Sugarcane for water-limited environments. Variation in stomatal conductance and its genetic correlation with crop productivity.

Stomatal conductance (g(s)) and canopy temperature have been used to estimate plant water status in many crops. The behaviour of g(s) in sugarcane indicates that the internal leaf water status is controlled by regular opening and closing of stomata. A large number of g(s) measurements obtained across varying moisture regimes, locations, and crop cycles with a diverse sugarcane germplasm composed of introgression, and commercial clones indicated that there is a high genetic variation for g(s) that can be exploited in a breeding programme. Regardless of the environmental influences on the expression of this trait, moderate heritability was observed across 51 sets of individual measurements made on replicated trials over 3 years. The clone×water status interaction (G×E) variation was smaller than the clone (G) variation on many occasions. A wide range of genetic correlations (r(g)= -0.29 to 0.94) between g(s) and yield were observed across test environments in all three different production regions used. Canopy conductance (g(c)) based on g(s) and leaf area index (LAI) showed a stronger genetic correlation than the g(s) with cane yield (tonnes of cane per hectare; TCH) at 12 months (mature crop). The regression analysis of input weather data for the duration of measurements showed that the predicted values of r(g) correlated with the maximum temperature (r=0.47) during the measurements and less with other environmental variables. These results confirm that the g(c) could have potential as a criterion for early-stage selection of clones in sugarcane breeding programmes.

Sugarcane for water-limited environments. Genetic variation in cane yield and sugar content in response to water stress.

Water limitation is a major production constraint for sugarcane worldwide. However, to date, there has been little investigation of patterns of genetic variation in the response to water stress in sugarcane. Field experiments were conducted over 3 years under fully irrigated and managed water stress conditions at two locations in Northern Queensland in Australia. Eighty-nine genetically diverse clones were evaluated for their yield performance and sugar attributes. Water stress treatments reduced cane yield [tonnes of cane per hectare (TCH)] and total dry matter (TDM) by 17-52% and 20-56%, respectively, compared with irrigated treatments in the same experiments. Nevertheless, there was little genotype×environment interaction variation for TCH, TDM, or commercial cane sugar (CCS), and hence high genetic correlations between the irrigated and water stress treatments across environments. Both commercial and unselected clones performed poorly under severe stress environments, while the commercial clones outperformed the unselected clones under mild and moderate stress conditions. The results presented here highlight the contribution of intrinsic potential yields (yield under well-irrigated conditions) of some selected and unselected clones to maintain relatively high productivity in a range of moderate stress conditions imposed. The physiological basis for the high genetic correlations is at present unclear, but some explanations are hypothesized. The choice of stress levels in selection trials would not appear to be a critical issue for sugarcane breeding programmes, at least for the early phases of selection, where similar ranking clones across a range of moderate water stresses may be expected.

Multicentre evaluation of intraoperative molecular analysis of sentinel lymph nodes in breast carcinoma.

BACKGROUND: Ideally, intraoperative sentinel lymph node (SLN) analysis in breast cancer should be automated, have high concordance with extensive histopathology, and be applicable in any hospital setting. A prospective multicentre evaluation of the one-step nucleic acid amplification (OSNA) automated molecular diagnostic system of SLN analysis was undertaken. METHODS: Intraoperative examination of SLNs from 204 patients with breast cancer was performed by OSNA at four sites in the UK. Half of each SLN was assessed by OSNA (for cytokeratin 19 mRNA) and the remaining half was paraffin embedded for intensive histological examination at ten levels. Discordant cases were reanalysed by further molecular biological techniques and by additional histological examination of all remaining nodal material to ascertain whether the discordance was due to an uneven distribution of metastases, known as tissue allocation bias (TAB). RESULTS: After exclusion of samples affected by TAB, the overall concordance rate for OSNA versus histopathology was 96.0 per cent, with a sensitivity of 91.7 per cent and a specificity of 96·9 per cent. The median time to process a single SLN was 32 (range 22-97) min, and that for two nodes 42 (30-73) min. CONCLUSION: OSNA enables accurate automated intraoperative diagnosis and can be used successfully in different UK hospitals. When the SLN is shown to be positive, the patient can undergo immediate axillary clearance under the same anaesthetic rather than having a delayed second procedure.

Acute and chronic effects of multivitamin/mineral supplementation on objective and subjective energy measures.

