Basidiomycetous Yeast, Glaciozyma antarctica, Forming Frost-Columnar Colonies on Frozen Medium.

The basidiomycetous yeast, Glaciozyma antarctica, was isolated from various terrestrial materials collected from the Sôya coast, East Antarctica, and formed frost-columnar colonies on agar plates frozen at -1 °C. Thawed colonies were highly viscous, indicating that the yeast produced a large number of extracellular polysaccharides (EPS). G. antarctica was then cultured on frozen media containing red food coloring to observe the dynamics of solutes in unfrozen water; pigments accumulated in frozen yeast colonies, indicating that solutes were concentrated in unfrozen water of yeast colonies. Moreover, the yeast produced a small quantity of ice-binding proteins (IBPs) which inhibited ice crystal growth. Solutes in unfrozen water were considered to accumulate in the pore of frozen colonies. The extracellular IBPs may have held an unfrozen state of medium water after accumulation in the frost-columnar colony.

Characteristics of Industry-Sponsored Drug Clinical Trials Registered in Japan Pharmaceutical Information Center Clinical Trials Information 2010-2018.

PURPOSE: To examine the trends and characteristics of industry-sponsored drug clinical trials registered in the JapicCTI (Japan Pharmaceutical Information Center Clinical Trials Information) in 2010-2018. METHODS: A data set of 3116 clinical trials registered from Jan. 2010 to Dec. 2018 were analyzed. Fundamental characteristics of the clinical trials were analyzed by 3-year time periods. The analysis was also focused on 3 therapeutic areas: cardiovascular, mental health, and oncology. RESULTS: Of all the trials (2010-2018), 74.7% were conducted in Japan only; the rate decreased from 82.8 to 65.3% over the 3 time periods. Most trials were phase 3 trials, which comprised 44.1% of the trials. Small trials (anticipated number of 1000 or fewer participants) made up 94.0% of the trials. Oncology trials (29.5%) were the most common type and involved more phase 1 trials than mental health and cardiovascular trials (33.6% vs 14.5% and 11.5%, respectively). Oncology trials composed the smallest proportion of trials conducted in "Japan only" at 57.3% vs 81.0% and 83.1% for mental health and cardiovascular trials, respectively (p < 0.001). The median of the anticipated number of participants in mental health trials were larger than those in cardiovascular and oncology trials (p = 0.001). Mental health trials were more likely to permit children under age 15 (10.9% vs 4.9% for cardiovascular and 1.2% for oncology). Oncology trials were more likely not to set an upper age limit (89.8% vs 51.4% for cardiovascular and 41.7% for mental health). Cardiovascular and mental health trials were more likely to be conducted as "double blind" (42.4% and 47.1%, respectively vs 16.7% for oncology). CONCLUSION: During this time, the majority of industry-sponsored trials in Japan were phase 3 trials, Japan only and small trials. There were differences in clinical trials among the 3 therapeutic areas: size of the trial, globalization, phase, age of participants, blinding.

Meta-analyses of individual versus group interventions for pre-school children with autism spectrum disorder (ASD).

