Search

Male penis is required to become erect during copulation. In the upper (dorsal) part of penis, the erectile tissue termed corpus cavernosum (CC) plays fundamental roles for erection by regulating the inner blood flow. When blood flows into the CC, the microvascular complex termed sinusoidal space is reported to expand during erection. A novel in vitro explant system to analyze the dynamic erectile responses during contraction/relaxation is established. The current data show regulatory contraction/relaxation processes induced by phenylephrine (PE) and nitric oxide (NO) donor mimicking dynamic erectile responses by in vitro CC explants. Two-photon excitation microscopy (TPEM) observation shows the synchronous movement of sinusoidal space and the entire CC. By taking advantages of the CC explant system, tadalafil (Cialis) was shown to increase sinusoidal relaxation. Histopathological changes have been generally reported associating with erection in several pathological conditions. Various stressed statuses have been suggested to occur in the erectile responses by previous studies. The current CC explant model enables to analyze such conditions through directly manipulating CC in the repeated contraction/relaxation processes. Expression of oxidative stress marker and contraction-related genes, Hypoxia-inducible factor 1-alpha (Hif1a), glutathione peroxidase 1 (Gpx1), Ras homolog family member A (RhoA), and Rho-associated protein kinase (Rock), was significantly increased in such repeated contraction/relaxation. Altogether, it is suggested that the system is valuable for analyzing structural changes and physiological responses to several regulators in the field of penile medicine.

Background Because of the high frequency of chronic edema formation in the current “aged” society, analyses and detailed observation of post-surgical edema are getting more required. Post-surgical examination of the dynamic vasculature including L.V. (Lymphatic Vasculature) to monitor edema formation has not been efficiently performed. Hence, procedures for investigating such vasculature are essential. By inserting transparent sheet into the cutaneous layer of mouse tails as a novel surgery model (theTailEdema bySilicone sheet mediatedTransparency protocol; TEST), the novel procedures are introduced and analyzed by series of histological analyses including video-based L.V. observation and 3D histological reconstruction of vasculatures in mouse tails. Results The dynamic generation of post-surgical main and fine (neo) L.V. connective structure during the edematous recovery process was visualized by series of studies with a novel surgery model. Snapshot images taken from live binocular image recording for TEST samples suggested the presence of main and elongating fine (neo) L.V. structure. After the ligation of L.V., the enlargement of main L.V. was confirmed. In the case of light sheet fluorescence microscopy (LSFM) observation, such L.V. connections were also suggested by using transparent 3D samples. Finally, the generation of neo blood vessels particularly in the region adjacent to the silicone sheet and the operated boundary region was suggested in 3D reconstruction images. However, direct detection of elongating fine (neo) L.V. was not suitable for analysis by such LSFM and 3D reconstruction procedures. Thus, such methods utilizing fixed tissues are appropriate for general observation for the operated region including of L.V. Conclusions The current surgical procedures and analysis on the post-surgical status are the first case to observe vasculatures in vivo with a transparent sheet. Systematic analyses including the FITC-dextran mediated snap shot images observation suggest the elongation of fine (neo) lymphatic vasculature. Post-surgical analyses including LSFM and 3D histological structural reconstruction, are suitable to reveal the fixed structures of blood and lymphatic vessels formation.

Intricate branching patterns emerge in internal organs because of the repetitive presence of simple deformations in epithelial tissues. During murine lung development, epithelial cells in distal tips of a single tube require fibroblast growth factor (FGF) signals generated by their surrounding mesenchyme to form repetitive tip bifurcations. However, it remains unknown how the cells employ FGF signaling to convert their behaviors to achieve the recursive branching processes. Here we show a self-sustained epithelial regulatory system during the murine lung branching morphogenesis, mediated by extracellular signal-regulated kinase (ERK), which acts as a downstream driver of FGF signaling. We found that tissue-scale curvature regulated ERK activity in the lung epithelium using two-photon live cell imaging and mechanical perturbations. ERK was activated specifically in epithelial tissues with a positive curvature, regardless of whether the change in curvature was attributable to morphogenesis or artificial perturbations. Moreover, ERK activation accelerated actin polymerization specifically at the apical side of cells, and mechanically contributed to the extension of the apical membrane, leading to a decrease in epithelial tissue curvature. These results indicate the existence of negative feedback loop between tissue curvature and ERK activity beyond scale. We confirmed that this regulation was sufficient to generate the recursive branching processes by a mathematical model. Taken together, we propose that ERK mediates the curvature feedback loop underlying the process of branching morphogenesis in developing lungs.

