Mitochondrial DNA polymorphism in Japanese

Summary The mitochondrial DNA (mtDNA) from 120 Japanese was analysed with 15 restriction enzymes that recognize six base pairs, of which 11 enzymes showed at least one atypical cleavage pattern. Digestion patterns with HincII and HaeII were highly polymorphic. The observed restriction enzyme morphs were classified into 22 types of distinct cleavage patterns. By pairwise comparison of each restriction type, the average number of nucleotide substitutions per nucleotide site () was estimated at 0.00417, which agreed with the values obtained from other human populations in previous studies. There were 11 site gains, of which seven were transitions and four were transversions. Phylogenetic analysis of the present data suggested that the Japanese population conceals a considerably high degree of mtDNA diversity.     

正常人群磁共振全身弥散加权成像特点的初步研究

目的：探讨健康志愿者不同性别各年龄组人体多脏器全身弥散加权成像的图像特征。方法：对50例健康志愿者行全身弥散加权成像扫描,其中,20．30岁、30．40岁、40—50、50．60、60岁以上各二十例,每组内男女各十例。所得图像均经nDeMR后处理工作站（AW4．3）行MIP重建和黑白反转显示技术,并重建ADC图,观察不同性别各年龄组人体多脏器弥散加权图像的特点,并测定不同部位的ADC值,测量的部位主要包括脑灰质、脑白质、尾状核头、丘脑、桥脑、腮腺、肝脏、脾脏、T8椎体、T8—9椎间盘、L2椎体、L2．3椎间盘、前列腺、乳腺、子宫颈等,每个部位重复测量三次取平均值,单位为×10^-3mm^2／S。结果：1、WB-DWI的正常表现。2、不同性别及年龄组间各组织器官ADC值的统计学分析脑白质、T8椎体ADC值在不同性别及年龄组之间存在统计学差异（P〈0．05）;T8．9椎间盘、L2．3椎间盘及女性乳腺ADC值在不同年龄组之间存在统计学差异（P〈0．05）;L2椎体ADC值在男女之间存在统计学差异（P〈0．05）。3、不同部位ADC值平均值。结论：正常人体不同组织器官弥散加权图像特点及ADC值受到性别、年龄的影响,不同性别各年龄组的正常WB-DWI表现存在差异。了解健康成人正常WB-DWI的表现对于准确判断各组织器官的疾病有着重要的意义。     

Multiple organ mutation in the lacZ transgenic mouse (Muta mouse) 6 months after oral treatment (5 days) with benzo[a]pyrene.

We have recently demonstrated that not all organs with high rates of mutation in the lacZ transgene develop tumors using the Muta Mouse. To better understand the role of in vivo mutation in carcinogenesis, we examined the mutant frequencies (MF) of the lacZ transgene in tumor-bearing and non tumor-bearing organs. MF, recovered after 2 weeks (the data taken from our previous study) and after 26 weeks following oral doses of 125 mg kg-1 day-1 benzo[a]pyrene (BP) for five days were compared. The organs examined included the target organs (forestomach, spleen, and lung) and non-target organs (colon, glandular stomach, and liver) for BP carcinogenesis. The data indicated that lacZ MF were markedly increased over spontaneous frequencies in the organs examined and that the organ which showed the highest MF was the colon, followed by the forestomach>spleen>glandular stomach, liver, and lung in that order. These findings indicate that the MF of the lacZ transgene in each organ, even 26 weeks after the start of the treatment does not fully correlate with the known target organs of BP. Furthermore, the lacZ MF in a non-papilloma region of a forestomach with a papilloma was equivalent to the two highest MF observed in the healthy colon (non-target organ) of mice at 26 weeks. These observations also indicate that the generation of tumors requires the induction of mutations as well as other factor(s) specific to the target organs. These results clearly suggest that highly mutated organs do not always progress to tumors in the transgenic mouse.     

Immunodiffusion analysis of water-soluble rat bone marrow antigens

Antisera were obtained against six electrophoretic fractions of the rat bone marrow extract. With their help, 18 tissular antigens and 11 antigens immunologically similar to the blood serum proteins were revealed in the rat bone marrow. All tissular antigens are divided in five groups by the degree of organ specificity: 1) bone marrow organospecific antigens (4 antigens), 2) antigens present in the bone marrow, spleen and lung extracts (2), 3) "granulocytic" antigens (4), 4) antigens common for many rat organs, but not found in the extracts of blood formed elements, skeletal muscle, heart, brain, eyes (3), 5) antigens present in all the organs studied (5). The bone marrow organospecific antigens may be specific antigens of hemopoietic cell precursors. The possibility of utilization of antisera against the bone marrow water soluble proteins for labelling hemopoietic cells of different lines of differentiation is discussed.     

