MutMapPlus identified novel mutant alleles of a rice starch branching enzyme IIb gene for fine-tuning of cooked rice texture.

Physicochemical properties of storage starch largely determine rice grain quality and food characteristics. Therefore, modification of starch property is effective to fine-tune cooked rice textures. To obtain new resources with modified starch property as breeding materials, we screened a mutant population of a japonica cultivar Nipponbare and found two independent mutant lines, altered gelatinization (age)1 and age2, with moderate changes in starch gelatinization property. A combination of conventional genetic analyses and the latest mapping method, MutMapPlus, revealed that both of these lines harbour novel independent mutant alleles of starch branching enzyme IIb (BEIIb) gene. In age1, amino acid substitution of Met-723 to Lys completely abolished BEIIb enzyme activity without significant reduction in its protein level. A transposon insertion in an intron of BEIIb gene reduced BEIIb protein level and activity in age2. Production of a series of the mutant lines by combining age alleles and indica-type starch synthase IIa allele established stepwise alteration of the physicochemical properties of starch including apparent amylose content, thermal property, digestibility by α-amylase and branched structures of amylopectin. Consistent with the alteration of starch properties, the results of a sensory evaluation test demonstrated that warm cooked rice of the mutants showed a variety of textures without marked reduction in overall palatability. These results suggest that a series of the mutant lines are capable of manipulation of cooked rice textures.

High Temperature-Induced Expression of Rice α-Amylases in Developing Endosperm Produces Chalky Grains.

Global warming impairs grain filling in rice and reduces starch accumulation in the endosperm, leading to chalky-appearing grains, which damages their market value. We found previously that high temperature-induced expression of starch-lytic α-amylases during ripening is crucial for grain chalkiness. Because the rice genome carries at least eight functional α-amylase genes, identification of the α-amylase(s) that contribute most strongly to the production of chalky grains could accelerate efficient breeding. To identify α-amylase genes responsible for the production of chalky grains, we characterized the histological expression pattern of eight α-amylase genes and the influences of their overexpression on grain appearance and carbohydrate components through a series of experiments with transgenic rice plants. The promoter activity of most α-amylase genes was elevated to various extents at high temperature. Among them, the expression of Amy1A and Amy3C was induced in the internal, especially basal to dorsal, region of developing endosperm, whereas that of Amy3D was confined near the ventral aleurone. These regions coincided with the site of occurrence of chalkiness, which was in clear contrast to conventionally known expression patterns of the enzyme in the scutellum and aleurone during seed germination. Furthermore, overexpression of α-amylase genes, except for Amy3E, in developing endosperm produced various degrees of chalky grains without heat exposure, whereas that of Amy3E yielded normal translucent grains, as was the case in the vector control, even though Amy3E-overexpressing grains contained enhanced α-amylase activities. The weight of the chalky grains was decreased due to reduced amounts of starch, and microscopic observation of the chalky part of these grains revealed that their endosperm consisted of loosely packed round starch granules that had numerous pits on their surface, confirming the hydrolysis of the starch reserve by α-amylases. Moreover, the chalky grains contained increased amounts of soluble sugars including maltooligosaccharides at the expense of starch. The integrated analyses proposed that expression of Amy1A, Amy3C, and Amy3D at the specific regions of the developing endosperm could generate the chalkiness. This finding provides the fundamental knowledge to narrow down the targets for the development of high temperature-tolerant premium rice.

An Activity-Staining Method on Filtration Paper Enables High-Throughput Screening of Temperature-Sensitive and Inactive Mutations of Rice α-Amylase for Improvement of Rice Grain Quality.

