Initial Experience of Laparoscopic Pectopexy for Apical Prolapse in South Korea.

Pelvic organ prolapse (POP) is the herniation or bulging of the pelvic organs to or beyond the vaginal introitus. POP is a common condition affecting more than half of parous women. We recently performed POP surgery and repair in 37 patients using laparoscopic pectopexy, a new technique for apical prolapse surgery, in women with advanced POP. The surgery was performed by a single surgeon at a single institute. The methods of operation were divided into total laparoscopic hysterectomy with pectopexy, supracervical hysterectomy with pectopexy or pectopexy alone, with additional anterior or posterior colporrhaphy selectively performed. All patients were analyzed in terms of age, body mass index, parity, estimated blood loss (EBL), operation time, intraoperative complications, and postoperative complications. The patients were followed up for at least 6 months after surgery, and the short-term clinical outcomes were analyzed. All operations were performed successfully and without severe intraoperative or postoperative complications. The mean EBL was 84 mL, and the mean operation time was 121 minutes. The operation satisfaction rates were high in most patients. All patients had no recurrence of apical prolapse, de novo urgency, frequency, incontinence, de novo constipation, or ileus. In our experience, laparoscopic pectopexy is a feasible method for apical prolapse repair as it does not have severe intraoperative or postoperative complications and de novo gastrointestinal complications. Considerable follow-up period for possible postoperative events is warranted.

Transcriptome Analysis of Diurnal Gene Expression in Chinese Cabbage.

Plants have developed timing mechanisms that enable them to maintain synchrony with daily environmental events. These timing mechanisms, i.e., circadian clocks, include transcriptional/translational feedback loops that drive 24 h transcriptional rhythms, which underlie oscillations in protein abundance, thus mediating circadian rhythms of behavior, physiology, and metabolism. Circadian clock genes have been investigated in the diploid model plant Arabidopsis thaliana. Crop plants with polyploid genomes-such as Brassica species-have multiple copies of some clock-related genes. Over the last decade, numerous studies have been aimed at identifying and understanding the function of paralogous genes with conserved sequences, or those that diverged during evolution. Brassicarapa's triplicate genomes retain sequence-level collinearity with Arabidopsis. In this study, we used RNA sequencing (RNAseq) to profile the diurnal transcriptome of Brassicarapa seedlings. We identified candidate paralogs of circadian clock-related genes and assessed their expression levels. These genes and their related traits that modulate the diurnal rhythm of gene expression contribute to the adaptation of crop cultivars. Our findings will contribute to the mechanistic study of circadian clock regulation inherent in polyploidy genome crops, which differ from those of model plants, and thus will be useful for future breeding studies using clock genes.

Reduction of GIGANTEA expression in transgenic Brassica rapa enhances salt tolerance.

KEY MESSAGE: Here we report the enhancement of tolerance to salt stress in Brassica rapa (Chinese cabbage) through the RNAi-mediated reduction of GIGANTEA ( GI ) expression. Circadian clocks integrate environmental signals with internal cues to coordinate diverse physiological outputs. The GIGANTEA (GI) gene was first discovered due to its important contribution to photoperiodic flowering and has since been shown to be a critical component of the plant circadian clock and to contribute to multiple environmental stress responses. We show that the GI gene in Brassica rapa (BrGI) is similar to Arabidopsis GI in terms of both expression pattern and function. BrGI functionally rescued the late-flowering phenotype of the Arabidopsis gi-201 loss-of-function mutant. RNAi-mediated suppression of GI expression in Arabidopsis Col-0 and in the Chinese cabbage, B. rapa DH03, increased tolerance to salt stress. Our results demonstrate that the molecular functions of GI described in Arabidopsis are conserved in B. rapa and suggest that manipulation of gene expression through RNAi and transgenic overexpression could enhance tolerance to abiotic stresses and thus improve agricultural crop production.

Mutagenesis Study Reveals the Rim of Catalytic Entry Site of HDAC4 and -5 as the Major Binding Surface of SMRT Corepressor.

Histone deacetylases (HDACs) play a pivotal role in eukaryotic gene expression by modulating the levels of acetylation of chromatin and related transcription factors. In contrast to class I HDACs (HDAC1, -2, -3 and -8), the class IIa HDACs (HDAC4, -5, -7 and -9) harbor cryptic deacetylases activity and recruit the SMRT-HDAC3 complex to repress target genes in vivo. In this regard, the specific interaction between the HDAC domain of class IIa HDACs and the C-terminal region of SMRT repression domain 3 (SRD3c) is known to be critical, but the molecular basis of this interaction has not yet been addressed. Here, we used an extensive mutant screening system, named the "partitioned one- plus two-hybrid system", to isolate SRD3c interaction-defective (SRID) mutants over the entire catalytic domains of HDAC4 (HDAC4c) and -5. The surface presentation of the SRID mutations on the HDAC4c structure revealed that most of the mutations were mapped to the rim surface of the catalytic entry site, strongly suggesting this mutational hot-spot region as the major binding surface of SRD3c. Notably, among the HDAC4c surface residues required for SRD3c binding, some residues (C667, C669, C751, D759, T760 and F871) are present only in class IIa HDACs, providing the molecular basis for the specific interactions between SRD3c and class IIa enzymes. To investigate the functional consequence of SRID mutation, the in vitro HDAC activities of HDAC4 mutants immuno-purified from HEK293 cells were measured. The levels of HDAC activity of the HDAC4c mutants were substantially decreased compared to wild-type. Consistent with this, SRID mutations of HDAC4c prevented the association of HDAC4c with the SMRT-HDAC3 complex in vivo. Our findings may provide structural insight into the binding interface of HDAC4 and -5 with SRD3c, as a novel target to design modulators specific to these enzymes.

Regulation of autoimmune germinal center reactions in lupus-prone BXD2 mice by follicular helper T cells.

BXD2 mice spontaneously develop autoantibodies and subsequent glomerulonephritis, offering a useful animal model to study autoimmune lupus. Although initial studies showed a critical contribution of IL-17 and Th17 cells in mediating autoimmune B cell responses in BXD2 mice, the role of follicular helper T (Tfh) cells remains incompletely understood. We found that both the frequency of Th17 cells and the levels of IL-17 in circulation in BXD2 mice were comparable to those of wild-type. By contrast, the frequency of PD-1+ CXCR5+ Tfh cells was significantly increased in BXD2 mice compared with wild-type mice, while the frequency of PD-1+ CXCR5+ Foxp3+ follicular regulatory T (Tfr) cells was reduced in the former group. The frequency of Tfh cells rather than that of Th17 cells was positively correlated with the frequency of germinal center B cells as well as the levels of autoantibodies to dsDNA. More importantly, CXCR5+ CD4+ T cells isolated from BXD2 mice induced the production of IgG from naïve B cells in an IL-21-dependent manner, while CCR6+ CD4+ T cells failed to do so. These results together demonstrate that Tfh cells rather than Th17 cells contribute to the autoimmune germinal center reactions in BXD2 mice.