Herbivory, plant traits and nectar chemistry interact to affect the community of insect visitors and pollination in common milkweed, Asclepias syriaca.

Herbivory can alter plant fitness directly through changing reproductive allocation and indirectly through changing pollinator identity or behavior. Common milkweed is a plant of conservation concern with an inducible chemical defense that is also an important nectar resource. In this study, we aim to understand how herbivory severity and plant traits, including morphology and nectar chemistry, interact to affect insect visitation and pod production in common milkweed. We conducted pollinator watches on plants with experimentally varied herbivory severity and quantified insect frequency and visit length as a response to nectar chemistry, ramet height, number of inflorescences, number of flowers per inflorescence and percent tissue removed. We also quantified pollinator effectiveness and importance. Increased herbivory severity reduced floral displays, including fewer inflorescences and fewer flowers per inflorescence. A reduced floral display was correlated with reduced sucrose, fructose and glucose and resulted in a reduced number and species richness of insect visitors. Fewer flowers per inflorescence reduced the frequency of bumble bee and fly visitors, which were two important pollinators. Although honeybees, flies, small bees, soldier beetles and bumble bees were equally effective pollinators, only bumble bee frequency was positively correlated with pod production. The differences in pollinator visitation have the potential to create diversifying selection on plant floral traits, many of which are also affected by herbivores. This research demonstrates potentially conflicting selection pressures between native and non-native pollinators as well as non-native herbivores.

Whole genome sequencing of Shigella sonnei through PulseNet Latin America and Caribbean: advancing global surveillance of foodborne illnesses.

OBJECTIVES: Shigella sonnei is a globally important diarrhoeal pathogen tracked through the surveillance network PulseNet Latin America and Caribbean (PNLA&C), which participates in PulseNet International. PNLA&C laboratories use common molecular techniques to track pathogens causing foodborne illness. We aimed to demonstrate the possibility and advantages of transitioning to whole genome sequencing (WGS) for surveillance within existing networks across a continent where S. sonnei is endemic. METHODS: We applied WGS to representative archive isolates of S. sonnei (n = 323) from laboratories in nine PNLA&C countries to generate a regional phylogenomic reference for S. sonnei and put this in the global context. We used this reference to contextualise 16 S. sonnei from three Argentinian outbreaks, using locally generated sequence data. Assembled genome sequences were used to predict antimicrobial resistance (AMR) phenotypes and identify AMR determinants. RESULTS: S. sonnei isolates clustered in five Latin American sublineages in the global phylogeny, with many (46%, 149 of 323) belonging to previously undescribed sublineages. Predicted multidrug resistance was common (77%, 249 of 323), and clinically relevant differences in AMR were found among sublineages. The regional overview showed that Argentinian outbreak isolates belonged to distinct sublineages and had different epidemiologic origins. CONCLUSIONS: Latin America contains novel genetic diversity of S. sonnei that is relevant on a global scale and commonly exhibits multidrug resistance. Retrospective passive surveillance with WGS has utility for informing treatment, identifying regionally epidemic sublineages and providing a framework for interpretation of prospective, locally sequenced outbreaks.

Not another type of potato: MC1R and the russet coloration of Burmese cats.

The Burmese is a breed of domestic cat that originated in Southeast Asia and was further developed in the United States. Variants in melanocortin 1 receptor (MC1R) causes common coat colour phenotypes in a variety of mammalian species but only limited colour variation in the domestic cat. Known as the extension (E) locus, melanocortin 1 receptor (MC1R) interacts with the agouti locus to produce the eumelanin and pheomelanin pigments. Recently, a novel reddish coloration, which is termed russet, was identified in the Burmese cat breed. Because this russet Burmese coloration changes with aging, MC1R was suggested as candidate gene. The similar colouration in specific lineages of Norwegian Forest cat known as amber (e) (c.250G>A; p.Asp84Asn) was excluded for this Burmese phenotype. The complete 954-bp coding region of MC1R was directly sequenced in russet Burmese and suspected carriers. A 3-bp deletion (c.439_441del) associated with the deletion of a phenyalanine (p.Phe146del) in the protein sequence was identified. All russet coloured cats were homozygous for the variant, and all obligate carriers were heterozygous, confirming that the deletion segregated concordantly with colouring in Burmese cats from the New Zealand foundation lineage. The variant was not identified in 442 cats from 26 different breeds and random-bred cats. Twenty-six Burmese from the USA did not have the variant. This MC1R variant defines a unique coloration and the second breed-specific MC1R variant in cats. The interactions of the two recessive feline MC1R alleles (E  >  e, er ) is unknown.

