Increase in urinary markers during the acute phase reflects the degree of chronic tubulointerstitial injury after ischemia-reperfusion renal injury.

CONTEXT: Acute kidney injury (AKI) could lead to progressive chronic kidney disease (CKD). OBJECTIVES: To demonstrate that urinary markers in AKI are associated with the degree of persistent renal injury. MATERIAL AND METHODS: Human L-FABP chromosomal transgenic (Tg) mice were subjected to ischemia-reperfusion (I/R) clamping renal pedicle for 20 min or 30 min. Kidneys were obtained at one and 40 days after I/R. RESULTS: Urinary L-FABP, NGAL, Kim-1 and albumin levels increased during the acute phase and were significantly correlated with the degree of tubulointerstitial fibrosis during the chronic phase. DISCUSSION AND CONCLUSION: These markers could detect higher risk of progression to CKD.

Renoprotective effect of the xanthine oxidoreductase inhibitor topiroxostat on adenine-induced renal injury.

The aim of the present study was to reveal the effect of a xanthine oxidoreductase (XOR) inhibitor, topiroxostat (Top), compared with another inhibitor, febuxostat (Feb), in an adenine-induced renal injury model. We used human liver-type fatty acid-binding protein (L-FABP) chromosomal transgenic mice, and urinary L-FABP, a biomarker of tubulointerstitial damage, was used to evaluate tubulointerstitial damage. Male transgenic mice (n = 24) were fed a 0.2% (wt/wt) adenine-containing diet. Two weeks after the start of this diet, renal dysfunction was confirmed, and the mice were divided into the following four groups: the adenine group was given only the diet containing adenine, and the Feb, high-dose Top (Top-H), and low-dose Top (Top-L) groups were given diets containing Feb (3 mg/kg), Top-H (3 mg/kg), and Top-L (1 mg/kg) in addition to adenine for another 2 wk. After withdrawal of the adenine diet, each medication was continued for 2 wk. Serum creatinine levels, the degree of macrophage infiltration, tubulointerstitial damage, renal fibrosis, urinary 15-F2t-isoprostane levels, and renal XOR activity were significantly attenuated in the kidneys of the Feb, Top-L, and Top-H groups compared with the adenine group. Serum creatinine levels in the Top-L and Top-H groups as well as renal XOR in the Top-H group were significantly lower than those in the Feb group. Urinary excretion of L-FABP in both the Top-H and Top-L groups was significantly lower than in the adenine and Feb groups. In conclusion, Top attenuated renal damage in an adenine-induced renal injury model.

Human liver-type fatty acid-binding protein protects against tubulointerstitial injury in aldosterone-induced renal injury.

To demonstrate the renoprotective function of human liver-type fatty acid-binding protein (hL-FABP) expressed in proximal tubules in aldosterone (Aldo)-induced renal injury, hL-FABP chromosomal transgenic (Tg) and wild-type (WT) mice received systemic Aldo infusions (Tg-Aldo and WT-Aldo, respectively) were given 1% NaCl water for 28 days. In this model, elevation of systolic blood pressure, monocyte chemoattractant protein-1 expression, macrophage infiltration in the interstitium, tubulointerstitial damage, and depositions of type I and III collagens were observed. Elevation of systolic blood pressure did not differ in WT-Aldo vs. Tg-Aldo animals, however, renal injury was suppressed in Tg-Aldo compared with WT-Aldo mice. Dihydroethidium fluorescence was used to evaluate reactive oxidative stress, which was suppressed in Tg-Aldo compared with WT-Aldo mice. Gene expression of angiotensinogen in the kidney was upregulated, and excretion of urinary angiotensinogen was increased in WT-Aldo mice. This exacerbation was suppressed in Tg-Aldo mice. Expression of hL-FABP was upregulated in proximal tubules of Tg-Aldo mice. Urinary excretion of hL-FABP was significantly greater in Tg-Aldo than in Tg-control mice. In conclusion, hL-FABP ameliorated the tubulointerstitial damage in Aldo-induced renal injury via reducing oxidative stress and suppressing activation of the intrarenal renin-angiotensin system.

