KNS4/UPEX1: A Type II Arabinogalactan ß-(1,3)-Galactosyltransferase Required for Pollen Exine Development.

Pollen exine is essential for protection from the environment of the male gametes of seed-producing plants, but its assembly and composition remain poorly understood. We previously characterized Arabidopsis (Arabidopsis thaliana) mutants with abnormal pollen exine structure and morphology that we named kaonashi (kns). Here we describe the identification of the causal gene of kns4 that was found to be a member of the CAZy glycosyltransferase 31 gene family, identical to UNEVEN PATTERN OF EXINE1, and the biochemical characterization of the encoded protein. The characteristic exine phenotype in the kns4 mutant is related to an abnormality of the primexine matrix laid on the surface of developing microspores. Using light microscopy with a combination of type II arabinogalactan (AG) antibodies and staining with the arabinogalactan-protein (AGP)-specific ß-Glc Yariv reagent, we show that the levels of AGPs in the kns4 microspore primexine are considerably diminished, and their location differs from that of wild type, as does the distribution of pectin labeling. Furthermore, kns4 mutants exhibit reduced fertility as indicated by shorter fruit lengths and lower seed set compared to the wild type, confirming that KNS4 is critical for pollen viability and development. KNS4 was heterologously expressed in Nicotiana benthamiana, and was shown to possess ß-(1,3)-galactosyltransferase activity responsible for the synthesis of AG glycans that are present on both AGPs and/or the pectic polysaccharide rhamnogalacturonan I. These data demonstrate that defects in AGP/pectic glycans, caused by disruption of KNS4 function, impact pollen development and viability in Arabidopsis.

Transplantation of adipose-derived mesenchymal stem cell sheets directly into the kidney suppresses the progression of renal injury in a diabetic nephropathy rat model.

AIMS/INTRODUCTION: Adipose-derived mesenchymal stem cell (ASC) transplantation is a promising therapy for diabetic nephropathy (DN). However, intravascular administration of ASCs is associated with low engraftment in target organs. Therefore, we considered applying the cell sheet technology to ASCs. In this study, ASC sheets were directly transplanted into the kidneys of a DN rat model, and therapeutic consequences were analyzed. MATERIALS AND METHODS: Adipose-derived mesenchymal stem cells were isolated from adipose tissues of 7-week-old enhanced green fluorescent protein rats, and ASC sheets were prepared using a temperature-responsive culture dish. A DN rat model was established from 5-week-old Spontaneously Diabetic Torii fatty rats. Seven-week-old DN rats (n = 21) were assigned to one of the following groups: sham-operated (n = 6); ASC suspension (6.0 × 106  cells/mL) administered intravenously (n = 7); six ASC sheets transplanted directly into the kidney (n = 8). The therapeutic effect of the cell sheets was determined based on urinary biomarker expression and histological analyses. RESULTS: The ASC sheets survived under the kidney capsule of the DN rat model for 14 days after transplantation. Furthermore, albuminuria and urinary tumor necrosis factor-α levels were significantly lower in the ASC sheets transplanted directly into the kidney group than in the sham-operated and ASC suspension administered intravenously groups (P < 0.05). Histologically, the ASC sheets transplanted directly into the kidney group presented mild atrophy of the proximal tubule and maintained the renal tubular structure. CONCLUSIONS: Transplantation of ASC sheets directly into the kidney improved transplantation efficiency and suppressed renal injury progression. Therefore, the ASC sheet technology might be a promising novel treatment for DN.

Development of the Mitsucal computer system to identify causal mutation with a high-throughput sequencer.

