Noriyuki Iwama

Characterization of lymphangiogenesis in various stages of idiopathic diffuse alveolar damage

In pulmonary fibrosis, an abnormal healing process, is believed to be involved in the damage to lung tissue. This process has not been correlated with lymphangiogenesis, which has garnered current interest in relation to wound healing. The aim of the present study was to clarify the characteristics of lymphangiogenesis in pulmonary fibrosis associated with idiopathic diffuse alveolar damage. Formalin-fixed and paraffin-embedded lung tissues from 13 autopsy cases with idiopathic diffuse alveolar damage were used. Antibodies specific to CD34 and D2-40 were used to detect blood vessels and lymphatics, respectively, and immunohistochemical examinations and morphometry analyses were performed. The standardized density of capillaries was increased significantly in the exudative stage of diffuse alveolar damage, whereas that of the lymphatics remained unchanged. In the proliferative stage, new lymphatics emerged, primarily in the intra-alveolar fibrotic lesions where capillaries were absent. In the fibrotic stage, in which the lung was shrunken, as revealed by the elevated density of pulmonary arteries, the standardized density of capillaries was reduced significantly. The standardized area density of the interstitium was elevated in the proliferative stage and subsequently reduced in the fibrotic stage. Three-dimensional reconstruction of images revealed that some new lymphatics lacked connection to existing lymphatics. During the progression of diffuse alveolar damage, lymphangiogenesis occurs independent of capillary angiogenesis.

Macrophages participate in lymphangiogenesis in idiopathic diffuse alveolar damage through CCL19-CCR7 signal

An abnormal healing process is believed to be involved in lung tissue damage in pulmonary fibrosis. Lymphangiogenesis has been determined to play a role in structural remodeling in diffuse alveolar damage. However, the mechanism of lymphangiogenesis remains unclear. The aim of this study is to investigate the cellular mechanisms of lymphangiogenesis in diffuse alveolar damage, focusing on the roles of macrophages. Formalin-fixed and paraffin-embedded lung tissues from 13 autopsy cases with idiopathic diffuse alveolar damage were used. Antibodies specific for D2-40 and CD68 were mainly used as the markers for lymphatics and macrophages, respectively, and immunohistochemical examinations and morphometric analyses were performed. Immunohistochemistry showed the aggregation of numerous CD68+ macrophages around newly formed lymphatics in the intraalveolar fibrotic lesions in the proliferative stage, and some of the CD68+ macrophages were colocalized with the lymphatic endothelium. These macrophages were characterized by the expression of vascular endothelial growth factor-C. Among the 3 stages of diffuse alveolar damage, the number of Ki-67+ cells on the lymphatic endothelium tended to increase in newly formed lymphatics of the proliferative stage. In addition, the number of CD68+ macrophages on the lymphatic endothelium was significantly increased in the newly formed lymphatics of the proliferative stage more than those of the fibrotic stage. The CD68+ macrophages around the newly formed lymphatics coexpressed CCR7. Dual immunostaining showed the coexpression of CCL19-a ligand for CCR7-on the lymphatic endothelium. Thus, macrophages may participate in lymphangiogenesis in diffuse alveolar damage, which is facilitated by CCL19 and CCR7.

The definition of fibrogenic processes in fibroblastic foci of idiopathic pulmonary fibrosis based on morphometric quantification of extracellular matrices

There is limited information regarding the process of tissue remodeling in fibroblastic foci associated with idiopathic pulmonary fibrosis. The aim of this study was to identify the different pathologic stages of tissue remodeling in fibroblastic foci based on the histopathologic differences in the glycosaminoglycan distribution and collagen deposition. In addition, we also aimed at clarifying the stage-specific characteristics by taking into consideration the expression pattern of matrix metalloproteinase and angiogenesis. Lung biopsies of 16 patients with idiopathic pulmonary fibrosis were used. The presence of glycosaminoglycans was detected by Alcian blue staining, and type I collagen was detected by immunohistochemical analysis with a primary antibody specific to the cross-linked carboxyterminal telopeptide of type I collagen. The fibroblastic foci characterized by the expression intensity of Alcian blue and telopeptide of type I collagen were divided into 3 groups, namely, Alcian blue(+)telopeptide of type I collagen(weak), Alcian blue(+)telopeptide of type I collagen(+), and Alcian blue(weak)telopeptide of type I collagen(+); consequently, 3 new stages were defined--stages I, II, and III, respectively. A significant inverse correlation was observed between the area densities of Alcian blue(+) and telopeptide of type I collagen(+) in fibroblastic foci. Stage I was characterized by the expression of matrix metalloproteinase-2 and tissue inhibitor of matrix metalloprotease-2 in fibroblasts and the overlying epithelium of fibroblastic foci, and also the absence of capillary angiogenesis. In contrast, the expression of these proteins was attenuated in stage III, except for that of matrix metalloproteinase-2 in fibroblasts. In stages II and III, capillary angiogenesis was observed. Lymphangiogenesis was undetected in all the 3 stages. Thus, pathologic staging helps understand the roles of the factors involved in tissue remodeling in idiopathic pulmonary fibrosis.