Infant feeding practice in medieval Japan: stable carbon and nitrogen isotope analysis of human skeletons from Yuigahama-minami.

A longer breastfeeding duration provides various positive effects in subadult health because of abundant immunological factors and nutrients in human breast milk, and decreases the natural fertility of a population through lactational amenorrhea. In this study, we measured stable carbon and nitrogen isotope ratios in the bone collagen of three adults and 45 subadults from the Yuigahama-minami site (from 12th to 14th century) in Kamakura, the early medieval capital of Japan. Marine foods, C3 -based terrestrial foods, and freshwater fish are the primarily protein sources for adults. The changes in the nitrogen isotope ratios of subadults suggest that the relative dietary protein contribution from breast milk started to decrease from 1.1 years of age and ended at 3.8 years. The age at the end of weaning in the Yuigahama-minami population was greater than that in the typical non-industrial populations, a premodern population in the Edo period Japan, and medieval populations in the UK. Skeletons of townspeople from medieval Kamakura indicate severe nutritional stress (e.g., enamel hypoplasia and cribra orbitalia), yet this longer duration of breastfeeding did not compensate adverse effects for nutritional deficiency. The longer breastfeeding period may have been a consequence of complementary food shortage and bad health of subadults. Kamakura experienced urbanization and population increase in the early medieval period. The younger age-at-death distribution and high nutritional stresses in the Yuigahama-minami population and later weaning, which is closely associated with longer inter-birth interval for mothers, suggests that Kamakura developed and increased its population by immigration during urbanization.

Renoprotective effect of renal liver-type fatty acid binding protein and angiotensin II type 1a receptor loss in renal injury caused by RAS activation.

The aim of this study was to assess the renoprotective effect of renal human liver-type fatty acid binding protein (hL-FABP) and angiotensin II (ANG II) type 1A receptor (AT1a) loss in renal injury caused by renin-angiotensin system (RAS) activation. We established hL-FABP chromosomal transgenic mice (L-FABP(+/-)AT1a(+/+)), crossed the L-FABP(+/-)AT1a(+/+) with AT1a knockdown homo mice (L-FABP(-/-)AT1a(-/-)), and generated L-FABP(+/-)AT1a hetero mice (L-FABP(+/-)AT1a(+/-)). After the back-cross of these cubs, L-FABP(+/-)AT1a(-/-) were obtained. To activate the renal RAS, wild-type mice (L-FABP(-/-)AT1a(+/+)), L-FABP(+/-)AT1a(+/+), L-FABP(-/-)AT1a(+/-), L-FABP(+/-)AT1a(+/-), L-FABP(-/-)AT1a(-/-), and L-FABP(+/-)AT1a(-/-) were administered high-dose systemic ANG II infusion plus a high-salt diet for 28 days. In the L-FABP(-/-)AT1a(+/+), RAS activation (L-FABP(-/-)AT1a(+/+)RAS) caused hypertension and tubulointerstitial damage. In the L-FABP(+/-)AT1a(+/+)RAS, tubulointerstitial damage was significantly attenuated compared with L-FABP(-/-)AT1a(+/+)RAS. In the AT1a partial knockout (AT1a(+/-)) or complete knockout (AT1a(-/-)) mice, reduction of AT1a expression led to a significantly lower degree of renal injury compared with L-FABP(-/-)AT1a(+/+)RAS or L-FABP(+/-)AT1a(+/+)RAS mice. Renal injury in L-FABP(+/-)AT1a(+/-)RAS mice was significantly attenuated compared with L-FABP(-/-)AT1a(+/-)RAS mice. In both L-FABP(-/-)AT1a(-/-)RAS and L-FABP(+/-)AT1a(-/-)RAS mice, renal damage was rarely found. The degrees of renal hL-FABP expression and urinary hL-FABP levels increased by RAS activation and gradually decreased along with reduction of AT1a expression levels. In conclusion, in this mouse model, renal hL-FABP expression and a decrease in AT1a expression attenuated tubulointerstitial damage due to RAS activation.