BACKGROUND: Vitamins and minerals play an essential role within many cellular processes including energy production and metabolism. Previously, supplementation with a multivitamin/mineral (MVM) for ≥28 days resulted in improvements to cognition and subjective state. We have also demonstrated shifts in metabolism during cognitively demanding tasks following MVM in females, both acutely and following 8-week supplementation. The current study aimed to assess these effects further in males and females using metabolically challenging exercise and cognitive tasks. METHODS: The current randomised, placebo-controlled, parallel groups study investigated the effects of a MVM complex in 82 healthy young (18-35y) exercisers. Subjective ratings and substrate metabolism were assessed during 30 min each of increasingly effortful incremental exercise and demanding cognitive tasks. Assessments took place on acute study days following a single dose (Day 1) of MVM, containing 3 times recommended daily allowance of water-soluble vitamins plus CoQ10, and following 4-week supplementation (Day 28). RESULTS: Energy expenditure (EE) was increased during cognitive tasks following MVM across Day 1 and Day 28, with greater effects in males. In males, MVM also increased carbohydrate oxidation and energy expenditure during exercise across Day 1 and Day 28. In females, mental tiredness was lower during exercise; increases in physical tiredness following 30 min of exercise were attenuated; and stress ratings following cognitive tasks were reduced following MVM. In males, MVM only lowered mental tiredness following 10 min of exercise. These effects were apparent irrespective of day, but effects on mental tiredness were greater on Day 28. Ferritin levels were also higher on Day 28 in those receiving MVM. CONCLUSION: These findings extend on existing knowledge, demonstrating increased carbohydrate oxidation and increased energy expenditure in males following MVM supplementation for the first time. Importantly, they show modulation of energy expenditure and subjective tiredness following a single dose, providing further evidence for acute effects of MVM. Differential effects in men and women suggest that sex may play an important role in the effects of MVM on energy metabolism and should be considered in future research. TRIAL REGISTRATION: ClinicalTrials.gov, NCT03003442. Registered 22nd November 2016 - retrospectively registered.

Evaluation of the dissemination and implementation of a nutritional guideline for pressure ulcer care.

OBJECTIVE: In 2004 the European Pressure Ulcer Advisory Panel nutritional working group developed a nutritional guideline for pressure ulcer prevention and treatment. This study investigated the degree to which the guideline was disseminated and implemented in clinical practice. METHOD: A cross-sectional study was undertaken in health-care organisations in The Netherlands, Germany and the UK. A printed, standardised questionnaire which followed Rogers' model of the innovation-decision process was developed, translated and distributed to 1087 health-care organisations. RESULTS: The response rate was 33% (n = 363). Sixty-one per cent of respondents knew of the guideline. Twenty-five per cent had applied it to their clinical practice and used it for nutritional screening. The main barrier to the provision of nutritional support appeared to be lack of knowledge and skills. CONCLUSION: One year after its dissemination, more than half of respondents knew of the guideline, with one in four applying it to their practice. The guideline was better disseminated and implemented in The Netherlands and UK than in Germany, where only 4% of participants had used it.

Effect of vagal cooling on the counterregulatory response to hypoglycemia induced by a low dose of insulin in the conscious dog.

We previously demonstrated, using a nerve-cooling technique, that the vagus nerves are not essential for the counterregulatory response to hypoglycemia caused by high levels of insulin. Because high insulin levels per se augment the central nervous system response to hypoglycemia, the question arises whether afferent nerve fibers traveling along the vagus nerves would play a role in the defense of hypoglycemia in the presence of a more moderate insulin level. To address this issue, we studied two groups of conscious 18-h-fasted dogs with cooling coils previously placed on both vagus nerves. Each study consisted of a 100-min equilibration period, a 40-min basal period, and a 150-min hypoglycemic period. Glucose was lowered using a glycogen phosphorylase inhibitor and a low dose of insulin infused into the portal vein (0.7 mU.kg(-1) min(-1)). The arterial plasma insulin level increased to 15 +/- 2 microU/ml and the plasma glucose level fell to a plateau of 57 +/- 3 mg/dl in both groups. The vagal cooling coils were perfused with a 37 degrees C (SHAM COOL; n = 7) or a -20 degrees C (COOL; n = 7) ethanol solution for the last 90 min of the study to block parasympathetic afferent fibers. Vagal cooling caused a marked increase in the heart rate and blocked the hypoglycemia-induced increase in the arterial pancreatic polypeptide level. The average increments in glucagon (pg/ml), epinephrine (pg/ml), norepinephrine (pg/ml), cortisol (microg/dl), glucose production (mg.kg(-1). min(-1)), and glycerol (micromol/l) in the SHAM COOL group were 53 +/- 9, 625 +/- 186, 131 +/- 48, 4.63 +/- 1.05, -0.79 +/- 0.24, and 101 +/- 18, respectively, and in the COOL group, the increments were 39 +/- 7, 837 +/- 235, 93 +/- 39, 6.28 +/- 1.03 (P < 0.05), -0.80 +/- 0.20, and 73 +/- 29, respectively. Based on these data, we conclude that, even in the absence of high insulin concentrations, afferent signaling via the vagus nerves is not required for a normal counterregulatory response to hypoglycemia.