α-Amylase is a starch-hydrolyzing enzyme (EC 3.2.1.1) indispensable for germination of cereal seeds, but it is also expressed during the ripening stage. Previous studies demonstrated that the enzyme is activated in developing rice seeds under extremely hot weather and triggers a loss of grain quality by hindering the accumulation of storage starch in the endosperm. Since inactive or, preferably, heat-labile α-amylases are preferable for breeding premium rice, we developed a method for rapid screening of inactive and temperature-sensitive mutants of the enzyme by combining the random mutagenesis by error-prone PCR and an on-filter activity test of the recombinant enzyme expressed by Escherichia coli. This technique was applied to a major α-amylase in the developing seed, Amy3D, and the activity of the isolated mutant enzymes was verified with both the bacteria-expressed recombinant proteins and the extract from the endosperm overexpressing each of them. Then, we identified several substitutions leading to loss of the activity of amino acid residues (Leu28, Asp112, Cys149, Trp201, Asp204, Gly295, Leu300 and Cys342), as well as a variety of heat-sensitive substitutions of Asp83, Asp187 and Glu252. Furthermore, variations of the heat-labile enzymes were created by combining these heat-sensitive mutations. The effects of the respective mutations and their relationship to the structure of the enzyme molecule are discussed.

Unmet supportive needs of cancer patients in an acute care hospital in Japan--a census study.

PURPOSE: Little research has been done on supportive needs of cancer patients in acute hospitals in Japan. This study aims to comprehensively assess the unmet supportive needs of hospitalized cancer patients, as well as literacy and utilization of appropriate professional care. METHODS: All cancer patients (aged 20 to 80 years) who were hospitalized in a university hospital in Tokyo during the designated 3-day period between September 1 and October 31, 2007 were recruited for participation in the study. The M.D. Anderson Symptom Inventory, Brief Cancer-Related Worry Inventory, and Hospital Anxiety and Depression Scale were administered. Patients' knowledge and use of relevant services were evaluated. The results were compared with those of non-cancer patients in the same treatment settings. RESULTS: A total of 125 cancer patients and 59 non-cancer patients were enrolled. Cancer patients and non-cancer patients equally suffered from physical symptoms (15-26% had severe appetite loss, 18-19% had severe dry mouth, and 16-22% had severe pain); however, psychological distress of cancer patients exceeded that of non-cancer patients (28.0% vs 8.5%; p ≤ 0.05). Severe psychological distress was associated with severe worry about future prospects or interpersonal and social issues and presence of two or more severe symptoms. Two thirds of the patients with severe psychological distress knew about the psychiatric division, but only one third actually sought treatment. CONCLUSIONS: Needs related to psychological issues were more prevalent among cancer patients than among non-cancer patients, despite a similar level of physical distress. Special attention should be paid to cancer patients who worry over future prospects or interpersonal and social issues, and those who have two or more severe symptoms.

Physician and nurse attitudes toward artificial hydration for terminally ill cancer patients in Japan: results of 2 nationwide surveys.

This study investigated physician and nurse attitudes toward artificial hydration in terminally ill cancer patients and compared differences in attitudes between these 2 professions and among clinical settings in Japan. The response rate was 53% (584/1,123) for physicians and 79% for nurses (3,328/4,210). More physicians answered that artificial hydration alleviates the sensation of thirst. More palliative care unit physicians and nurses answered that withholding artificial hydration alleviated several physical symptoms. Oncologists answered that artificial hydration alleviated the sensation of thirst and fatigue. Discussion among patient-centered teams and individualized decision making are important. Because the differences identified here are attributable to differences in knowledge of artificial hydration for terminal cancer patients, oncologists should place greater emphasis on the opinion of palliative care specialists. Medical practitioners caring for terminal cancer patients should consider a broader range of views on hydration therapy, with a focus on effective hydration techniques and alternative interventions.

Nurse views of the adequacy of decision making and nurse distress regarding artificial hydration for terminally ill cancer patients: a nationwide survey.