A notable example of spiral architecture in organs is the mammalian cochlear duct, where the morphology is critical for hearing function. Genetic studies have revealed necessary signaling molecules, but it remains unclear how cellular dynamics generate elongating, bending, and coiling of the cochlear duct. Here, we show that extracellular signal-regulated kinase (ERK) activation waves control collective cell migration during the murine cochlear duct development using deep tissue live-cell imaging, Förster resonance energy transfer (FRET)-based quantitation, and mathematical modeling. Long-term FRET imaging reveals that helical ERK activation propagates from the apex duct tip concomitant with the reverse multicellular flow on the lateral side of the developing cochlear duct, resulting in advection-based duct elongation. Moreover, model simulations, together with experiments, explain that the oscillatory wave trains of ERK activity and the cell flow are generated by mechanochemical feedback. Our findings propose a regulatory mechanism to coordinate the multicellular behaviors underlying the duct elongation during development., うずまき管の伸⻑を司る分子活性と細胞群の波を発見 --綱引きによる細胞群の流れと臓器の成長--. 京都大学プレスリリース. 2021-03-09.

Tracheal cartilage provides architectural integrity to the respiratory airway, and defects in this structure during embryonic development cause severe congenital anomalies. Previous genetic studies have revealed genes that are critical for the development of tracheal cartilage. However, it is still unclear how crosstalk between these proteins regulates tracheal cartilage formation. Here we show a core regulatory network underlying murine tracheal chondrogenesis from embryonic day (E) 12.5 to E15.5, by combining volumetric imaging of fluorescence reporters, inhibitor assays, and mathematical modeling. We focused on SRY-box transcription factor 9 (Sox9) and extracellular signal-regulated kinase (ERK) in the tracheal mesenchyme, and observed a synchronous, inverted U-shaped temporal change in both Sox9 expression and ERK activity with a peak at E14.5, whereas the expression level of downstream cartilage matrix genes, such as collagen II alpha 1 (Col2a1) and aggrecan (Agc1), monotonically increased. Inhibitor assays revealed that the ERK signaling pathway functions as an inhibitory regulator of tracheal cartilage differentiation during this period. These results suggest that expression of the cartilage matrix genes is controlled by an incoherent feedforward loop via Sox9 and ERK, which is supported by a mathematical model. Furthermore, the modeling analysis suggests that a Sox9-ERK incoherent feedforward regulation augments the robustness against the variation of upstream factors. The present study provides a better understanding of the regulatory network underlying the tracheal development and will be helpful for efficient induction of tracheal organoids.

A notable example of spiral architecture in organs is the mammalian cochlear duct, where the duct morphology is critical for hearing function. Molecular genetics has revealed the necessary signaling molecules for the formation of spirals in organs, but it remains unclear how cellular dynamics generate bending and coiling of the cochlear duct during development. Here we show two modes of multicellular dynamics underlying the morphogenetic process by combining deep tissue live-cell imaging, Förster resonance energy transfer (FRET)-based quantitation, and mathematical modeling. First, surgical separation of the cochlear duct revealed that bending forces reside primarily in the medial side of the duct. In the medial pseudostratified epithelium, we found that nuclei stall at the luminal side during interkinetic nuclear migration, which would cause the extension of the luminal side, thereby bending the duct. Second, long-term organ-scale FRET imaging of extracellular signal-regulated kinase (ERK) activity showed that helical ERK activation waves propagate from the duct tip concomitant with the reverse multicellular flow in the lateral side of the duct, resulting in advection-based duct elongation. We propose an interplay of distinct multicellular behaviors underpinning spiral morphogenesis in the developing cochlear duct.

Tracheal cartilage provides architectural integrity to the respiratory airway, and defects in this structure during embryonic development cause severe congenital anomalies. Previous genetic studies have revealed genes that are critical for the development of tracheal cartilage. However, it is still unclear how crosstalk between these proteins regulates tracheal cartilage formation. Here we show a core regulatory network underlying murine tracheal chondrogenesis from embryonic day (E) 12.5 to E15.5, by combining volumetric imaging of fluorescence reporters, inhibitor assays, and mathematical modeling. We focused on SRY-box transcription factor 9 (Sox9) and extracellular signal-regulated kinase (Erk) in the tracheal mesenchyme, and observed a synchronous, inverted U-shaped temporal change in both Sox9 expression and Erk activity with a peak at E14.5, whereas the expression level of downstream cartilage matrix genes, such as collagen II alpha 1 (Col2a1) and aggrecan (Agc1), monotonically increased. Inhibitor assays revealed that the Erk signaling pathway functions as an inhibitory regulator of tracheal cartilage differentiation during this period. These results suggest that expression of the cartilage matrix genes is controlled by an incoherent feedforward loop via Sox9 and Erk, which is supported by a mathematical model. Furthermore, the modeling analysis suggests that a Sox9-Erk incoherent feedforward regulation augment the robustness against the variation of upstream factors. The present study provides a better understanding of the regulatory network underlying the tracheal development and will be helpful for efficient induction of tracheal organoids.

