Effect of porcine placental extract on the mild menopausal symptoms of climacteric women.

OBJECTIVES: This study assessed the effects of oral porcine placental extract (PPE) on the mild menopausal symptoms of climacteric women. METHODS: In this 12-week, multicenter, randomized, double-blind, placebo-controlled, parallel-group study, 50 climacteric Japanese women were randomized 1 : 1 to oral PPE (300 mg/day) or placebo. Menopausal symptoms were evaluated by using the Simplified Menopausal Index (SMI), as were serum estradiol (E2) and follicle stimulating hormone (FSH) levels. Blood biochemical and cellular and urinary tests were done to evaluate safety aspects of repeated oral administration of PPE. RESULTS: The total SMI score of the PPE group was significantly more improved after 12 weeks than that of the placebo group (p = 0.031). This score and three subscores (vasomotor, psychological, and somatic symptoms) were significantly improved at 8 and/or 12 weeks compared with the initial values in the PPE group (p < 0.05). E2 and FSH levels were not improved in either group. No adverse events were observed. CONCLUSIONS: Oral PPE at 300 mg/day improved the mild menopausal symptoms of climacteric women. Since oral PPE did not improve serum E2 and FSH levels, PPE is thought not to ameliorate hormonal balance itself but to improve subjective feelings of climacteric women.

Randomized phase III study of bevacizumab plus FOLFIRI and bevacizumab plus mFOLFOX6 as first-line treatment for patients with metastatic colorectal cancer (WJOG4407G).

BACKGROUND: FOLFIRI and FOLFOX have shown equivalent efficacy for metastatic colorectal cancer (mCRC), but their comparative effectiveness is unknown when combined with bevacizumab. PATIENTS AND METHODS: WJOG4407G was a randomized, open-label, phase III trial conducted in Japan. Patients with previously untreated mCRC were randomized 1:1 to receive either FOLFIRI plus bevacizumab (FOLFIRI + Bev) or mFOLFOX6 plus bevacizumab (mFOLFOX6 + Bev), stratified by institution, adjuvant chemotherapy, and liver-limited disease. The primary end point was non-inferiority of FOLFIRI + Bev to mFOLFOX6 + Bev in progression-free survival (PFS), with an expected hazard ratio (HR) of 0.9 and non-inferiority margin of 1.25 (power 0.85, one-sided α-error 0.025). The secondary end points were response rate (RR), overall survival (OS), safety, and quality of life (QoL) during 18 months. This trial is registered to the University Hospital Medical Information Network, number UMIN000001396. RESULTS: Among 402 patients enrolled from September 2008 to January 2012, 395 patients were eligible for efficacy analysis. The median PFS for FOLFIRI + Bev (n = 197) and mFOLFOX6 + Bev (n = 198) were 12.1 and 10.7 months, respectively [HR, 0.905; 95% confidence interval (CI) 0.723-1.133; P = 0.003 for non-inferiority]. The median OS for FOLFIRI + Bev and mFOLFOX6 + Bev were 31.4 and 30.1 months, respectively (HR, 0.990; 95% CI 0.785-1.249). The best overall RRs were 64% for FOLFIRI + Bev and 62% for mFOLFOX6 + Bev. The common grade 3 or higher adverse events were leukopenia (11% in FOLFIRI + Bev/5% in mFOLFOX6 + Bev), neutropenia (46%/35%), diarrhea (9%/5%), febrile neutropenia (5%/2%), peripheral neuropathy (0%/22%), and venous thromboembolism (6%/2%). The QoL assessed by FACT-C (TOI-PFC) and FACT/GOG-Ntx was favorable for FOLFIRI + Bev during 18 months. CONCLUSION: FOLFIRI plus bevacizumab was non-inferior for PFS, compared with mFOLFOX6 plus bevacizumab, as the first-line systemic treatment for mCRC. CLINICAL TRIALS NUMBER: UMIN000001396.

Randomised phase II trial of S-1 plus oxaliplatin vs S-1 in patients with gemcitabine-refractory pancreatic cancer.

