Exploring types of conversational agents for resolving cancer patients' questions and concerns: Analysis of 100 telephone consultations on breast cancer.

OBJECTIVE: This study was conducted to investigate the types of conversational agents (CA) that can help address questions and concerns ("lay topics" [LTs]). METHODS: We analyzed audio recordings of telephone consultations with 100 breast cancer patients and their families. (1) We identified the content and mode of expression of LTs about breast cancer raised during actual telephone consultations. (2) We checked for the presence of clue information (CI) that can help patients resolve their LTs. RESULTS: None of the 805 LTs of the 100 callers were the same. Treatment-related questions occurred in 70 of the 100 consultations. CIs were present in 52.5% of the LTs. CONCLUSION: The results suggest that chatbots (a type of CA) that offer CIs are more feasible than chatbots that answer each question directly in cancer consultations. Moreover, it is difficult to answer questions directly because preparing answers to all LTs in a breast cancer consultation is challenging owing to LT differences. Therefore, preparing high-quality CIs focused on treatments is required. PRACTICE IMPLICATIONS: An increasing number of cancer patients are seeking information to resolve their LTs. CAs can help supplement the limited human resources available if they are supplied with appropriate CIs.

Serum cell-free DNA concentration as a possible prognostic marker in newly diagnosed diffuse large B-cell lymphoma.

Cell-free DNA (cfDNA) is a fragment of DNA circulating in the blood, and its concentration is often elevated in cancer patients. To investigate the relationships between serum cfDNA concentration and clinical characteristics, including prognosis, we measured serum cfDNA concentration in 114 newly diagnosed lymphoma patients. The cfDNA concentrations in diffuse large B cell lymphoma (DLBCL) (62.5 ng/mL) and follicular lymphoma patients (51.6 ng/mL) were significantly elevated compared to healthy individuals (7.5 ng/mL, P < 0.001). In DLBCL, patients with elevated serum cfDNA (> 38.9 ng/mL) at diagnosis had significantly shorter time-to-progression compared to those without (P = 0.033). The addition of cfDNA concentration to the international prognostic index showed improved predictive power for time-to-progression. Moreover, cfDNA added significant prognostic value to other inflammatory markers such as B symptoms and sIL2R. There was a trend towards shorter progression-free survival and overall survival in patients with elevated cfDNA. Furthermore, B symptoms (P = 0.038), bulky masses (P = 0.031), non-GCB subtype (P = 0.012), and serum sIL-2R levels > 2,000 U/mL (P = 0.012) were associated with higher cfDNA levels. Our study showed that serum cfDNA concentration at diagnosis was associated with certain clinicopathological characteristics, and may be predictive of survival outcomes in DLBCL patients.

Users' evaluation of Japan's cancer information services: process, outcomes, satisfaction and independence.

BACKGROUND: Cancer information service (CIS) programmes are becoming increasingly important because patients need to obtain appropriate information and take an active role in their treatment decisions. Programme evaluation is required to determine the level of satisfaction and quality of experiences of users. The purpose of this study is (1) to identify users' evaluation of CIS programmes by both satisfaction and outcomes that reflect the quality of experience and impact of using the CIS, (2) to examine the related factors of these evaluation outcomes and (3) to analyse the differences of those relations between patients and families. METHOD: The self-reported questionnaire was answered by 447 patients and 216 families of patients who used Cancer Information Support Centres (CISCs) at 16 designated cancer hospitals from January 2016 to April 2016. We developed 12 evaluation items, including satisfaction, experience and the impact of using CISC. RESULTS: Respondents evaluated the CISC highly, especially in terms of overall satisfaction, followed by the counselling process. Immediate access to CISC was the strongest factor affecting outcomes. Patients who wanted to consult about 'disease or symptoms' or 'had no specific problem' tended to provide high scores for some outcomes, but those who wanted to consult about a 'financial problem' or 'discharge or care at home' provided negative scores. These trends were also observed in families but to a more limited extent. CONCLUSION: Users' evaluation of CISCs was sufficiently high in terms of overall satisfaction, showing reasonable scores in outcome levels. Immediate access was the strongest factor affecting outcomes and topics of consultation more directly affected evaluation by patients than by families. The distribution of the scores of the measures and related factors was reasonable. The 12-item measurement tool employed in this study seems to be useful for quality monitoring of the CIS.

Creation and provision of a question and answer resource for esophageal cancer based on medical professionals' reports of patients' and families' views and preferences.

