Promotion of Intestinal Peristalsis by Bifidobacterium spp. Capable of Hydrolysing Sennosides in Mice

BACKGROUND:While there are a variety of identifiable causes of constipation, even idiopathic constipation has different possible mechanisms. Sennosides, the main laxative constituents of Daio, an ancient Kampo medicine, are prodrugs that are converted to an active principle, rheinanthrone, by intestinal microbiota. In this study, we aimed to determine the sennoside hydrolysis ability of lactic acid bacterial strains and bifidobacteria in the intestine and to investigate their effect on intestinal peristalsis in mice. METHODOLOGY/PRINCIPAL FINDINGS:A total of 88 lactic acid bacterial strains and 47 bifidobacterial strains were evaluated for their ability to hydrolyze sennosides. Our results revealed that 4 strains, all belonging to the genus Bifidobacterium, had strong sennoside hydrolysis ability, exhibiting a decrease of >70% of sennoside content. By thin-layer chromatography analysis, rheinanthrone was detected in the medium cultured with B. pseudocatenulatum LKM10070 and B. animalis subsp. lactis LKM512. The fecal sennoside contents significantly (P<0.001) decreased upon oral administration of these strains as compared with the control. Intestinal peristalsis activity was measured by the moved distance of the charcoal powder administered orally. The distance travelled by the charcoal powder in LKM512-treated mice was significantly longer than that of control (P<0.05). Intestinal microbiota were analysed by real-time PCR and terminal-restriction fragment length polymorphism. The diversity of the intestinal microbiota was reduced by kanamycin treatment and the diversity was not recovered by LKM512 treatment. CONCLUSION/SIGNIFICANCE:We demonstrated that intestinal peristalsis was promoted by rheinanthrone produced by hydrolysis of sennoside by strain LKM512 and LKM10070

Ability of lactic acid bacterial strains and bifidobacterial strains to hydrolyse Sennoside A and B.

<p>Sennoside-hydrolysing values were expressed as the ratio of sennoside content in the medium without bacterial strain to that in the medium with bacterial strain.</p

Sennoside hydrolysis in the intestine by bifidobacterial strain LKM10070 (A) and LKM512 (B) administered orally after kanamycin treatment.

<p>Feces were collected every 2 hours up to 8 hours. The total sennoside contents in feces discharged up to 8 hours after administration of sennoside were significantly higher in kanamycin-treated mice than in the blank (no kanamycin) (<i>p</i><0.001). These tests for the blank group were performed before kanamycin treatment. The amount of sennoside collected from the feces of mice administered LKM10070 (n = 8) or LKM512 (n = 9) was significantly lower than that collected from the feces of control mice (<i>p</i><0.001, Mann-Whitney <i>U</i> test). Bars indicate the average value. The times of dosages are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031700#s2" target="_blank">Materials and Methods</a> section and in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031700#pone-0031700-g001" target="_blank">Figure 1</a>.</p

Detection of rheinanthrone in sennoside-containing medium cultivated by bifidobacterial strain LKM10070 or LKM512 by thin-layer chromatography (TLC).

<p>Spots of rheinanthrone were coloured yellow, and the Rf value was 0.68.</p

Effects of <i>B. animalis</i> subsp. <i>lactis</i> LKM512 on intestinal peristalsis of mice administered sennoside after kanamycin treatment.

<p>(A) Picture of the intestinal tract. The activity of intestinal peristalsis was expressed as the distance travelled by the charcoal powder from the end of the ileum (i.e. the base of the caecum or start of the colon) to the top of the charcoal powder, measured using a ruler. (B) Comparison of the length of charcoal powder from the end of the ileum. This value was significantly longer in LKM512-adminstered mice (n = 5) than in the control (n = 5) (<i>p</i><0.05), modified Wilcoxon signed-rank test). Error bars show the standard error of the mean.</p

Time schedule of animal assay.

<p>These tests were performed individually (one mouse/cage). (A) Sennoside-hydrolysing test by <i>Bifidobacterium</i> spp. Feces were collected every 2 hours. (B) Intestinal peristalsis promotion test by LKM512. This test was performed using a pair of mice: 1 LKM512-administered mouse and 1 control mouse.</p

Effects of kanamycin and LKM512 administration after kanamycin treatment on intestinal microbiota.

<p>(A) Change in the number of total bacteria and <i>B. animalis</i> subsp. <i>lactis</i> LKM512 in feces. Since the detection limit of the standard curve is 10³ cells, bacterial counts below this limit were assigned a value of 10³ cells/g of feces. (B) Change in the T-RFLP profile after kanamycin treatment (middle) and LKM512 administration after kanamycin treatment (lower) on digestion with <i>Hae</i>III. Arrow indicates T-RF predicted to be derived from LKM512.</p