BACKGROUND: This randomised, open-label, multicenter phase II study compared progression-free survival (PFS) of S-1 plus oxaliplatin (SOX) with that of S-1 alone in patients with gemcitabine-refractory pancreatic cancer. METHODS: Patients with confirmed progressive disease following the first-line treatment with a gemcitabine-based regimen were randomised to receive either S-1 (80/100/120 mg day(-1) based on body surface area (BSA), orally, days 1-28, every 6 weeks) or SOX (S-1 80/100/120 mg day(-1) based on BSA, orally, days 1-14, plus oxaliplatin 100 mg m(-2), intravenously, day 1, every 3 weeks). The primary end point was PFS. RESULTS: Between January 2009 and July 2010, 271 patients were randomly allocated to either S-1 (n=135) or SOX (n=136). Median PFS for S-1 and SOX were 2.8 and 3.0 months, respectively (hazard ratio (HR)=0.84; 95% confidence interval (CI), 0.65-1.08; stratified log-rank test P=0.18). Median overall survival (OS) was 6.9 vs 7.4 months (HR=1.03; 95% CI, 0.79-1.34; stratified log-rank test P=0.82). The response rate (RR) was 11.5% vs 20.9% (P=0.04). The major grade 3/4 toxicities (S-1 and SOX) were neutropenia (11.4% and 8.1%), thrombocytopenia (4.5% and 10.3%) and anorexia (12.9% and 14.7%). CONCLUSIONS: Although SOX showed an advantage in RR, it provided no significant improvement in PFS or OS compared with S-1 alone.

Comparison of preoperative chemotherapy using docetaxel, cisplatin and fluorouracil with cisplatin and fluorouracil in patients with advanced carcinoma of the thoracic esophagus.

We retrospectively compared preoperative docetaxel, cisplatin, and fluorouracil (DCF) with cisplatin and fluorouracil (CF) in patients with esophageal cancer. The study included patients with advanced thoracic esophageal carcinoma (excluding T4 tumors) receiving preoperative chemotherapy. In the DCF group, five patients received two courses of treatment every 4 weeks, and 33 patients received three courses every 3 weeks. In the CF group, 38 patients received two courses of treatment every 4 weeks. Patients underwent curative surgery 4-5 weeks after completing chemotherapy. Patient demographic characteristics did not differ between the two study groups. The incidence of a grade 3 or 4 hematologic toxicity was significantly higher in the DCF group (33 patients) than in the CF group (five patients; P < 0.001). Curative resection was accomplished in 79% of patients in the DCF group and 66% in the CF group (P = 0.305). There were no in-hospital deaths. The incidence of perioperative complications did not differ between the groups. A grade 2 or 3 histological response was attained in a significantly higher proportion of patients in the DCF group (63%) than in the CF group (5%; P < 0.001). Progression-free survival and overall survival were significantly higher in the DCF group (P = 0.013, hazard ratio 0.473; P = 0.001, hazard ratio 0.344). In conclusion, a grade 3 or 4 hematologic toxicity was common in the DCF group but was managed by supportive therapy. Histological response rate, progression-free survival, and overall survival were significantly higher in the DCF group compared with the CF group.

Local conductance measurements of double-layer graphene on SiC substrate.

The microscopic structural and electrical properties of few-layer graphene grown on an SiC substrate were characterized by low-energy electron microscopy, transmission electron microscopy and scanning probe microscopy measurements of local conductance. The double-layer graphene sheet was confirmed to be continuous across the atomic steps on the buried SiC substrate surface, and the measured local conductance was clearly modified in the vicinity of the steps. The conductance decreased (slightly increased) at the lower (upper) side of the steps, suggesting deformation-induced strain is the origin of the conductance modification. From the contact force dependence of the conductance images, the effective contact areas for both nanogap-probe and point-probe measurements were estimated.

Evidence for involvement of mast cells in tumor suppression in mice.

Mast cell-deficient W/Wv mice had an increased tumor incidence after subcutaneous treatment with 3-methylcholanthrene, compared with that in normal congenic mice treated in the same way. This increased tumor incidence was suppressed to the normal level when the carcinogen was given after the mast cell deficiency had been overcome by transplantation of bone marrow cells from normal congenic mice. The W/Wv mice, however, were not defective in natural killer and T-cell-mediated cytotoxic activities. These results support the hypothesis that mast cells are involved in tumor suppression.