Non-thermal irreversible electroporation (N-TIRE) and adjuvant fractionated radiotherapeutic multimodal therapy for intracranial malignant glioma in a canine patient.

Non-thermal irreversible electroporation (N-TIRE) has shown promise as an ablative therapy for a variety of soft-tissue neoplasms. Here we describe the therapeutic planning aspects and first clinical application of N-TIRE for the treatment of an inoperable, spontaneous malignant intracranial glioma in a canine patient. The N-TIRE ablation was performed safely, effectively reduced the tumor volume and associated intracranial hypertension, and provided sufficient improvement in neurological function of the patient to safely undergo adjunctive fractionated radiotherapy (RT) according to current standards of care. Complete remission was achieved based on serial magnetic resonance imaging examinations of the brain, although progressive radiation encephalopathy resulted in the death of the dog 149 days after N-TIRE therapy. The length of survival of this patient was comparable to dogs with intracranial tumors treated via standard excisional surgery and adjunctive fractionated external beam RT. Our results illustrate the potential benefits of N-TIRE for in vivo ablation of undesirable brain tissue, especially when traditional methods of cytoreductive surgery are not possible or ideal, and highlight the potential radiosensitizing effects of N-TIRE on the brain.

Physical analysis of the complex rye (Secale cereale L.) Alt4 aluminium (aluminum) tolerance locus using a whole-genome BAC library of rye cv. Blanco.

Rye is a diploid crop species with many outstanding qualities, and is important as a source of new traits for wheat and triticale improvement. Rye is highly tolerant of aluminum (Al) toxicity, and possesses a complex structure at the Alt4 Al tolerance locus not found at the corresponding locus in wheat. Here we describe a BAC library of rye cv. Blanco, representing a valuable resource for rye molecular genetic studies, and assess the library's suitability for investigating Al tolerance genes. The library provides 6 x genome coverage of the 8.1 Gb rye genome, has an average insert size of 131 kb, and contains only ~2% of empty or organelle-derived clones. Genetic analysis attributed the Al tolerance of Blanco to the Alt4 locus on the short arm of chromosome 7R, and revealed the presence of multiple allelic variants (haplotypes) of the Alt4 locus in the BAC library. BAC clones containing ALMT1 gene clusters from several Alt4 haplotypes were identified, and will provide useful starting points for exploring the basis for the structural variability and functional specialization of ALMT1 genes at this locus.

A preliminary assessment of whole-body radiotherapy interposed within a chemotherapy protocol for canine lymphoma.

Six dogs with spontaneously occurring, previously untreated lymphoma were treated with half-body radiation therapy (RT) doses interposed in a CHOP-based 25-week chemotherapy regimen. Chemotherapy-related toxicities were as expected and were mildly increased in severity post-RT compared with pre-RT. Treatment was delayed by 1-2 weeks per delay in four dogs due to chemotherapy-related neutropenia. Radiation therapy was administered in two consecutive day fractions of 4 Gray to the cranial and caudal halves of the body 4 weeks apart. Radiation-related toxicities consisted of lethargy, alopecia, diarrhoea of less than 2-day duration and average decreases in neutrophil counts of 50%. Late effects from RT were not evident. Median remission and survival times for the six dogs were 455 and 560 days, respectively. The protocol was well tolerated and should be studied further to evaluate the potential therapeutic gain of the addition of RT to chemotherapy for the treatment of canine lymphoma.

Randomized trial of paclitaxel plus supportive care versus supportive care for patients with advanced non-small-cell lung cancer.