BACKGROUND: In the rapidly-progressing healthcare environment, it is essential to improve treatment quality through continuous clarification of the needs and concerns of esophageal cancer patients and their families. Effective collaboration between information providers and academic associations could help make such clarified information available. METHODS: We analyzed esophageal cancer patients' views and preferences (PVPs) using data that were previously obtained from medical staff in Japan. Based on these PVPs, we created a question and answer (Q&A) resource through collaboration with the Cancer Information Service in Japan (CISJ) and the Japan Esophageal Society (JES). RESULTS: Regarding esophageal cancer, "diet and eating behavior" was the most frequent PVP mentioned by patients and their families, followed by "treatment-related symptoms and adverse effects" and "daily life, recuperation, and survivorship." These PVPs were noted by a wide variety of medical specialties. By analyzing the PVPs, the CISJ developed 11 proposed questions and sent them to the JES, which then created answers based on evidence and clinical-practice-associated consensus. The resultant Q&A resource was uploaded to the CISJ website with mutual linkage to the JES website. CONCLUSIONS: This study showed the usefulness of collecting esophageal-cancer-related PVPs from medical staff and fostering successful collaboration between a cancer-information provider and an academic association. This arrangement may represent a model case for developing a sustainable system that can satisfactorily respond to PVPs regarding other cancers and/or issues.

New Approach for Collecting Cancer Patients' Views and Preferences Through Medical Staff.

PURPOSE: It is crucial for health professionals to understand patients' and families' views and preferences (PVPs) to enhance their adherence to treatments and subsequent satisfaction. Regularly and consistently collecting comprehensive information on the needs and concerns of patients/families and utilizing the information is vital for improving clinical practice and the healthcare environment. As an initial approach, this study aimed to develop a new system for appropriately collecting PVPs regarding cancer from nationwide medical staff and consider the potential utilization of PVPs in clinical practice. METHODS: Web-based anonymous surveys were conducted with medical staff in nationwide cancer care hospitals in Japan. The surveys queried the questions, values, desires, and experiences expressed by cancer patients or their families on five topics, namely two cancer sites (colorectal and esophageal cancers) and three symptoms and signs (lymphedema, urinary symptoms, and tingling/numbness/pain) within the past year. The PVPs were compared to the five topics and staff medical specialties, and those on tingling/numbness/pain were analyzed qualitatively. RESULTS: Among the 904 medical staff who responded to this survey, the PVPs encountered by the staff differed according to the topic and staff medical specialty. Tingling/numbness/pain was the most frequently encountered symptom, and urinary symptoms were the least encountered. Only half or fewer of the medical staff had information available regarding urinary symptoms and tingling/numbness/pain. Further, qualitative content analysis of the expressed PVPs regarding tingling/numbness/pain raised clinical questions on this topic that led to the construction of a "Questions & Answers" section on a public website in Japan. CONCLUSION: This study suggests that collecting PVPs through nationwide cancer-related medical staff might be an efficient way to understand the specific requirements of patients/families. It would also be possible to document PVP trends according to changes in the environments of patients/families by nationwide, consistent, and continuous PVP collection.

Employing multiple-attribute utility technology to evaluate publicity activities for cancer information and counseling programs in Japan.

BACKGROUND: Cancer Information and Support Centers (CISCs), which are set up at designated cancer care hospitals (DCCHs) according to the national cancer control programs in Japan, are expected to provide cancer patients and their families with information and counseling, but their public recognition remains low. Therefore, it is necessary to evaluate the publicity activities of CISCs. METHODS: In this study, we evaluated two types of activities for publicizing CISCs conducted inside and outside one of the DCCHs ('Inside Activity' and 'Outside Activity', respectively) by multi-attribute utility technology (MAUT). Seven types of stakeholders, including physicians (n = 5), nurses (n = 4), CISC cancer counselors (n = 5), executives (n = 5), office department staff (n = 5), and licensed social insurance consultants in the hospital (n = 5), as well as patients and citizens (n = 19) were involved in the evaluation. RESULTS: Eight goals of the program for publicizing CISCs were determined. The utility score for each goal was separately calculated for Inside Activity and Outside Activity, and those of the eight goals were summed up to obtain an overall utility score for each activity. Finally, no substantial difference in overall utility score was found between the activity types (Inside Activity = 82.00 vs. Outside Activity = 81.18). CONCLUSIONS: It is implied that omitting or reducing either activity should be avoided in streamlining publicity activities at the CISCs. Despite the limited external validity of the finding, the methodology presented in this study is widely applicable to evaluate other multifaceted programs under an increasingly tight budget in the oncology setting. As the MAUT approach enables us to obtain numerical data for priority settings, it will contribute to rationalized healthcare management, especially in the oncology setting in which objective evaluations of the national cancer-related control programs are lacking and a broad range of stakeholders is involved.