During collective migration of epithelial cells, the migration direction is aligned over a tissue-scale expanse. Although the collective cell migration is known to be directed by mechanical forces transmitted via cell-cell junctions, it remains elusive how the intercellular force transmission is coordinated with intracellular biochemical signaling to achieve collective movements. Here, we show that intercellular coupling of extracellular signal-regulated kinase (ERK)-mediated mechanochemical feedback yields long-distance transmission of guidance cues. Mechanical stretch activates ERK through epidermal growth factor receptor (EGFR) activation, and ERK activation triggers cell contraction. The contraction of the activated cell pulls neighboring cells, evoking another round of ERK activation and contraction in the neighbors. Furthermore, anisotropic contraction based on front-rear polarization guarantees unidirectional propagation of ERK activation, and in turn, the ERK activation waves direct multicellular alignment of the polarity, leading to long-range ordered migration. Our findings reveal that mechanical forces mediate intercellular signaling underlying sustained transmission of guidance cues for collective cell migration., 分子活性の波が細胞集団に伝わる制御機構を解明 --細胞同士の綱引きが情報を遠くに伝える--. 京都大学プレスリリース. 2020-06-04., Cells communicate by doing the 'wave'. 京都大学プレスリリース. 2020-07-22.

Summary The in vivo niche and basic cellular properties of nephron progenitors are poorly described. Here we studied the cellular organization and function of the MAPK/ERK pathway in nephron progenitors. Live-imaging of ERK activity by a Förster resonance energy transfer biosensor revealed a dynamic activation pattern in progenitors, whereas differentiating precursors exhibited sustained activity. Genetic experiments demonstrate that MAPK/ERK activity controls the thickness, coherence, and integrity of the nephron progenitor niche. Molecularly, MAPK/ERK activity regulates niche organization and communication with extracellular matrix through PAX2 and ITGA8, and is needed for CITED1 expression denoting undifferentiated status. MAPK/ERK activation in nephron precursors propels differentiation by priming cells for distal and proximal fates induced by the Wnt and Notch pathways. Thus, our results demonstrate a mechanism through which MAPK/ERK activity controls both progenitor maintenance and differentiation by regulating a distinct set of targets, which maintain the biomechanical milieu of tissue-residing progenitors and prime precursors for nephrogenesis., Graphical Abstract, Highlights • Embryonic kidneys exhibit heterogeneous activation of MAPK/ERK pathway • Nephron progenitor maintenance depends on constant MEK-induced ERK activation • MAPK/ERK activity regulates nephron progenitor niche via PAX2 and ITGA8 • Nephron precursors require MAPK/ERK to propel differentiation, In this article, Ihermann-Hella and colleagues show that MAPK/ERK activity, typically altered in several cancers, plays a dual function in regulation of embryonic nephron progenitors. MAPK/ERK activity maintains nephron progenitors within their in vivo niche by regulating PAX2 and ITGA8, and fosters nephrogenesis in early differentiating nephrons. Utilization of these findings may substantially improve prospective iPSC-derived kidney organoid differentiation.

In vitro tubulogenesis has been employed as an experimental model system to study tissue morphogenesis of internal organs. It has been previously shown that Madin-Darby canine kidney (MDCK) cells form tubes in the presence of hepatocyte growth factor in 3D cultures. Although these cells are expected to form tube structures in some microenvironments independent of chemical stimulation, little is known about the cellular mechanisms in organizing such an anisotropic multicellular structure. Here, we report 3D culture conditions that induce MDCK tubulogenesis without growth factor stimulation. We found that the cells spontaneously form elongated tube structures through aggregation processes in a specific range of both constituent cell number and scaffold gel concentration, while they form spherical aggregates in other conditions. We then examined cellular activities affecting tubulogenesis and showed that cell proliferation is not required for the tube elongation. Furthermore, we revealed that cells in the tube tips generate traction forces and pull the surrounding scaffold gel to migrate, resulting in the tube elongation. Our results suggest that the constituent cells during the aggregation process interact each other via mechanical forces transmitted in the scaffold gel, leading to the spontaneous tube formation.

OBJECTIVES Lung transplantation is the only effective therapy for patients with end-stage lung disease but an organ shortage crisis necessitates the development of alternative therapies. Recent studies have highlighted the potential of foetal tissue transplantation to facilitate the regeneration of vital organs such as liver that have been damaged by lethal diseases. Herein, with the aim of restoring pulmonary function, we hypothesized that allogenic foetal lung tissue implantation would attenuate severe respiratory failure. METHODS Lung tissue from the foetuses of pregnant green fluorescent protein-C57BL/6 mice at 13.5 days of gestation was injected into the left lungs of recipient mice. Severe lung injury was induced by paraquat, and we analysed the survival rate and pathohistological findings after 1 month. RESULTS The survival rate of the therapy group was 39%, which was significantly higher than the vehicle group at 5.9% (P = 0.034). Immunochemical staining showed that positive cytoplasmic stained cells with anti-interleukin-10 antibody were identified in the gland-like structure of embryonic day 13.5 foetal lung. At 4 weeks after orthotopic implantation, haematoxylin and eosin staining showed reduced lung inflammatory cells, reduced lung oedema and increased active cell proliferation of foetal lung cells. Lung injury score showed that the airway septal thickening revealed statistically significant differences between vehicle and foetal lung therapy (P CONCLUSIONS Immature foetal lungs improved the survival rate of mice with paraquat-induced severe lung injury, establishing the need for systematic follow-up studies. The anti-inflammatory cytokine in the tissue from embryonic day 13.5 foetal lung might suppress severe lung injury.

