Composition of Vaginal Microbiota in Pregnant Women With Aerobic Vaginitis.

Vaginal dysbiosis, such as bacterial vaginosis (BV) and aerobic vaginitis (AV), is an important cause of premature birth in pregnant women. However, there is very little research on vaginal microbial distribution in AV compared to that in BV. This study aimed to analyze the composition of the vaginal microbiota of pregnant women with AV using microbial community analysis and identify the causative organism using each criterion of the AV scoring system. Also, we compared the quantification of aerobic bacteria using quantitative polymerase chain reaction (qPCR) and their relative abundances (RA) using metagenomics. This prospective case-control study included 228 pregnant Korean women from our previous study. A wet mount test was conducted on 159 women to diagnose AV using the AV scoring system. Vaginal samples were analyzed using metagenomics, Gram staining for Nugent score determination, conventional culture, and qPCR for Staphylococcus spp., Streptococcus spp., and Enterobacteriaceae. The relative abundances (RAs) of eleven species showed significant differences among the three groups (Normal flora (NF), mild AV, and moderate AV). Three species including Lactobacillus crispatus were significantly lower in the AV groups than in the NF group, while eight species were higher in the AV groups, particularly moderate AV. The decrease in the RA of L. crispatus was common in three criteria of the AV scoring system (Lactobacillary, WBC, and background flora grades), while it did not show a significant difference among the three grade groups of the toxic leukocyte criterion. Also, the RAs of anaerobes, such as Gardnerella and Megasphaera, were higher in the AV groups, particularly moderate AV, while the RAs of aerobes were very low (RA < 0.01). Therefore, qPCR was performed for aerobes (Staphylococcus spp., Streptococcus spp., and Enterobacteriaceae); however, their quantification did not show a higher level in the AV groups when compared to that in the NF group. Therefore, AV might be affected by the RA of Lactobacillus spp. and the main anaerobes, such as Gardnerella spp. Activation of leukocytes under specific conditions might convert them to toxic leukocytes, despite high levels of L. crispatus. Thus, the pathogenesis of AV can be evaluated under such conditions.

A rare case of ovarian vein thrombosis in a gestational trophoblastic neoplasia patient.

Ovarian vein thrombosis (OVT) is a rare disease with complications that can be life-threatening. An ovarian vein thrombus in a gestational trophoblastic neoplasia (GTN) is an extremely rare condition that has not been previously reported in the literature. We report the case of a 23-year-old woman who presented with symptoms of amenorrhea for 15 weeks and 6 days along with intermittent lower abdominal pain. She was diagnosed with a hydatidiform mole, and a metastatic workup was scheduled. Abdominal computed tomography showed a right ovarian vein thrombus. She received methotrexate chemotherapy combined with oral anticoagulants. Complete radiological remission was obtained. During the 12-month follow-up period, no disease progression or recurrence was noted. Early recognition and detection of the condition are of the utmost importance. The differential diagnosis of OVT must be considered when there is unexplained abdominal pain, fever, and leukocytosis during the diagnosis and treatment of GTN. A high level of suspicion is required for prompt diagnosis of OVT.

Postoperative outcomes of natural orifice transluminal endoscopic surgery-assisted vaginal hysterectomy and conventional laparoscopic-assisted vaginal hysterectomy: a comparative study.