Development of a New Tool for Better Social Recognition of Cancer Information and Support Activities Under the National Cancer Control Policy in Japan.

Under the recent trend of an increasing number of cancer survivors, there is a need to devise measures for visualization of medical care and public health programs related to cancer control, which will enable better overview of the activities at hospitals and local communities and allow various stakeholders to share the information about such activities. The aim of this study was to propose a new tool for proper implementation of cancer information and support programs provided under the national cancer control policy in Japan. Considering 5 elements reported by Handler et al (macro context, mission, structural capacity, processes, and outcomes), we conducted the focus group discussions to confirm the goals of activities of Cancer Information and Support Centers. Eventually, 2 final goals ("reduction in the number of patients/families having difficulties related to cancer" and "being able to live at ease even after diagnosis of cancer") were identified, accompanied by 5 semifinal goals and 16 prerequisite conditions needed to achieve the final goals, as well as the necessary states and the activity indicators corresponding to them. This tool was utilized by 180 (42.7%) of 422 cancer care hospitals designated by the government of Japan (designated cancer care hospitals [DCCHs]) in 2016 and by 336 (77.1%) of 436 DCCHs in 2018, which were the data at 6 months and 3 years after introduction of the tool, respectively. Thus, the tool for evaluating the Cancer Information and Support Centers' activity presented here is expected to stimulate the stakeholders involved in providing supports in various fields of each local community, to share the final goals, to evaluate the status of their achievement, and to further advance their own activities.

An exploratory study of the efficacy of medical illustration detail for delivering cancer information.

Medical illustrations are an effective means of conveying complex information about cancer to patients and laypersons. However, there are no standard criteria for creating these illustrations. This study used online exploratory research focussed on laypersons to identify the aspects of an illustration that convey the optimal degree of detail and understand these findings. Six illustrations depicting pancreatic cancer and nine depicting cervical cancer, with high, medium, and low levels of detail, were created. A total of 420 participants (male = 210, female = 210, aged 20 years and above) answered an online questionnaire about these illustrations. The results indicated that female participants tended to prefer the same illustration level that they also found most comprehensible. Younger participants (under 40 years) tended to prefer illustrations with a medium level of detail for both cancers. Older participants (40 years and above) tended to prefer illustrations with a high level of detail for both cancers. This study was unable to determine the reasons behind these preferences. However, the researchers hypothesise that older people's preference for higher levels of detail may be due to their greater concerns about cancer and because they find the illustrations to be more effective in conveying relevant information.

A multicenter survey of temporal changes in chemotherapy-induced hair loss in breast cancer patients.

PURPOSE: Many breast cancer patients suffer from chemotherapy-induced hair loss. Accurate information about temporal changes in chemotherapy-induced hair loss is important for supporting patients scheduled to receive chemotherapy, because it helps them to prepare. However, accurate information, on issues such as the frequency of hair loss after chemotherapy, when regrowth starts, the condition of regrown hair, and the frequency of incomplete hair regrowth, is lacking. This study aimed to clarify the long-term temporal changes in chemotherapy-induced hair loss using patient-reported outcomes for chemotherapy-induced hair loss. METHODS: We conducted a multicenter, cross-sectional questionnaire survey. Disease-free patients who had completed adjuvant chemotherapy consisting of anthracycline and/or taxanes for breast cancer within the prior 5 years were enrolled from 47 hospitals and clinics in Japan. Descriptive statistics were obtained in this study. The study is reported according to the STROBE criteria. RESULTS: The response rate was 81.5% (1511/1853), yielding 1478 questionnaires. Hair loss occurred in 99.9% of patients. The mean time from chemotherapy until hair loss was 18.0 days. Regrowth of scalp hair occurred in 98% of patients. The mean time from the completion of chemotherapy to the beginning of regrowth was 3.3 months. Two years after chemotherapy completion, the scalp-hair recovery rate was <30% in approximately 4% of patients, and this rate showed no improvement 5 years after chemotherapy. Eighty-four percent of the patients initially used wigs, decreasing to 47% by 1 year after chemotherapy and 15.2% after 2 years. The mean period of wig use was 12.5 months. However, a few patients were still using wigs 5 years after completing chemotherapy. CONCLUSIONS: Our survey focused on chemotherapy-induced hair loss in breast cancer patients. We believe these results to be useful for patients scheduled to receive chemotherapy.

