Phenolic and short-chained aliphatic organic acid constituents of wild oat (Avena fatua L.) seeds.

The objective of this research was to identify and quantify the phenolic and short-chained aliphatic organic acids present in the seeds of three wild-type populations of wild oat and compare these results to the chemical composition of seeds from two commonly utilized wild oat isolines (M73 and SH430). Phenolic acids have been shown to serve as germination inhibitors, as well as protection for seeds from biotic and abiotic stress factors in other species, whereas aliphatic organic acids have been linked to germination traits and protection against pathogens. Wild oat populations were grown under a "common garden" environment to remove maternal variation, and the resulting seeds were extracted to remove the readily soluble and chemically bound phenolic and aliphatic organic acid components. Compounds were identified and quantified using gas chromatography-mass spectrometry. Ferulic and p-coumaric acid comprised 99% of the total phenolic acids present in the seeds, of which 91% were contained in the hulls and 98% were in the chemically bound forms. Smaller quantities of OH benzoic and vanillic acid were also detected. Soluble organic acids concentrations were higher in the M73 isoline compared to SH430, suggesting that these chemical constituents could be related to seed dormancy. Malic, succinic, fumaric and azelaic acid were the dominant aliphatic organic acids detected in all seed and chemical fractions.

Characterization of an 18,166 EST dataset for cassava (Manihot esculenta Crantz) enriched for drought-responsive genes.

Cassava (Manihot esculenta Crantz) is a staple food for over 600 million people in the tropics and subtropics and is increasingly used as an industrial crop for starch production. Cassava has a high growth rate under optimal conditions but also performs well in drought-prone areas and on marginal soils. To increase the tools for understanding and manipulating drought tolerance in cassava, we generated expressed sequence tags (ESTs) from normalized cDNA libraries prepared from dehydration-stressed and control well-watered tissues. Analysis of a total of 18,166 ESTs resulted in the identification of 8,577 unique gene clusters (5,383 singletons and 3,194 clusters). Functional categories could be assigned to 63% of the unigenes, while another approximately 11% were homologous to hypothetical genes with unclear functions. The remaining approximately 26% were not significantly homologous to sequences in public databases suggesting that some may be novel and putatively specific to cassava. The dehydration-stressed library uncovered numerous ESTs with recognized roles in drought-responses, including those that encode late-embryogenesis-abundant proteins thought to confer osmoprotective functions during water stress, transcription factors, heat-shock proteins as well as proteins involved in signal transduction and oxidative stress. The unigene clusters were screened for short tandem repeats for further development as microsatellite markers. A total of 592 clusters contained 646 repeats, representing 3.3% of the ESTs queried. The ESTs presented here are the first dehydration stress transcriptome of cassava and can be utilized for the development of microarrays and gene-derived molecular markers to further dissect the molecular basis of drought tolerance in cassava.

5 a day fruit and vegetable intervention improves consumption in a low income population.

OBJECTIVE: This study evaluated the Michigan Farmers' Market Nutrition Program in one Michigan county to determine its effect on fruit and vegetable consumption behavior. SUBJECTS/SETTING: Subjects were selected from WIC and Community Action Agency populations: 564 low income women completed the pretest; 455 completed the posttest. Attrition rate was 19.3%. INTERVENTION: Subjects were assigned to one of 4 interventions: education about the use, storage and nutritional value of fruits and vegetables, distribution of farmers' market coupons, both education and coupons, or no intervention. DESIGN: Education-only and coupon and education groups were randomly assigned; clinic appointment timing determined assignment to no-intervention and coupon-only groups. MAIN OUTCOME MEASURES: A self-administered questionnaire before and after intervention measured attitudes about fruit and vegetable consumption and intake of fruits and vegetables. WIC records documented redemption of coupons. STATISTICAL ANALYSES: Data analysis included 2-way multivariate analysis of covariance, univariate analysis of covariance, logistic regression, and covariance structure modeling. RESULTS: Both the education interventions and the coupon interventions had positive effects. Coupons had a direct effect on increasing fruit and vegetable consumption behavior but no effect on attitudes. Education had a direct effect on attitudes and seemed to exert an effect on consumption behavior through attitudes. The maximum impact of the intervention was achieved through a combination of education and coupons. APPLICATIONS: This study demonstrated that a low-income population may be more likely to increase its fruit and vegetable consumption behavior when incentives such as coupons improve affordability.