OBJECTIVE: The present study aimed to determine the differences in outcomes between natural orifice transluminal endoscopic surgery-assisted vaginal hysterectomy (NAVH) and conventional laparoscopy-assisted vaginal hysterectomy (LAVH). METHODS: We retrospectively reviewed the charts of patients who between July 2012 and September 2015, were diagnosed as having benign uterine disease such as uterine myoma, endometriosis, or adenomyosis and managed via NAVH or LAVH in a single-center (Eulji University Hospital). Data such as age, body weight, height, parity, operation time, intra/post-operative complications, and uterus weight were obtained from the clinical charts. NAVH and LAVH recipients were matched 1:3 in terms of baseline characteristics, and the 2 groups were compared regarding surgical outcomes. RESULTS: Of the 160 patients with benign uterine disease included in the present study. Forty received NAVH and remaining 120 received LAVH. There were significant differences between the groups regarding operation time and hemoglobin change. Notably, although the operation time was shorter for LAVH, hemoglobin change was lower for NAVH. Additionally, although maximum hospitalization duration was shorter for LAVH, the average length of hospitalization was similar between NAVH and LAVH. There were no significant differences between the groups in terms of other variables. CONCLUSION: NAVH may become a new alternative surgical method of choice for hysterectomy, as it represents a clinically feasible and safe approach; moreover is superior to LAVH in terms of bleeding loss.

Predominant Lactobacillus species types of vaginal microbiota in pregnant Korean women: quantification of the five Lactobacillus species and two anaerobes.

OBJECTIVE: To investigate the predominant Lactobacillus species types (LSTs) of vaginal microbiota in pregnant Korean women by quantifying five Lactobacillus species and two anaerobes. METHODS: In all, 168 pregnant Korean women under antenatal care at Eulji University Hospital and local clinics were enrolled in the prospective cohort study during pregnancy (10-14 weeks). Vaginal samples were collected with Eswab for Quantitative polymerase chain reaction (qPCR) and stored in a -80 °C freezer. qPCR was performed for five Lactobacillus species and two anaerobes. To identify the predominant LSTs, quantifications were analyzed by the Cluster and Tree View programs of Eisen Lab. Also the quantifications were compared among classified groups. RESULTS: L. crispatus and L. iners were most commonly found in pregnant Korean women, followed by L. gasseri and L. jensenii; L. vaginalis was nearly absent. Five types (four predominant LSTs and one predominant anaerobe type without predominant Lactobacillus species) were classified. CONCLUSIONS: Five predominant LSTs were identified in vaginal microbiota of pregnant Korean women. L. crispatus and L. iners predominant types comprised a large proportion.

Identification of a novel cellulose-binding domain within the endo-ß-1,4-xylanase KRICT PX-3 from Paenibacillus terrae HPL-003.

The model 3-D structure of xylanase KRICT PX3 (JF320814) identified by DNA sequence analysis revealed a catalytic domain and CBM4-9 which functions as a xylan binding domain (XBD). To identify its role in xylan hydrolysis, six expression plasmids were constructed encoding the N-terminal CBM plus the catalytic domain or different glycosyl hydrolases, and the biochemical properties of the recombinant enzymes were compared to the original structure of PX3 xylanase. All six of the recombinant xylanases with the addition of CBM in the pIVEX-GST expression vector showed no improved PX3 hydrolytic activity. However, the absence of the CBM domain resulted in a decrement of 40% in thermostability, movement of the optimal temperature from 55°C to 45°C, alteration of the optimal pH range from 5-10 to 6-8, and reduction of the enzymatic activity to one-second under the same condition, respectively. The putative XBD in PX3 comprises a new N-terminal domain homologous to the catalytic thermostabilizing domains from other xylanases. Analysis of the main products released from xylan indicate that the recombinant enzymes act as endo-1,4-ß-xylanases but differ in their hydrolysis of xylan from beech wood, birch wood, and oat spelt.

Triazolyl phenyl disulfides: 8-Amino-7-oxononanoate synthase inhibitors as potential herbicides.

The chemical validation of a potential herbicide target was investigated with 8-amino-7-oxononanoate synthase (AONS, also known as 7-keto-8-aminopelargonate synthase, KAPAS) and triazolyl phenyl disulfide derivatives in vitro and in vivo. AONS activity was completely inhibited by these synthesized compounds, with an IC50 of 48 to 592µM in vitro. Forty five-day old Arabidopsis thaliana plants were completely killed by representative compound KHG23844 {N-(2-fluorophenyl)-3-(phenyldisulphanyl)-1H-1,2,4-triazole-1-carboxamide} at the application rate of 250gha(-1) of foliar treatment in greenhouse conditions. Foliar application of 1000gha(-1) KHG23844 induced 2.3-fold higher l-alanine accumulation in the treated A. thaliana plants. Foliar supplement of 1mM biotin at 1 and 2days before KHG23844 application effectively recovered the growth inhibition of A. thaliana plant treated with KHG23844. The results strongly suggested that representative compound KHG23844 and its derivatives are potential AONS inhibitors.