During cochlear development, hair cells (HCs) and supporting cells differentiate in the prosensory domain to form the organ of Corti, but how one row of inner HCs (IHCs) and three rows of outer HCs (OHCs) are organized is not well understood. Here, we investigated the process of HC induction by monitoring Atoh1 expression in cochlear explants of Atoh1-EGFP knock-in mouse embryos and showed that only the cells that express Atoh1 over a certain threshold are selected for HC fate determination. HC induction initially occurs at the medial edge of the prosensory domain to form IHCs and subsequently at the lateral edge to form OHCs, while Hedgehog signaling maintains a space between IHCs and OHCs, leading to formation of the tunnel of Corti. These results reveal dynamic Atoh1 expression in HC fate control and suggest that multi-directional signals regulate OHC induction, thereby organizing the prototype of the organ of Corti.

Size control in biological tissues involves multicellular communication via mechanical forces during development. Although fundamental cellular behaviours in response to mechanical stimuli underlie size maintenance during morphogenetic processes, the mechanisms underpinning the cellular mechano-response system that maintains size along an axis of a polarized tissue remain elusive. Here, we show how the diameter of an epithelial tube is maintained during murine epididymal development by combining quantitative imaging, mechanical perturbation and mathematical modelling. We found that epithelial cells counteract compressive forces caused by cell division exclusively along the circumferential axis of the tube to produce polarized contractile forces, eventually leading to an oriented cell rearrangement. Moreover, a mathematical model that includes the polarized mechano-responsive regime explains how the diameter of proliferating tubes is maintained. Our findings pave the way for an improved understanding of the cellular response to mechanical forces that involves collective multicellular behaviours for organizing diverse tissue morphologies., Summary: Polarized cellular constriction responding to mechanical stress controls the diameter of a developing epithelial tube during murine epididymal development.

Cellular behaviors responding to mechanical forces control the size of multicellular tissues as demonstrated in isotropic size maintenance of developing tissues. However, how mechanoresponse systems work to maintain anisotropic tissue size including tube radial size remains unknown. Here we reveal the system underlying radial size maintenance of the murine epididymal tubule by combining quantitative imaging, mathematical modeling, and mechanical perturbations. We found that an oriented cell intercalation making the tubule radial size smaller counteracts a cell tension reduction due to neighbor cell division along the tubule circumferential axis. Moreover, we demonstrated that the tubule cells enhance actomyosin constriction driving the cell intercalation in response to mechanical forces anisotropically applied on the cells. Our results suggest that epididymal tubule cells have endogenous systems for responding as active cell movement to mechanical forces exclusively along the circumferential axis, and the anisotropic cellular mechanoresponse spontaneously controls the tubule radial size.

SummaryA single epithelial tubule undergoes morphogenesis to form a functional shape during the development of internal organs; however, the mechanical processes that are directed by the molecular signals regulating tubular morphogenesis are poorly understood. Here, axial tubular buckling triggered by cell proliferation is shown to drive the morphogenesis of murine epididymal tubules through mechanical interactions between the developing epithelial tubule and its surrounding tissues. Through immunofluorescence labeling and mathematical modeling, epididymal tubule shape formation is found to depend on two factors: cell proliferation area in the tubule and mechanical resistance from the tissues surrounding the tubule. Moreover, experimental perturbations of these two factors alter the shape of the epididymal tubule as predicted by the mathematical model, suggesting that the shape of the epididymal tubule spontaneously emerges through mechanical coupling between developing tissues instead of by growing according to a predetermined fate.