Primary canine and feline nervous system tumors: intraoperative diagnosis using the smear technique.

The recent application of neuroimaging techniques in veterinary neurology has led to the accurate localization of many types of intracranial lesions but has also created a clinical need, particularly with brain tumors, for a specific intraoperative diagnosis. For human brain tumors, a smear technique has been used successfully for many years to provide an extremely rapid, highly accurate intraoperative diagnosis. In similar smear preparations of intracranial lesions, obtained either by computed tomography (CT)-guided stereobiopsy or from a craniotomy, we have described distinguishing cytologic features of some primary spontaneous nervous system tumors in 80 dogs and 13 cats. A final diagnosis was confirmed by evaluation of paraffin-embedded sections from the same sample and, when appropriate, by immunocytochemical staining. Preliminary findings indicate that, in dogs and cats, this procedure is useful for rapid, accurate intraoperative diagnosis of many primary nervous system tumors. The distinguishing features of the canine and feline tumors bear a remarkably close resemblance to their human counterparts.

Atrial natriuretic peptide in type 2 diabetes mellitus: response to a physiological mixed meal and relationship to renal function.

Relatively few data exist on atrial natriuretic peptide (ANP) characteristics in Type 2 diabetes mellitus (DM). Therefore, plasma immunoreactive ANP concentrations were measured before and for 4 h following the ingestion of a physiological mixed meal in 8 newly diagnosed, normotensive, normoalbuminuric, patients with Type 2 DM and 6 normotensive, non-diabetic controls. In patients with Type 2 DM, basal plasma ANP concentrations were 4.0 +/- 2.0 and not significantly changed following ingestion of the meal, with peak levels of 4.9 +/- 2.8 pmol l(-1). Non-diabetic controls had higher basal plasma ANP concentrations, 8.7 +/- 3.4 pmol l(-1) (p < 0.05), significantly increasing to a peak of 11.9 +/- 6.3 pmol l(-1) at 30 min post meal. Extracellular fluid volume (ECV) was not different between diabetic patients and controls (15877 +/- 2679 vs 13668 +/- 1792 ml3). Glomerular filtration rate (GFR) (isotopic clearance corrected for body surface area) was elevated in diabetic patients (mean +/- SD) 130 +/- 39 vs 98 +/- 10 ml min(-1), p < 0.05). For the DM subjects, basal ANP levels were negatively correlated with GFR (rs - 0.74, p < 0.05) and effective renal plasma flow (ERPF) (rs - 0.8, p < 0.05). We conclude that patients with Type 2 DM demonstrate reduced basal plasma ANP concentrations which are inversely correlated to renal function. In contrast to non-diabetic controls, ANP in Type 2 DM does not rise in response to feeding.

Position of the American Dietetic Association: the role of nutrition in health promotion and disease prevention programs.

A healthful diet and wise food choices are critical components of promoting health and reducing the risk of chronic disease. A substantial amount of health care resources could be saved by expanding health promotion and disease prevention programs that target dietary change among Americans. To effectively reduce health care costs, the emphasis and delivery of health care must promote health as well as deliver treatment and rehabilitative services to the sick. Prevention measures, such as nutrition interventions that also encourage physical activity, can help prevent or halt progression of full-blown chronic disease and thus decrease chronic disease disability. Health promotion and disease prevention need to be integral parts of all health care, community, public health, and worksite programs across the life cycle. Correspondingly, such programs must be culturally competent and address the specific needs of vulnerable or underserved populations. Dietetics professionals in all areas of practice should play an integral role in health promotion and disease prevention programs. Achieving this goal will require expansion of training programs and active learning by dietetics professionals that includes theory and practice in using team approaches, developing coalitions, and managing complex systems. Dietetics professionals also need to amplify their understanding of politics, administration, health care financing, and reimbursement. Attention must also be expanded to include social and behavioral sciences and to address program evaluation, outcomes, and cost-benefit and cost-effectiveness in nutrition-focused health promotion and disease prevention programs. Continued training in program development, research, and evaluation will help build the body of evidence that supports ongoing inclusion of prevention in a rapidly changing health care environment.

