Modelling catchment management impact on in-stream phosphorus loads in northern Victoria.

Phosphorus pollution severely impairs the water quality of rivers in Australia and worldwide. Conceptual models have proved useful to assess management impact on phosphorus loads, particularly in data-sparse environments. This paper develops and evaluates the coupling of a point-scale model (HowLeaky2008) to a catchment scale model (CatchMODS) to enhance modelling of farm management impacts on in-stream phosphorus loads. The model was tested in two adjacent catchments in northern Victoria (Avon-Richardson and Avoca), Australia. After calibration of the in-stream attenuation parameter against measurements at gauging stations, the model simulated specific annual phosphorus loads across the catchments well (Nash-Sutcliffe model efficiency of 0.52 in the Avon-Richardson and 0.83 for the Avoca catchment). Phosphorus loads at both catchment outlets under current conditions were estimated at 7 t y(-1) and were dominated by field exports. Changes to farm management practices, i.e. the use of perennial pastures in grazing systems and zero-tillage in cropping systems were estimated to reduce phosphorus load by 31% in the Avon-Richardson catchment and 19% in the Avoca catchment, relative to current practices (annual pasture and minimum tillage). The model afforded a major improvement in conceptual modelling by explicit simulation of the impacts of soil and climatic conditions on field-scale exports and by placing them in the context of landscape processes.

PCR and molecular detection for differentiating Vibrio species.

Vibriosis is an economically important disease of fish, marine invertebrates (particularly penaeid shrimps), and large marine mammals and is responsible for high mortality rates in aquaculture worldwide. Some Vibrio species are also responsible for zoonoses, whereas others are relatively nonpathogenic. Using 16S- and 23S-based PCR reactions, we obtained species-specific patterns and a 470-bp band, respectively. DNA sequences obtained on the 23S rRNA gene allowed us to identify species-specific probes for Vibrio parahaemolyticus, V. alginolyticus, V. anguillarum and for a cluster of taxonomically related species: V. carchariae/harveyi/campbelii. A phylogenetic tree based on the 23S sequences confirmed previous results obtained by Western blotting.

Weight gain and quality of life in women treated with adjuvant chemotherapy for early-stage breast cancer.

PURPOSE/OBJECTIVES: To document weight gain in women treated with adjuvant chemotherapy for early-stage breast cancer and to examine the relationship of weight gain and perceived quality of life (QOL). DESIGN: Descriptive, correlational study. SETTING: Data collected in three settings: an ambulatory oncology service of a university teaching hospital, a private oncology office, and a university-affiliated health clinic, all located in southern New England. SAMPLE: Women with stage I or II breast cancer with primary treatment of simple or modified mastectomy or breast-conserving surgery with radiotherapy scheduled to receive adjuvant chemotherapy. METHODS: Weight data collected through retrospective chart review. QOL data collected prospectively using the Linear Analog Self Assessment Symptom Distress Scale for Breast Cancer and the Functional Assessment of Cancer Therapy for Breast Cancer Scale. MAIN RESEARCH VARIABLES: QOL and weight gain of five pounds or more. FINDINGS: One year after treatment began, 62.5% of the study participants experienced weight gain (X = 10.44 lb), with a range of 5-27 pounds. After two and three years, 68% and 40%, respectively, maintained a clinically significant weight gain. A greater weight gain occurred over time in premenopausal women. No correlation between overall QOL and weight gain existed, but selected items were significantly positively correlated with weight gain. CONCLUSIONS: This study documented a significant weight gain in women treated with adjuvant chemotherapy for early-stage breast cancer. A large percentage of those women maintained this weight gain. Women premenopausal at diagnosis had a greater tendency to gain weight. Although weight gain was not correlated with overall QOL it was distressing for these women. IMPLICATIONS FOR NURSING PRACTICE: Nurses can incorporate the possibility of weight gain into the plan of care for women with breast cancer. Nurses should include this information in education about side effects of treatment and in the ongoing nursing assessment of patients with breast cancer.

Sequence and comparative structural analysis of the murine leukaemia virus amphotropic strain 4070A RNase H domain.