Purpose Lung transplantation is an effective therapy for end-stage patients with chronic respiratory failure due to various benign lung diseases. The organ shortage crisis is a serious problem, so it is required to develop alternative therapies. There have been reports of organ organoids ameliorating organ function but there have been few reports of lung organoids being capable of restoring respiratory function. We therefore hypothesized that immature mouse fetal lungs would ameliorate severe respiratory failure and connect the airways between recipient and donor lungs. Methods Lungs from the fetuses of pregnant GFP-C57BL/6 mice at 13.5 days gestation were injected into the left lungs of recipient mice. Severe lung injury was induced by paraquat, and we analyzed survival rate, pulmonary function, PaO2/FiO2, lung cytokine levels and pathohistological findings with light-sheet microscopy after 1 month. Results At 4 weeks after orthotopic implantation histopathological findings showed that there was no airway connection between recipient lung and immature fetal lungs of mice. However, hematoxylin and eosin staining showed reduced lung inflammatory cells, reduced lung edema, and active cell proliferation of fetal lung cells. Maximum airway pressures of the sham, vehicle and therapy groups (n=3, 1 and 6 respectively) were 8.91±1.47, 9.75, and 9.21±1.22 mmHg after one month. The airway compliance of the sham, vehicle and therapy groups were 0.041±0.017, 0.036 and 0.037±0.008 mmHg. The arterial partial blood pressures in the left ventricle of the sham, vehicle and therapy groups under FiO2 1.0 were 449±13.4, 516 and 427±28.4 mmHg. The survival rate of the therapy group increased from 5.9% to 39%, compared with that of the vehicle group (p=0.0338 by log-rank test). There was no difference of lung cytokine levels between the vehicle and therapy groups. The low number of mice and the high standard deviation affected the results of lung cytokine levels. The level of cytokines of some treated mice showed severe lung injury, however, other mice showed decreased cytokine levels. Conclusion Immature fetal lungs did not self-organize airway structures, however, they improved the survival rate of mice with paraquat-induced severe lung injury.

Extracellular signal-regulated kinase (ERK) is a key factor in the widely used signaling cascade of phosphorylation-dephosphorylation cycles and plays pivotal roles in many aspects of biological processes. Experimental studies in yeast and in Drosophila embryo have suggested that the phosphorylation and spatial localization of ERK are influenced by the level of its downstream substrates. However, the mechanism, through which these substrates control properties of ERK signaling, has been unclear. I propose a mass-action kinetic model of ERK cycle with its substrate, and demonstrate that the substrate can modulate the ERK activity by directly interacting with ERK. The model shows that the addition of substrate controls the level of ERK phosphorylation positively or negatively, depending on the balance between dissociation constants of ERK-substrate interaction and properties of ERK cyclic signaling in the absence of the substrate. In addition, by considering cellular compartments, cytosol and nucleus, the substrate can lead to nuclear accumulation of ERK, suggesting that the substrate can act as a nuclear anchor of ERK. The model gives a possible mechanism that can account for substrate-mediated modulation of ERK signaling.

Whole-tissue quantification at single-cell resolution has become an inevitable approach for further quantitative understanding of morphogenesis in organ development. The feasibility of the approach has been dramatically increased by recent technological improvements in optical tissue clearing and microscopy. However, the series of procedures required for this approach to lead to successful whole-tissue quantification is far from developed. To provide the appropriate procedure, we here show tips for each critical step of the entire process, including fixation for immunofluorescence, optical clearing, and digital image processing, using developing murine internal organs such as epididymis, kidney, and lung as an example. Through comparison of fixative solutions and of clearing methods, we found optimal conditions to achieve clearer deep-tissue imaging of specific immunolabeled targets and explain what methods result in vivid volume imaging. In addition, we demonstrated that three-dimensional digital image processing after optical clearing produces objective quantitative data for the whole-tissue analysis, focusing on the spatial distribution of mitotic cells in the epididymal tubule. The procedure for the whole-tissue quantification shown in this article should contribute to systematic measurements of cellular processes in developing organs, accelerating the further understanding of morphogenesis at the single cell level.

In early lung development, epithelial tubes (lung buds) intrude into mesenchyme covered with pleural cells (lung border), and form tree-like networks, by means of repeated use of morphogenetic processes: “elongation,” “terminal bifurcation,” and “lateral budding.” When a bud is elongating, a peak of Fgf10 expression is formed in the mesenchyme near the tip; whereas when terminal bifurcation and lateral budding occur, two separate peaks are formed instead. To explain the spatial pattern of Fgf10 expression, we developed a mathematical model for the regulation of Fgf10 expression with geometrical conditions including shapes of the lung buds and the lung border. Different localization patterns of Fgf10 expression can be explained by the geometrical conditions. Fgf10 expression has a single peak when a length between the tip of lung bud and the lung border is large. When the length is small, Fgf10 expression has two peaks, whose location depends on the curvature of lung border. Developmental Dynamics 238:2813–2822, 2009. © 2009 Wiley-Liss, Inc.