A genetic factor for age-related cataract: identification and characterization of a novel galactokinase variant, "Osaka," in Asians

Galactokinase (GALK) deficiency is an autosomal recessive disorder characterized by hypergalactosemia and cataract formation. Through mass screening of newborn infants, we identified a novel and prevalent GALK variant (designated here as the "Osaka" variant) associated with an A198V mutation in three infants with mild GALK deficiency. GALK activity and the amount of immunoreactive protein in the mutant were both 20% of normal construct in expression analysis. The K(m) values for galactose and ATP-Mg(2+) in erythrocytes with homozygous A198V were similar to those of the healthy adult control subjects. A population study for A198V revealed prevalences of 4.1% in Japanese and 2.8% in Koreans, lower incidence in Taiwanese and Chinese, no incidence in blacks and whites from the United States, and a significantly high frequency (7.8%; P < .023) in Japanese individuals with bilateral cataract. This variant probably originated in Japanese and Korean ancestors and is one of the genetic factors that causes cataract in elderly individuals.     

Isolation and Characterization of Circulating Tumor Cells in Squamous Cell Carcinoma of the Lung Using a Non-EpCAM-Based Capture Method

Introduction

The exclusion of circulating tumor cells (CTCs) that have lost epithelial antigens during the epithelial-to-mesenchymal transition (EMT) process by using Epithelial Cell Adhesion Molecule (EpCAM) based capture methods is still a matter of debate. In this study, cells obtained after depletion procedure from blood samples of squamous cell lung cancer (SQCLC) patients were identified based on morphology and characterized with the combination of FISH assessment and immunophenotypic profile.

Materials and Methods

Five mL blood samples, collected from 55 advanced SQCLC patients, were analyzed by a non-EpCAM-based capture method. After depletion of leukocytes and erythroid cells, the negative fraction was characterized by both FISH using a fibroblast growth factor receptor 1 (FGFR1) probe and by immunocytochemistry. Thirty healthy donors were also tested.

Results

Based on morphology (nuclear dimension ≥10 μm, shape and hypercromatic aspect) suspicious circulating cells clearly distinguishable from contaminant leukocytes were observed in 49/55 (89%) SQCLC patients. Thirty-four of the 44 (77%) samples evaluable for FGFR1 FISH showed ≥ 6 FGFR1 gene copy number on average per cell. Vimentin expression involved 43% (18/42) of pooled circulating SQCLC cells, whereas only 29% (14/48) were EpCAM positive. Confocal microscopy confirmed the localization of FGFR1 probe in suspicious circulating cells. Suspicious circulating elements were also observed in healthy donors and did not show any epithelial associated antigens. A significantly lower number of suspicious circulating cells in healthy donors compared to SQCLC patients was found.

Conclusions

Among the heterogeneous cell population isolated by depletion procedure, the coexistence of cells with epithelial and/or mesenchymal phenotype suggests that EMT may participate to transendothelial invasion and migration of tumor cells in advanced SQCLC. The finding of cells with neither EpCAM or EMT phenotype, retrieved after non-EpCAM-based systems, underlines the presence of suspicious elements in the blood of both SQCLC patients and healthy donors. Further phenotyping and molecular analyses are necessary to fully characterize these circulating elements.     

Exhaled Breath Analysis for Lung Cancer Detection Using Ion Mobility Spectrometry

Background

Conventional methods for lung cancer detection including computed tomography (CT) and bronchoscopy are expensive and invasive. Thus, there is still a need for an optimal lung cancer detection technique.

Methods

The exhaled breath of 50 patients with lung cancer histologically proven by bronchoscopic biopsy samples (32 adenocarcinomas, 10 squamous cell carcinomas, 8 small cell carcinomas), were analyzed using ion mobility spectrometry (IMS) and compared with 39 healthy volunteers. As a secondary assessment, we compared adenocarcinoma patients with and without epidermal growth factor receptor (EGFR) mutation.