Stable isotopic reconstructions of adult diets and infant feeding practices during urbanization of the city of Edo in 17th century Japan.

The urbanization of the city of Edo, the capital of premodern Japan, has been assumed to be not as a result of natural increase but that of in-migration although this assumption has never been verified. To obtain information on natural fertility in Edo, we analyzed stable carbon and nitrogen isotopes in 46 adult and 84 subadult human skeletons excavated from the Hitotsubashi site (1657-1683 AD: the early Edo period), Tokyo, Japan and reconstructed their breastfeeding period, one of the most important determinants of fertility. Adult females are significantly more depleted in (15) N by 0.7‰ than adult males, suggesting a dietary differentiation between sexes and/or the effect of pregnancy. The changes in the nitrogen isotope ratios of subadults suggest that supplementary foods were introduced around the age of 0.2 years and weaning ended around 3.1 years, which agrees with descriptions in various historical documents of the period. The duration of breastfeeding in the Hitotsubashi population was relatively longer than those in modern industrial and traditional societies and four previously reported populations in medieval and in the industrial England. As later weaning closely associates with longer inter-birth interval for mothers, our data suggest a lower natural fertility for the Hitotsubashi population. Assuming that the proportion of married people was also lower in the major cities of the earlier Edo period, our results support the assumption that Edo developed and increased its population by attracting immigrants during urbanization.

Estimation of fetal age at death from the basilar part of the occipital bone.

The purposes of this study are to examine documented fetal skeletal remains of Japanese, to measure the basilar part of the occipital bone, and to develop diagnostic standards for estimating fetal age at death which can be applied to poorly preserved skeletons. The sample is composed of 272 Japanese individuals of the early to middle twentieth century, whose ages were recorded in months from gestations of 5 to 11 months. The measurement items used here are the length, breadth, and index of the basilar part. The regression equations of gestational age in months for one or two variables were calculated. The results indicated that it is possible to use the regression equations to estimate the age at death of fetuses directly from the basilar part measurements. Another indicator for estimating age at death from the basilar part is the ratio of the width to the length, which was here expressed as the index of the basilar part. The width exceeded the length at 7 months and the basilar part changed with age from an anteriorly posteriorly long shape to a bilaterally wide one. It is concluded that the basilar part is a good indicator for estimating the fetal age at death.

Using multidetector row computed tomography to evaluate baggy eyelid.

BACKGROUND: Baggy eyelids, formed by intraorbital fat herniation in the lower eyelids, are a sign of aging observed in the midface. This study aimed to identify the cause of baggy eyelids by evaluating the relationship between orbicularis oculi muscle thickness, orbital fat prolapse length, and age using multidetector row computed tomography (MDCT). METHODS: The 34 patients in the study ranged in age from 20 to 79 years. The patients were divided into three age groups: 20-29 years, 30-54 years, and 55-79 years. Orbicularis oculi muscle thickness and orbital fat prolapse length were measured using reconstructed computed tomography (CT) images. RESULTS: The orbicularis oculi muscle was found to be significantly thinner in the oldest group. Likewise, orbital fat prolapse was found to be significantly more prominent in the oldest group. A strong and significant negative correlation was found between orbicularis oculi muscle thickness and orbital fat prolapse length (r = -0.78; P < 0.001) and between orbicularis oculi muscle thickness and age (r = -0.77; P < 0.001). A strong and significant positive correlation was found between orbital fat prolapse length and age (r = 0.78; P < 0.001). CONCLUSIONS: The use of MDCT provides insight into the detailed changes associated with aging that take place within the lower eyelids. This study objectively demonstrated that the major factors associated with baggy eyelids include orbicularis oculi muscle thickness, orbital fat prolapse length, and age. Decreased orbicularis oculi muscle thickness leads to the orbital fat prolapse.

Insulin-like growth factor binding protein-4 gene silencing in lung adenocarcinomas.