Preparation of human antibodies to insulin.

Insulin was directly coupled and also indirectly coupled by a side chain to Sepharose 4B to form immunoabsorbents for affinity chromatography of insulin antibodies. The two direct derivatives were insulin lysyl and insulin phenylalanyl sepharose. The indirect derivative, insulin carboxyhexyl sepharose, was prepared in two ways: by a carbodiimide procedure and by an N-hydroxysuccinimide ester method. Insulin carboxyhexyl sepharose prepared by the latter method was found to be the most satisfactory material for affinity chromatography and was used to isolate insulin antibodies from human serum.

A comparison of large block macrosectioning and conventional techniques in breast pathology.

Two techniques for the histological handling of breast specimens, namely conventional sampling using small blocks (SB) and a large block (LB) macrosectioning technique have been compared, with special emphasis on tumour size and in situ carcinoma, in an attempt to objectively demonstrate the advantages of the latter method. This is considered to be of particular importance in guiding the clinicians in their use of the many treatment modalities available for breast carcinoma. All cases were from the routine surgical caseload; 100 examined by the LB technique and 111 using conventional SB. The LB technique gave a reliable measurement of invasive carcinoma in 100% of cases compared to only 63% of SB cases. Ductal carcinoma in situ (DCIS), was found more frequently (80% versus 64%) and its extent was more easily and reliably measured in the large blocks. The extent of DCIS was significantly greater in all cases using large block techniques. Concurrent carcinomas were found more frequently in the LB series and these tumours were smaller than those in the SB series. Similar differences were noted with radial scars, and other proliferative lesions also had a higher incidence in the LB series. We conclude that the LB technique has sufficient advantages to recommend it as a standard technique in breast pathology.

Relation between stage, grade, proliferation, and expression of p53 and CD44 in adenomas and carcinomas of the colorectum.

AIMS: To investigate the changes in and relations among p53, CD44 and MIB-1 expression in adenocarcinomas of the colorectum and to determine whether these changes are progressive across the adenoma-carcinoma sequence. METHODS: Expression of p53 protein, CD44 adhesion molecule and MIB-1 proliferation antigen was detected using immunohistochemistry in 68 colorectal carcinomas and 32 colorectal adenoma. The staining characteristics were compared with degree of dysplasia in adenomas, and differentiation and Dukes' stage in carcinomas. Results were analysed and assessed using Spearman's rank correlation and independent t tests. RESULTS: p53 staining was present in som adenomas and correlated with the degree of dysplasia. There was significantly more staining in carcinomas than adenomas and significant correlation between staining and Dukes' stage. CD44 staining was maximal in adenomas, diminished in carcinomas and was minimal in metastasising carcinomas. There was inverse correlation between p53 and CD44 expression across the adenoma-carcinoma-metastasising carcinoma sequence. MIB-1 expression was highest in carcinomas but did not correlate with either p53 or CD44 expression. CONCLUSIONS: There are progressive changes in p53, CD44 and MIB-1 expression in adenomas and carcinomas. A combination of these tests may prove useful in assessing which patients with adenomas are at greatest risk of progressing to carcinoma.

Portal glucose infusion increases hepatic glycogen deposition in conscious unrestrained rats.