Cancer information needs according to cancer type: A content analysis of data from Japan's largest cancer information website.

The provision of information about cancer is an important aspect of cancer care. Cancer information provided online is expected to meet the needs of information seekers. Whether information needs vary according to tumor site is largely unknown. We aimed to examine similarities and differences in informational needs by cancer type. Data were collected using a questionnaire administered on Japan's largest cancer information website, "Ganjoho service". A total of 2782 free descriptive responses in the period from April 2012 to December 2017 were analyzed using text-mining software. We identified the top 10 informational need contents, in order of appearance frequency, for eight tumor sites: gastric, colorectal, esophageal, lung, pancreatic, breast, cervical, and prostate cancer. Frequent information needs common to all tumor sites included symptoms, disease stages, treatments, chance of cure, recovery, metastasis, and recurrence. A need for information about diet, pain, side effects of treatments, complementary and alternative medicine was frequent for some tumor sites. Tumor site-specific information should include the following, according to cancer type: information of scirrhous carcinoma for gastric cancer; unusual feces for colorectal cancer; lung X-ray images for lung cancer; early detection for pancreatic cancer; adenocarcinoma, sexual activity, pregnancy, and childbirth for cervical cancer; breast conservation or reconstruction and triple negative cancer for breast cancer; test values and diagnosis and urinary problems for prostate cancer; and hormone therapy for breast and prostate cancer. Cancer information provided online should meet these frequent informational needs, considering similarities and differences of the information required according to tumor site.

Detection of HER2 Amplification in Circulating Tumor Cells of HER2-Negative Gastric Cancer Patients.

A key to the successful use of targeted cancer therapy is the ability to preselect patients who are likely to benefit from the treatment according to molecular markers. Assessment for predicting therapy response is mostly done using tumor biopsies. However, these might not truly represent all of the patient's malignant cells because of tumor heterogeneity and/or clonal evolution during disease progression. One potential strategy that can complement primary tumor biopsy is the analysis of circulating tumor cells (CTCs). In this study, we analyzed CTCs of patients with gastric cancer (GC) to find those who were likely to benefit from trastuzumab therapies. We developed an imaging-based method that enabled CTC identification simultaneously with evaluation of HER2 gene amplification (the 3D-IF-FISH method). Then we performed a study enrolling 101 GC patients in whom we analyzed CTCs by both 3D-IF-FISH and an FDA-approved CellSearch system. As compared with the CellSearch system, 3D-IF-FISH methods identified a higher number of patients whose primary tumors were HER2- but who had HER2+ CTCs, suggesting that the 3D-IF-FISH method is effective in preselecting patients for trastuzumab therapies. To demonstrate this, we performed an exploratory clinical study to evaluate the clinical benefits of trastuzumab treatment for advanced GC patients (n = 15) whose primary tumors were HER2-, but whose CTCs showed HER2 amplification. An interim evaluation after the first stage showed that these preselected patients had response rates comparable to those reported in the trastuzumab-plus-chemotherapy arm of the ToGA study. The present study offers a new, non-invasive strategy to select patients who are likely to benefit from trastuzumab-based therapies, despite their primary biopsy being HER2-negative. (UMIN ID: UMIN000008622).

Stereoselective hydroxylation by CYP2C19 and oxidation by ADH4 in the in vitro metabolism of tivantinib.

1. In prior studies, it has been shown that tivantinib is extensively metabolized in humans to many oxidative metabolites and glucuronides. In order to identify the responsible enzymes, we investigated the in vitro metabolism of tivantinib and its four major circulating metabolites. 2. The primary isoforms involved in the elimination of tivantinib were CYP2C19 and CYP3A4/5. CYP2C19 showed catalytic activity for the formation of M5 (hydroxylated metabolite), but not for M4 (a stereoisomer of M5), whereas CYP3A4/5 catalyzed the formation of both metabolites. For the elimination of M4, M5 and M8 (keto-metabolite), CYP3A4/5 was the major cytochrome P450 isoform and UGT1A9 was mainly involved in the glucuronidation of M4 and M5. 3. ADH4 was identified as one of the major alcohol dehydrogenase isoforms contributing to the formation of M6 (sequential keto-metabolite of M4 and M5) and M8. The substrate preference of ADH for M4, and not M5, was observed in the formation of M6. 4. In conclusion, CYP2C19, CYP3A4/5, UGT1A9 and ADH4 were the primary drug metabolizing enzymes involved in the in vitro metabolism of tivantinib and its metabolites. The stereoselective hydroxylation by CYP2C19 and substrate stereoselectivity of ADH4-catalyzed oxidation in the in vitro metabolism of tivantinib was discovered.

