Protein interaction networks characterizing the A549 cells Klotho transfected are associated with activated pro-apoptotic Bim and suppressed Wnt/ß-catenin signaling pathway.

Invasive assays and lung tumor-bearing mice models using a human lung adenocarcinoma cell line A549 cells transfected with the Klotho (KL) gene, A549/KL cells, have confirmed that KL suppresses invasive/metastatic potential. This study aimed to identify the co-expression protein networks and proteomic profiles associated with A549/KL cells to understand how Klotho protein expression affects molecular networks associated with lung carcinoma malignancy. A two-step application of a weighted network correlation analysis to the cells' quantitative proteome datasets of a total of 6,994 proteins, identified by mass spectrometry-based proteomic analysis with data-independent acquisition (DIA), identified one network module as most significantly associated with the A549/KL trait. Upstream analyses, confirmed by western blot, implicated the pro-apoptotic Bim (Bcl-2-like protein 11) as a master regulator of molecular networks affected by Klotho. GeneMANIA interaction networks and quantitative proteome data implicated that Klotho interacts with two signaling axes: negatively with the Wnt/ß-catenin axis, and positively by activating Bim. Our findings might contribute to the development of future therapeutic strategies.

4D Flow MR Imaging of the Left Atrium: What is Non-physiological Blood Flow in the Cardiac System?

Most cardiac diseases cause a non-physiological blood flow pattern known as turbulence around the heart and great vessels, which further worsen the disease itself. However, there is no consensus on how blood flow can be defined in disease conditions. Especially, in the left atrium, the fact that vortex flow already exists makes this debate more complicated. 3D time-resolved phase-contrast (4D flow) MRI is expected to be able to capture blood flow patterns from multiple aspects, such as blood flow velocity, stasis, and vortex quantification. Previous studies have confirmed that physiological vortex flow is predominantly induced by the higher-volume flow from the superior left pulmonary vein. In atrial fibrillation, 4D flow MRI reveals a non-physiological blood flow pattern, which information may add value to well-established clinical risk factors. Currently, the research target of LA analysis has also widened to lung surgeons, pulmonary vein stump thrombosis after left upper lobectomy. 4D flow MRI is expected to be utilized for many more variable diseases that are currently unimaginable.

New Technique for Introducing a Surgical Stapler during Robot-Assisted Lobectomy for Lung Cancer.

BACKGROUND: The da Vinci Si version robot lacks a vascular stapler that can be controlled by the operating surgeon at the surgical console when dividing pulmonary vessels. Therefore, to initiate and safely perform robotic anatomical lobectomy for lung cancer, it is important to develop a safe method for introducing a surgical stapler. METHODS: We performed a retrospective study of the first 42 consecutive patients who underwent robotic lobectomy for lung cancer at Nippon Medical School Hospital between January 2019 and December 2020. RESULTS: Up to case 18, we performed robot-assisted thoracoscopic surgery (RATS) lobectomy by using a four-arm approach with two assistant ports. For dividing pulmonary vessels, the surgical stapler was introduced through the assist ports. However, since this is not the port position usually used in video-assisted thoracoscopic surgery (VATS), there were many difficult situations. For RATS lobectomy case 19 and all subsequent cases, we utilized a total port approach that uses three robotic arms and two assistant ports. To resect the pulmonary vessels or bronchi with endoscopic staplers, the port for the robotic arm was removed and the endoscopic staplers were placed through a 12-mm Xcel bladeless port. This change reduced operation time, blood loss, and robotic arm interference. No patient developed intraoperative complications during RATS lobectomy. CONCLUSION: The present total port approach, with three robotic arms, appears to be feasible for introducing surgical staplers during RATS with the da Vinci Si robotic system.

Dual-VENC 4D Flow MRI Can Detect Abnormal Blood Flow in the Left Atrium That Potentially Causes Thrombosis Formation after Left Upper Lobectomy.