BACKGROUND: In phase II trials, paclitaxel has been shown to have antitumor activity in patients with advanced non-small-cell lung cancer (NSCLC). However, the survival and quality-of-life (QOL) benefits of paclitaxel used as a single agent compared with supportive care alone have not been assessed in a randomized clinical trial. METHODS: A total of 157 patients with stage IIIB or IV NSCLC who had received no prior chemotherapy were randomly assigned to receive either best supportive care alone (78 patients) or paclitaxel plus supportive care (79 patients). Paclitaxel was administered as a 3-hour intravenous infusion every 3 weeks. Supportive care included palliative radiotherapy and supportive therapy with corticosteroids, antibiotics, analgesics, antiemetics, transfusions, and other symptomatic therapy as required. The primary end point of the study was survival. Time to disease progression, response rate, adverse events, and QOL were secondary end points. RESULTS: Pretreatment characteristics were evenly distributed between the two arms. Survival was statistically significantly better in the paclitaxel plus supportive care arm than in the supportive care alone arm (two-sided P =.037) (median survival = 6.8 months versus 4.8 months). Cox multivariate analysis showed paclitaxel plus supportive care to be statistically significantly associated with improved survival (two-sided P =.048). QOL was similar for both treatment arms, except for the functional activity score of the Rotterdam Symptom Checklist, where QOL data statistically significantly favored the paclitaxel plus supportive care arm (two-sided P =.043). CONCLUSION: The addition of paclitaxel to best supportive care significantly improved survival and time to disease progression compared with best supportive care in patients with advanced NSCLC and may improve some aspects of QOL.

Impact of dietary fat and fiber intake on nutrient intake of adolescents.

OBJECTIVE: To evaluate the impact of fat and fiber intake on energy and nutrient intake of 15-year-old adolescents. STUDY DESIGN: Twenty-four-hour dietary recalls were collected on a random sample of 15-year-olds. Subjects were then categorized into groups based on fat and fiber intake, with 319 students meeting criteria for 1 of 4 dietary intake quadrants: low fat, low fiber; low fat, high fiber; high fat, low fiber; and high fat, high fiber. Students with medium fat or fiber intakes were excluded from the study. Low-fat and high-fat intake were defined as <30% and >40% of total energy intake, respectively. Low- and high-fiber intake were defined as <15 g/day and >20 g/day, respectively. RESULTS: Low-fat and high-fiber intake had a minimal impact on energy intake and did not adversely affect nutrient intake. High-fiber intake was associated with greater likelihood of adequate intake of vitamins A, B6, B12, and C; niacin; thiamin; riboflavin; folacin; magnesium; iron; zinc; phosphorus; and calcium. High-fat intake was associated with greater likelihood of adequate vitamin B12 intake. Significant differences in fat and fiber intake distributions were found for ethnic background and gender, with more non-white than white students in the high-fat groups and more males than females in the high-fiber groups. CONCLUSIONS: A low-fat and high-fiber diet meeting current nutrition recommendations does not adversely affect energy or nutrient intake, increases nutrient density of the diet, and increases the likelihood of adequate intake for several key nutrients.

Paclitaxel is safe and effective in the treatment of advanced AIDS-related Kaposi's sarcoma.

PURPOSE: Liposomal anthracyclines are the present standard treatment for advanced AIDS-related Kaposi's sarcoma (KS). No effective therapies have been defined for use after treatment failure of these agents. A phase II trial was thus conducted with paclitaxel in patients with advanced KS to assess safety and antitumor activity. MATERIALS AND METHODS: A regimen of paclitaxel at a dose of 100 mg/m(2) was given every 2 weeks to patients with advanced AIDS-related KS. Patients were treated until complete remission, disease progression, or unacceptable toxicity occurred. RESULTS: Fifty-six patients with advanced AIDS-related KS were accrued. Tumor-associated edema was present in 70% of patients and visceral involvement in 45%. Forty patients (71%) had received prior systemic therapy; 31 of these were resistant to an anthracycline. The median entry CD4(+) lymphocyte count was 20 cells/mm(3) (range, 0 to 358). A median of 10 cycles (range, 1 to 54+) of paclitaxel was administered. Fifty-nine percent of patients showed complete (n = 1) or partial response (n = 32) to paclitaxel. The median duration of response was 10.4 months (range, 2.8 to 26.7+ months) and the median survival was 15.4 months. The main side effects of therapy were grade 3 or 4 neutropenia in 61% of patients and mild-to-moderate alopecia in 87%. CONCLUSION: Paclitaxel at 100 mg/m(2) given every 2 weeks is active and well tolerated in the treatment of advanced and previously treated AIDS-related KS. The median duration of response is among the longest observed for any regimen or single agent reported for AIDS-related KS. Paclitaxel at this dosage and schedule is a treatment option for patients with advanced AIDS-related KS, including those who have experienced treatment failure of prior systemic therapy.