There is little evidence regarding the effects of individual and group intervention for children with autism spectrum disorder (ASD) on important outcomes. We performed meta-analyses using a random effects model to investigate the effectiveness of the individual and group intervention studies and to compare the effectiveness of these two types if possible. The main analysis which excluded studies at a high risk of bias (Analysis I) included 14 randomised controlled trials targeting children with ASD≤6 years of age (594 children). The results suggested that both individual and group interventions showed significant effects compared to the control condition on "reciprocity of social interaction towards others" (standard mean difference[SMD] [95%confidence interval{CI}] = 0.59[0.25, 0.93], p = 0.16; 0.45[0.02, 0.88], p = 0.39, respectively). Only individual interventions showed significant effects compared to the control condition on "parental synchrony" (SMD [95%CI] = 0.99 [0.70, 1.29], p<0.01). Our results showed no significant differences between individual and group interventions in effects on "autism general symptoms" (no study available for group intervention), "developmental quotient" (no study available for group intervention), "expressive language" (p = 0.56), "receptive language" (p = 0.29), "reciprocity of social interaction towards others" (p = 0.62), or "adaptive behaviour" (p = 0.43). We also performed sensitivity analyses including studies that had been excluded due to being at a high risk of potential bias (Analysis II). The results suggested that "reciprocity of social interactions towards others" showed significant effects for individual intervention compared to the control condition (0.50[0.31,0.69], p<0.001) but not for group intervention (0.23[-0.33, 0.78], p = 0.42). Individual intervention also showed significant effects on "parental synchrony" (0.98[0.30,1.66], p = 0.005) in the sensitivity analysis. The results also suggested no significant difference on all the outcomes between the individual and group interventions. We also reanalysed the data using cluster-robust standard errors as sensitivity analyses (Analysis III). Analysis III showed no significant effects in the intervention condition compared to the control condition on all the outcomes for both individual and group interventions. When Analysis II was reanalysed using cluster-robust standard errors (Analysis IV), individual interventions showed significant effects compared to the control condition on "reciprocity of social interaction towards others" and "parental synchrony" (mean estimate[95%CI], robust standard error, p = 0.50[0.20, 0.81], 0.13, 0.006; and 1.06[0.08, 2.05], 0.42, 0.04, respectively), and none of the outcomes showed significant effects under the intervention condition compared to the control condition for group interventions. The discrepancies in the results between the main analysis (Analysis I) and the sensitivity analyses (Analyses II, III, and IV) may be due to the small number of included studies. Since the outcome of "reciprocity of social interaction towards others" can be a dependent variable that is usually measured in a context-bound setting with the child's parent, we cannot conclude that individual interventions for pre-school children with ASD have significant effects on generalised skills for engaging in reciprocal interactions with others, even if the interventions have significant effects on the outcome. However, the outcomes of "reciprocity of social interaction towards others" may be promising targets for both individual and group interventions involving pre-school children with ASD. "Parental synchrony" may also be a promising target for individual interventions. TRIAL REGISTRATION: (CRD42011001349).

Sinonasal NUT carcinoma: clinicopathological and cytogenetic analysis with autopsy findings.

Nuclear protein in testis (NUT) carcinoma is a rare malignant neoplasm with an undifferentiated morphology. Its diagnosis is often difficult, especially as the sinonasal tract gives rise to many tumors with undifferentiated morphologies. Not many cases of sinonasal NUT carcinomas have been reported, and its clinicopathological features have not been sufficiently clarified. In this study, we performed a clinicopathological study of 4 patients with sinonasal NUT carcinoma, including wide-ranging immunohistochemical tests and cytogenetic analyses using fluorescence in situ hybridization and DNA sequencing. Autopsy findings were obtained from 2 patients. Patients' ages ranged from 9 months to 66 years (median, 37 years). Three cases involved the nasal cavity; of these, 2 also involved the ethmoid sinus. One case only involved the frontal sinus. Histologically, all cases revealed undifferentiated small round cell morphology and necrosis with indistinct cell borders, vesicular chromatin, and distinct nucleoli. All patients received chemoradiotherapy; 3 died of disease 10 to 15 months after their diagnoses, while one was lost to follow-up. The 2 autopsied patients showed multiorgan metastases; interestingly, one showed cartilaginous differentiation in a metastatic lesion. Immunohistochemically, all cases were diffusely positive for NUT, p63, and Myc, and were focal for p40. The cells variably expressed epithelial markers, and CD34 was positive in one patient. Cytogenetically, all showed BRD4-NUT fusion genes, but one had a different breakpoint in each exon. Finally, a literature review indicated that sinonasal NUT carcinoma tends to involve frontal and ethmoidal sinuses more frequently than other sinonasal cancers.

A systematic review and meta-analysis of comprehensive interventions for pre-school children with autism spectrum disorder (ASD).