Experimental radiation therapy and apparent radioresistance of autochthonous tumors subcutaneously induced with 3-methylcholanthrene in mice.

The radiation response of autochthonous tumors induced by s.c. injection of 3-methylcholanthrene in ICR/JCL mice was studied. The tumors were mostly fibrosarcomas and grew with an average volume-doubling time of 2.6 days, independently of the time tumor appearance or dose of the chemical used. The tumors were locally and singly irradiated with 6-MV X-rays through a filter. Autochthonous tumors were similar to their transplants in postirradiation regression and gross cellular radiosensitivity (Do, 400 rads), as estimated from regrowth time. However, most of the autochthonous tumors irradiated with single doses of up to 7.4 kilorads recurred within 120 days, whie in identically irradiated tumor transplants a complete cure was obtained with doses of 6.5 kilorads or more. The 50% tumor control dose with no recurrence within 120 days was 4.4 kilorads for the transplants. Transplantation of 26 irradiated autochthonous fibrosarcomas produced only a few tumors in autochthonous hosts, and they were completely rejected in other, previously tumor-free mice; a 48% recurrence was noted even after resection of the irradiated tumor. Two possibilities for the apparent radioresistance of autochthonous tumors were suggested: (a) existence of radioresistant cells in autochthonous tumors; and (b) induction of a second new tumor by the additive effect of the chemical and radiation. Although our results are preliminary, the second possibility is not favored since autochthonous tumors induced with low doses of 3-methylcholanthrene recurred with high frequency after irradiation. Moreover, irradiation of tumor-free mouse skin that had been treated with the carcinogen produced tumors at a very low frequency.

Involvement of ovarian innervation in steroid secretion.

The effects of electrical stimulation of the brain on the ovarian venous plasma concentrations of estradiol (E2) and progesterone (P) were investigated in female rats hypophysectomized and adrenalectomized at 1000 h on the day of proestrus. Stimulation was applied during the proestrous critical period under pentobarbital anesthesia, and contralateral ovarian venous blood was collected from 105-120 min after the stimulation E2 and P concentrations, estimated by RIA, were significantly increased by stimulation of the medial basal prechiasmatic area (PVA), ventromedial hypothalamus, and areas in the mesencephalon and decreased by stimulation of the dorsal hippocampus, lateral amygdala, and mesencephalic areas. Ovarian blood flow was not affected by these stimulations. Ovarian nervotomy in the proestrous rats abolished E2 secretion induced by PVA stimulation. In intact proestrous rats, electrochemical stimulation of the PVA but not the preoptic suprachiasmatic area increased E2 and P concentrations, although stimulation of both areas commonly potentiated gonadotropin secretion. Present findings suggest that the efferent neural system from the brain to the ovaries is supplementary to the brain-pituitary-ovarian hormonal mechanisms in the regulation of ovarian steroid secretion, and the system may be required for the adjustment of ovarian responsiveness and sensitivity to gonadotropins. In addition, evidence for neural feedback on basal FSH secretion in the ovarian-denervated proestrous rats is presented in this study.

Role of natriuretic peptide receptor type C in Dahl salt-sensitive hypertensive rats.

The natriuretic peptide system is suggested to be involved in the pathogenesis of salt-sensitive hypertension; a recent report indicated that disruption of the atrial natriuretic peptide precursor gene caused salt-sensitive hypertension. However, natriuretic peptide receptor (NPR)-A knockout mice did not show enhanced salt sensitivity of blood pressure. The aim of the present study was to investigate the role of NPR-C, the other receptor for atrial natriuretic peptide, in increased salt sensitivity of blood pressure. Dahl salt-sensitive (DS) and salt-resistant (DR) rats were placed on a 0.3% or 8% NaCl diet for 4 weeks. Blood pressure was elevated by salt loading only in DS rats. RNase protection assay demonstrated that NPR-C transcript level in the kidney was reduced by chronic salt loading in both DR and DS rats, whereas expression of NPR-A and NPR-B was not altered. The reduction of NPR-C mRNA in response to salt loading was enhanced in DS compared with DR rats. In situ hybridization indicated that the salt-induced NPR-C change was attributed mainly to suppressed expression of NPR-C in the podocytes. NPR-C gene expression was regulated by salt loading in a tissue-specific manner; the marked decrease in NPR-C mRNA by salt loading was seen only in the kidney. These data suggest that the exaggerated salt-induced reduction of NPR-C in the kidney of DS rats may play an important role in the pathogenesis of salt hypertension in this animal, possibly related to impaired renal sodium excretion.