Results

A decision tree algorithm could separate patients with lung cancer including adenocarcinoma, squamous cell carcinoma and small cell carcinoma. One hundred-fifteen separated volatile organic compound (VOC) peaks were analyzed. Peak-2 noted as n-Dodecane using the IMS database was able to separate values with a sensitivity of 70.0% and a specificity of 89.7%. Incorporating a decision tree algorithm starting with n-Dodecane, a sensitivity of 76% and specificity of 100% was achieved. Comparing VOC peaks between adenocarcinoma and healthy subjects, n-Dodecane was able to separate values with a sensitivity of 81.3% and a specificity of 89.7%. Fourteen patients positive for EGFR mutation displayed a significantly higher n-Dodecane than for the 14 patients negative for EGFR (p<0.01), with a sensitivity of 85.7% and a specificity of 78.6%.

Conclusion

In this prospective study, VOC peak patterns using a decision tree algorithm were useful in the detection of lung cancer. Moreover, n-Dodecane analysis from adenocarcinoma patients might be useful to discriminate the EGFR mutation.     

Novel Fluorescein Angiography-Based Computer-Aided Algorithm for Assessment of Retinal Vessel Permeability

Purpose

To present a novel method for quantitative assessment of retinal vessel permeability using a fluorescein angiography-based computer algorithm.

Methods

Twenty-one subjects (13 with diabetic retinopathy, 8 healthy volunteers) underwent fluorescein angiography (FA). Image pre-processing included removal of non-retinal and noisy images and registration to achieve spatial and temporal pixel-based analysis. Permeability was assessed for each pixel by computing intensity kinetics normalized to arterial values. A linear curve was fitted and the slope value was assigned, color-coded and displayed. The initial FA studies and the computed permeability maps were interpreted in a masked and randomized manner by three experienced ophthalmologists for statistical validation of diagnosis accuracy and efficacy.

Results

Permeability maps were successfully generated for all subjects. For healthy volunteers permeability values showed a normal distribution with a comparable range between subjects. Based on the mean cumulative histogram for the healthy population a threshold (99.5%) for pathological permeability was determined. Clear differences were found between patients and healthy subjects in the number and spatial distribution of pixels with pathological vascular leakage. The computed maps improved the discrimination between patients and healthy subjects, achieved sensitivity and specificity of 0.974 and 0.833 respectively, and significantly improved the consensus among raters for the localization of pathological regions.

Conclusion

The new algorithm allows quantification of retinal vessel permeability and provides objective, more sensitive and accurate evaluation than the present subjective clinical diagnosis. Future studies with a larger patients’ cohort and different retinal pathologies are awaited to further validate this new approach and its role in diagnosis and treatment follow-up. Successful evaluation of vasculature permeability may be used for the early diagnosis of brain microvascular pathology and potentially predict associated neurological sequelae. Finally, the algorithm could be implemented for intraoperative evaluation of micovascular integrity in other organs or during animal experiments.     

Rapid microwave fixation--a comparative morphometric study

Summary Tissue blocks taken from healthy human lung tisues, from primary bronchial carcinoma and from mediastinal and hilar lymph nodes were placed in the following solutions. Tris buffer; buffered formalin (0.5%, 1%, 7%); 0.1 mol NaCl; distilled water; DMSO (1%, 10%, 20%); acetone (10%); methanol (50%, 80%, 100%); glutaraldehyde (2.5%), and fixed by use of a commercial microwave oven.Tissue blocks obtained from the same surgical specimens were fixed in 7% buffered formalin for 24 h for comparison. Conventional and microwave-fixed tissue was embedded in paraffin, and stained with haematoxylin and eosin. Fifteen specimens of each group and each solution were examined by light microscopy. Minimum diameter and area of 100 nuclei of each specimen were measured interactively.Histomorphological sections fixed with Tris buffer in a microwave oven revealed best morphological results showing more contrast in chromatin distribution of nuclei and opening of interstitial lung capillaries in comparison to conventional formalin-fixed specimens. No statistically significant differences in area and minimum diameter of nuclei between the different groups were found. Microwave fixation using Tris buffer is a time-saving fixation method at least comparable to conventional formalin fixation. It is not accompanied by hazards to the environment that are unavoidable by formalin fixation.     

Soil chemical properties affect the concentration of elements (N,P, K,Ca, Mg,As, Cd,Cr, Cu,Fe, Mn,Ni, Pb,and Zn) and their distribution between organs of Rumex obtusifolius

Background and aims

The ionome (elemental composition) of grassland species has rarely been studied at the level of individual organs and little is known about effects of soil chemical properties on the ionome. Using the model oxalate plant Rumex obtusifolius, we asked how its biomass production and the distribution of elements between its organs is affected by soil chemical properties.