症例は79歳の女性で, 急性の腹痛, 嘔気, 嘔吐を主訴として腸閉塞の診断で入院した.腹部の単純X線超音波, computed tomography検査などから腸重積症を疑い, 保存的にイレウス管による減圧をはかったが改善しないため, 5日後に緊急手術を行った.開腹所見では回盲部腸重積症で, 用手整復が困難なため回盲部切除と端々吻合術を施行した.切除標本では回盲弁から約20cmの回腸に3.5×3.0×2.5cmのポリープ状粘膜下腫瘤が存在し, 重積先進部となっていた.組織学的には, 粘膜下組織原発のまれな発育形式を呈した腹腔内デスモイド腫瘍と診断された.本症例では大腸腺腫症の併存はなく, 腹腔内デスモイド腫瘍単独症例としては文献上本邦第16例目, さらに腸重積症をきたした症例としては本邦第1例目と思われる.術後経過は順調で, 第31病日に軽快退院した.腸重積症の術前画像診断, 特に腹部超音波検査の有用性を強調したい.

Morbidly obese patients undergoing vertical banded gastroplasty were studied preoperatively and/or postoperatively to characterize its manometric pattern. The esophageal manometry using station pull through technique involved 14 preoperative patients with a mean age of 28 +/- 7 years who were a obesity index of 220 +/- 32% and 14 postoperative patients with a mean age of 30 +/- 7 years who were a obesity index of 158 +/- 23%. Preoperative lower esophageal sphincter (LES) pressure using gastric base line as O was 16.1 +/- 8.5 cmH2O and a LES length of 3.4 +/- 0.9 cm. Postoperatively, the vertical banded gastroplasty channel had a pressure of 17.7 +/- 7.1 cmH2O and a length of 8.2 +/- 1.7 cm which was higher than gastric base line. No difference was seen between preoperative LES pressure and postoperative channel pressure, however postoperative channel length was significantly (p less than 0.01) larger than preoperative LES length. We concluded that vertical banded gastroplasty for morbid obesity created a longer high pressure zone in accordance with channel which would inhibit reflux of gastric juice.

The clinical features of 86 cases of idiopathic portal hypertension, the equivalent of hepatoportal sclerosis in the United States and of noncirrhotic portal fibrosis in India, are presented. This disease is characterized by overt splenomegaly with pancytopenia, portal hypertension, and relatively mild abnormalities in liver function tests. Although differential diagnosis from liver cirrhosis is not always easy, liver histology, laparoscopy, portography, hepatic venography, and measurement of wedged hepatic vein pressure are useful in diagnosis. Prognosis is relatively benign if variceal bleeding is controlled or prevented, and the disease does not progress to cirrhosis. The etiology is still undetermined, but the liver pathology characterized by occlusive changes of the intrahepatic portal radicles, portal and peripheral fibrosis, and irregularly distributed parenchymal atrophies suggests some sort of portal venopathy that causes decreased portal perfusion of peripheral liver parenchyma. These patients with idiopathic portal hypertension were compared with 63 cases of cirrhosis with splenomegaly and 80 cases of cirrhosis without splenomegaly. There was some similarity in hematologic findings between idiopathic portal hypertension and cirrhosis with splenomegaly, but the basic disease process seemed distinctly different. The cause of marked splenomegaly does not seem to be simply congestion, and remains an enigma.

教室では十二指腸潰瘍に対する外科的治療法の一つとして迷走神経切離兼幽門成形術を90例に行った.本法術後の減酸効果は全体としてほぼ満足すべき結果であった.とくに完全迷切例では術後早期 (術後3-4週) と遠隔時 (術後1-10年) のいずれ場合も, 減酸率は60-80%で良好であった.しかしながら不完全迷切例では, 術後早期には満足すべき灘効果を示したが, 遠隔時では基礎分泌とインスリン胃酸分泌が上昇回復して, 術後早期よりかなり高値となり, 減酸効果は不十分で潰瘍再発の可能性が生じる.再発潰瘍例は90例中5例に発生し, 5.5%の頻度となる.再発潰瘍例の遠隔時胃酸分泌の特長は, 不完全迷切例と同様に刺激後胃酸分泌が高値を示す一方, 基礎分泌が不完全迷切例より有意に高いことであった.

Transabdominal mucosal transection of the esophagus was applied to 20 patients with esophageal varices. The operative procedure involves a longitudinal incision of the muscular layers which follows transection and reanastomosis of the denuded mucosa. With the preservation the muscular structure, postoperative complications such as anastomotic leakage, stenosis or regurgitation, have been minimized. The effect of this procedure on the varices proved satisfactory.

Serum levels of L-thyroxine (T4) and L-triiodothyronine (T3) of the masu salmon, Oncorhynchus masou, were measured by radioimmunoassay during the months from March to September. Serum levels of T4 and T3 varied from 0.06 to 1.44 microgram and from 0.06 to 0.35 microgram per 100 ml serum respectively, although T4 content could not be detected in some specimens. T4 levels showed a seasonal fluctuation in which a higher value was observed in April when parr-smolt transformation was proceeding. T4 level in spring, particularly in April and June, was significantly higher in smolt than in parr. T3 content showed a small variation during the observed months and did not increase at smoltification, although the content tended to decrease to a minimum level in September when T4 level showed also the lowest level.