PURPOSE: The purpose of the current study was to clarify the blood flow pattern in the left atrium (LA), potentially causing the formation of thrombosis after left upper lobectomy (LUL). The blood flow in the LA was evaluated and compared between LUL patients with and without thrombosis. For the evaluation, we applied highly accelerated 4D flow MRI with dual-velocity encoding (VENC) scheme, which was expected to be able to capture slow flow components in the LA accurately. METHODS: Eight volunteers and 18 patients subjected to LUL underwent dual-VENC 4D Flow MRI. Eight patients had a history of thrombosis. We measured the blood flow velocity and stasis ratio (proportion in the volume that did not exceed 10 cm/s in any cardiac phase) in the LA and left superior pulmonary vein (LSPV) stump. For visual assessment, the presence of each collision of the blood flow from pulmonary veins and vortex flow in the LA were evaluated. Each acquired value was compared between healthy participants and LUL patients, and in LUL patients with and without thrombosis. RESULTS: In LUL patients, blood flow velocity near the inflow part of the left superior pulmonary vein (Lt Upp) and mean velocity in the LA were lower, and stasis ratio in the LA was higher compared with healthy volunteers (Lt Upp 9.10 ± 3.09 vs.13.23 ± 14.19 cm/s, mean velocity in the LA 9.81 ± 2.49 vs. 11.40 ± 1.15 cm/s, and stasis ratio 25.28 ± 18.64 vs. 4.71 ± 3.03%, P = 0.008, 0.037, and < 0.001). There was no significant difference in any quantification values between LUL patients with and without thrombosis. For visual assessment, the thrombus formation was associated with no collision pattern (62.5% vs. 10%, P = 0.019) and not with vortex flow pattern (50% vs. 30%, P = 0.751). CONCLUSION: The net blood flow velocity was not associated with the thrombus formation. In contrast, a specific blood flow pattern, the absence of blood flow collision from pulmonary veins, correlates to the thrombus formation in the LA.

Usefulness of simultaneous type image-enhanced endoscope system in photodynamic therapy for centrally located lung cancer.

BACKGROUND: Photodynamic therapy (PDT) is established as one of the standard treatment options for centrally located early lung cancer. In order to improve the effectiveness of PDT, it is very important to accurately diagnose the extent of the tumor and focus the laser irradiation accurately. With the use of the conventional video-endoscope system, which adopts the frame-sequential (RGB-based) display method, mainly used in Japan, for PDT laser irradiation, the system only recognizes the strong white light, and color information is lost. Therefore, it is difficult to irradiate the lesion while simultaneously observing the lesion. In this study, we investigated the usefulness of a new type of video-endoscope system during PDT. METHODS: We used ELUXEO 7000® (FUJIFILM, Japan), which is a simultaneous-type video-endoscope system that has been in use at Nippon Medical School Hospital since October 2018. We analyzed the clinical usefulness of the ELUXEO® system for PDT as compared to other endoscope systems, such as EVIS LUCERA ELITE® (Olympus, Japan), an autofluorescence imaging (AFI) system. RESULTS: After the administration of talaporfin sodium for PDT, the tumor lesion was not visualized in magenta color with AFI, yielding false-negative results. On the other hand, no false-negative results after the administration of talaporfin sodium were obtained with the use of ELUXEO®. Using the ELUXEO® system in the blue light imaging (BLI) mode, we were able to deliver a red laser light while observing the extent of the tumor. Missed laser exposure was avoided and the accuracy of PDT was improved with the use of this system. CONCLUSIONS: ELUXEO® is useful for accurate evaluation of the extent of centrally located lung cancer and therefore, for accurate laser irradiation of the tumor lesion.

Antiaging gene Klotho regulates epithelial-mesenchymal transition and increases sensitivity to pemetrexed by inducing lipocalin-2 expression.