It has been demonstrated in the conscious dog that portal glucose infusion creates a signal that increases net hepatic glucose uptake and hepatic glycogen deposition. Experiments leading to an understanding of the mechanism by which this change occurs will be facilitated if this finding can be reproduced in the rat. Rats weighing 275-300 g were implanted with four indwelling catheters (one in the portal vein, one in the left carotid artery, and two in the right jugular vein) that were externalized between the scapulae. The rats were studied in a conscious, unrestrained condition 7 days after surgery, following a 24-h fast. Each experiment consisted of a 30- to 60-min equilibration, a 30-min baseline, and a 120-min test period. In the test period, a pancreatic clamp was performed by using somatostatin, insulin, and glucagon. Glucose was given simultaneously either through the jugular vein to clamp the arterial blood level at 220 mg/dl (Pe low group) or at 250 mg/dl (Pe high group), or via the hepatic portal vein (Po group; 6 mg. kg(-1). min(-1)) and the jugular vein to clamp the arterial blood glucose level to 220 mg/dl. In the test period, the arterial plasma glucagon and insulin levels were not significantly different in the three groups (36 +/- 2, 33 +/- 2, and 30 +/- 2 pg/ml and 1.34 +/- 0.08, 1. 37 +/- 0.18, and 1.66 +/- 0.11 ng/ml in Po, Pe low, and Pe high groups, respectively). The arterial blood glucose levels during the test period were 224 +/- 4 mg/dl for Po, 220 +/- 3 for Pe low, and 255 +/- 2 for Pe high group. The liver glycogen content (micromol glucose/g liver) in the two Pe groups was not statistically different (51 +/- 7 and 65 +/- 8, respectively), whereas the glycogen level in the Po group was significantly greater (93 +/- 9, P < 0.05). Because portal glucose delivery also augments hepatic glycogen deposition in the rat, as it does in the dogs, mechanistic studies relating to its function can now be undertaken in this species.

A review of individual psychotherapy outcomes for adult survivors of childhood sexual abuse.

This review examines the current literature on individual psychotherapy outcomes with adult survivors of childhood sexual abuse. As compared to the literature on treatment with victims of sexual trauma in adulthood, fewer researchers have explored the field of psychotherapy outcome with this population, particularly with regard to individual treatments. For this reason, a review of psychotherapy outcome for adult survivors of child sexual abuse is needed. This review first presents issues salient to the study of treatment with this population, such as prevalence, short-term impact, and long-term sequelae of childhood sexual trauma. Next, the eight studies that have been published on individual psychotherapy for this population are presented and evaluated according to both efficacy and effectiveness criteria for a methodologically sound study. Last, directions for future research with this population include continued integration of efficacy and effectiveness methodology, use of multi-method/multi-rater assessment data, as well as further investigation of interpersonal variables such as the therapeutic alliance.

Lobular carcinoma of the breast: cytological features supporting the diagnosis of lobular cancer.

Lobular carcinomas have a distinct natural history with a better response to endocrine therapy and a higher incidence of local recurrence and are more often bilateral. The cytological diagnosis of lobular carcinoma permits a discriminating therapeutic approach with pre-operative Tamoxifen, more generous resection margins, and assessment of the contralateral breast. The cytological features of lobular cancer however are not well defined and the low cell yield from such neoplasms can result in a high false negative rate. To determine whether we could improve the pre-operative diagnosis, we reviewed the cytological features of 112 lobular carcinomas. They had small uniform sized nuclei with irregular outlines and inconspicuous nucleoli. The degree of dissociation was similar to duct carcinomas and the incidence of inadequate aspirates was no higher. We found no features that confidently diagnosed lobular cancer or its sub-types. However, using a combination of features, typing of lobular cancer on aspirated material is possible and should be attempted.

Clinical indications for echocardiography.

Echocardiography is an important adjunct to clinical diagnosis. Evaluation of chamber size, estimation of left ventricular function, valvular defects, and tricuspid atresia or stenosis are among the many disorders in which it can be helpful.

Efficacy of arthroscopic surgery and midlaser treatments for chronic temporomandibular joint articular disc derangement following motor vehicle accident.

As a result of motor vehicle accident soft-tissue injury, temporomandibular joint articular disc derangement may develop and persist despite symptomatic treatment and medication. This study reports the effectiveness of management directed at controlling the TMJ and masticatory neuromuscular pain dysfunction with a TMJ/interocclusal stabilization appliance, specific biofeedback and ultrasound therapy. Following these conservative measures residual articular disc derangement was present in some subjects who were offered arthroscopic surgery and infrared midlaser with TMJ/occlusal stabilization. Twenty subjects with residual disc derangement were randomly selected into two groups with and without arthroscopic surgery, and analyses of variance made before treatment, 12 months after conservative procedures, 3 months following arthroscopic surgery and midlaser therapy and 3 years since commencement of management. Dependent variables compared were pain-discomfort, Clinical Dysfunction Index, articular disc derangement and maximal voluntary jaw opening. Conservative management alone provided significant reduction of pain-discomfort and clinical dysfunction, while arthroscopic surgery resulted in significant reduction in articular disc derangement. The midlaser with TMJ/occlusal stabilization maintained significant improvement in the variables (p < 0.01) for both groups. The common articular deviations in form found at arthroscopy were soft tissue alteration with hyperaemia, synovitis, synovial membrane and posterior attachment folding with connective tissue hyperplasia, and disc displacement with fibrous adhesions. The Global Status Score of pain behaviour compared with residual function, confirmed the presence of greater pain before treatment commenced.