Prevalence of Helicobacter pylori in acromegalic patients during treatment with octreotide.

BACKGROUND: Octreotide, a synthetic long-acting analogue of somatostatin, now has an established role in the treatment of acromegaly. In acromegalic patients treated with octreotide there is an increased incidence of gallstones and possibly gastritis. OBJECTIVES: (1) To compare the seroprevalence of Helicobacter pylori (H. pylori) infection, in acromegalic patients treated with octreotide to that in patients given other treatment modalities. (2) To study retrospectively the temporal relation between H. pylori acquisition and octreotide treatment. PATIENTS: Three groups of acromegalic patients were studied; 35 (20 M) had been treated with octreotide, 17 (10 M) with bromocriptine and 19 (12 M) had received no pharmacological intervention (untreated, surgically treated or treated with radiotherapy). DESIGN/MEASUREMENTS: The presence of H. pylori infection was assessed serologically (Bio-Rad GAP test for IgG), using stored serum, on the most recent sample from each patient and on serial samples from patients treated with octreotide. RESULTS: The prevalence of H. pylori seropositivity was similar in each treatment group, 34, 35 and 37%, respectively. Mean age and duration of acromegaly were similar in the first two groups. Patients who had never received medical treatment were slightly younger. GH levels were similar in all three groups. Patients on octreotide who were seropositive for H. pylori did not differ from those with negative serology with respect to age, duration of acromegaly, duration of octreotide treatment or serum GH level. Serial samples in octreotide treated patients showed a change in status in only one patient; 18 patients continued with negative serology during a mean period of 30 (range 4-62) months. In each of the 6 patients with persistently positive serology during octreotide treatment, stored samples predating octreotide therapy were shown to have already been positive. CONCLUSIONS: The seroprevalence of H. pylori infection in acromegalic patients does not appear to be increased in a manner dependent on the type or duration of medical treatment. In particular, octreotide therapy, while causing the development of histological gastritis in some patients, does not appear to induce the development of H. pylori infection.

Spinach leaf 70-kilodalton heat-shock cognate stabilizes bovine adrenal glucose-6-phosphate dehydrogenase in vitro without apparent stable binding.

Spinach (Spinacia oleracea L.) leaf tissue 70-kilodalton heat-shock cognate was purified by ATP-agarose affinity and gel filtration. Gel filtration of the affinity-purified protein resolved it into three forms: monomer, dimer, and oligomer. In the absence of ATP, the majority of the heat-shock cognate existed as a monomeric form with lesser amounts of dimer and oligomer. Addition of 3 mM ATP to the purified protein, containing all three forms, converted the dimeric and monomeric forms to a high-molecular-weight complex. Removal of ATP from the complex by dialysis resulted in the reappearance of the dimeric and monomeric forms. Addition of ATP to the highly purified monomer had no effect on its gel-filtration migration. Neither purified monomeric or dimeric forms showed stable binding to denatured proteins; however, both forms of the purified heat-shock cognate were able to stabilize the enzymatic activity of bovine adrenal glucose-6-phosphate dehydrogenase over a 48-h period at 25 degrees C. In addition, the activity of glucose-6-phosphate dehydrogenase in the presence of purified heat-shock cognate dimer or monomer could be rapidly decreased in an ATP-dependent fashion depending on the order of the substrate addition to the reaction mixture. Circular-dichroism studies indicated that addition of ATP to the spinach 70-kDa heat-shock cognate caused a conformation change from alpha-helical to a greater beta-sheet content. How conformational character may influence the stabilizing activity of the heat-shock cognate in a mechanism which does not require stable peptide binding is discussed.

Structural organization of the spinach endoplasmic reticulum-luminal 70-kilodalton heat-shock cognate gene and expression of 70-kilodalton heat-shock genes during cold acclimation.