A new bi-modular endo-ß-1,4-xylanase KRICT PX-3 from whole genome sequence of Paenibacillus terrae HPL-003.

A new bi-modular, wide pH spectrum and highly active xylanase KRICT PX3 (JF320814) isolated from Paenibacillus terrae HPL-003 (KCTC11987BP) has been cloned and expressed in Escherichia coli. Purified recombinant xylanase KRICT PX-3 (1,620 bp, 540aa, NCBI accession number JF320814) showed highly active at 55°C in pH 4.0-11.0, and stability for at least 24h at 50°C, and exhibited Km and Vmax of 0.2mg/mL and 153.8 U/mg on birchwood xylan. Most common ions did not affect the enzyme activity at 1mM concentration. This enzyme could belong to glycoside hydrolase family 10 because hydrolyzed glucuronoxylan and arabinoxylan substrate to xylobiose, xylotriose, and some traces of xylose as hydrolysis products. Model 3-D structure was composed of two domains, the catalytic domain of a (ß/α)8 barrel fold while the small domain probably functions as a xylan binding domain, and the two domains are connected by a flexible linker peptide (PPLAIEKDIPSL). However, sequence alignment between xylan-binding module in this xylanase KRICT PX3 and CBM22 showed 21% of identity and 35% of similarity. This xylanase structure showed a distinctive group of enzyme cluster separately from the rest of GH10 xylanases, and seems to constitute a new type of xylanases.

An efficient and heterogeneous recyclable silicotungstic acid with modified acid sites as a catalyst for conversion of fructose and sucrose into 5-hydroxymethylfurfural in superheated water.

Acidity modified silver exchanged silicotungstic acid (AgSTA) catalyst was prepared and characterized by X-ray diffraction, FT-IR spectroscopy, Raman spectroscopy, FT-IR pyridine adsorption, SEM imaging, EDX mapping, and antimicrobial activity was also tested. The catalytic activity was evaluated for the dehydration of fructose and sucrose in superheated water. As a result, 98% conversion of fructose with 85.7% HMF yield and 87.4% HMF selectivity in 120 min reaction time at 120 °C reaction temperature using 10 wt.% of AgSTA catalyst was achieved. While, 92% sucrose conversion with 62.5% of HMF yield was obtained from sucrose at uniform condition in 160 min. The effect of reaction parameters, such as reaction temperature, time, catalyst dosage, and effect acidity on HMF yield was also investigated. The AgSTA catalyst was separated from the reaction mixture by filtration process at end of the reaction and reused eight times without loss of catalytic activity.

Genome sequence of Paenibacillus terrae HPL-003, a xylanase-producing bacterium isolated from soil found in forest residue.

This article reports on the full genome sequence of Paenibacillus terrae HPL-003, which is a gram-positive, endospore-forming, xylanase-producing bacterium isolated from soil found in forest residue on Gara Mountain. The strain HPL-003 contains 6,083,395 bp with a G+C content of 46.77 mol%, 2,633 protein-coding genes, and 117 structural RNAs.

A facile synthesis of highly functionalized 4-arylcoumarins via Kostanecki reactions mediated by DBU.

An efficient synthesis of 4-arylcoumarins has been accomplished via Kostanecki reactions of 2-hydroxybenzophenones with acetic anhydride employing DBU at ambient temperature. Using the same strategy, several 2-acyloxybenzophenone derivatives were readily converted to 3,4-difunctionalized coumarins. This protocol offers a notable improvement in reaction conditions for coumarin synthesis and takes advantage of its synthetic capability, especially for highly functionalized 4-arylcoumarins with structural diversity.