Epithelial-mesenchymal transition (EMT) is considered to serve an important role in the metastatic/invasive ability of cancer cells, in the acquisition of drug resistance, and in metabolic reprogramming. In the present study, it was hypothesized that the Klotho gene is involved in the metastatic/invasive ability of lung cancer. We previously reported an association between Klotho expression and overall survival in patients with small cell lung cancer and large cell neuroendocrine cancer. We also found that Klotho expression was associated with EMT-related molecules in lung squamous cell carcinoma. The present study aimed to analyze the function of the Klotho gene and to elucidate its relevance to the regulation of the EMT. For this purpose, GFP-Klotho plasmids were transfected into lung adenocarcinoma cells (A549) and cell lines with stable expression (A549/KL-1 and A549/KL-2) were established. A549/KL-1 cells expressed higher levels of Klotho protein by western blot analysis compared with A549/KL-2 cells. In western blotting of A549 and A549/KL-1 cells, the expression of the mesenchymal marker N-cadherin was found to be completely inhibited in A549/KL-1 cells suggesting that Klotho expression may regulate the EMT in cancer cells via the inhibition of N-cadherin. The results of the sensitivity tests demonstrated that A549/KL-1 cells were significantly more sensitive to pemetrexed compared with A549 cells (IC50 A549/KL-1 vs. A549 cells, 0.1 µM vs. 0.7 µM). The results of the microarray analysis demonstrated that a very high level of lipocalin-2 (LCN2) expression was induced in the A549/KL-1 cells. Klotho overexpression completely suppressed the expression of mesenchymal markers, such as N-cadherin and Snail1 (Snail). The results of the present study suggested that there may be a new mechanism of action for the antitumor effects of pemetrexed, namely, LCN2-mediated modulation of N-cadherin expression. Klotho expression during cancer treatment has great potential as a predictor for efficacy of pemetrexed and as a factor in the selection of personalized medicine for postoperative adjuvant chemotherapy.

[Therapeutic Strategies for Metachronous Multiple Primary Lung Cancer].

BACKGROUNDS: It is not uncommon to encounter metachronous primary lung cancer after surgical treatment along with the increase in the elderly patients. In consideration of increasing number of such patients, it is necessary to take various treatment strategies. METHODS: In order to establish a treatment strategy for multiple lung cancer, we retrospectively examined multiple lung cancer cases operated for primary lung cancer in our department from January 2013 to December 2019, and the future treatment strategy was examined. RESULTS: Of 821 patients who underwent surgery for primary lung cancer, 61 were multiple lung cancers, 31 were synchronous multiple lung cancers, and 30 were metachronous multiple lung cancers. Among the cases of metachronous multiple lung cancer, 28 cases had undergone lobectomy or more in the first operation, 1 case of segmental resection, and 1 case of partial resection. As for the treatment of secondary lung cancer lesions, 21 lesions were performed surgery( lobectomy;2, segmental resection;2, partial resection;17), 6 lesions of photodynamic therapy (PDT), and 3 lesions of stereotactic body radiation therapy (SBRT). Among the surgical cases, there were three cases on the same side as the first cancer and 18 cases on the opposite side. The three cases underwent partial resection. Among 21 patients, postoperative home oxygen therapy was introduced in 2 patients. Regarding the prognosis, three patients who had surgical resection, died of recurrence of the first lesion. One of the six PDT patients died of recurrence of the first lung cancer and another died of other disease. All three patients who underwent SBRT are alive without recurrence. CONCLUSIONS: If early detection and early diagnosis are made as a treatment strategy for metachronous multiple lung cancer, it may be possible to preserve lung function by reducing surgery or SBRT, PDT, and to cure without damaging the quality of life.

4D Flow MR Imaging Reveals a Decrease of Left Atrial Blood Flow in a Patient with Cardioembolic Cerebral Infarction after Pulmonary Left Upper Lobectomy.

We present the case of a 76-year-old woman who underwent left upper lobectomy for lung adenocarcinoma. Three days after the surgery, a thrombus was observed in the pulmonary vein (PV) stump. Four months after the surgery, we performed 4D flow MRI, which revealed decreased blood flow, not only in the left superior PV stump, but also over a widespread region around the left atrium-left superior PV junction. 4D flow MRI can clarify the altered hemodynamics underlying thrombus formation.

Expression Profile of Intravitreous Cytokines, Chemokines and Growth Factors in Patients with Fuchs Heterochromic Iridocyclitis.