Women and men with hypercholesterolemia respond similarly to an American Heart Association step 1 diet.

OBJECTIVE: To compare serum lipid level responses of women and men with hypercholesterolemia to an American Heart Association (AHA) step 1 diet. DESIGN: Sixty-three women and 99 men with varying degrees of hypercholesterolemia were instructed on an AHA step 1 diet. Subjects were followed up on a biweekly basis with individual visits and group classes for 8 weeks. SETTING: Outpatient clinic facility of the Metabolic Research Group, University of Kentucky, Lexington. SUBJECTS/SAMPLES: Through community cholesterol screenings, we recruited 76 women and 108 men aged 30 to 70 years who were within 80% to 130% of their ideal body weight. Serum cholesterol levels were between 5.17 mmol/L and 8.99 mmol/L and serum triglyceride levels when subjects were fasting were less than 5.08 mmol/L. Sixty-three women and 99 men completed the study. INTERVENTION: Subjects followed an AHA step 1 diet (30% of energy from fat, 50 to 60% of energy from carbohydrate, 10 to 20% of energy from protein, and less than 300 mg cholesterol per day) for 8 weeks. MAIN OUTCOME MEASURES: Serum lipid levels, nutrient intake, and body weight. STATISTICAL ANALYSES PERFORMED: Subjects were divided into three groups according to initial serum cholesterol levels (mild = 5.17 to 6.17 mmol/L; moderate = 6.20 to 6.95 mmol/L; severe = > or = 6.98 mmol/L). Within-individual changes in nutrient intakes, body weights, and serum lipid levels were analyzed using dependent t tests. Between-group comparisons were made using analysis of variance (ANOVA). When significant differences were found using ANOVA, differences between groups were evaluated with the Tukey test. RESULTS: All subjects tolerated the diet well and average dietary adherence was good, as assessed by a food frequency questionnaire and analysis of 3-day diet records. Serum total cholesterol levels decreased 9.2% overall for women (P < .001) and 7.2% for men (P < .001); serum low-density lipoprotein cholesterol levels decreased 9.2% for women and 9.8% for men; and serum high-density lipoprotein cholesterol levels decreased 3.6% for women and 2.8% for men. Mean serum triglyceride levels decreased significantly for women but not for men. No significant differences were found in the responses of women and men in the corresponding groups. Women and men with higher initial serum cholesterol values showed significantly greater hypocholesterolemic responses to diet than those with lower initial serum cholesterol values. APPLICATIONS/CONCLUSIONS: The findings of this study confirm the beneficial role of dietary intervention for reducing atherogenic serum lipid levels in women and men.

Food-containing hypocaloric diets are as effective as liquid-supplement diets for obese individuals with NIDDM.

OBJECTIVE: This study was designed to determine if food-containing hypocaloric diets are as effective as liquid-supplement diets in promoting weight loss for obese individuals with non-insulin-dependent diabetes mellitus (NIDDM). RESEARCH DESIGN AND METHODS: Forty NIDDM subjects with body mass indexes (BMIs) of 30-40 kg/m2 were randomized to one of two 800-kcal diets for 12 weeks. Group A received liquid supplement only, and group B received supplement plus an evening meal. Both groups received an intensive behavioral education program. RESULTS: Weight loss and improvements in glycemic, blood lipid, and blood pressure parameters were similar for the two groups. Weight loss averaged 15.7 kg for the entire group. The need for insulin, anti-diabetes, and anti-hypertensive medication decreased significantly. No serious side effects were observed. CONCLUSIONS: Both food-containing and supplement diets providing 800 kcal a day effectively promote weight loss for obese individuals with NIDDM.

Recombinant human superoxide dismutase (h-SOD) fails to improve recovery of ventricular function in patients undergoing coronary angioplasty for acute myocardial infarction.