We evaluated nurse views on the adequacy of decision-making discussion among nurses and physicians regarding artificial hydration for terminally ill cancer patients and nurse distress arising from artificial hydration issues, as well as factors related to this distress. A self-administered questionnaire consisting of 4 questions about nurse views of discussions regarding artificial hydration and 6 questions about nurse distress arising from artificial hydration issues was distributed in participating institutions in October 2002 and returned by mail. A total of 3328 responses (79%) were analyzed. Almost half of the nurses felt that discussion of terminal hydration issues was insufficient. Among responses, 39% of oncology nurses and 78% of palliative care unit (PCU) nurses agreed that patients and medical practitioners discuss the issue of artificial hydration adequately, and 49% and 79%, respectively, agreed that medical practitioners discuss the issue of artificial hydration with other physicians adequately. As for distress on behalf of patients and families who refuse artificial hydration, 44% of oncology nurses and 57% of PCU nurses experienced such distress for patients, and 19% and 28% did so for families, respectively. Furthermore, 48% of oncology nurses and 47% of PCU nurses experienced distress arising from disagreements among medical practitioners about withholding artificial hydration, whereas 44% and 43% experienced distress about medical practitioners refusing artificial hydration, respectively. Discussion among care providers regarding artificial hydration is insufficient, particularly in general wards. Medical practitioners caring for terminally ill cancer patients should engage in greater discussion among patient-centered teams and facilitate individualized decision making.

The current status of palliative care teams in Japanese university hospitals: a nationwide questionnaire survey.

GOAL: Although the importance of the palliative care team (PCT) to university hospitals is widely accepted, the issues of palliative care at the national level have not been clarified. We conducted a nationwide survey of the current status of PCTs in all (123) Japanese university hospitals. MATERIALS AND METHODS: In 2003, 2004 and 2005, the authors conducted a self-reporting cross-sectional survey. Questionnaires were mailed to nursing directors and selected PCT members of all Japanese university hospitals. RESULTS: Of 123 hospitals in 2005, 99 (80%) returned the questionnaire; 33% used PCTs, and 11% used certified PCTs. Our findings include: annual number of patients treated by PCTs (83/70 +/- 64, mean/median +/- SD), daily number of patients treated by PCTs (12/11 +/- 14), and days of PCT care per patient (30/30 +/- 22). Certified PCTs treated more patients per year (p = 0.004) and more patients per day (p < 0.001) compared to noncertified PCTs. Over the 3-year period, the number of hospitals utilizing PCTs only slightly increased (2003: 27%, 2004: 29%, 2005: 33%), as did those using certified PCTs (2003: 3%, 2004: 9%, 2005: 11%). In 2005, the reasons for noncertification of PCTs included "lack of physicians who specialize in palliative care (82%)" and "lack of nurses who specialize in palliative care (56%)." CONCLUSIONS: The entire system of palliative care in Japanese university hospitals is currently insufficient. The lack of physicians and nurses who specialize in palliative care is a significant barrier, and therefore, the initiation of a formal training system for these health care professionals is a high priority issue.

Pain management and related factors in advanced cancer patients who initiated opioid therapy in an outpatient setting.

OBJECTIVE: The aim of this study was to clarify the state of pain management in Japanese patients with advanced cancer who initiated opioid therapy in an outpatient setting. METHODS: Interview surveys using questionnaires were conducted and medical records were reviewed. Pain relief was defined as >33% decrease in worst pain intensity score, and significance of early pain relief was investigated in terms of changing self-efficacy for activities of daily living (ADL). Factors related to early pain relief were also investigated. RESULTS: The study was conducted between June and December 2003, on 20 patients (13 women, 7 men; mean age, 59 years). Compared to score at initiation of opioid administration (Numerical Rating Scale, 8.3 +/- 1.3), pain relief was generally insufficient at 2 weeks (early pain relief ratio was only 42%). Patients with sufficient pain relief at 2 weeks displayed significant improvements in numerous ADL functions and symptom-coping efficacy (p = 0.037), confirming the importance of early pain relief. Early pain relief was associated with high frequency of hospital visits before opioid administration and absence of sudden excavation within the first 2 weeks. SIGNIFICANCE OF RESULTS: Result of this study indicated insufficient pain relief at an outpatients setting with advanced cancer patients. In the meantime, patients had who their pain decrease after 2 weeks also had improved ADL and self-efficacy at the same time. These findings suggest that to achieve early pain relief in an outpatient setting, clinical staff must assess pain earlier and inform patients of possible breakthrough pain following opioid administration and available treatments for such occurrences.