Methods

We established a pot experiment with R. obtusifolius planted in acidic non-contaminated control and in slightly acidic and alkaline soils anthropogenically contaminated by the risk elements As, Cd, Pb, and Zn. Both contaminated soils were untreated and treated by lime and superphosphate. We determined biomass production and the concentrations of elements in its organs.

Results

Biomass production was negatively related to the mobility of micro- and risk elements. Restricted transport of micro- and risk elements from belowground organs into leaves was recorded in untreated contaminated soils. In both lime-treated soils and in superphosphate-treated alkaline soil, elevated transport of micro- and risk elements from belowground organs into leaves was recorded in comparison to untreated contaminated soils. The lowest concentrations of micro- and risk elements were recorded in stems and seeds, followed by belowground organs and leaves.

Conclusions

R. obtusifolius is an As-, Cd-, Pb-, and Zn-excluder and is sensitive to high availability of micro- and risk elements in the soil. Soil chemical properties affect the distribution of essential elements within the plant greatly.     

Polymorphism analysis of the CYP1A1 locus in Koreans: presence of the solitary m2 allele

This study determined the complete genotype and the frequencies of all four mutations [T6235C (m1), A4889G (m2), T5639C (m3) and C4887A (m4)] of the CYP1A1 from 48 healthy Koreans and 17 Korean lung cancer patients. The mutations were analyzed by polymerase chain reaction (PCR)/restriction fragment length polymorphism (RFLP) and single stand conformation polymorphism (SSCP) simultaneously in order to improve accuracy as well as to screen for possible new alleles. Previously, the m2 mutation has always been linked to the m1 mutation. Also, the m1m2 double mutant allele (*2B) was thought to have a positive correlation with lung cancer susceptibility. Here we report the presence of the solitary m2 mutant allele without the m1 mutation (m1+m2) for the first time. This would be an evidence to support the theory of intragenic recombination in the CYP1A1 locus. The m1 mutation frequencies of healthy Koreans and lung cancer patients were 38.5% and 29.4%, respectively. The m2 mutation frequencies of healthy Koreans and lung cancer patients were 25.0% and 14.7%, respectively. Unlike the case for both Japanese and Caucasian lung cancer patients, neither m1 nor m2 mutations were overrepresented in Korean lung cancer patients. The m2 mutation frequency in Korean patients was significantly higher than those for Caucasians (2.7%) and the Japanese (19.8%). The African-American specific m3 mutation and m4 mutation found in Caucasians were not discovered in this study. The CYP1A1 allele with novel mutation was also not present.     

Assessment of Metal Contaminants in Non-Small Cell Lung Cancer by EDX Microanalysis

Human cardio-respiratory diseases are strongly correlated to concentrations of atmospheric elements. Bioaccumulation of heavy metals is strictly monitored, because of its possible toxic effects. In this work, we utilized the EDX microanalysis in order to identify the potential heavy metal accumulation in the lung tissue. To this aim, we enrolled 45 human lung biopsies: 15 non-small cell lung cancers, 15 lung benign lesions and 15 control biopsies. Lung samples were both paraffin embedded for light microscopy study and epon-epoxid embedded for transmission electron microscopy. EDX microanalysis was performed on 100 nm thick unstained ultrathin-sections placed on specific copper grids. Our results demonstrated that the EDX technology was particularly efficient in the study of elemental composition of lung tissues, where we found heavy metals, such as Cobalt (Co), Chromium (Cr), Manganese (Mn) and Lead (Pb). Furthermore, in malignant lesions we demonstrated the presence of multiple bio-accumulated elements. In fact, a high rate of lung cancers was associated with the presence of 3 or more bio-accumulated elements compared to benign lesions and control tissue (91.7%, 0%, 8.3%, respectively). The environmental impact on pulmonary carcinogenesis could be better clarified by demonstrating the presence of polluting agents in lung tissues. The application of EDX microanalysis on biological tissues could shed new light in the study of the possible bioaccumulation of polluting agents in different human organs and systems.Key words: EDX microanalysis, transmission electron microscopy, non-small cell lung cancer, heavy metals, environmental pollution     