Comprehensive assessment of cancer patients' concerns and the association with quality of life.

OBJECTIVE: Comprehensive assessment of perceived concerns can be used to guide supportive care appropriate to individual cancer patients. This study sought to determine the prevalence of cancer patients' concerns and the degree to which these concerns contribute to patients' quality of life. METHODS: Participants were patients with all types of cancer, who completed an Internet survey questionnaire regarding comprehensive concerns about physical, psychological, psychosocial and economic aspects of having cancer. The questionnaire was based on the newly developed Comprehensive Concerns Assessment Tool and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire. RESULTS: We obtained complete data from 807 patients. Factors related to 'self-management' concerns were the most common (61.2%), followed by concerns about 'psychological symptoms' (48.5%), 'medical information' (46.2%), 'daily living' (29.9%), 'pain' (17.6%), 'constipation' (15.6%) and other 'physical symptoms' (15.2%). Multiple regression analysis revealed that all concerns except those about 'medical information' significantly contributed to quality of life. CONCLUSIONS: Cancer patients' concerns were shown to be multidimensional and significantly associated with quality of life. Thus, assessment of patients' concerns should be multidimensional in nature, and a multidisciplinary care team should help patients improve their quality of life.

Characteristics of elderly cancer patients' concerns and their quality of life in Japan: a Web-based survey.

OBJECTIVE: Cancer incidence and the number of cancer patients are increasing in today's aging society. The purpose of this study was to investigate the characteristics of elderly cancer patients' concerns and examine the association between their concerns and quality of life. METHODS: This was a cross-sectional web-based survey completed by ambulatory cancer patients aged 20 years or older. The questionnaire on cancer patients' concerns, comprehensive concerns assessment tool and the European Organization for Research and Treatment of Cancer QLQ-C30 were distributed to the subjects. Multiple regression analysis was conducted to determine which patients' concerns significantly contributed to their quality of life. RESULTS: The final study population consisted of 807 cancer patients, among whom 243 (30%) were elderly (65 years or older). Elderly cancer patients had particular difficulty with self-management, psychological symptoms and medical information, and the prevalence of their concerns was generally lower than that of younger patients, with the exception of physical symptoms. Multiple types of elderly patients' concerns were independently associated with quality of life. CONCLUSIONS: We found that elderly cancer patients suffered from various concerns, thus multidisciplinary intervention is important for providing them with optimal care. The results of this study suggest that elderly cancer patients' quality of life will improve if their concerns are properly handled.

Rheinanthrone, a metabolite of sennoside A, triggers macrophage activation to decrease aquaporin-3 expression in the colon, causing the laxative effect of rhubarb extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Aquaporin-3 (AQP3) is expressed in mucosal epithelial cells in the colon and is important for regulating fecal water content. We examined the role of AQP3 in the laxative effect of rhubarb extract. METHODS: After orally administering rhubarb extract or its major component (sennoside A) to rats, the fecal water content, AQP3 expression and prostaglandin E2 (PGE2) concentrations in the colon were examined. The mechanism by which sennoside A decreases the expression of AQP3 was examined using the human colon cancer HT-29 cells and macrophage-derived Raw264.7 cells. RESULTS: During diarrhea by rhubarb extract administration, the PGE2 levels in the colon increased while the AQP3 expression significantly decreased. Similar changes were also observed when sennoside A was administered. When sennoside A or its metabolites, rheinanthrone and rhein were added to Raw264.7 cells, a significant increase in the PGE2 concentration was observed only in cells treated with rheinanthrone. Fifteen minutes after adding PGE2 to the HT-29 cells, the AQP3 expression decreased to approximately 40% of the control. When pretreated with indomethacin, sennoside A neither decreased the AQP3 expression nor induced diarrhea. CONCLUSIONS: Sennoside A may decrease AQP3 expression in the colon to inhibit water transport from the luminal to the vascular side, leading to a laxative effect. The decreases in the levels of AQP3 are caused by rheinanthrone, which is a metabolite of sennoside A, this metabolite activates the macrophages in the colon and increases the secretion of PGE2; PGE2 acts as a paracrine factor and decreases AQP3 expression in colon mucosal epithelial cells.