The 70-kD heat-shock proteins (HSP70s) are encoded by a multigene family in eukaryotes. In plants, the 70-kD heat-shock cognate (HSC70) proteins are located in organellar and cytosolic compartments of cells in most tissues. Previous work has indicated that HSC70 proteins of spinach (Spinacia oleracea) are actively synthesized during cold-acclimating conditions. We have isolated, sequenced, and characterized cDNA and genomic clones for the endoplasmic reticulum (ER) luminal HSC70 protein (immunoglobulin heavy chain-binding protein; BiP) of spinach. The spinach ER-luminal HSC70 is a constitutively expressed gene consisting of eight exons. Spinach BiP mRNA appears to be up-regulated during cold acclimation but is not expressed during water stress or heat shock. In contrast to the differential regulation of mRNA, the ER-luminal HSC70 protein levels remain constant in response to various environmental stresses. Two other members of the spinach 70-kD heat-shock (HS70) multigene family also show differential expression in response to a variety of environmental stresses. A constitutively expressed cytosolic HSC70 protein in spinach appears also to be up-regulated in response to both cold-acclimating and heat-shock treatments. Spinach also contains a cold-shock-induced HS70 gene that is not expressed during heat shock or water stress. Since HSP70s are considered to be involved with the chaperoning and folding of proteins, the data further support the concept that they may be important for maintaining cellular homeostasis and proper protein biogenesis during cold acclimation of spinach.

Differential influence of ATP on native spinach 70-kilodalton heat-shock cognates.

A constitutively expressed class of 70-kD heat-shock cognate (HSC70) proteins from spinach leaf tissue was purified based on their affinity for ATP-agarose. The affinity-purified spinach proteins were resolved into at least three different forms on two-dimensional gels. Under native conditions, and iN the absence of ATP, the affinity-purified proteins were separated into three molecular mass classes by gel-filtration chromatography; a monomer of 85 kD, a multimer of 280 kD, and a large molecular mass oligomer of > 650 kD. All molecular mass forms contained a major protein that migrated at 79 kD on sodium dodecyl sulfate-polyacrylamide gels. N-terminal sequencing of the 79-kD purified monomer showed the highest homology to the endoplasmic reticulum-luminal HSC70. Addition of Mg-ATP to monomeric HSC70 did not alter its migration during gel filtration. Addition of Mg-ATP to the dimer converted it to monomer and oligomeric forms, whereas the presence of ATP converted a fraction of the large molecular mass oligomeric form of HSC70 to dimeric and monomeric forms. Only the large molecular mass oligomeric HSC70 appears to autophosphorylate in vitro in the presence of [gamma-32P]-ATP. Dimers and monomers can bind ATP by a nonhydrolytic mechanism and undergo a conformational change in the presence of Mg-ATP. In this paper we discuss the effects that ATP may have on the regulation of plant HSC70.

Antigen binding of a mouse monoclonal IgG1 is inactivated by heating but not by freeze/thaw cycling.

A mouse monoclonal antibody in ascitic fluid or in purified form was subjected to heating or repeated freeze/thaw cycles. Heating the antibody at 80 degrees C for 10 min completely destroyed its ability to bind to antigen. Unpurified antibody in ascitic fluid or purified antibody in buffer was frozen to -196 degrees C in liquid nitrogen and then thawed at room temperature. Purified antibody was also slowly frozen at -4 degrees C followed by thawing at room temperature. Regardless of the manner of freeze/thaw treatment or whether the antibody was purified or not, immunologic competence for its antigen was not lost. The results call into question the belief that freezing and thawing in all cases are necessarily deleterious to antibody structure and function.

A work-site weight management program to reinforce behavior.

This pilot weight management project addresses the efficacy of reinforcing dietary behavior change versus weight loss. This 6-month program served professional and support staff participating in the work-site wellness program at a midwestern university. Behavior-contingent program data were compared with data from the previous model where contracts were made for weight loss. In the behavior-contingent program, dropout rate and satisfaction with the program compared favorably with the old model weight loss-contingent program. Contract adherence was 93% compared with 74% in the weight loss-contingent program. Actual pounds of weight lost were lower in the behavior-contingent program, however, long-term weight management must still be studied with this population.

Malignant prolactinoma with extracranial metastases: a report of three cases.