BACKGROUND: There has an increasing number of published trials on psychosocial intervention programmes for pre-school children with autism spectrum disorder (ASD). To achieve better quality of unbiased evidence for the effectiveness of ASD interventions, it is necessary to conduct a comprehensive review that covers studies with adequate quality standards, such as randomised controlled trials (RCTs), and different types of intervention In this study, we categorize interventions for ASD as behavioural, social-communication focused, and multimodal developmental based on Howlin's classification of early interventions for children with ASD. The aim of this study was to compare these three models and investigate the strengths and weaknesses of each type of intervention and to identify the approaches that contribute to a successful outcome for children with autism. METHODS: We performed a systematic review and meta-analysis. We included RCTs targeting children with ASD 6 years old or younger. A random effects model was used to present the effect estimate for the outcomes. This study also performed combined meta-analyses of all the three models to investigate the overall effectiveness of the intervention programmes. RESULTS: 32 randomized controlled studies were found to be eligible for inclusion. The synthesized data included 594 children from 14 RCTs. There was no statistically significant difference in the effects on autism general symptoms between the social-communication-focused model and the multimodal developmental model (p = 0.83). The results suggest that there is evidence of an effect on 'reciprocity of social interaction towards others' (standard mean difference [95% confidential interval] = 0.53[0.29,0.78], p<0.01) and 'parental synchrony' (SMD = 0.99[0.70,1.29], p<0.01). CONCLUSION: The small number of studies included in the present study limited the ability to make inferences when comparing the three models and investigating the strengths and weaknesses of each type of intervention with respect to important outcomes. Since the outcome of 'reciprocity of social interaction towards others' could be a dependent variable that might be context-bound to interactions with the child's parent, we cannot conclude the interventions for pre-school children with ASD have significant effects on a generalized skill to engage in reciprocal interactions with others. However, the outcomes of 'reciprocity of social interaction towards others' and 'parental synchrony' may be promising targets for interventions involving pre-school children with ASD. TRIAL REGISTRATION: Prospero CRD42011001349.

Quantitative Assessment of Premium Rates for Clinical Usefulness in New Drug Price Calculation in Japan.

BACKGROUND: In Japan, the reimbursement price of a newly approved drug is calculated according to the rule established by the government. As an incentive to innovative Research & Development, a new drug that meets certain criteria obtains premiums on its price. To quantify the clinical value of new drugs in pricing, a point-based system was proposed by an academic study group. This paper gives the background to and overview of the system, and reviews the drugs that gained the premiums after its introduction. METHODS: For drugs to which premiums for innovativeness/usefulness were applied between April 2008 and August 2013, detailed information including the grounds for the premiums was identified. Then, subdivided factors for the requirement of the premium were set and points were allocated to them inductively depending on the degree of clinical impact. Finally, consistency between the rate actually applied and that calculated based on the system were reviewed for new drugs that gained the premiums after its introduction. RESULTS: Forty-seven drugs gained the premium for usefulness between April 2008 and August 2013. Based on the grounds for the premium, a point-based system was established. After its introduction, 11 drugs gained premium for innovativeness/usefulness. The applied rates of premium were consistent with the calculated rate by the system in most cases. CONCLUSIONS: Predictability of future drug price is expected to be enhanced by the point-based system. As the control of health expenditure becomes strict, the importance of considering drug pricing policy that properly reflects the drug's clinical value increases.

Establishment and characterization of two 5-fluorouracil-resistant hepatocellular carcinoma cell lines.