The sequence of a 900-nucleotide segment (encoding part of the reverse transcriptase, including the entire RNase H domain) of the pol gene of the murine leukaemia virus (MLV) amphotropic strain 4070A is presented. Alignment of the inferred 4070A RNase H amino acid sequence (157 residues) with other MLV RNase H sequences revealed only minor differences compared with the divergence between other retroviral and prokaryotic or eukaryotic RNase H sequences. Only 10 residues were invariant across the entire sample set, but secondary structure predictions for the enzymes from E. coli, yeast, human liver and diverse retroviruses (HIV, Rous sarcoma virus, foamy viruses) supported, in every case, the five beta-strands (1 to 5) and four or five alpha-helices (A, B/C, D, E) that have been identified by crystallography in the RNase H domain of HIV-1 reverse transcriptase and in E. coli RNase H. In the case of MLV, analysis of the RNase H domain sequences inferred from 10 different strains (including the amphotropic 4070A) predicted all five alpha-helices (A-E), as well as beta-strands 4 and 5. However, the N-terminal segment (residues 1-40) was predicted, without exception and with high probability, to fold uniquely into one (or two adjacent) alpha-helix(es) encompassing residues 13-37, instead of the three beta-strands known to exist in the HIV-1 and E. coli enzymes. The unerring consistency between the known and predicted structures of the HIV-1 and E. coli enzymes, and the prediction of the same structural elements (including beta-strands 1-3 within the N-terminal segment) for all other (non-MLV) RNase H proteins examined in this study, suggests that the N-terminal segment of the MLV RNase H domain assumes a conformation distinct from that of other retroviral and cellular RNase H molecules. An additional (sixth) beta-strand was also predicted uniquely within the C-terminal region of foamy virus RNase H domains.

De novo reverse transcription of HTLV-1 following cell-to-cell transmission of infection.

Analogous to transmission of human T-cell leukemia virus type 1 (HTLV-1) in vivo, an in vitro cell-to-cell infection model was established by coculturing MT-2 cells as virus donors and HUT78 cells as recipients. At a donor:recipient ratio of 1:2, cell fusion occurred and a new round of HTLV-1 genome replication was initiated in the cocultured cells. Newly synthesized unintegrated viral DNA was detected by Southern blot within 4-8 h and then increased between 8 and 48 h following cell mixing. The most dominant species of unintegrated viral DNA was 3.7 kb in size which hybridized to a full-length HTLV-1 DNA probe but not to a Kpnl viral DNA fragment that is absent from a defective proviral genome that has been previously identified in MT-2 cells. Northern blot analysis showed large amounts of viral RNA in the virus donor cells and in the cocultured cells, with a 3.4-kb species being the most abundant. This 3.4-kb RNA gave a pattern identical to that of the 3.7-kb unintegrated viral DNA in hybridization studies using the two probes. It seems likely that the unspliced RNA transcript from the defective proviral genome in MT-2 cells was effectively reverse transcribed upon initiation of cell-to-cell viral transmission to susceptible HUT78 cells. Despite active de novo reverse transcription, however, viral RNA levels remained unchanged following cell-to-cell transmission of HTLV-1 infection and no viral antigen production could be attributed to the newly initiated round of viral genome replication. As an abortive infection model this simple cell-to-cell infection system warrants more detailed study as it has the potential to provide reliable information regarding the early events in HTLV-1 transmission and infection.

Nucleotide sequence of the murine leukaemia virus amphotropic strain 4070A integrase (IN) coding region and comparative structural analysis of the inferred polypeptide.

The complete nucleotide sequence of the integrase (IN) protein coding region of the murine leukaemia virus (MLV) amphotropic strain 4070A is presented. The sequence comprises 1,224 nucleotides, encoding a 408-residue polypeptide of M(r) 46,312. Alignment of the inferred 4070A IN amino acid sequence with the IN proteins of other MLV showed that substitutions are confined largely to segments within the N- and C-terminal domains. In the N-terminal domain the majority of substitutions occur as contiguous 2- to 6-residue blocks, whereas in the C-terminal domain they occur as isolated entities except within a short segment characterized by deletions/insertions. Selection appears to act on the C-terminal 19 residues of IN rather than on the N-terminal residues of ENV (encoded by overlapping reading frames), suggesting a functional role for this segment. Phylogenetic analyses grouped the sequences into two clusters, one comprising IN from the amphotropic strain 4070A and three ecotropic MLV (CAS-BR-E, Moloney and Friend), the other consisting of IN from three ecotropic MLV (two radiation-induced viruses and AKV) and a mink cell focus-forming (MCF) MLV virus. The same dichotomy and cluster composition was obtained from analysis of the long terminal repeat (LTR) regions from these viruses (consistent with the functional interrelationship of IN and LTR) but not from analysis of envelope protein sequences (consistent with the functional independence of ENV proteins from both IN and LTR). Secondary structure predictions supported features determined from the catalytic domain of human immunodeficiency virus and avian sarcoma virus IN, and identified probable structures within the relatively long N- and C-terminal domains of MLV IN proteins.