Jewish inmates: an atypical prison group.

12 Jewish inmates scored significantly higher than 1497 non-Jewish inmates on a measure of intelligence, the Raven's Standard Progressive Matrices. Interview findings indicated that the present Jewish inmates were never very involved in the practice of Judaisn and were not strongly integrated into their local Jewish communities.

A differential intra-operative molecular biological test for the detection of sentinel lymph node metastases in breast carcinoma. An extended experience from the first U.K. centre routinely offering the service in clinical practice.

INTRODUCTION: One-Step Nucleic acid Amplification (OSNA) is a molecular biological assay of cytokeratin-19 (a breast epithelial marker) mRNA. It can be employed intra-operatively for detection of lymph node metastases in breast carcinoma. Patients with positive sentinel nodes may proceed to axillary lymph node dissection (ALND) level I or higher dependent upon the OSNA quantitative result, during the same surgical procedure, avoiding a second operation and eliminating the technical difficulties possibly associated with delayed ALND. AIMS: Our Breast Unit was the first in the UK to implement this novel technique in routine practice. This study reviews our first 44-month data following introduction of OSNA "live" on whole sentinel nodes following an extensive validation study (Snook et al.).(9) METHODS: Data was collected prospectively from the period of introduction 01/12/2008 to 30/08/2012. All patients eligible for sentinel node biopsy were offered OSNA and operations were performed by five consultant breast surgeons. On detection of macro-metastasis a level II/III and for a micro-metastasis a level I ALND was performed. RESULTS: A total of 859 patients (1709 sentinel lymph nodes) were analysed. All except one were females. The majority underwent wide local excision (73.4%, n = 631) or mastectomy 25% (n = 215) and 1.6% (13) underwent SLN biopsy alone. IDC was seen in 79% (n = 680) of the patients and 53.5% (n = 460) had grade II tumours. One-third (30.8%, n = 265) had positive sentinel nodes and had further axillary surgery at the time of SLN biopsy. Of these, 47% (n = 125/265) had macro-metastases, 38% (n = 101/265) had micro-metastases and 14.7% (n = 39/265) had "positive but inhibited" results. Positive non-sentinel lymph nodes (NSLN) were seen in 35% (44/125) of those with macro-metastases; 17.8% (18/101) of the patients with micro-metastases and 10.2% (4/39) of the "positive but inhibited" group. CONCLUSION: In our series over a third of our patients had positive lymph nodes detected with OSNA allowing them to proceed directly to axillary surgery at the same operation. This technique eliminates the need for a second operation in sentinel lymph node positive patients and avoids the anxiety waiting for histological results.

Genetic control of yield related stalk traits in sugarcane.

A major focus of sugarcane variety improvement programs is to increase sugar yield, which can be accomplished by either increasing the sugar content of the cane or by increasing cane yield, as the correlation between these traits is low. We used a cross between an Australian sugarcane variety Q165, and a Saccharum officinarum accession, IJ76-514, to dissect the inheritance of yield-related traits in the complex polyploid sugarcane. A population of 227 individuals was grown in a replicated field trial and evaluated over 3 years for stalk weight, stalk diameter, stalk number, stalk length and total biomass. Over 1,000 AFLP and SSR markers were scored across the population and used to identify quantitative trait loci (QTL). In total, 27 regions were found that were significant at the 5% threshold using permutation tests with at least one trait; individually, they explained from 4 to 10% of the phenotypic variation and up to 46% were consistent across years. With the inclusion of digeneic interactions, from 28 to 60% of the variation was explained for these traits. The 27 genomic regions were located on 22 linkage groups (LGs) in six of the eight homology groups (HGs) indicating that a number of alleles or quantitative trait alleles (QTA) at each QTL contribute to the trait; from one to three alleles had an effect on the traits for each QTL identified. Alleles of a candidate gene, TEOSINTE BRANCHED 1 (TB1), the major gene controlling branching in maize, were mapped in this population using either an SSR or SNP markers. Two alleles showed some association with stalk number, but unlike maize, TB1 is not a major gene controlling branching in sugarcane but only has a minor and variable effect.