Gene silencing by promoter hypermethylation plays an important role in molecular pathogenesis. We previously reported that insulin-like growth factor (IGF) binding protein-4 (IGFBP-4), which inhibits IGF-dependent growth, is expressed via early growth response-1 (EGR-1) and is often silenced in cultivated lung cancer cells. The purpose of the present study was to clarify clinicopathological factors associated with IGFBP-4 gene silencing in lung adenocarcinomas. Seventy-six surgically resected adenocarcinomas (20 well-, 35 moderately-, and 21 poorly-differentiated) were subjected to methylation-specific polymerase chain reaction (PCR) analysis for EGR-1-binding sites located in the IGFBP-4 promoter and immunohistochemistry for IGFBP-4, EGR-1, and Ki-67. Thirty-two adenocarcinomas (42%) revealed IGFBP-4 promoter hypermethylation, and the severity inversely correlated with the level of IGFBP-4 expression (P < 0.0001) and tumor differentiation (well versus poor, P = 0.0278; well/moderate versus poor, P = 0.0395). Furthermore, there was a negative correlation between Ki-67 labeling index and IGFBP-4 expression (P = 0.0361). These findings suggest that the expression of IGFBP-4 in adenocarcinoma cells in vivo is downregulated by epigenetic silencing in association with tumor differentiation, resulting in disruption of the mechanism of IGFBP-4-mediated growth inhibition.

Anatomical Study of the Cuboid and Its Ligamentous Attachments and Its Implications for a Cuboid Osteotomy.

BACKGROUND: Lateral column lengthening (LCL) for flexible flatfoot is an effective surgery with powerful correction of deformity because it tightens only the lateral third of the long plantar ligament (LPL). However, LCL has been associated with joint damage at the osteotomy site and loss of foot flexibility owing to joint fixation. We focused on the cuboid and investigate a novel anatomical LCL osteotomy site that effectively tightens the LPL without damaging any joints. METHODS: We studied 24 feet of 12 cadavers (mean age, 80.8 years). The lengths of the LPL and short plantar ligament, locations of the attachments, and shape and location of the cuneocuboid joint on the medial side of the cuboid were studied. ImageJ software was used to measure the osteotomy angle. RESULTS: The lateral cuboid attachment of the LPL on average was located 4.6 mm from the calcaneocuboid joint, and the cuneocuboid joint on average was located 6.7 mm from the cuboid-metatarsal joint on the medial surface of the cuboid. The direct line connecting the anterior cuneocuboid joint and the oblique crest of the cuboid on average was at a 10.3-degree inclination posterior to the cuboid-metatarsal joint. CONCLUSION: A straight line must be selected between a point 4 mm from the calcaneocuboid joint laterally and 6 mm from the cuboid-metatarsal joint medially at a 10-degree posterior tilt to the cuboid-metatarsal joint to perform a cuboid osteotomy LCL without damaging the articular surface. CLINICAL RELEVANCE: We investigated a potential novel cuboid osteotomy method for LCL.

Oral administration of melatonin contained in drinking water increased bone strength in naturally aged mice.

Melatonin has recently been found to be a possible new regulator of bone metabolism. However, the influence of melatonin in natural age-related osteoporosis has not been fully elucidated yet, although there have been some reports regarding postmenopausal osteoporosis with melatonin treatments. The present study investigated the effects of long-term melatonin administration during the aging process on bone metabolism. Using quantitative computed tomography methods, we found that the total bone density of both the femur metaphysis and diaphysis decreased significantly in 20-month-old male mice. In the metaphysis, both trabecular bone mass and Polar-Strength Strain Index (SSI), which is an index of bone strength, decreased significantly. Judging from bone histomorphometry analysis, trabecular bone in 20-month-old male mice decreases significantly with age and is small and sparse, as compared to that of 4-month-old male mice. Loss of trabecular bone is one possible cause of loss of bone strength in the femoral bone. In the metaphysis, the melatonin administration group had significantly higher trabecular bone density than the non-administration group. The Polar-SSI, cortical area, and periosteal circumference in the diaphysis was also significantly higher with melatonin treatments. Since the melatonin receptor, MT2, was detected in both osteoblasts and osteoclasts of the femoral bone of male mice, we expect that melatonin acts on osteoblasts and osteoclasts to maintain the bone strength of the diaphysis and metaphysis. Thus, melatonin is a potential drug for natural age-related osteoporosis.