Priming effect of hydroxyapatite on the chemiluminescence response in human polymorphonuclear leukocytes.

In order to determine whether hydroxyapatite modulates the response of polymorphonuclear leukocyte (PMN) to oxidative stimuli, human PMNs were incubated with a non-activating concentration (1 or 10 micrograms/ml) of hydroxyapatite prior to stimulation with N-formyl-methionyl-leucyl-phenylalanine (FMLP; 0.1 or 1 microM), phorbol 12-myristate 13-acetate (PMA; 100 pg/ml), sodium fluoride (50 microM), zymosan (1 microgram/ml), or the calcium ionophore A23187 (0.1 microM). Chemiluminescence was measured with an automatic microcomputer-controlled luminescence analyzer at 37 degrees C. Hydroxyapatite alone did not stimulate chemiluminescence at concentrations below 10 micrograms/ml. Levels 300-400% higher than 'stimulus only' controls without preincubation with hydroxyapatite have been recorded. This synergism between hydroxyapatite and subsequent stimuli reveals a new activity of hydroxyapatite and suggests that particulate material may prepare PMNs for an exaggerated inflammatory response to other phlogistic mediators. This is the first report demonstrating PMNs primed with particulate material.

The effect of crystallinity on hydroxyapatite-induced production of reactive oxygen metabolites by polymorphonuclear leukocytes.

To assess the role of crystallinity in biological response, we quantitated the generation of reactive oxygen metabolites in human polymorphonuclear leukocytes (PMN) with a chemiluminescence assay using three hydroxyapatite preparations with sintering temperatures of 1,200 degrees C and 900 degrees C and a drying temperature of 110 degrees C on the basis of equal weights (1 mg/ml). These crystals have almost the same average diameters and similar average zeta potentials. The crystals prepared at higher temperatures have higher crystallinity, or larger domain sizes, which were calculated by X-ray diffraction line broadening. The production of reactive oxygen metabolites by PMN in hydroxyapatite of 1,200 degrees C was 10-times that by PMN in hydroxyapatite of 900 degrees C and more than 50-times greater than that in hydroxyapatite of 110 degrees C. A single linear correlation was observed for a plot of log (peak chemiluminescence levels) vs. a plot of log (domain sizes). These results clearly show that the maximal effect of crystallinity on hydroxyapatite-induced production of reactive oxygen metabolites by human PMN was seen at higher crystallinity.

Tissue distribution and localization of natriuretic peptide receptor subtypes in stroke-prone spontaneously hypertensive rats.

OBJECTIVE: To investigate tissue distribution and localization of the natriuretic peptide receptor (NPR) subtypes' messenger RNA (mRNA) and to compare their expression between stroke-prone spontaneously hypertensive rats (SHR-SP) and Wistar-Kyoto (WKY) rats. METHODS: Total RNA was extracted from organs of SHR-SP and WKY rats aged 13 weeks. The mRNA level was examined by RNase protection assay. The localization of the transcripts was determined by in-situ hybridization. RESULTS: In SHR-SP aged 13 weeks, NPR-A was expressed most abundantly in the adrenal gland, lung and aorta, in that order. NPR-B was expressed highly in the uterus and ovary, and also in the lung, adrenal, and brain. NPR-C was expressed predominantly in the atrium and mesentery, less so in the lung, vein, and kidney. In the adrenal gland, NPR-A was expressed mainly in zona glomerulosa cells. In the atrium, NPR-C was expressed throughout the wall. In the mesentery, NPR-C mRNA was detected mainly in adipocytes. In the kidney, NPR-C was found predominantly in podocytes. Whereas the levels of expression of NPR subtypes in most tissues examined did not differ between SHR-SP and WKY rats, the NPR-C mRNA level was significantly greater in the kidneys of SHR-SP than it was in those of WKY rats. CONCLUSIONS: These results indicated that each NPR subtype had a distinct tissue distribution pattern and that the expression of NPR-C in the kidneys of SHR-SP was greater than that in the kidneys of WKY rats.