最近, 食道癌の治療成績は著しく向上したとはいえ, いまだ進行癌が多く, また合併症も他の消化管手術に比べて多く発生し, 遠隔成績もけつして満足されるものではない. これは, 高齢者が多いこと, また癌の占居部位上早期に食事摂取制限に陥り, 患者のリスクを悪くする要因が多いことによる.最近, 経中心静脈栄養法が行われるようになり, 従来考えられなかつたほどの高カロリー輸液が可能となつた. われわれも本法を消化器外科手術前後の栄養管理にとり入れて以来, とくに食道癌の術前, 従来の胃瘻造設による栄養改善法も, この高カロリー輸液に変り得るものが多く, いままで単なる栄養状態から分割手術を行わざるを得なかつた症例の中にも, 一期的に再建術まで行えるものが多くなり適応が拡大されたといえる.また, 本法の適応で第一のものは, 種々の消化管術後縫合不全の治療であろう. 過去3年間, 食道外科その他で, 術後縫合不全のために, 従来行つて来た空腸瘻等をあらためて造設したものは1例もなく, すべて本法のみで治癒させている.われわれの方法は, 持続注入ポンプも不要で, 単に注入回路に450mμのフィルターを挿入するのみで全く普通の点滴のように行い特別な手技も必要としなく, 合併症も少ない安全な方法といえる.

Result of operative treatment in 196 cases of achalasia was reviewed. The procedures employed include cardiolysis, Heller's extramucosal myotomy, Wendel's cardioplasty, Heyrovsky's esophagocardiostomy, with and without pyloroplasty, cardiac resection and esophagocardioplasty with gastric patch. Follow-up study on 166 cases revealed that the result was classified as good in 99 cases or 59.6 per cent and improved in 46 cases or 27.7 per cent. Overall satisfactory result was obtained in 87.3 per cent. When the result was broken down to groups following three classifications, i. e. according to X-ray, endoscopic and manometric findings, the interesting correlation emerged. Heller's myotomy and esophagocardioplasty with gastric patch gave best results in early stage of achalasia, while in later stage the latter procedure seems to be the operation of choice.

食道静脈瘤における食道の動態を, 食道機能検査法としての食道内圧・pH測定を用いて正常43例と食道静脈瘤47例について比較検討した. 本症の特徴は, 静止圧曲線では下部昇圧帯圧の低下と食道胃圧差の増大であり, pH曲線では食道胃pH差の減少である. また内圧変化では陽性波の伝達機序は保たれているが, 食道各部の陽性波の波高の平低化と食道胃接合部における陰性波の波高の平低化である. これらの特徴は本症における食道の逆流防止機序の抑制や食道蠕動運動の減弱および食道胃接合部における開口機序の抑制を示している. これら本症の病態生理学的な所見は, 食道静脈瘤の破綻に影響を及ぼすものと考えられる.

肝硬変症69例,特発性門脈圧亢進症29例を対象として,食道静脈瘤に対する直達手術が,門脈血行動態へいかなる影響を及ぼすか検討した.門脈血流量は,2～5年前に直達手術を受けた肝硬変症14例,特発性門脈圧亢進症9例共に,非手術例の肝硬変症49例,特発性門脈圧亢進症17例に較べ有意に減少していた.肝硬変症5例,特発性門脈圧亢進症3例で,術前,術直後の門脈圧を測定し,またこれらの症例と肝硬変症1例を加えた計9例で術前と術後2週間目の門脈血流量の変動を検討した.術直後測定した門脈圧は,肝硬変症5例中2例で上昇,2例は下降,1例が不変であった.特発性門脈圧亢進症では,2例で減少し,巨大脾腎短絡路及び胃腎短絡路を有する1例は増加していた.術後2週間目の門脈血流量は,肝硬変症全例で減少し,特発性門脈圧亢進症では,門脈圧が術後下降した例では減少し,上昇した1例では増加していた.

We performed the esophagocardioplasty with gastric patch in the device of SATO and HIRASHIMA on 44 cases of achalasia of esophagus. We evaluated the post-operative results of these cases with esophago-cardiac manometry and pH measurement, which would be the best indicators on the esophago-cardiac function.We comfirmed that this method improved the passage prominently even in the progressed cases, judging the resting pressure and manometric configurations, and that this method preserved very well the anti-reflex function of the cardia, judging from the pH measurement.On the selection of the operative methods, we believe that, on the early cases, Heller's method and esophagocardioplasty with gastric patch are the choice and, on the progressed cases, esophagocardioplasty with gastric patch is to be performed.