Blood Pressure Measured in the Clinic and at Home During Pregnancy Among Nulliparous and Multiparous Women: The BOSHI Study

BACKGROUND Hypertension during pregnancy can cause serious problems during delivery, such as stroke, premature delivery, or low birthweight. Nulliparity is believed to be a risk factor for hypertension during pregnancy. However, the relationship between parity and blood pressure determined at home during pregnancy is still unknown. METHODS We assessed the incidence of gestational hypertension or preeclampsia in 575 nulliparous and multiparous women. Also, we examined blood pressure measured in the clinic and at home among 530 normotensive pregnant women who received antenatal care at a maternity hospital in Japan. Clinic blood pressures (CBPs) were obtained by duplicate measurement at each antenatal care visit. The participants were also required to measure their own blood pressures every morning at home while they were pregnant. A linear mixed model was used for analysis of the blood pressure course throughout pregnancy. RESULTS A total of 315 nulliparous and 215 multiparous women were entered into this study (mean age, 30.1 ± 4.6 years and 33.0 ± 4.1 years, respectively). CBP levels during pregnancy among nulliparous women were significantly higher than among multiparous women (P = 0.02/P <0.0001 for systolic/diastolic blood pressure), whereas there were no significant differences in home blood pressure (HBP) levels during pregnancy between the two groups (P = 0.4/P = 0.2 for systolic/diastolic blood pressure). CONCLUSIONS HBP levels during pregnancy were shown not to differ between nulliparous and multiparous women, while CBP levels during pregnancy were higher among nulliparous than among multiparous women.

Architectural pattern of gastric adenocarcinoma - A 3-dimensional reconstruction study

The 3-D microstructure of various adenocarcinomas was studied by graphic reconstruction of tubules and lumina from serial sections in four gastrectomy specimens in order to contribute to better diagnosis of the lesion by establishing the morphology of atypical glands. The cell nests in the moderately differentiated type had multiple anastomoses with one another forming a network, a pattern different from arborescent normal glands, while the lumina were separated into many small vesicles, giving the nests a porous character. Well differentiated tumors had more connections between the lumina forming a luminal network, whereas in the poorly differentiated lesions the nests also began to split into fragments. These findings provided a new viewpoint from which to establish an architectural basis for the discrimination of dysplastic from overtly malignant lesions.

Atypical glands in gastric adenoma. Three-dimensional architecture compared with carcinomatous and metaplastic glands.

The pathogenetic relationship of gastric adenoma to carcinoma remains unsettled, partly due to the difficulty in discriminating between the atypical tubules of adenoma and those of adenocarcinoma. Although it has been said that this discrimination should depend not only on cellular changes but also on disorganization of glands, the latter has not been described in accurate morphological terms. In view of this, gastrectomy specimens from three patients with tubular adenoma were submitted to graphic reconstruction of atypical glands from serial sections, and were compared with well differentiated adenocarcinoma and metaplastic mucosa. Reconstruction disclosed that in adenoma, unlike in metaplastic mucosa, atypical tubules had multiple connections with adjacent ones, forming a network. At some sites of anastomosis the lumen was also connected. Though this pattern was similar to that of well differentiated adenocarcinoma, the meshes of the network were much more coarse than in the latter, showing that adenoma was a mere miniature of adenocarcinoma. The porous structure, the commonest architecture of adenocarcinoma, was never found in adenoma. There were in addition giant glands with complicated branching which, together with microcysts forming at mucosal bottom, caused convolution and twisting of tubules, producing those abnormal patterns in section on which too much stress is placed.

Daily Serial Hemodynamic Data During Pregnancy and Seasonal Variation: The BOSHI Study

Although there are some reports that low plasma volume or increased cardiac output is associated with developing preeclampsia, there are few reports of daily serial hemodynamic data during pregnancy. A total of 37 092 home blood pressure (BP) and heart rate (HR) measurements were obtained from 425 normal pregnant women. Heart rate and shock index (SI) gradually increased by gestational week 32 and then decreased, whereas double product (DP) increased linearly during pregnancy. Although systolic BP and DP were consistently and negatively correlated with daily minimum outside temperature, HR and SI were positively correlated with minimum outside temperature in summer.