Reliability of metric determination of sex based on long-bone circumferences: perspectives from Yuigahama-minami, Japan.

The purpose of this study is to develop new standards to determine the sex of fragmentary human skeletal remains from archaeological sites in Japan. In order to accomplish this, we measured the long-bone circumferences of Japanese skeletons from the medieval period and provided metric diagnosis of sex using discriminant function analysis. We discuss whether the osteometric approach provides the criterion for sex assessment of human skeletal remains. The materials comprised human skeletal remains from the Yuigahama-minami site, Kamakura, Japan. The sample size used in this study was 68 males and 62 females excavated from individual burial graves. The accuracy of sex classification is more than 80% for discriminant functions with only one variable and reaches 90% for those with a combination of multiple variables. The multivariate functions provide better results than the univariate functions. Another improvement in sex diagnosis which this study contributes to is that new standards enabling reliable diagnosis with small numbers of variables are developed from well-preserved parts of the skeletons. This paper provides new standards, focusing on the diaphysial circumferences of limb bones from a medieval population, and will contribute to the advancement of medieval studies of skeletal remains from archaeological sites in Japan.

Amelioration of diabetic tubulointerstitial damage in liver-type fatty acid-binding protein transgenic mice.

BACKGROUND: Renoprotection of liver-type fatty acid binding protein (L-FABP) was demonstrated in a streptozotocin (STZ)-induced diabetic mouse model. METHODS: Established human L-FABP (hL-FABP) transgenic (Tg) mice and wild-type (WT) mice were divided into two groups: diabetic mice were uninephrectomized and injected with STZ; control mice were uninephrectomized and injected with a citrate buffer alone. Although mouse L-FABP was not expressed in WT mice, hL-FABP was expressed in the proximal tubules of the diabetic Tg mice and in the control Tg mice at 8 and 14 weeks after these injections. RESULTS: The expression of renal hL-FABP increased significantly in diabetic Tg mice compared to control Tg mice. A number of macrophages (F4/80) infiltrating the interstitium, the gene expressions of MCP-1, MCP-3, TGF-beta, Fas, Bax and RAGE were significantly lower in diabetic Tg kidneys compared with diabetic WT kidneys. In the diabetic Tg kidneys, the degree of the tubulointerstitial injury and the deposition of type IV collagen were significantly lower than that of diabetic WT kidneys. The expressions of catalase and glutathione peroxidase-1 were significantly lower in diabetic Tg kidneys compared with diabetic WT kidneys. CONCLUSIONS: Renal L-FABP ameliorated the tubulointerstitial damage of type 1 diabetic mice.

A health crisis during the Japanese Medieval Period: A new paleodemographic perspective.

OBJECTIVE: To test the hypothesis that the "Little Ice Age" (LIA) (in Japan, ˜1440 - 1730 CE) co-occurred with unique age-at-death patterns. MATERIALS: 810 adult human skeletons from the early Medieval Period (EMP) of Japan, which are contemporaneous with the Medieval Warm Period (10th - mid 13th century AD), and the late Medieval Period (LMP) and Edo Period, which are contemporary with the LIA. METHODS: Age at death and sex was determined for each skeleton and demographic profiles of the Yayoi Period (5th century BC - 3rd century AD), EMP, LMP, and Edo site samples were compared. Paleopathological data from previously published reports were evaluated. RESULTS: The EMP had the highest mortality among young adults. Longevity increased in the samples (LMP and Edo) contemporaneous with the LIA. CONCLUSIONS: EMP early age-at-death was the result of poor community health, violent death, and frequent large-scale natural catastrophes. The LMP and Edo Period samples have an older age-at-death pattern and higher frequency of stress markers, argued to be a consequence of a colder climate. SIGNIFICANCE: This study is the first to synthesize paleodemographic and paleopathological data on a large scale to assess the possible effects of the Little Ice Age in Japan. LIMITATIONS: Varying skeletal preservation and focus on adult skeletons reduces the ability to evaluate health throughout the life span. SUGGESTIONS FOR FURTHER RESEARCH: Analysis of nonadult remains and multiple health indicators will likely shed more light on the effects of the Little Ice Age in Japan.