Kaposi's sarcoma-associated herpesvirus infection of endothelial progenitor cells impairs angiogenic activity in vitro.

A recent study reported that endothelial progenitor cells (EPCs0 are one of the reservoirs of Kaposi's sarcoma associated herpesvirus (KSHV). Although EPCs are closely linked to angiogenesis and vasculogenesis, little is known about the angiogenic potential of KSHV in EPCs. In this study, we used EPCs isolated from human umbilical cord blood to show that early infection by KSHV in vitro impaired the neovascularization of EPCs in matrigel. Our results suggest that KSHV may disrupt the angiogenic potential of EPCs and that the disseminated infection of KSHV could be associated with EPC dysfunction.

Vascular endothelial growth factor, fms-like tyrosine kinase-1 (Flt-1) and soluble Flt-1 gene expressions in Korean pre-eclamptic placentas.

AIM: To assess the expressions of vascular endothelial growth factor (VEGF), fms-like tyrosine kinase-1 (Flt-1), and soluble Flt-1 (sFlt-1) genes in healthy normotensive and pre-eclamptic placentas of Korean women. METHODS: We investigated 12 healthy normotensive pregnant women and 10 pre-eclamptic pregnant women at Eulji University Hospital. The obtained placental tissues were analyzed using reverse transcription polymerase chain reaction and real-time quantitative polymerase chain reaction. RESULTS: The sFlt-1 messenger ribonucleic acid (mRNA) level was elevated 2.6 times more in pre-eclamptic placentas than in normal control placentas. However, the VEGF mRNA level of pre-eclamptic placentas was decreased. There was no difference in the Flt-1 mRNA level between control and pre-eclamptic placentas. CONCLUSIONS: Our study showed that expressions of genes relating to angiogenesis were altered in Korean pre-eclamptic placentas. These results suggest that the alteration in expressions of sFlt-1 and VEGF genes might be associated with the pathogenesis of pre-eclampsia.

Cloning and characterization of a xylanase, KRICT PX1 from the strain Paenibacillus sp. HPL-001.

The KRICT PX1 gene (GB: FJ380951) consisting of 996bp encoding a protein of 332 amino acids (38.1kDa) from the recently isolated Paenibacillus sp. strain HPL-001 (KCTC11365BP) has been cloned and expressed in Escherichia coli. The xylanase KRICT PX1 showed high activity on birchwood xylan, and was active over a pH range of 5.0 to 11.0, with two optima at pH 5.5 and 9.5 at 50 degrees C with K(m) value of 5.35 and 3.23, respectively. The xylanase activity was not affected by most salts, such as NaCl, LiCl, KCl, NH(4)Cl, CaCl(2), MgCl(2), MnCl(2), and CsCl(2) at 1mM, but affected by CuSO(4), ZnSO(4), and FeCl(3). One mM of EDTA, 2-mercaptoethanol, and PMSF did not affect the xylanase activity. TLC analysis of the catalyzed products after reaction with birchwood xylan revealed that xylobiose was the major product with smaller amounts of xylotriose and xylose. A similarity analysis of the amino acids in KRICT PX1 resulted 72% identity with xylanase from Geobacillus stearothermophilus (GB: ZP_03040360), 70% identity with intracellular xylanase from an uncultured bacterium (GB: AAP51133), 68% identity with endo-1-4-xylanse from Paenibacillus sp. (GB: ZP_02847150). In addition, the amino acid alignment of KRICT PX1 with glycosyl hydralase (GH) family 10 xylanases revealed a high degree of homology in highly conserved regions including the catalytic sites, and this was confirmed through PROSITE scan. These results imply that KRICT PX1 is a new xylanase gene, and this alkaline xylanase belongs to GH family 10.

Laparoscopic treatment of early retroperitoneal abdominal pregnancy implanted on inferior vena cava.