肺癌转移与中医脏象理论相关性分析

目的:分析肺癌转移与中医脏象理论的相关性。方法:回顾性选择我院收治的102例晚期肺癌转移患者为研究对象,统计患者的转移灶所属部位,并参照《中医内科学》对患者的中医临床证候进行判定。分别统计各证型患者肺癌转移的单器官及首发转移器官的分布、肺癌转移各单器官及首发转移器官的证型分布情况。结果:各证型患者肺癌转移的单器官及首发转移器官分布率比较,差异具有统计学意义(P0.05)。同时,肺癌转移各单器官及首发转移器官的证型分布率比较,差异具有统计学意义(P0.05)。结论:根据肺癌患者的中医临床证候可判断其病灶转移方向,同时,根据肺癌患者的转移灶所属部位可判断其中医临床证型。     

Trace Elements in Scalp Hair Samples from Patients with Relapsing-Remitting Multiple Sclerosis

Background

Epidemiological studies have suggested a possible role of trace elements (TE) in the etiology of several neurological diseases including Multiple Sclerosis (MS). Hair analysis provides an easy tool to quantify TE in human subjects, including patients with neurodegenerative diseases.

Objective

To compare TE levels in scalp hair from patients with MS and healthy controls from the same geographic area (Sicily).

Methods

ICP-MS was used to determine the concentrations of 21 elements (Ag, Al, As, Ba, Cd, Co, Cr, Cu, Fe, Li, Mn, Mo, Ni, Pb, Rb, Sb, Se, Sr, U, V and Zn) in scalp hair of 48 patients with relapsing–remitting Multiple Sclerosis compared with 51 healthy controls.

Results

MS patients showed a significantly lower hair concentration of aluminum and rubidium (median values: Al = 3.76 μg/g vs. 4.49 μg/g and Rb = 0.007 μg/g vs. 0.01 μg/g;) and higher hair concentration of U (median values U: 0.014 μg/g vs. 0.007 μg/g) compared to healthy controls. The percentages of MS patients showing hair elemental concentrations greater than the 95th percentile of controls were 20% for Ni, 19% for Ba and U, and 15% for Ag, Mo and Se. Conversely, the percentages of MS patients showing hair elemental concentrations lower than the 5th percentile of healthy controls were 27% for Al, 25% for Rb, 22% for Ag, 19% for Fe, and 16% for Pb. No significant association was found between levels of each TE and age, disease duration or Expanded Disability Status Scale (EDSS) score. After stratification by gender, healthy subjects did not show any significant difference in trace element levels, while MS patients showed significant differences (p<0.01) for the concentrations of Ag, Cr, Fe, Ni and Sr. No significant differences were also found, at p<0.01, in relation to the use of cigarettes, consume of water, vegetables and place of living.

Conclusion

The different distributions of TE in hair of MS patients compared to controls provides an additional indirect evidence of metabolic imbalance of chemical elements in the pathogenesis of this disease. The increase in U and decrease in Al and Rb levels in MS compared to controls require further assessments as well as the observed different distributions of other elements.     

Concentrations of essential elements after repeated administrations of tin and selenium

To study effects of simultaneous administration of tin (Sn) and selenium (Se) on concentrations of several essential elements, mice were injected with either SnCl2 (ip) or Na2SeO3 (sc), alone or both compounds at a daily dose of 5 mumol/kg each for 12 consecutive days. Mice were sacrificed at 20 h after the last injection and concentrations of Sn, Se, Na, Ca, Zn, P, Fe, K, and Mg in the liver, kidney, spleen, pancreas, testis, seminal vesicle, lung, femoral muscle, and femoral bone were determined. In the control mice, Sn and Se concentrations were the highest in bone (0.69 micrograms Sn and 6.93 micrograms Se/g dry wt). Administered Sn was found to accumulate in all organs except the testis. Among the essential elements determined, Na was the most affected in terms of concentration in the organs and Mg was the least affected element in these organs. Among the organs tested, each elemental concentration in the pancreas was most affected. Simultaneous injections of Sn and Se appeared to keep the correlation coefficients between elements similar to those found in the control mice.     

Bayesian probit regression model for the diagnosis of pulmonary fibrosis: proof-of-principle

Background

The accurate diagnosis of idiopathic pulmonary fibrosis (IPF) is a major clinical challenge. We developed a model to diagnose IPF by applying Bayesian probit regression (BPR) modelling to gene expression profiles of whole lung tissue.