Effects of unoprostone on phosphorylated extracellular signal-regulated kinase expression in endothelin-1-induced retinal and optic nerve damage.

Endothelin-1 (ET-1), a potent vasoconstrictor peptide, has been implicated in the development of normal- and high-tension glaucoma. We investigated the effects of unoprostone on extracellular signal-regulated kinase (ERK) in ET-1-induced retinal ganglion cell (RGC) death and optic nerve injury. Our morphometric study showed that intravitreal injection of ET-1 led to cell loss in the RGC layer (RGCL) in 28 days. Western blot analysis showed decreased neurofilament (NF) protein in the optic nerve 28 days after ET-1 injection. In this in vivo model, increased phosphorylated ERK (p-ERK) was observed in the retina on 1 day and subsequently in the optic nerve from 7 days after ET-1 injection. Simultaneous injection of M1, as a metabolite of unoprostone, showed further increased p-ERK levels compared with ET-1 injection alone. Our morphometric study of flat-mount preparations stained with cresyl violet or retrograde labeling with a neuro-tracer and Western blot analysis of NF showed that inhibition of ERK phosphorylation led to acceleration of ET-1-induced RGC death and optic nerve damage. In addition, M1 significantly attenuated both RGC loss and the decrease in NF protein induced by ET-1. The protective effects of M1 were significantly inhibited by U0126, an ERK inhibitor. These results suggest that unoprostone has neuroprotective effects against ET-1-induced neuronal injury through ERK phosphorylation.

Anatomical study of arthroscopic surgery for lateral epicondylitis.

We conducted an anatomical study of arthroscopic surgery for lateral epicondylitis. Currently, the pathogenesis suggests that lateral epicondylitis is caused by extra-articular lesions or intra-articular lesions. We anatomically and histologically examined the relationship between extensor carpi radialis brevis (ECRB), articular capsules, and the lateral collateral ligament complex (LCLC). In addition, we examined the number and location of synovial fringes. This study will be useful for less-invasive treatment of lateral epicondylitis.

Clinical anatomical study of pedicled vascularised scapular bone graft using the angular branch.

This study is an anatomical investigation of the angular branch of thoracodorsal artery, and examines the possible range of clinical targets for pedicled vascularised scapular bone graft. Forty-six cadavers were studied. The blood vessel length was calculated, and the distance required to reach the distal humerus from the lower end of the articular surface of the humeral head as reference point was compared with bone length. The length from reference point was an average of 121.7 mm. It was thought that the maximum distance to arrive in reference point to the distal humerus was a total of blood vessel length and transplantation bone length, which was an average of 246.3 mm. This was longer than the average of 240.8 mm of bone length from reference point to humeral medial condyle. This study had confirmed that a pedicled vascularised scapular bone graft using the angular branch could be transplanted to the distal humerus.

Proteomic profiling of archaeological human bone.

Ancient protein analysis provides clues to human life and diseases from ancient times. Here, we performed shotgun proteomics of human archeological bones for the first time, using rib bones from the Hitotsubashi site (AD 1657-1683) in Tokyo, called Edo in ancient times. The output data obtained were analysed using Gene Ontology and label-free quantification. We detected leucocyte-derived proteins, possibly originating from the bone marrow of the rib. Particularly prevalent and relatively high expression of eosinophil peroxidase suggests the influence of infectious diseases. This scenario is plausible, considering the overcrowding and unhygienic living conditions of the Edo city described in the historical literature. We also observed age-dependent differences in proteome profiles, particularly for proteins involved in developmental processes. Among them, alpha-2-HS-glycoprotein demonstrated a strong negative correlation with age. These results suggest that analysis of ancient proteins could provide a useful indicator of stress, disease, starvation, obesity and other kinds of physiological and pathological information.