The case of an early retroperitoneal abdominal pregnancy implanted on inferior vena cava successfully resected through laparoscopic surgery is reported here. A 28-year-old woman presented with vaginal spotting and amenorrhrea for 7(+5) weeks. Although a diagnosis of right cornual ectopic pregnancy by transvaginal ultrasound was made and a laparoscopic wedge resection of the left uterine cornus was performed, the beta-human chorionic gonadotropin level remained high. Abdominal ultrasonography and computerized tomography of the abdomen demonstrated a 3.4x2.6 cm sized hypervascular cystic mass along the anterior aspect of the Inferior vena cava. A diagnosis of retroperitoneal abdominal pregnancy was made and a laparoscopic resection was performed. This is the first retroperitoneal abdominal pregnancy that has been treated through laparoscopy.

A novel germline mutation of hMLH1 in a Korean hereditary non-polyposis colorectal cancer family.

Hereditary non-polyposis colorectal cancer (HNPCC) is an inherited disease caused by a germline mutation of the mismatch repair (MMR) genes, and the distinctive feature is that colorectal and extracolonic malignancies occur early in life. We report on the case of a Korean HNPCC family with endometrial cancer, with the goal of elucidating the involvement of an MMR deficiency. Although the family history did not fulfill the Amsterdam criteria II, blood samples were subjected to genetic testing by the revised Bethesda guidelines. Immunohistochemistry and direct sequencing of the genomic DNA identified a C insertion at the 1780th base in exon 16 of hMLH1, a pathogenic mutation that has not been reported before. By this mutation, premature termination at codon 592 resulted with an estimated deletion of 21% of the C-terminus of the hMLH1 protein. For early detection of the disease, the family was examined by colonoscopy and a gynecologic examination. The expression of hMLH1 in colon tissues was analyzed by Western blot analysis. We observed that the C-terminus portion of the hMLH1 protein was truncated in the HNPCC family members. Two young family members with no clinical symptoms were newly diagnosed with colorectal cancer by colonoscopy and a pathological examination. Hereby, we identified a novel pathogenic germline mutation of hMLH1 in a Korean HNPCC family. The loss of C-terminus of hMLH1 protein was thus considered to possibly play a role in the development of HNPCC with other tumors. Our findings might be useful for early diagnosis and management of the HNPCC family.

Evaluation of TMJ sound on the subject with TMJ disorder by Joint Vibration Analysis.

STATEMENT OF PROBLEM: Qualitative and semi-quantitative methods have been developed for TMJ sound classification, but the criteria presented are completely inhomogeneous. Thus, to develop more objective criteria for defining TMJ sounds, electroacoustical systems have been developed. We used Joint vibration analysis in the BioPAK system (Bioresearch Inc., Milwaukee, USA) as the electrovibratography. PURPOSE: The aim of this study was to examine the TMJ sounds with repect to frequency spectra patterns and the integral > 300 Hz /< 300 Hz ratios via six-months follow-up. MATERIAL AND METHODS: This study was done before and after the six-months recordings with 20 dental school students showed anterior disk displacement with reduction. Joint vibrations were analyzed using a mathematical technique known as the Fast Fourier Transform. RESULTS: In this study Group I and Group II showed varied integral > 300 /< 300 ratios before and after the six-months recordings. Also, by the comparative study between the integral > 300 /< 300 ratios and the frequency spectrums, it was conceivable that the frequency spectrums showed similar patterns at the same location that the joint sound occurred before and after the six-months recordings. while the frequency spectrums showed varied patterns at the different locations that the joint sound occurred before and after six-month recordings, it would possibly be due to the differences in the degree of internal derangement and/or in the shape of the disc. CONCLUSIONS: It is suggested that clinicians consider the integral > 300 /< 300 ratios as well as the frequency spectrums to decide the starting-point of the treatment for TMJ sounds.

Dual priming oligonucleotide system for the multiplex detection of respiratory viruses and SNP genotyping of CYP2C19 gene.