5-Fluorouracil (5-FU) chemotherapy is the first choice treatment for advanced hepatocellular carcinoma (HCC), and resistance is the major obstacle to successful treatment. Recent studies have reported that epithelial-to-mesenchymal transition (EMT) is associated with chemoresistance in cancers. We speculated that EMT and 5-FU metabolism are related to the mechanism of 5-FU resistance. First, two 5-FU-resistant cell lines, HLF-R4 and HLF-R10, were established from the HLF undifferentiated human HCC cell line. Whereas cell growth was similar in the HLF and HLF-R cell lines, HLF-Rs are about 4- and 10-fold more resistant compared with the HLF cells; thus, we named these cell lines HLF-R4 and HLF-R10, respectively. The terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling assay also showed a dramatically decreased number of apoptotic cells in the HLF-Rs after treatment with 5-FU. We next assessed the characteristics of the HLF, HLF-R4 and HLF-R10 cells. Consistent with our hypothesis, the HLF-Rs had typical morphologic phenotypes of EMT, loss of cell-cell adhesion, spindle-shaped morphology and increased formation of pseudopodia. Real-time quantitative reverse transcriptase polymerase chain reaction data showed downregulated E-cadherin and upregulated Twist-1 and also indicated that EMT changes occurred in the HLF-Rs. We also found decreased ribonucleotide reductase and increased multidrug resistance protein 5 genes in the HLF-R cells. Our results suggested that the metabolism of EMT and 5-FU has important roles in 5-FU chemoresistance in the HLF-R cells, and that the HLF-R cells would be useful in vitro models for understanding the 5-FU-resistant mechanisms in HCC.

Doubling of serum creatinine: is it appropriate as the endpoint for CKD? Proposal of a new surrogate endpoint based on the reciprocal of serum creatinine.

BACKGROUND: The evaluation of the progression of renal insufficiency, or decline in glomerular filtration rate (GFR), has been approached more simply and precisely by converting measured serum creatinine value into the reciprocal of serum creatinine, estimated GFR, or other parameters. Doubling of serum creatinine (simple doubling) is conveniently used as a surrogate endpoint for progression of renal disease but is thought to be biased unfairly by the initial value of serum creatinine (Scr(Int)). We proposed the definite decline in the reciprocal of serum creatinine (2-4 doubling) as a surrogate endpoint, comparing simple doubling with this new endpoint to verify the effect of Scr(Int) on the endpoint. METHODS: For the purpose of comparison between endpoints, 194 patients in a historical cohort of chronic glomerulonephritis were investigated. Kaplan-Meier survival analysis was performed with the composite endpoint of need for dialysis and either simple doubling or 2-4 doubling. Then, the distribution of Scr(Int) was compared between total patients and patients developing each endpoint. RESULTS: The endpoint value of serum creatinine (Scr(End)) with 2-4 doubling was lower than that with simple doubling at Scr(Int) <2.00 mg/dl, and the difference of Scr(End) between simple doubling and 2-4 doubling was larger, as Scr(Int) became lower. In patients reaching simple doubling, Scr(Int) was higher than that of the total patients (1.66 vs. 1.07 mg/dl in median, respectively; p < 0.001). In patients reaching 2-4 doubling, there was no significant difference in Scr(Int). CONCLUSION: Patients with low serum creatinine concentration at baseline had a tendency of prolonged development into simple doubling. In contrast, with 2-4 doubling, there was no bias of Scr(Int).

Patients' attitudes towards generic drug substitution in Japan.

OBJECTIVE: The aim of this study was to evaluate the understanding and attitude of Japanese patients towards generic drug substitutions. METHOD: The subjects were male and female patients, who purchased their prescription medications at a pharmacy. A questionnaire was created to assess their attitudes towards generic drugs. RESULTS: Of 1215 respondents, 68.4% knew the term "generic drugs." The majority of them had the correct understanding only on the following two points: generic drugs are less expensive than the brand name drugs (86.0%) and generic drugs contain the same active ingredients as brand name drugs (71.1%). However, their understanding was poor in other aspects of generic substitution: the availability and accessibility of generic drugs, etc. Only the experience of a previous generic drug substitution was significantly associated with the increased willingness for generic substitution (OR=2.93, CI 1.93-4.44). The main reasons for accepting generic substitutions were recommendations by physicians (48.6%) and by pharmacists (33.1%). CONCLUSION: The public awareness program on generic drugs should be expanded to include more detailed information so that patients obtain the correct understanding of generic substitution. It is critical that physicians and pharmacists have the proper understanding of generic drug substitution and provide the correct information to patients.

Public involvement in pharmacogenomics research: a national survey on patients' attitudes towards pharmacogenomics research and the willingness to donate DNA samples to a DNA bank in Japan.