PURPOSE: To report the postoperative courses of 2 patients with Fuchs heterochromic iridocyclitis (FHI) and the concentrations of various cytokines, chemokines and growth factors in vitreous fluid samples to obtain insights into pathobiochemical aspects. Subjects: The patients were a 27- and a 47-year-old woman. Phacoemulsification and aspiration, intraocular lens (IOL) implantation, and pars plana vitrectomy were performed to treat their cataracts and vitreous opacities. During their early postoperative periods, inflammatory cells precipitated on the IOL and intraocular pressure was increased in both patients. METHODS: At the time of surgery, undiluted vitreous fluid specimens were collected. The concentrations of multiple cytokines, chemokines and growth factors were measured by a bead array immunodetection system. RESULTS: The levels of interleukin-1ra, -5, -6, -8, -10 and -13, interferon-inducible 10-kDa protein, monocyte chemoattractant protein 1, macrophage inflammatory protein 1beta, and regulated upon activation, normal T-cell expressed and secreted (RANTES) were significantly elevated in vitreous fluid in both patients. CONCLUSION: Although the postoperative course was generally favorable in patients with FHI, steroid instillation was necessary for a few months postoperatively, as precipitates easily formed on the IOL surface and elevated intraocular pressure. The profiles of intravitreal concentrations of cytokines, chemokines and growth factors may characterize postoperative inflammatory reactions.

Enhancement effect of p-menthane-3,8-diol on in vitro permeation of antipyrine and indomethacin through Yucatan micropig skin.

The enhancing effect of p-Menthane-3,8-diol (MDO) on skin permeation of antipyrine (ANP) and indomethacin (IM) through Yucatan micropig skin in vitro was compared with 1-menthol. p-Menthane-3,8-diol is a metabolite of 1-menthol and has little odor. It is easy to combine the vehicle because of lower lipophilicity than 1-menthol. All formulations contained 40% (v/v) ethanol. The permeation of ANP increased with MDO about three times that without enhancer by increasing ANP concentration in the skin. However, the MDO effect was about a quarter that of 1-menthol. The permeation of IM with MDO was about 15 times that with no enhancer and it was almost the same as that with 1-menthol. The lag time of permeation was not significantly changed by MDO, which was not so in the case of 1-menthol. Skin concentration of IM increased about 11 times and six times with MDO and 1-menthol, respectively. MDO and 1-menthol partitioned to the skin relatively high concentrations, 5.9 and 2.5 mg/ cm3, respectively. The solubility of IM in the skin was improved by MDO, and consequently, the permeation of IM was enhanced.

Comparison of skin permeation enhancement by 3-l-menthoxypropane-1,2-diol and l-menthol: the permeation of indomethacin and antipyrine through Yucatan micropig skin and changes in infrared spectra and X-ray diffraction patterns of stratum corneum.

3-l-Menthoxypropane-1,2-diol (MPD) is a derivative of l-menthol, which has an enhancement effect on drug permeation through skin. In this study, the effect of MPD on drug permeation through skin was compared with that of l-menthol. MPD or l-menthol at final concentrations of 3% in 40% ethanol was added to the drugs indomethacin or antipyrine and each mix then applied to Yucatan micropig skin in vitro. Drug concentrations in the skin were higher in the presence of either MPD or l-menthol, however, only l-menthol shortened the lag time of permeation. MPD enhanced the skin permeation of the drugs only by increasing the skin concentration of the drugs. In contrast, l-menthol enhanced the skin permeation of the drugs by increasing both the skin concentration and the diffusion rate in skin. The infrared (IR) spectra and X-ray diffraction patterns of stratum corneum after treatment with MPD did not differ from those of intact stratum corneum. A change in the IR spectra of stratum corneum after treatment with l-menthol was observed at the CH band, and the peaks representative of the lipid structure in the X-ray diffraction patterns decreased in intensity. These results suggest that l-menthol, but not MPD, disrupts the intercellular lipid structure of stratum corneum. Thus, MPD is expected to be a moderate skin permeation enhancer.