the discipline in terms of the clinical problem content. Taking a different approach, a study group analyzed the care given to patients by family physicians irrespective of the specific clinical problems. Working with a reference group of family physicians in private practice, ten central elements were identified: (1) comprehensiveness of care, (2) anticipation of problems and continuity of care, (3) personal relationships with a patient, (4) medical knowledge and skills characteristic of family medicine, (5) values and attitudes that enhance family medicine, (6) problem definition and medical decision making, (7) problem management and resource coordination, (8) care of the individual within the family context, (9) involvement with the community, and (10) attentiveness to practice organization. This study provides a different point of departure for the design and evaluation of educational programs in family practice.

Improved production of chymosin in Aspergillus by expression as a glucoamylase-chymosin fusion.

We have extended the work on chymosin production in Aspergillus by constructing an expression vector in which the cDNA encoding bovine prochymosin B was fused in frame immediately following the codon for the last amino acid of the A. awamori glucoamylase (glaA) gene. Transformation of A. awamori with this plasmid led to the secretion of considerably higher amounts of chymosin than obtained with previous chymosin expression vectors. We present evidence that mature chymosin is autocatalytically released from the glucoamylase-chymosin fusion protein after secretion.