OBJECTIVE: Prolactin-secreting tumours of the pituitary almost never metastasize extracranially. However, we have recently seen three such patients, whose presentation, clinical course and response to treatment are reviewed in order to determine whether they possess unique clinical features, and to assess optimum therapy. DESIGN AND PATIENTS: Three patients with extracranial metastases from prolactinomas have been retrospectively reviewed. MEASUREMENTS: Clinical courses, biochemical parameters and imaging modalities (CT, MR and radiolabelled somatostatin analogue scanning) and histopathology were assessed before and during different therapeutic regimens. RESULTS: All three patients' presentations were initially unremarkable, but later each had persistent and relentlessly increasing serum prolactin concentrations and evidence of distant spread of their tumours. The first patient presented aged 32 and was treated with repeated transsphenoidal resections of the tumour, pituitary radiotherapy, high dose bromocriptine therapy, the somatostatin analogue octreotide and chemotherapy. Nine years after the original diagnosis a liver biopsy revealed carcinoma cells positively immunostaining for prolactin. At autopsy further microscopically similar metastases were seen in both lungs and the left hilar lymph nodes. The second patient presented aged 48 and was treated with external beam pituitary radiotherapy, bromocriptine, transsphenoidal hypophysectomy and chemotherapy. Fifteen years after the original diagnosis bone biopsy revealed carcinoma cells positively immunostaining for prolactin; the patient developed interstitial nephritis and died in renal failure. The third patient presented aged 48 with hypogonadism and bitemporal hemianopia due to a macroadenoma. This was removed transfrontally and followed by external beam radiotherapy. After recurrence, further surgery and radiotherapy were undertaken, but the tumour continued to grow and the patient died suddenly of a pulmonary embolus. At autopsy, tumour was found in thoracic lymph nodes and lung capillaries. CONCLUSIONS: These three patients may be added to the previous two reported cases of malignant prolactinoma with extracranial spread. Several different therapies were tried but were of limited value in controlling the progression of the disease, although chemotherapy is an important modality to be considered. No particular factor or combination of factors at presentation differentiated these patients from more typical patients whose disease is much less aggressive.

Effect of octreotide on gall stone prevalence and gall bladder motility in acromegaly.

Octreotide therapy in acromegaly is associated with an increased prevalence of gall stones, which may be the result of an inhibition of gall bladder motility. Gall stone prevalence in untreated acromegalic patients relative to the general population is unknown, however, and the presence of gall stones and gall bladder motility in these patients and in acromegalic patients receiving octreotide was therefore examined. Thirty four percent of 39 patients who had taken octreotide for a mean of 20 months had gall stones compared with 16% of 38 patients who had not been treated with octreotide (p < 0.005). In a subgroup of 21 patients studied prospectively over 4 to 18 months, two patients developed stones. No patient had symptoms referrable to their gall stones. In 31 untreated acromegalic patients, the mean fasting gall bladder volume was similar to that in normal subjects. Maximal percentage emptying, however, was impaired (34 v 64%, p < 0.001) and the mean postprandial residual gall bladder volume increased (21.7 v 9.0 ml, p < 0.001). Treatment with octreotide increased the mean postprandial residual volume further to 36.8 ml (p < 0.001). Gall bladder emptying in untreated acromegalic subjects is impaired. Octreotide further increases postprandial residual gall bladder volume and this may be a factor in the increased gall stone prevalence seen in these patients.

Possible mechanism and treatment of o,p'DDD-induced hypercholesterolaemia.

Ortho,para,dichlorodiphenyl dichloroethane (o,p'DDD, Mitotane (Roussell)) is used as an adrenolytic drug to reduce adrenocortical mass and circulating cortisol levels in Cushing's syndrome but has the unwanted side-effect of inducing hypercholesterolaemia. This paper examined the mechanism of that effect in 30 patients with Cushing's syndrome treated with o,p'DDD during the past 10 years. o,p'DDD increased serum cholesterol by 68 per cent, mainly by increasing LDL-cholesterol. The latter effect was not due to impaired binding of LDL to its receptor, as shown in vitro using cultured fibroblasts. Increases in plasma mevalonic acid during o,p'DDD administration were suggestive of increased cholesterol synthesis, this effect being reversed by simvastatin. These findings suggest that o,p'DDD causes hypercholesterolaemia by increasing cholesterol synthesis. It is proposed that this effect is due to the drug's known ability to block cytochrome P450-mediated reactions, thus impairing the formation of oxysterols responsible for down-regulating hepatic cholesterol synthesis. Treatment with simvastatin, an inhibitor of cholesterol synthesis, reverses the hyperlipidaemia and enables o,p'DDD therapy to be maintained without increasing cardiovascular risk.