Protection of alpha(3) integrin-mediated podocyte shape by superoxide dismutase in the puromycin aminonucleoside nephrosis rat.

Because reactive oxygen species (ROS) are involved in the development of puromycin aminonucleoside nephrosis (PAN), we examined whether superoxide dismutase (SOD) could ameliorate this condition. Phosphatidyl choline-bound SOD (PC-SOD) has higher affinity for the cell membrane than recombinant human SOD (rhSOD). In this study, PC-SOD had a longer half-life in the circulation and also higher affinity to renal fractions (glomerulus, brush border, and tubulus) than rhSOD. PAN was induced in rats with single injections of puromycin aminonucleoside. Rats were divided into four groups: group P, PAN rats without treatment; group PC-T and group rh-T, PAN rats treated with 30,000 U/kg PC-SOD and rhSOD, respectively; and group C, normal controls. The effect of PC-SOD versus rhSOD on PAN was evaluated by morphological podocyte changes (podocyte density along the GBM) and alpha(3) integrin expression at days 4 and 10. Proteinuria was measured over time until day 14. Distribution and quantitation of alpha(3) integrin were studied by confocal laser scan microscopy. On day 4, glomerular ROS was measured by chemiluminescence without stimulation. PC-SOD decreased proteinuria to the control level, but rhSOD only decreased proteinuria by 31%. PC-SOD significantly improved podocyte density (P < 0.05 versus group P). Total alpha(3) integrin expression decreased in the P and rh-T groups at day 4 and then had recovered by day 10, but the polarity of the site of expression did not recover. PC-T preserved both the amount and polarity of integrin expression on days 4 and 10. PC-SOD significantly suppressed ROS generation in PAN (P < 0.05). These findings suggest that alpha(3) integrin regulates glomerular permeability by maintaining podocyte shape and adhesion, which is disrupted by ROS overproduction.

Increased serum and urinary neopterin in nephrotic syndrome indicate cell-mediated immune dysfunction.

T-cell-mediated immune disturbances are likely but not certain to cause the nephrotic syndrome. Because neopterin (NP) production is closely related to activation of cell-mediated immunity, we addressed the question by measuring serum NP concentrations and urinary NP/creatinine (Cr) ratios, as well as by assessing interstitial lymphocyte and monocyte infiltration in the kidney and activation of the same cell types in peripheral blood. Finally, we observed whether urinary NP/Cr ratios in nephrotic syndrome are changed by steroid therapy. Seventy-four patients with primary glomerulonephritis were divided into 4 groups based on presence or absence of nephrotic syndrome and presence or absence of mesangial proliferation and expansion. Serum and urinary NP concentrations were measured chromatographically. Infiltrating cells in the kidney were identified by immunohistochemistry, and activation of peripheral blood cells was examined by fluorescent surface marker antibodies and flow cytometry. Irrespective of the pathohistology of glomeruli, nephrotic groups showed significantly higher urinary NP/Cr ratios and serum NP concentrations. Nephrotic groups also exhibited more activation of T cells in peripheral blood than did nonnephrotic groups or a healthy control group. Serum NP did not correlate with extent of interstitial renal infiltrates. Steroid therapy decreased urinary NP/Cr ratios in steroid-responsive patients, but not in steroid-resistant patients. Increased serum NP concentrations and urinary NP/Cr ratios may reflect disordered cell-mediated immunity in the nephrotic syndrome, irrespective of glomerular histology or interstitial cell infiltration.