Chronological Change of Resistance to ß-Lactams in Salmonella enterica serovar Infantis Isolated from Broilers in Japan.

Epidemiologic surveillance study was conducted in southern Japan to determine the antimicrobial resistance phenotypes and characterize the ß-lactamase genes and the plasmids harboring these genes in Salmonella enterica serovar Infantis (S. Infantis) isolates from broilers. Between January, 2007 and December, 2008, a total of 1,472 fecal samples were collected and examined at the Laboratory of Veterinary Public Health, Kagoshima University, Japan. In 93 (6.3%) isolates recovered, 33 (35.5%) isolates showed resistance to cefotaxime, an extended-spectrum cephalosporin (ESC), conferred by TEM-20, TEM-52 and CTX-M-25 extended-spectrum ß-lactamases (ESBLs). In addition to ESC-resistance, eight (8.6%) isolates exhibited resistance to cefoxitin mediated by CMY-2 AmpC ß-lactamase. Plasmid analysis and polymerase chain reaction replicon typing revealed the bla TEM-20 and bla CMY-2 genes were associated with IncP plasmids, bla TEM-52 was linked with a non-typable plasmid and bla CTX-M-25 was carried by an IncA/C plasmid. Non-ß-lactam resistance to streptomycin, sulfamethoxazole, and oxytetracycline encoded by the aadA1, sul1, and tet(A) genes, respectively, was found in 86 (92.5%) isolates. Resistance to kanamycin and ofloxacin was exhibited in 12 (12.9%) and 11 (11.8%) isolates, respectively, the former was mediated by aphA1-Iab. These data indicate that S. Infantis isolates producing ESBLs and AmpC ß-lactamase have spread among broiler farms in Japan. These data demonstrated that the incidence of ESC-resistant S. Infantis carrying bla TEM-52 remarkably increased and S. Infantis strains harboring bla CMY-2, bla TEM-20, or bla CTX-M-25 genes emerged from broilers in Japan for the first time in 2007 and 2008.

Embryo development after intracytoplasmic sperm injection can be predicted by assessment of sperm nuclear chromatin.

PURPOSE: To assess the influence of structural differences in sperm nuclei on embryo development in intracytoplasmic sperm injection (ICSI). METHODS: Semen obtained from forty-four infertile patients who underwent ICSI was examined. In assessing blastocyst development, only those patients who had successfully obtained over five fertilized eggs were included to exclude any oocyte factors (n = 22). Spermatozoa were assessed using excitation fluorescence flow cytometry with acridine orange and the sperm chromatin dispersion (SCD) test. RESULTS: There was a significant positive correlation between the COMP values obtained from flow cytometry and blastocyst formation. (r = 0.477, p = 0.025). There was a significant negative correlation between the SCD values representing DNA fragmentation and blastocyst formation. (r = 0.796, p < 0.001). COMP values and SCD values were independent parameters to assess sperm nuclear quality regarding embryo development in vitro (r = 0.224, p = 0.080). CONCLUSION: Results suggest that injection of spermatozoa with fewer disulfide bonds and less nuclear DNA fragmentation could achieve better blastocyst formation in human ICSI. Assessment of sperm chromatin should help to predict embryo development after ICSI.

Human protein factory for converting the transcriptome into an in vitro-expressed proteome,.

Appropriate resources and expression technology necessary for human proteomics on a whole-proteome scale are being developed. We prepared a foundation for simple and efficient production of human proteins using the versatile Gateway vector system. We generated 33,275 human Gateway entry clones for protein synthesis, developed mRNA expression protocols for them and improved the wheat germ cell-free protein synthesis system. We applied this protein expression system to the in vitro expression of 13,364 human proteins and assessed their biological activity in two functional categories. Of the 75 tested phosphatases, 58 (77%) showed biological activity. Several cytokines containing disulfide bonds were produced in an active form in a nonreducing wheat germ cell-free expression system. We also manufactured protein microarrays by direct printing of unpurified in vitro-synthesized proteins and demonstrated their utility. Our 'human protein factory' infrastructure includes the resources and expression technology for in vitro proteome research.