Coordinated elevation of membrane type 1-matrix metalloproteinase and matrix metalloproteinase-2 expression in rat uterus during postpartum involution

BACKGROUND: The changes occurring in the rodent uterus after parturition can be used as a model of extensive tissue remodeling. As the uterus returns to its prepregnancy state, the involuting uterus undergoes a rapid reduction in size primarily due to the degradation of the extracellular matrix, particularly collagen. Membrane type-I matrix metalloproteinase (MT1-MMP) is one of the major proteinases that degrades collagen and is the most abundant MMP form in the uterus. Matrix metalloproteinase-2(MMP-2) can degrade type I collagen, although its main function is to degrade type IV collagen found in the basement membrane. To understand the expression patterns of matrix metalloproteinases (MMPs) in the rat uterus, we analyzed their activities in postpartum uterine involution. METHODS: We performed gelatin zymography, northern blot analysis and immunohistochemistry to compare the expression levels of MT1-MMP, MMP-2, matrix metalloproteinase-9 (MMP-9) and the tissue inhibitors of MMPs-1 and 2 (TIMP-1 and TIMP-2) in the rat uterus 18 h, 36 h and 5 days after parturition with their expression levels during pregnancy (day 20). RESULTS: We found that both MT1-MMP and MMP-2 localized mainly in the cytoplasm of uterine interstitial cells. The expression levels of MT1-MMP and MMP-2 mRNAs and the catalytic activities of the expressed proteins significantly increased 18 h and 36 h after parturition, but at postpartum day 5, their mRNA expression levels and catalytic activities decreased markedly. The expression levels of MMP-9 increased 18 h and 36 h after parturition as determined by gelatin zymography including the expression levels of TIMP-1 and TIMP-2. CONCLUSION: These expression patterns indicate that MT1-MMP, MMP-2, MMP-9, TIMP-1 and TIMP-2 may play key roles in uterine postpartum involution and subsequent functional regenerative processes

Proteomic analysis of mechanisms of hypoxia-induced apoptosis in trophoblastic cells

Preeclampsia is often accompanied by hypoxia of the placenta and this condition induces apoptosis in trophoblastic cells. The aim of this study was to characterize global changes of apoptosis-related proteins induced by hypoxia in trophoblastic cells so as to clarify the mechanism of hypoxia-induced apoptosis by using the PoweBlot, an antibody-based Western array. Human choriocarcinoma cell line JAR was cultured for 24 hours under aerobic and hypoxic conditions. Hypoxia induced apoptosis accompanied by increased expression of Bcl-x, Caspase-3 and -9, Hsp70, PTEN, and Bag-1. Bad, pan-JNK/SAPK-1, Bcl-2, Bid, and Caspase-8 showed decreased expression. Hypoxia-induced apoptosis was increased with the transfection of a bag-1 antisense oligonucleotide. The bag-1 antisense oligonucleotide affected the expression of Bid, Bad, Bcl-2, JNK, and phosphorylated JNK, although expression of PTEN and Bcl-X did not change. Bag-1 may inhibit apoptosis by suppressing the expression of Bid and Bad. It may also enhance apoptosis by inhibiting the expression of Bcl-2 and by modulating phosphorylation of JNK. Both mitochondrial and stress-activated apoptosis pathways played important roles in the hypoxia induced cell death of trophoblastic cells. These findings will contribute to establish new approach to detect hypoxic stress of the placenta, which leads to preeclampsia and other hypoxia-related obstetrics complications.</p

Effect of multimodal comprehensive communication skills training with video analysis by artificial intelligence for physicians on acute geriatric care: a mixed-methods study

Objectives: To quantitatively analyse by artificial intelligence (AI) the communication skills of physicians in an acute care hospital for geriatric care following a multimodal comprehensive care communication skills training programme and to qualitatively explore the educational benefits of this training programme. Design: A convergent mixed-methods study, including an intervention trial with a quasi-experimental design, was conducted to quantitatively analyse the communication skills of physicians. Qualitative data were collected via physicians\u27 responses to an open-ended questionnaire administered after the training. Setting: An acute care hospital. Participants: A total of 23 physicians. Interventions: In a 4-week multimodal comprehensive care communication skills training programme, including video lectures and bedside instruction, from May to October 2021, all the participants examined a simulated patient in the same scenario before and after their training. These examinations were video recorded by an eye-tracking camera and two fixed cameras. Then, the videos were analysed for communication skills by AI. Main outcome measures: The primary outcomes were the physicians\u27 eye contact, verbal expression, physical touch and multimodal communication skills with a simulated patient. The secondary outcomes were the physicians\u27 empathy and burnout scores. Results: The proportion of the duration of the participants\u27 single and multimodal types of communication significantly increased (p\u3c 0.001). The mean empathy scores and the personal accomplishment burnout scores also significantly increased after training. We developed a learning cycle model based on the six categories that changed after training from the physicians\u27 perspective: multimodal comprehensive care communication skills training; increasing awareness of and sensitivity to changes to geriatric patients\u27 condition; changes in clinical management; professionalism; team building and personal accomplishments. Conclusions: Our study showed that multimodal comprehensive care communication skills training for physicians increased the proportions of time spent performing single and multimodal communication skills by video analysis through AI. Trial registration number: UMIN Clinical Trials Registry (UMIN000044288; https://center6.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000050586)