The electromyogram of the esophagus has been studied by means of intramuscularly implanted electrodes to investigate pathophysiological changes seen in experimentally produced achalasia-like condition in dogs. These studies have been attempted to elucidate the functional mechanism of the esophagogastric junction.Electrodes are implanted into the esophageal muscle at 4 different points; the upper, middle, lower parts of the esophagus and the esophagogastric junction respectively and the electrodes have been kept chronically. In dogs, the electromyogram is taken during spontaneous swallowing without anesthesia or restraint while the studies are done.After the electrodes have been connected with the multi-channel amplifier, the electromyogram can be easily recorded until 3 weeks postoperatively.One day after the operation, dogs can eat and the electromyographical bursts move down to the lower part of the esophagus, judging from the electromyographical patterns at the 4 different points. The bursts are high in the maximum amplitude and long in the duration at the lower part of the esophagus as compared with the one from the upper part. Changes are slight in amplitude at the esophagogastric junction.

The pressures in the esophagus and gastro-esophageal sphincter in 22 healthy unanesthetized dogs were studied by the use of three small open tipped tubes with internal diameter of 1.4 mm. The tubes were introduced and manipulated without anesthesia through the previously made esophageal fistula. To record the resting pressure, an open tipped tube was withdrawn slowly from the stomach back to the esophagus. The swallowing action was induced by an injection of water into the esophagus.The resting pressure in the esophagus was less than that in the f undus of the stomach.A band of elevated pressure was recorded at the gastro-esophageal junctional area in thedogs while at rest. The mean maximal pressure in this area was 9.5 cm H2O and themean width was 2.6 cm H2O.As for the changes in pressure during swallowing, the peristaltic waves were seen to sweep downward in the esophagus. In the gastro-esophageal junctional area, the negative wave (relaxation) occurred 4 seconds after the onset of water injection inducing swallowing, lasting about 3 seconds, and was followed by the positive wave (contraction) lasting 3.3 seconds.

Esophageal achalasia-like phenomena have been produced experimentally on 18 dogs after the administration of the anticholinesterase agent, phthalim-idomethyl O, O-dimethylphosphorodithioate (PMP). The agent was administered intramuscularly three times a week for 40 weeks. The dose to each group was varied from 0.5, 10, 20, 25, and as much as 50mg per kg of body weight. In the group with the dose of 20mg per kg of body weight, the symptom of vomiting or dysphagia appeared most markedly. Some dogs showed diarrhea, bloody stool and hoarseness. Upon x-ray examination, dogs showed the failure of passage at the esophago-cardiac junction and the dilatation of the lower esophagus. Intraluminal pressure recording showed the abnormal simultaneous rises of the swallowing pressure wave. Histological examination around the Auerbach's plexus showed pycnotic nuclei, swelling of ganglion cells, fibrosis and round cell infiltration. The MAO stain showed no changes. The acethylcholine-esterase stain suggested the suppression of the enzyme activity.

As one of the analyses of gastric emptying mechanism, clinical observa tions and experimental observations using dogs and rats have been performed. The inucosa of the pyloric area was topically anesthetized by ingestion of 4% Lidocaine through naso-gastric tube under the right recumbent position. X-ray contrast medium was ingested into the stomach through the tube five minutes later.(1) Topical anesthesia accelerated the emptying of the gastric content under the above described condition.(2) The movement of gastric content to the duodenum was continuous without periodicity. In controls, without topical anesthetic, the movement was periodic and rhythmical.(3) Concerning the mechanism of gastric emptying, it is suggested that a pressure stimulation above a certain level to the pyloric area induces the movement of the content to the duodenum. The topical anesthesia of this area looses the control of this mechanism.(4) A possible clinical application of this method, for example, as a treatment in the cases of antrospasm or pylorospasim is suggested.

The physiology of motility of the esophago-cardiac junction, pathophysiological changes of this area due to various esophageal diseases and their clinical applications were studied by our new method of obtaining electric internal pressure curve and several interesting findings were obtained. These examinations were carried out on 91 clicnial cases (including several controls).(1) Order of food passage through the normal esophago-cardiac junction is usually controlled and carried out mainly by pressure of the esophagocardiac junction at rest, positive pressure of the esophagus, and negative and positive waves of the gastric cardia which arise simultaneously.(2) Failure in presence of this negative pressure at the gastric cardia, is noted in all 17 cases of achalasia patients. By various values of the internal pressure at rest, this disease could be divided into two groups; the flaccid type (type A) and accelerated type (type B).(3) As to the surgical indications, cardioplasty type of surgery for group A, and esophago-gastrostomy type for group B, are better suited and effective.(4) Very specific inconsistent internal pressure was obtained in 8 out of 9 clinical patients with carcinoma of the gastric cardia. These specific curves were divided into three groups; the serrated type, step ladder type and canopy type.We have concluded therefore that this method has a certain advantage in utilizing diagnostic measure for various esophageal diseases.