MPO-ANCA-positive crescentic necrotizing glomerulonephritis and tubulointerstitial nephritis with renal eosinophilic infiltration and peripheral blood eosinophilia.

We present the case of a 67-year-old woman with myeloperoxidase (MPO)-antineutrophil cytoplasmic antibody (ANCA)-positive pauci-immune crescentic necrotizing glomerulonephritis and tubulointerstitial nephritis with renal eosinophilic infiltration and peripheral blood eosinophilia. Staining for eosinophil cationic protein indicated that activated eosinophils were involved in the tubulitis, as well as in the glomerular injury. Marked peripheral blood eosinophilia is uncommon in ANCA-positive crescentic necrotizing glomerulonephritis associated with tubulointerstitial nephritis, except in Churg-Strauss syndrome. However, our patient had no clinical history or signs of asthma, no other signs suggestive of allergic diseases, and no histologic findings of granulomas in the kidney, thus failing to fulfill the criteria for Churg-Strauss syndrome.

Increased transforming growth factor-beta2 expression in the glomerular arteriole of the juxtaglomerular apparatus in a Bartter's-like syndrome.

Although transforming growth factor-beta (TGF-beta) has been shown to participate in regulating hormone synthesis and release, little is known about involvement of individual human TGF-beta isoforms, TGF-beta1, -beta2, and -beta3, in renin synthesis and release. We examined expression of these TGF-beta isoforms in a 50-year-old man with a Bartter's-like syndrome whose renal biopsy specimen showed hyperplasia of the juxtaglomerular apparatus (GA), mild mesangial hypercellularity, focal tubular atrophy, and interstitial fibrosis. Immunoreactivity for renin and marked expression of TGF-beta2 mRNA were noted in the glomerular arteriole of the JGA, whereas mRNA expression for TGF-beta1 was only slight and that for TGF-beta3 was still more faint. Expression of mRNAs for all 3 TGF-beta isoforms was increased in the fibrotic interstitium. This expression pattern suggests that TGF-beta2 may be involved in inducing renin synthesis and/or release in the glomerular arteriole of the JGA.

Toxicity of silica-containing calcium phosphate glasses demonstrated in mice.

Suspensions of calcium phosphate glass containing various concentrations of silica (glass composition (moles): 100 Ca(PO3)2 to x SiO2,x = 0, 5, 10, 15 or 40) dispersed in normal saline were injected intraperitoneally into C57BL/6 mice to determine the mortality within 30 days. The mortality was 0/10, 3/10, 9/10, 10/10 and 10/10 at x = 0, 5, 10, 15 and 40 mol of silica, respectively. By means of inductively coupled plasma analysis, the amount of dissolved silica (Si4+) in water at 37 degrees C from the calcium phosphate glass depended on the amount of silica in the glasses. The mortality of mice was directly proportional to the silica content of the glass injected intraperitoneally. These results clearly show that the dissolved silica (Si4+) from the glass, monomeric or low molecular silicic anion, is highly toxic. The SiO2 component in biomaterials has toxic potential when dissolved in the body.

Salt sensitivity of blood pressure in patients with 17 alpha-hydroxylase deficiency.

To clarify the salt sensitivity of blood pressure in 17 alpha-hydroxylase deficiency (17 alpha-HD), a mineralocorticoid-induced hypertension, we examined the responses of blood pressure to salt loading (250 mEq/day for 6 days) after salt restriction (25 mEq/day for 3 days) in two 17 alpha-HD patients who had markedly high plasma deoxycorticosterone (DOC) (5.27 and 6.59 ng/mL; normal, 0.08 to 0.28) and corticosterone (B) (93 and 357 ng/mL; normal, 0.35 to 8.42). In case 1, moreover, the same study was repeated when plasma DOC and B restored to almost normal with the dexamethasone treatment (1 mg/day) (0.34 and 0.45 ng/mL). Salt loading elevated mean blood pressure markedly in both patients (14% and 20%), associated with cumulative sodium retention and body weight gain. Hemodynamic study showed marked increases in cardiac output (25% and 69%), but only slight decreases in systemic vascular resistance with salt loading. On the other hand, the treatment with dexamethasone could attenuate the salt-induced increase in blood pressure (2%), accompanied by lesser sodium retention, body weight gain, and elevation of cardiac output (7%) in case 1. The slope of the renal function curve for sodium excretion was decreased in these patients, and was restored toward normal by dexamethasone in case 1. We conclude that the patients with 17 alpha-HD showed salt-sensitive hypertension due to excessive DOC and B, through the impaired renal function for sodium excretion, disproportionate increase in cardiac output, and inadequate vasodilation.