Successful PCR starts with proper priming between an oligonucleotide primer and the template DNA. However, the inevitable risk of mismatched priming cannot be avoided in the currently used primer system, even though considerable time and effort are devoted to primer design and optimization of reaction conditions. Here, we report a novel dual priming oligonucleotide (DPO) which contains two separate priming regions joined by a polydeoxyinosine linker. The linker assumes a bubble-like structure which itself is not involved in priming, but rather delineates the boundary between the two parts of the primer. This structure results in two primer segments with distinct annealing properties: a longer 5'-segment that initiates stable priming, and a short 3'-segment that determines target-specific extension. This DPO-based system is a fundamental tool for blocking extension of non-specifically primed templates, and thereby generates consistently high PCR specificity even under less than optimal PCR conditions. The strength and utility of the DPO system are demonstrated here using multiplex PCR and SNP genotyping PCR.

5-(2,6-difluorobenzyl)oxymethyl-5-methyl-3-(3-methylthiophen-2-yl)- 1,2-isoxazoline as a useful rice herbicide.

5-(2,6-difluorobenzyl)oxymethyl-5-methyl-3-(3-methylthiophen-2-yl)-1,2-isoxazoline derivative was synthesized, and its herbicidal activity was assessed under glasshouse and flooded paddy conditions. 5-(2,6-Difluorobenzyl)oxymethyl-5-methyl-3-(3-methylthiophen-2-yl)-1,2-isoxazoline demonstrated good rice selectivity and potent herbicidal activity against annual weeds at 125 g of a.i. ha(-1) under greenhouse conditions. Soil application of this compound showed complete control of barnyard-grass to the fourth leaf stage at 250 g of a.i. ha(-1). Field trials indicated that this compound controlled annual weeds rapidly with a good tolerance on transplanted rice seedlings by post-emergence and soil application. This compound showed a low mammalian and environmental toxicity in various toxicological tests.

New 2-phenyl-4,5,6,7-tetrahydro-2H-indazole derivatives as paddy field herbicides.

A series of 3-chloro-2-(4-chloro-2-fluorophenyl)-4,5,6,7-tetrahydro-2H-indazole derivatives containing various substituted isoxazolinylmethoxy groups at the 5-position of the benzene ring were synthesized and their herbicidal activities assessed under greenhouse and flooded paddy conditions. Among them, compounds having a phenyl or cyano substituent at the 3-position of the 5-methyl-isoxazolin-5-yl structure demonstrated good rice selectivity and potent herbicidal activity against annual weeds at 16-63 g AI ha(-1) under greenhouse conditions. Field trials indicated that these two compounds controlled a wide range of annual weeds rapidly with a good tolerance on transplanted rice seedlings by pre-emergence application. They showed a low mammalian and environmental toxicity in various toxicological tests.

EK-2612, a new cyclohexane-1,3-dione possessing selectivity between rice (Oryza sativa) and barnyardgrass (Echinochloa crus-galli).

A newly synthesized experimental compound, EK-2612 is one of the class of cyclohexane-1,3-diones which are commonly known to be grasskillers. A greenhouse study was conducted to evaluate the herbicidal performances of EK-2612 on several grass species in comparison with tralkoxydim, a commercialized cyclohexanedione derivative. Like tralkoxydim, the compound EK-2612 showed excellent control efficacy on most grass weeds tested through foliar application rates between 250 and 63 g AI ha(-1). Unlike tralkoxydim, however, EK-2612 showed a good rice safety, and there was no rice damage observed at the level below 125 g AI ha(-1), while rice injury developed at the same application rates of tralkoxydim. With this rice safety, EK-2612 controlled barnyardgrass effectively up to the two-leaf stage under both submerged and dried paddy conditions. An in vitro ACCase assay indicated that EK-2612 is a strong ACCase inhibitor; however, the dose-response was not substantially different in rice and barnryardgrass, showing I50 values of 0.1 and 0.12 microM, respectively. These results suggest that the compound EK-2612 is targeting plant ACCase, but the whole-plant rice safety is not attributable to a different inhibition of the target site in rice from that in barnyardgrass.