To assess the attitude of Japanese patients towards pharmacogenomics research and a DNA bank for identifying genomic markers associated with adverse drug reactions (ADRs) and their willingness to donate DNA samples, we conducted a survey of 550 male and female patients. The majority of the respondents showed a positive attitude towards pharmacogenomics research (87.6%) and a DNA bank (75.1%). The willingness to donate DNA samples when experiencing severe ADRs (55.8%) was higher than when taking medications (40.4%). Positive attitudes towards a DNA bank and organ donation were significantly associated with an increased willingness to donate. Though the level of positive attitude in the patient population was higher than that in the general public in our former study (81.0 and 70.4%, respectively), the level of the willingness of patients to donate was 40.4% when taking medications and 55.8% when experiencing severe ADRs which was lower than that of the general public in our former study (45.3 and 61.7%). The results suggested that the level of true willingness in the patient population was lower than that of the general public considering the fictitious situation presented to the public (to suppose that they were patients receiving medication). It is important to assess the willingness of patients who are true potential donors, not the general public.

Expression of Drug-Resistant Factor Genes in Hepatocellular Carcinoma Patients Undergoing Chemotherapy with Platinum Complex by Arterial Infusion.

This study investigated gene expression of drug resistance factors in biopsy tissue samples from hepatocellular carcinoma (HCC) patients undergoing chemotherapy by platinum complex. Liver biopsy was performed to collect tissue from the tumor site (T) and the non-tumor site (NT) prior to the start of treatment. For drug-resistant factors, drug excretion transporters cMOAT and MDR-1, intracellular metal binding protein MT2, DNA repair enzyme ERCC-l and inter-nucleic cell transport protein MVP, were investigated. The comparison of the expression between T and NT indicated a significant decrease of MT2 and MDR-1 in T while a significant increase in ERCC-1 was noted in T. Further, expression was compared between the response cases and non-response cases using the ratios of expression in T to those in NT. The response rate was significantly low in the high expression group when the cutoff value of cMOAT and MT2 was set at 1.5 and 1.0, respectively. Furthermore, when the patients were classified into A group (cMOAT ≧ 1.5 or MT2 ≧ 1.0) and B group (cMOAT < 1.5 and MT2 < 1.0), the response rate of A group was significantly lower than B group when we combined the cutoff values of cMOAT and MT2. It is considered possible to estimate the therapeutic effect of platinum complex at a high probability by combining the expression condition of these two genes.

Public involvement in pharmacogenomics research: a national survey on public attitudes towards pharmacogenomics research and the willingness to donate DNA samples to a DNA bank in Japan.

To assess the attitudes of the Japanese general public towards pharmacogenomics research and a DNA bank for identifying genomic markers associated with ADRs and their willingness to donate DNA samples, we conducted a national survey for 1,103 Japanese adults from the general public, not a patient population. The response rate was 36.8%. The majority of the respondents showed a positive attitude towards pharmacogenomics research (81.0%) and a DNA bank (70.4%). Considering fictitious clinical situations such as taking medications and experiencing ADRs, the willingness to donate DNA samples when experiencing ADRs (61.7%) was higher than when taking medications (45.3%). Older generations were significantly associated with a decreased willingness to donate (OR = 0.45, CI 0.28-0.72 in 50s. OR = 0.49, CI: 0.31-0.77 in 60s). Positive attitudes towards pharmacogenomics research, a DNA bank, blood/bone marrow/organ donation were significantly associated with an increased willingness. However, the respondents had the following concerns regarding a DNA bank: the confidentiality of their personal information, the manner by which research results were utilized and simply the use of their own DNA for research. In order to attain public understanding to overcome these concerns, a process of public awareness should be put into place to emphasize the beneficial aspects of identifying genomic markers associated with ADRs and to address these concerns raised in our study. Further study is needed to assess the willingness of actual patients taking medications in real situations, since the respondents in our study were from the general public, not a patient population, and their willingness was assessed on the condition of assuming that they were patients taking medications.