The effect of angiostatic steroids and beta-cyclodextrin tetradecasulfate on corneal neovascularization in the rat.

Folkman and coworkers have described angiostatic steroids that markedly inhibit neovascularization of the rabbit cornea when given topically with beta-cyclodextrin tetradecasulfate (beta-CD), yet have minimal or no glucocorticoid or mineralocorticoid activity. Our objective was to extend these observations to another species, the rat. We induced neovascularization by cauterizing rat corneas with silver nitrate/potassium nitrate; drugs were applied topically four times per day for 4 days in most experiments. Submicron sized emulsions of lipid-soluble dexamethasone and the angiostatic steroids 17 alpha-hydroxyprogesterone (1 or 10 mg ml-1) and cortexolone (1 or 10 mg ml-1) were prepared by lecithin encapsulation of drug microcrystals. The vehicle for water-soluble hydrocortisone 21-phosphate (HCP) +/- beta-CD (Molecusol; Pharmatec, Inc) was 10% Tween 20 in Tris-buffered 0.9% saline. Angiogenesis was significantly inhibited only by 1 mg ml-1 dexamethasone (-63.2% when compared with controls), 0.5 mg ml-1 HCP + 1 mg ml-1 beta-CD (-33.4%), and 1 mg ml-1 HCP (-40.2%). HCP (0.5 mg ml-1) or beta-CD (1 or 2 mg ml-1) alone had no significant effect on neovascularization; the inhibition by 1.0 mg ml-1 HCP was not potentiated by 2 mg ml-1 beta-CD. We also tested HCP and tetrahydro-S (TH-S) using 1.5% hydroxypropyl methylcellulose vehicle and beta-CD from Takeda Chemical Industries, Ltd., to simulate the procedure of Folkman and coworkers.(ABSTRACT TRUNCATED AT 250 WORDS)

Genital Chlamydia trachomatis infection in Australia.

OBJECTIVE: To review available data on genital chlamydial infections in Australia, to determine trends and to make recommendations for improving future data collection. DATA SOURCES: Data are presented from representative laboratories around Australia for the 11-year period 1980-1990. Studies of the prevalence of chlamydial infection in Australia and overseas for the same period were selected for review where relevant epidemiological information (such as patient population, reason for test, demographic data) was available. RESULTS: Overall rates of chlamydial infection from the surveyed laboratories ranged from 3.2%-14.6% for the period 1980-1982 and 2.7%-5.5% for 1990. Rates of diagnosis of Chlamydia appeared to decline in some centres while remaining relatively stable at others. Between 2800 and 4000 cases per year have been reported to the CDI (Communicable Diseases Intelligence) scheme since 1985. Substantial increases in the number of tests performed were seen in nearly all laboratories, reflecting a shift towards screening asymptomatic lower-risk women, however detailed data on the populations tested were unavailable. Studies of prevalence of genital chlamydial infection in Australia revealed highest infection rates among sexually transmitted disease clinic clients (2.5%-14%) and a prevalence of more than 5% in women attending family planning clinics. Studies from overseas showed wide variations in prevalence of infection, from 6% to 28% depending on the populations studied. CONCLUSION: Improved data collection is imperative for assessing the impact of intervention programs for chlamydial infection, which has potentially serious but largely preventable sequelae in women. Although diagnosis of genital chlamydial infection appears to be declining or at least stable in Australia, possibly due to intervention programs, it remains a relatively common sexually transmitted infection. Comparison of rates and interpretation of the figures is made difficult by changes in screening practices, lack of standard case definitions, denominator information and probable under-reporting. Recommendations proposed for improving data collection for Chlamydia include establishing sentinel sites, standardising the collection of data, and ensuring standard case definitions between sites.

A controlled evaluation of continuous passive motion in patients undergoing total knee arthroplasty.