BACKGROUND: Animal studies have demonstrated a burst of oxygen free radical generation after reperfusion of ischemic myocardium that could be blocked by administration of the free radical scavenger recombinant human superoxide dismutase (h-SOD). A multicenter, randomized, placebo-controlled clinical trial was designed to test the hypothesis that free radical-mediated reperfusion injury could be reduced by intravenous administration of h-SOD begun before percutaneous transluminal coronary angioplasty (PTCA) in patients with acute transmural myocardial infarction. METHODS AND RESULTS: One hundred twenty patients were randomized to receive placebo (n = 59) or h-SOD (n = 61) given as a 10-mg/kg intravenous bolus followed by a 60-minute infusion of 0.2 mg.kg-1.min-1. Left ventricular function was analyzed via paired contrast left ventriculograms performed before PTCA and after 6 to 10 days and paired radionuclide ventriculograms performed within 24 hours of PTCA and after 4 to 6 weeks. Both h-SOD- and placebo-treated patients showed improvement in global and regional left ventricular function after successful reperfusion. Compared with the placebo group, no additional improvement was observed in the patients treated with h-SOD. CONCLUSIONS: The results of this clinical trial failed to demonstrate a beneficial effect of h-SOD on global or regional left ventricular function in patients who underwent successful PTCA for treatment of acute myocardial infarction.

Health benefits and practical aspects of high-fiber diets.

Over the past 20 y dietary fiber has emerged as a leading dietary factor in the prevention and treatment of chronic diseases. High fiber intakes are associated with lower serum cholesterol concentrations, lower risk of coronary heart disease, reduced blood pressure, enhanced weight control, better glycemic control, reduced risk of certain forms of cancer, and improved gastrointestinal function. Dietary fiber can be categorized into water-soluble and water-insoluble components. Dried beans, oat products, and certain fruits and vegetables are good sources of soluble fiber. Most plant foods are good sources of insoluble fiber and wheat bran is a concentrated form of insoluble fiber. Current guidelines advise a doubling of dietary fiber intake for Americans. Inclusion of ample servings of fruits and vegetables, whole grains, and dried beans and peas will help individuals meet these guidelines.

Relation between plasma concentrations of didanosine and markers of antiviral efficacy in adults with AIDS or AIDS-related complex.

The relation between the average steady-state plasma concentration (Cpss) of didanosine and selected measures of efficacy, such as CD4 cell count, p24 antigenemia, and weight gain, was evaluated in patients participating in a phase 1 safety and pharmacokinetics study. All patients were diagnosed as having AIDS or severe AIDS-related complex. These individuals first received intravenous didanosine for 2 weeks at doses of 0.8-33 mg/(kg.d) and then took the drug orally at twice the intravenous dose. Cpss values were calculated on the basis of apparent oral clearance after 4 weeks of oral administration and average daily dose over the first 12 weeks of the study. These data were available for 61 patients enrolled at three clinical sites. High values for Cpss were strongly correlated with an increase in CD4 count (P = .006), a decrease in serum levels of p24 antigen (P = .006), and weight gain (P = .0001) at week 12. Logistic regression analysis was used to assess the influence of Cpss on response (as judged by the three criteria just mentioned) after adjustment for other potential factors related to infection with human immunodeficiency virus. The baseline CD4 cell count and the status with regard to prior zidovudine therapy were related to the CD4 response. However, the odds that a response would include all three parameters were nearly twice as high when the Cpss value increased by twofold.(ABSTRACT TRUNCATED AT 250 WORDS)

Cholesterol-lowering effects of psyllium-enriched cereal as an adjunct to a prudent diet in the treatment of mild to moderate hypercholesterolemia.

Psyllium hydrophilic mucilloid, rich in soluble fiber, significantly lowers serum cholesterol concentrations. To investigate the lipid-lowering effects of a ready-to-eat cereal enriched with psyllium, we studied 44 hypercholesterolemic ambulatory individuals for 7 wk, using a randomized, double-blind, parallel design. After a 1-wk baseline period, subjects were randomly assigned to consume 114 g/d of a psyllium-flake or wheat-bran-flake cerearl for 6 wk adjunctive to an American Heart Association Step I diet. Serum high-density-lipoprotein cholesterol and triglycerides remained unchanged throughout the study; however, serum cholesterol and low-density lipoprotein (LDL) concentrations were significantly lower on psyllium than on wheat-bran cereal. Additionally, serum total and LDL cholesterol values were significantly lower than baseline values in the psyllium group. The dietary nutrient intake of the two groups did not differ except for soluble fiber. Adding psyllium-enriched cereal to a prudent low-fat diet may enhance dietary management of hypercholesterolemia.