初回治療から6年後に肺転移にて再発した子宮頸部中腎管腺癌の一例

子宮頸部に発生する中腎管腺癌は中腎管の遺残組織から発生する稀な子宮頸部腫瘍である。今回，初回治療から6年経過後に肺転移にて再発した子宮頸部中腎管腺癌の一例を経験したので文献的考察を加え報告する。症例は初発時年齢69歳女性，3妊2産。腰痛を主訴に近医産婦人科を受診。子宮腫大を認め，子宮頸部細胞診は異常を認めず，子宮内膜細胞診が陽性（腺癌）のため当科紹介となった。経腟超音波断層検査で子宮頸部に4cm大の充実性腫瘤を認め，子宮鏡検査では腫瘍は頸管内に突出していた。この部位から生検したところ，中腎管腺癌が疑われた。子宮頸部中腎管腺癌の診断にて広汎子宮全摘術と両側付属器切除を施行した。術後の病理組織検査でも中腎管腺癌であることが確認され，腫瘍は子宮頸部から腟壁に及び，右傍子宮組織にも浸潤していた。進行期は子宮頸部中腎管腺癌IIB期であり，術後補助療法として同時化学放射線療法を行った。子宮頸癌手術より6年目に左肺野の病変とCA19-9の上昇を認めた。左肺上葉部分切除を行ったところ中腎管腺癌の再発であった。左肺上葉部分切除18か月後に，右肺野腫瘤の増大を認め中腎管腺癌の再々発の診断となった。これに対してCPT-11の単剤治療3コース実施したところ，右肺野の腫瘤は縮小した。現在，子宮頸癌術後108か月時点でパフォーマンスステータス0の担癌生存の状態である。子宮頸部中腎管腺癌はその発生学的な特徴より術前診断が困難であり，子宮頸部細胞診に異常を認めない場合でも，子宮頸部に充実性病変を認める場合には，本疾患の存在を念頭に置く必要がある。また，本症例のように術後6年経過しても再発する場合があり，長期間のフォローアップが必要である。In this case report, we present a rare case of mesonephric adenocarcinoma of the cervix that recurred with pulmonary metastases six years after initial treatment. At first presentation, the patient was a 69-year-old woman, gravida 3 para 2. She presented to a clinic with a chief complaint of lower back pain. She was referred to our hospital because she had an enlarged uterus, cervical cytology showed no abnormalities and endometrial cytology was positive. Transvaginal ultrasound revealed a substantial cervical mass measuring 4 cm, and a biopsy from this site suggested mesonephric adenocarcinoma. A radical hysterectomy and bilateral adnexectomy were performed, and histopathology confirmed the presence of mesonephric adenocarcinoma with extension from the cervix to the vaginal wall and invasion of the right parametrium. The final clinical stage was stage IIB mesonephric adenocarcinoma of the cervix, and concurrent chemoradiotherapy was given as adjuvant therapy. Six years after surgery for cervical cancer, lesions in the left lung field and elevated CA19-9 were observed. Partial resection of the upper lobe of the left lung revealed a recurrence of mesonephric adenocarcinoma. 18 months later, an enlarged mass was found in the right lung field and a diagnosis of recurrent mesonephric adenocarcinoma was made. After three courses of CPT-11 chemotherapy, the mass in the right lung field shrank and the patient is currently alive with a performance status of 0, 108 months after surgery for cervical cancer. Mesonephric adenocarcinoma of the cervix is a rare and challenging disease to diagnose preoperatively due to its embryological features. The presence of a solid lesion on the cervix, even in the absence of abnormal cervical cytology, should raise suspicion for this disease. Long-term follow-up is crucial as recurrence can occur several years after surgery

Detection of Atrial Fibrillation Using Insertable Cardiac Monitors in Patients With Cryptogenic Stroke in Japan (the LOOK Study): Protocol for a Prospective Multicenter Observational Study

BackgroundParoxysmal atrial fibrillation (AF) is a probable cause of cryptogenic stroke (CS), and its detection and treatment are important for the secondary prevention of stroke. Insertable cardiac monitors (ICMs) are clinically effective in screening for AF and are superior to conventional short-term cardiac monitoring. Japanese guidelines for determining clinical indications for ICMs in CS are stricter than those in Western countries. Differences between Japanese and Western guidelines may impact the detection rate and prediction of AF via ICMs in patients with CS. Available data on Japanese patients are limited to small retrospective studies. Furthermore, additional information about AF detection, including the number of episodes, cumulative episode duration, anticoagulation initiation (type and dose of regimen and time of initiation), rate of catheter ablation, role of atrial cardiomyopathy, and stroke recurrence (time of recurrence and cause of the recurrent event), was not provided in the vast majority of previously published studies. ObjectiveIn this study, we aim to identify the proportion and timing of AF detection and risk stratification criteria in patients with CS in real-world settings in Japan. MethodsThis is a multicenter, prospective, observational study that aims to use ICMs to evaluate the proportion, timing, and characteristics of AF detection in patients diagnosed with CS. We will investigate the first detection of AF within the initial 6, 12, and 24 months of follow-up after ICM implantation. Patient characteristics, laboratory data, atrial cardiomyopathy markers, serial magnetic resonance imaging findings at baseline, 6, 12, and 24 months after ICM implantation, electrocardiogram readings, transesophageal echocardiography findings, cognitive status, stroke recurrence, and functional outcomes will be compared between patients with AF and patients without AF. Furthermore, we will obtain additional information regarding the number of AF episodes, duration of cumulative AF episodes, and time of anticoagulation initiation. ResultsStudy recruitment began in February 2020, and thus far, 213 patients have provided written informed consent and are currently in the follow-up phase. The last recruited participant (May 2021) will have completed the 24-month follow-up in May 2023. The main results are expected to be submitted for publication in 2023. ConclusionsThe findings of this study will help identify AF markers and generate a risk scoring system with a novel and superior screening algorithm for occult AF detection while identifying candidates for ICM implantation and aiding the development of diagnostic criteria for CS in Japan. Trial RegistrationUMIN Clinical Trial Registry UMIN000039809; https://tinyurl.com/3jaewe6a International Registered Report Identifier (IRRID)DERR1-10.2196/3930