KEY MESSAGE: Development of Mitsucal. Recent advances in DNA sequencing technology have facilitated whole-genome sequencing of mutants and variants. However, the analyses of large sequence datasets using a computer remain more difficult than operating a sequencer. Forward genetic approach is powerful even in sexual reproduction to identify key genes. Therefore, we developed the Mitsucal computer system for identifying causal genes of mutants, using whole-genome sequence data. Mitsucal includes a user-friendly web interface to configure analysis variables, such as background and crossed strains. Other than configuration, users are only required to upload short reads. All results are presented through a web interface where users can easily obtain a short list of candidate mutations. In the present study, we present three examples of Arabidopsis mutants defective in sexual reproduction in which Mitsucal is used to identify causal mutation. One mutant was screened from seeds of a transgenic line with a reporter gene to elucidate the mechanisms involved in the regulation of seed oil storage. The identified gene codes for a protein may be involved in mRNA splicing. Other two mutants had defects in the surface walls on pollen termed exine. Both causal genes were identified, and mutants were found to be allele of known mutants. These results show that Mitsucal could facilitate identification of causal genes.

Quality of life in Japanese patients with type 1 diabetes and end-stage renal disease undergoing simultaneous pancreas and kidney transplantation.

We conducted this cross-sectional study to assess quality of life (QOL) in Japanese patients with type 1 diabetes mellitus (T1DM) and end-stage renal disease (ESRD) undergoing simultaneous pancreas and kidney transplantation (SPK). Japanese patients with T1DM without diabetic nephropathy (N = 10), and those undergoing chronic dialysis (N = 52), kidney transplantation alone (KTA, N = 25), and SPK (N = 16) were studied. Comprehensive health-related QOL was assessed using the Short Form 36 version 2 (SF-36v2). Emotional functioning in diabetes was measured by the Problem Area In Diabetes (PAID) scale. Severity of impaired hypoglycemic awareness was assessed using the Clarke hypoglycemic score. SPK patients had significantly higher (or tended to have higher) subscale and summary SF-36 scores than dialysis patients and KTA patients. PAID scores were significantly lower in SPK patients than in dialysis patients and KTA patients. Clarke hypoglycemic scores were also significantly lower in SPK patients than dialysis patients. In KTA and dialysis patients, there were no significant differences in the SF-36 subscale/summary scores, PAID scores, or Clarke hypoglycemic scores. In conclusion, QOL for Japanese patients receiving SPK may be superior to that of dialysis patients and KTA patients. Whether SPK actually improves QOL needs to be clarified in longitudinal studies.

Comparative Effects of Statins on the Kidney Function in Patients with Type 2 Diabetes.

AIM: Whether there are differences among statins in their effect on the kidney function in diabetic patients remains controversial. In this report, we aimed to examine the comparative effects of statins on the kidney function in a long-term follow-up study. METHODS: This was a single-center longitudinal observational historical cohort study. We enrolled 326 Japanese adult ambulatory patients with type 2 diabetes who were newly prescribed one of four statins (pravastatin, rosuvastatin, atorvastatin and pitavastatin) and who had an estimated glomerular filtration rate (eGFR) of ≥30 mL/min/1.73m(2). The outcome measurement was the annual rate of change in eGFR. We used the standardized inverse probability of treatment weighted (IPTW) method based on the propensity score to adjust for the effects of confounding factors. Furthermore, in order to take into account the variety in the number and spacing of eGFR measurements and the duration of the follow-up period for each individual, we conducted a linear mixed-effects model regression analysis. RESULTS: The median follow-up period was 4.3years (range, 3.0-7.1years). In an analysis using the IPTW method, the mean (±standard error) annual rate of change in eGFR among the patients treated with pravastatin (-0.86±0.28 mL/min/1.73m(2)/year) was significantly lower than that observed among the patients treated with rosuvastatin (-1.80±0.27, p=0.02), atorvastatin (-1.99± 0.28, p=0.004) and pitavastatin (-2.23±0.49, p=0.02). Similar results were obtained in the linear mixed-effects model regression analysis. CONCLUSIONS: Pravastatin may be superior to rosuvastatin, atorvastatin and pitavastatin in preserving the kidney function in patients with type 2 diabetes.