Physician- and nurse-reported effects of intravenous hydration therapy on symptoms of terminally ill patients with cancer.

PURPOSE: To clarify physician- and nurse-reported effects of intravenous hydration therapy on symptoms of terminally ill patients with cancer. METHODS: A cross-sectional questionnaire survey of Japanese physicians and nurses. The respondents were requested to report their clinical observations about improvement or deterioration of seven symptoms of terminally ill patients with lung or gastric cancer receiving 0.5-1 L/d intravenous hydration therapy, 1.5-2 L/d intravenous hydration therapy, and reduction of intravenous hydration volume from 1.5-2 L/d to 0.5-1 L/d. RESULTS: The responses from a total of 413 oncologists, 88 palliative care physicians, 2735 oncology nurses, and 593 palliative care nurses were analyzed (response rates, 53% in physicians and 83% in nurses). Fewer than 30% of the respondents in all specialties reported that they often or very often observed improvement of dehydration symptoms with 0.5-1 L/d or 1.5-2 L/d intravenous hydration therapy. Deterioration of fluid retention symptoms was reported by 5.8%-13% of the oncologists and 20%-50% of the other specialists with 0.5-1 L/d intravenous hydration therapy for patients with lung cancer, and by 9.3%-24% of the oncologists and 16%-68% of the other specialties with 1.5-2.0 L/d hydration for patients with gastric cancer. By reducing intravenous hydration volume, 20%-70% of the palliative care physicians and nurses reported that they often or very often observed improvement of fluid retention symptoms, while less than 7.0% of all specialists reported that they often or very often observed deterioration of dehydration symptoms. CONCLUSIONS: The physicians and nurses in both oncology and palliative care settings frequently observed deterioration of fluid retention symptoms with limited benefits in alleviating dehydration symptoms by intravenous hydration therapy for terminally ill patients with cancer. It is suggested that routine use of artificial hydration therapy should not be recommended, and individualized treatment policy based on the comprehensive assessment of each patient's needs is strongly required.

Emotional burden of nurses in palliative sedation therapy.

BACKGROUND: Palliative sedation therapy is often required in terminally ill cancer patients, and may cause emotional burden for nurses. The primary aims of this study were 1) to clarify the levels of nurses' emotional burden related to sedation, and 2) to identify the factors contributing to the burden levels. METHODS: A questionnaire survey of 3187 nurses, with a response rate of 82%. RESULTS: Eighty-two percent of the nurses (n = 2607) had clinical experience in continuous-deep sedation. Thirty per cent reported that they wanted to leave their current work situation due to sedation-related burden (answering occasionally, often, or always). Also, 12% of the nurses stated that being involved in sedation was a burden, 12% that they felt helpless when patients received sedation, 11% that they would avoid a situation in which they had to perform sedation if possible, and 4% that they felt what they had done was of no value when they performed sedation. The higher nurse-perceived burden was significantly associated with shorter clinical experience, nurse-perceived insufficient time in caring for patients, lack of common understanding of sedation between physicians and nurses, team conference unavailability, frequent experience of conflicting wishes for sedation between patient and family, nurse-perceived inadequate interpersonal skills, belief that it was difficult to diagnose refractory symptoms, belief that sedation would hasten death, belief that sedation was ethically indistinguishable from euthanasia, nurse-perceived inadequate coping with their own grief, and nurses' personal values contradictory to sedation therapy. CONCLUSIONS: A significant number of nurses felt serious emotional burden related to sedation. To relieve nurses' emotional burden, we encourage 1) management efforts to reduce work overload, 2) a team approach to resolving conflicting opinions, especially between physicians and nurses, 3) co-ordination of early patient-family meetings to clarify their preferred end-of-life care, 4) education and training about sedation specifically focused on interpersonal skills, systematic approaches to diagnosing refractory symptoms, minimum life-threatening potency in sedation, and ethical principals differentiating sedation from euthanasia, and 5) exploring nurses' personal values through the patient-centered principle.