OBJECTIVE: To evaluate the efficacy of continuous passive motion (CPM) in the postoperative management of patients undergoing total knee arthroplasty. DESIGN: A randomized controlled single-blind trial of CPM plus standardized rehabilitation vs standard rehabilitation alone. SETTING: A referral hospital for arthritis and musculoskeletal care. PATIENTS: Consecutive patients with end-stage osteoarthritis or rheumatoid arthritis undergoing primary total knee arthroplasty who had at least 90 degrees of passive knee flexion. One hundred fifty-four patients were eligible and 102 patients agreed to participate and were randomized. Ninety-three patients completed the study protocol. INTERVENTION: Continuous passive motion machines programmed for rate and specified arc of motion within 24 hours of surgery with range increased daily as tolerated with standardized rehabilitation program compared with standardized rehabilitation program alone. MAIN OUTCOME MEASURES: Primary outcomes were pain, active and passive knee range of motion, swelling (or circumference), quadriceps strength at postoperative day 7, as well as complications, length of stay, and active and passive range of motion and function at 6 weeks. RESULTS: Use of CPM increased active flexion and decreased swelling and the need for manipulations but did not significantly affect pain, active and passive extension, quadriceps strength, or length of hospital stay. At 6 weeks there were no differences between the two groups in either range of motion or function. In this series, use of CPM resulted in a net savings of $6764 over conventional rehabilitation in achieving these results. CONCLUSION: For the average patient undergoing total knee arthroplasty, CPM is more effective in improving range of motion, decreasing swelling, and reducing the need for manipulation than is conventional therapy and lowers cost.

The outcome of decompressive laminectomy for degenerative lumbar stenosis.

The outcome of laminectomy for the relief of symptoms resulting from degenerative lumbar stenosis is not well established. Eighty-eight consecutive patients who had had a laminectomy for degenerative lumbar stenosis between 1983 and 1986 were studied. Eight of the patients had had a concomitant arthrodesis. The follow-up evaluation included a review of charts and standardized questionnaires that were completed by the patients. One year postoperatively, five patients (6 per cent) had had a second operation and five still had severe pain. By the time of the latest follow-up, in 1989, fifteen (17 per cent) of the original eighty-eight patients had had a repeat operation because of instability or stenosis; twenty-one (30 per cent) of the seventy patients who were evaluated by questionnaire in 1989 had severe pain. The factors found to be associated with a poor long-term outcome, defined as severe pain or the need for a repeat operation, or both, included co-existing illnesses (such as osteoarthrosis, cardiac disease, rheumatoid arthritis, or chronic pulmonary disease) (p = 0.004), the duration of follow-up (p = 0.01), and an initial laminectomy involving a single interspace (p = 0.04). We concluded that the long-term outcome of decompressive laminectomy is less favorable than has been previously reported, and that co-morbidity and a single-interspace laminectomy are risk factors for a poor outcome.

Nonradioactive, photobiotin-labelled DNA probes for routine diagnosis of viroids in plant extracts.

Avocado sunblotch viroid (ASBV), coconut cadang cadang viroid (CCCV), chrysanthemum stunt viroid (CSV) and potato spindle tuber viroid (PSTV) were detected in plant extracts by dot-blot hybridization using nonradioactive photobiotin-labelled nucleic acid probes. Recombinant DNA probes, containing full-length monomer viroid inserts in the plasmid vectors pSP64 or pUC9, were biotinylated with photobiotin and used as sonicated double-stranded DNA fragments. Using fresh leaf material, a general method (suitably modified for avocado tissue) was developed for the rapid preparation of purified nucleic acid extracts. Plant extracts from a range of field samples were spotted onto nitrocellulose, subjected to hybridization and the biotin-labelled DNA bound to the target nucleic acid was detected with an avidin-alkaline phosphatase conjugate. Under the stated hybridization and washing conditions, each individual viroid probe was specific. Each viroid was readily detected with a sensitivity similar to that obtained with the same (or a like) probe labelled with 32P. Healthy plant extracts gave colourless spots.

Visual field differences in the processing of numerical stimuli.

Twenty-four right-handed subjects received random presentations of the numbers 1-6 in the form of words, digits, and dot patterns, to the left and right visual fields. Accuracy and reaction time were recorded for an odd-even judgment requiring a manual response. A significant stimulus type of visual field interaction was obtained, with words showing a left-hemisphere advantage and digits and dot patterns showing a right-hemisphere advantage. This pattern supports Coltheart's (1980, Deep dyslexia: A right hemisphere hypothesis, In M. Coltheart, K. Patterson, & J.C. Marshall (Eds.), Deep dyslexia, London: Routledge & Kegan Paul) right hemisphere reading hypothesis, which suggests that the left hemisphere's general advantage in processing linguistic material may be specific to stimuli which involve phonological processing. When phonological processing is not possible (e.g., for arabic digits and other ideographic orthographies), the right hemisphere may have an advantage because of its superior visuospatial processing capabilities.