Maternal clinic and home blood pressure measurements during pregnancy and infant birth weight: the BOSHI study

This prospective cohort study compared measurements of maternal home blood pressure (HBP) with clinic blood pressure (CBP) before 20 weeks’ gestation to determine associations with the risk of delivering a lower birth weight infant. A total of 605 Japanese women were included. Exposures were initial CBP, made between 10 weeks 0 days and 19 weeks 0 days, and HBP for comparison made within 1 week of CBP. Outcome was infant’s birth weight, categorized and ranked as follows: ⩾3500 g, 3000–3499 g, 2500–2999 g and <2500 g. The proportional odds model with possible confounding factors was applied to compare the associations between CBP and HBP on infant birth weight. When both CBP and HBP were included simultaneously, the adjusted odds ratios (ORs) per 1 standard deviation (1s.d.) increase in clinic and home diastolic BP (DBP) were 1.06 (95% confidence interval (CI): 0.87–1.30) and 1.28 (95% CI: 1.04–1.58), respectively. The adjusted ORs per 1s.d. increase in clinic and home mean arterial pressure (MAP) were 1.02 (95% CI: 0.83–1.24) and 1.29 (95% CI: 1.04–1.59), respectively. Systolic BP measurement was not associated with infant birth weight. In conclusion, high maternal home DBP and MAP before 20 weeks’ gestation was associated with a higher risk of lower infant birth weight than clinic DBP and MAP. Therefore, in addition to CBP, it may be worth having pregnant women measure HBP to determine the risk of lower infant birth weight.

Parity as a factor affecting the white-coat effect in pregnant women: the BOSHI study.

Parity has previously been reported to affect the difference in blood pressure (BP) measured in the office and at home, also known as the white-coat effect, during pregnancy. The objective of this study was to identify possible factors that cause the white-coat effect during pregnancy, focusing on parity. In total, 530 pregnant women (31.3±4.7 years old) who delivered at a maternal clinic were eligible for the study. The association between parity and the white-coat effect (clinic BP compared with home BP) was investigated for each trimester of pregnancy by multivariate analysis of covariance adjusted for age, body mass index, family history of hypertension and smoking habits. The magnitudes of the white-coat effect for systolic BP in the first, second and third trimesters were 4.1±9.8, 3.4±7.1 and 1.8±6.0 mm Hg, respectively and those for diastolic BP were 3.8±7.4, 1.6±5.8 and 2.4±4.9 mm Hg, respectively. Parity was significantly and negatively associated with the white-coat effect for systolic BP in the first trimester of pregnancy (nulliparous women: 5.07±0.61 mm Hg and multiparous women: 2.78±0.74 mm Hg, P=0.02) as well as for diastolic BP in the second and third trimesters of pregnancy. Age, body mass index, family history of hypertension and smoking were not significantly associated with the white-coat effect in any trimester of pregnancy. Parity may have an influence on the white-coat effect in pregnancy; however, the observed effect, on average 1–2 mm Hg, was small.

Expression of steroidogenic enzymes and their transcription factors in cortisol-producing adrenocortical adenomas: immunohistochemical analysis and quantitative real-time polymerase chain reaction studies

Adrenal Cushing syndrome (CS) is caused by the overproduction of cortisol in adrenocortical tumors including adrenal cortisol-producing adenoma (CPA). In CS, steroidogenic enzymes such as 17α-hydroxylase/17, 20-lase (CYP17A1), 3β-hydroxysteroid dehydrogenase (HSD3B), and 11β-hydroxylase (CYP11B1) are abundantly expressed in tumor cells. In addition, several transcriptional factors have been reported to play pivotal roles in the regulation of these enzymes in CPA, but their correlations with those enzymes above have still remained largely unknown. Therefore, in this study, we examined the status of steroidogenic enzymes and their transcriptional factors in 78 and 15 CPA cases by using immunohistochemistry and quantitative real-time polymerase chain reaction (qPCR), respectively. Immunoreactivity of HSD3B2, CYP11B1, CYP17A1, steroidogenic factor-1 (SF1[NR5A1]), GATA6, and nerve growth factor induced-B (NGFIB[NR4A1]) was detected in tumor cells. Results of qPCR analysis revealed that expression of HSD3B2 mRNA was significantly higher than that of HSD3B1, and CYP11B1 mRNA was significantly higher than CYP11B2. In addition, the expression of CYP11B1 mRNA was positively correlated with those of NR5A1, GATA6, and NR4A1. These results all indicated that HSD3B2 but not HSD3B1 was mainly involved in cortisol overproduction in CPA. In addition, NR5A1, GATA6, and NR4A1 were all considered to play important roles in cortisol overproduction through regulating CYP11B1 gene transcription.