Glomerular handling of immune complex in the acute phase of active in situ immune complex glomerulonephritis employing cationized ferritin in rats. Ultrastructural localization of immune complex, complements and inflammatory cells.

The ultrastructural localization of immune complex (IC) and inflammatory mediator systems in the glomerulus was investigated in active in situ IC glomerulonephritis employing cationized ferritin in rats. Glomerulonephritis was induced by unilateral renal perfusion of cationized ferritin as antigen (Ag) in preimmunized rats, and anti-ferritin antibody (Ab), C3 and the rat C5b-9 complex were localized by means of immunogold electron microscopy. Ag-Ab complexes were initially formed subendothelially, associated with C3, and attracted platelets, polymorphonuclear leucocytes (PMN) and monocytes. Then Ag-Ab complexes, without C3, passed across the glomerular basement membrane to re-aggregate subepithelially accompanied by C3 deposition after 1 day. Ag-Ab complexes without C3 accumulated in the inter-podocyte space within 1 day and were seen in the epithelial cells at 6 h. C5b-9 complexes were found in subepithelial immune deposits and in membrane vesicles of the epithelial cells, but only in very small amounts in subendothelial immune deposits. Accumulated platelets, PMN, and monocyte were in direct contact with endothelial cells or subendothelial IC. PMN and monocytes contained Ag, Ab and C3 in intracytoplasmic vacuoles. Ag-Ab complexes were also found in the mesangial matrix adjacent to the subendothelial region after 2 h and increased slightly in number, with expansion of the mesangial area thereafter. Most ICs formed in the subendothelial space rapidly formed lattices of a size that activated C3 and were then translocated to the subepithelial space. The potential ability of C3 to solubilize ICs in the subendothelial region may be important in this process. Endocytosis of subendothelial ICs by PMN and/or monocytes and the movement of ICs to the mesangial matrix may also contribute to the removal of IC from the subendothelial space.

A high-fat diet aggravates tubulointerstitial but not glomerular lesions in obese Zucker rats.

BACKGROUND: Despite a large body of evidence that manipulation of dietary fat alters glomerular lesions, reports regarding the effects of dietary fat on tubulointerstitial lesions are limited. Obese Zucker rats (OZR) spontaneously develop glomerular and tubulointerstitial lesions in association with hyperlipidemia. We sought to elucidate the effects of dietary fat on glomerular and tubulointerstitial lesions in OZR versus lean Zucker rats (LZR) and to assess the involvement of macrophages in the development of these lesions. METHODS: We fed LZR and OZR either a low- (1%) or high-fat (20%) diet. After 30 weeks of the specified diet, the creatinine clearance (Ccr) and renal histology as well as plasma lipid concentrations were examined. For morphological evaluation, glomerular sclerosis (GSI) and tubulointerstitial indices (TII) were each determined by a point-counting method. Infiltrating macrophages were stained immunohistochemically using an avidin-biotin complex technique. RESULTS: The high-fat diet increased the plasma low-density lipoprotein concentration in OZR. Both low- and high-fat OZR groups had higher GSI and TII than LZR receiving either diet. The high-fat diet aggravated TII but not GSI or Ccr in OZR; conversely, high fat intake worsened GSI and Ccr but not TII in LZR. Tubulointerstitial macrophages were most prominent in the high-fat OZR group, followed by the low-fat OZR group. Glomerular macrophages were similar in number in all groups. CONCLUSIONS: The manipulation of dietary fat has diverse effects on the kidney. A high-fat diet aggravated macrophage-mediated tubulointerstitial lesions in OZR, whereas in LZR, the diet induced glomerulosclerosis.