Lack of effect of peroral acyclovir for the treatment of acute infectious mononucleosis.

Perorally administered acyclovir was evaluated in the therapy of acute infectious mononucleosis in a multicentered, randomized, double-blind, placebo-controlled trial. A total of 120 patients received 600 mg of acyclovir or placebo five times daily for 10 days. All patients were entered into the study within 7 days of symptom onset and had a positive Monospot test. Analysis of mean values and time to resolution of fever, lymphadenopathy, weight change, hepatomegaly, splenomegaly, liver function tests, atypical lymphocytes, hours of bed rest, sense of well-being, and return to normal activities revealed no significant differences. There was a trend toward suppression of Epstein-Barr virus excretion in the oropharynx in acyclovir recipients. No toxicity was detected in patients treated with acyclovir. Under the conditions of the study, there was no evidence that treatment with perorally administered acyclovir affected the course of infectious mononucleosis.

Serum lipid response of hypercholesterolemic men to single and divided doses of canned beans.

Dried beans lower serum lipid concentrations in healthy and hyperlipidemic subjects. To determine the effects of canned beans on serum lipid concentrations, 24 hyperlipidemic men ate one of three bean diets for 21 d in a metabolic ward. Diets A and B included 227 g canned beans (120 g beans with 107 g tomato sauce) daily, in a single dose for diet A and in a divided dose for diet B. Diet C included 182 g canned beans (162 g beans with 20 g tomato sauce) daily in a divided dose. All bean diets combined lowered serum cholesterol and triglyceride concentrations 10.4% (p less than 0.001) and 10.8% (p less than 0.025), respectively. Diet B was as effective as diet C, and the groups on those diets had greater cholesterol reductions than did the group on diet A. Serum cholesterol reduction was positively correlated (p less than 0.01) with intake of total dietary fiber and soluble fiber. The ratios of low- to high-density-lipoprotein cholesterol remained constant for all groups. Body weight decreased 1.0-1.5 kg for all groups despite constant energy intakes. Canned beans may make an important contribution to hyperlipidemia management.

Adherence to high-carbohydrate, high-fiber diets.

Successful diabetes management demands a high degree of adherence. Adherence to medical prescriptions, particularly special diets, is notoriously poor. A study of 40 individuals with IDDM and NIDDM who were followed an average of 23 months found good or excellent dietary adherence in 70% of subjects; only 5% demonstrated poor adherence. Adherence was assessed objectively based on carbohydrate, fat, and fiber intakes reported in 24-hour food recalls, 7-day food frequency surveys, and home food records. High dietary fiber intake may independently enhance adherence, perhaps by increasing satiety or because of the simplicity of the concept. Good communication between the health professional and patient is also fundamental to all stages of dietary adherence. Use of adherence-enhancing techniques throughout adoption of new diet behaviors promotes long-term adherence to high-carbohydrate, high-fiber diets.

Effect of a cancer chemotherapy prescription form on prescription completeness.

Completeness of physicians' orders for cancer chemotherapy was compared before and after implementation of a preprinted form for these orders. When an oncology pharmacy service (OPS) in which pharmacy prepared all drugs for cancer chemotherapy was implemented in 1985 at an 879-bed teaching hospital, the pharmacists began educational sessions for house-staff physicians on writing orders for cancer chemotherapy. Pharmacists assessed the effectiveness of these sessions by comparing completeness of orders written before implementation of the OPS with orders during a four-week period starting two months after implementation. Orders were checked for nine components: patient's diagnosis, height, weight, and body surface area and drug regimen, dose, dosage, frequency, and route. Inclusion of the same components was assessed after implementation of a form that physicians were required to use for prescribing all antineoplastic agents. During the baseline period, orders for 143 patients were evaluated. Only two prescription components, dose and route, were present is more than 90% of the orders. Educational intervention led to some improvement in order completeness, but only dose and route appeared in at least 90% of the 87 orders evaluated. The components necessary to verify physicians' calculations for body surface area and dose--height, weight, and dosage--were absent in 29 of the orders, and a pharmacist spent 420 minutes clarifying them. After the order form was implemented, orders for 77 patients were reviewed. Compliance exceeded 90% for eight of the nine components, and 12 medication errors were prevented by the form. A pharmacist spent 70 minutes clarifying five orders.(ABSTRACT TRUNCATED AT 250 WORDS)