Stretch-induced spinal accessory nerve palsy.

Left spinal accessory nerve palsy occurred in a young man when he quickly turned his head to the right while his shoulders were pulled down by heavy hand-held objects. Electrophysiologic studies demonstrated partial axonotmesis of the spinal accessory nerve branches innervating the sternocleidomastoid and upper and middle trapezius and complete axonotmesis of spinal accessory branches to the lower trapezius. There was a separate, although functionally minor, cervical plexus innervation of the lower trapezius.

Improved separation of multi-locus hypervariable DNA restriction fragments by field inversion gel electrophoresis and fragment detection using a biotinylated probe.

DNA probes to multiple hypervariable human genomic loci are potentially highly informative as regards individual genome identification and parentage studies. However the resultant Southern blot patterns are generally highly complex and their interpretation difficult, with one important limitation being the extent of fragment separation and resolution. The separation of large DNA restriction fragments in agarose gels may be improved, in comparison to separations obtained by conventional electrophoresis, by the use of field inversion techniques. This is demonstrated by the analysis of the complex human Satellite III related DNA polymorphism. Detection of the Satellite III related restriction fragments is achieved either by using a [35S]-labelled probe (228S) or by using the same probe in a convenient non-isotopic form constructed by the photobiotin process. In addition, the probe 228S is useful for sexing the human genome, by the identification of a Y-specific restriction fragment.

Photobiotin-labeled DNA and RNA hybridization probes.

Nucleic acid hybridization, the formation of a duplex between two complementary nucleotide sequences, is being increasingly utilized in the research laboratory and for the routine diagnosis of disease. Biotin-labeled nucleic acid hybridization probes have advantages over radioactively labeled probes in terms of stability, safety, and time of detection. The hybridization of a biotin-labeled probe to a target nucleic acid is carried out on a nitrocellulose or nylon filter; the bound probe is usually detected by the binding of an avidin-enzyme conjugate or a streptavidin-enzyme complex (both avidin and streptavidin have a high affinity for biotin), followed by a colorimetric reaction in which the bound enzyme converts a colorless substrate into a colored product.

Nonradioactive, photobiotin-labelled DNA probes for the routine diagnosis of barley yellow dwarf virus.

Photobiotin was used to prepare biotinylated, nonradioactive nucleic acid probes for the detection of the RNA of barley yellow dwarf virus (BYDV) in plant extracts. A 1.7-kb cDNA of the PAV isolate of BYDV in the plasmid pUC8 vector was biotinylated and then used intact or as sonicated double-stranded DNA fragments. Simple methods were developed for the preparation of partially purified nucleic acid extracts of cereals and their spotting, after formaldehyde treatment, onto nitrocellulose membranes. After hybridization, biotin-labelled DNA bound to BYDV RNA on the nitrocellulose was detected with an avidin-alkaline phosphatase conjugate. BYDV RNA was readily detected with a sensitivity similar to that found with the same probe labelled with 32P by nick translation. Healthy plant extracts gave colourless spots.

Non-radioactive hybridization probes prepared by the chemical labelling of DNA and RNA with a novel reagent, photobiotin.

A photo-activatable analogue of biotin, N-(4-azido-2-nitrophenyl)-N'-(N-d-biotinyl-3-aminopropyl)-N'-methyl-1,3- propanediamine (photobiotin), has been synthesized and used for the rapid and reliable preparation of large amounts of stable, non-radioactive, biotin-labelled DNA and RNA hybridization probes. Upon brief irradiation with visible light, photobiotin formed stable linkages with single- and double-stranded nucleic acids yielding probes which were purified from excess reagent by 2-butanol extraction and ethanol precipitation. Using single-stranded phage M13 DNA probes chemically labelled with one biotin per 100-400 residues and dot-blot hybridization reactions on nitrocellulose, as little as 0.5 pg (6 X 10(-18) mol) of target DNA was detected colorimetrically by avidin or streptavidin complexes with acid or alkaline phosphatase from three commercial sources. The sensitivity of detection of target RNA in dot-blots and Northern blots was equivalent to that obtained with 32p-labelled DNA probes. Photobiotin was also used for the labelling of proteins with biotin.