Methods

Whole lung tissue was obtained from patients with idiopathic pulmonary fibrosis (IPF) undergoing surgical lung biopsy or lung transplantation. Controls were obtained from normal organ donors. We performed cluster analyses to explore differences in our dataset. No significant difference was found between samples obtained from different lobes of the same patient. A significant difference was found between samples obtained at biopsy versus explant. Following preliminary analysis of the complete dataset, we selected three subsets for the development of diagnostic gene signatures: the first signature was developed from all IPF samples (as compared to controls); the second signature was developed from the subset of IPF samples obtained at biopsy; the third signature was developed from IPF explants. To assess the validity of each signature, we used an independent cohort of IPF and normal samples. Each signature was used to predict phenotype (IPF versus normal) in samples from the validation cohort. We compared the models' predictions to the true phenotype of each validation sample, and then calculated sensitivity, specificity and accuracy.

Results

Surprisingly, we found that all three signatures were reasonably valid predictors of diagnosis, with small differences in test sensitivity, specificity and overall accuracy.

Conclusions

This study represents the first use of BPR on whole lung tissue; previously, BPR was primarily used to develop predictive models for cancer. This also represents the first report of an independently validated IPF gene expression signature. In summary, BPR is a promising tool for the development of gene expression signatures from non-neoplastic lung tissue. In the future, BPR might be used to develop definitive diagnostic gene signatures for IPF, prognostic gene signatures for IPF or gene signatures for other non-neoplastic lung disorders such as bronchiolitis obliterans.

     

Genetic analysis of twelve X-chromosomal STRs in Japanese and Chinese populations

We investigated the genetic markers of twelve X-STR loci in 670 healthy, unrelated Japanese (438 men and 232 women) from Tokyo and 488 Chinese (263 men and 225 women) from Shenyang, using the Investigator Argus X-12 kit. Allele and haplotype analyses of twelve X-STRs clustered into four linkage groups indicated that they are highly informative for forensic applications in Japanese and Chinese populations. Hardy–Weinberg equilibrium tests demonstrated no significant deviations in the two populations. Among the four closely linked X-STR trios, some haplotype unique to Japanese or Chinese population were detected. Haplotype diversity for each linkage group ranged from 0.9861 to 0.9968, showing high values in each of the study populations. The genetic distances between populations based on the 12 X-STR loci and the phylogenetic tree revealed long genetic distances between Asian and Caucasian populations and between Asian and African population (Moroccan). These results suggest that the twelve X-STR loci will contribute to forensic casework in Japanese and Chinese populations.     

Comparative electrophoretic study of phosphoglucose isomerase,phosphoglucose mutase and tetrazolium oxidase in some hydatid cysts of Echinococcus granulosus collected from different mammal hosts

Hydatid cyst scolexes of lung and liver from sheep and water buffalo were electrophoreticaily examined for phosphoglucose isomerase (PGI), phosphoglucose mutase (PGM) and tetrazolium oxidase (Te) activity. One each of phosphoglucose isomerase, phosphoglucose mutase and tetrazolium oxidase activities were found in all of the hydatid cysts studied. PGI and PM in ail hydatid cysts studied appeared te be controlled by two codominant alleles. PGI and PGM proved to be a good taxonomic criterion to differentiate the hydatid cysts of the water buffalo from those from sheep. On the other hand, TO failed to differentiate between the sheep and water buffalo hydatid cysts. The hydatid cysts that parasitise the sheep's organs can be considered as arising from a different strain from the cysts that parasitise the water buffalo organs, while lung and liver hydatid cysts of the sheep appeared to be a similar strain.     

The increased ecto-5'-nucleotidase activity in muscle, heart and liver of laminin alpha2-deficient mice is not caused by an elevation in the mRNA content

We have previously shown that mouse muscle and liver contain catalytically active and inactive ecto-5'-nucleotidase (eNT) variants and that eNT activity in these tissues increases in laminin alpha2 (merosin)-deficient Lama2dy mice. These results prompted us to study whether: (1) the increase of eNT activity depends on the change in the content of merosin between healthy and dystrophic organs; (2) the active and inactive eNT variants arise from the same or distinct mRNAs; (3) the enhancement of the activity is caused by an increase in the eNT mRNA content. Compared to healthy organs, the activity in dystrophic organs increased four-fold in muscle, 1.7-fold in liver, 1.4-fold in heart and not at all in kidney and lung. The level of immunolabelled eNT protein per unit of activity suggested a similar ratio of inactive to active eNT in healthy liver, kidney, heart and muscle, which increased greatly in lung. The size of the eNT subunit in liver, kidney, heart and muscle (72 kDa) decreased to 66 kDa in lung. The identification of a single RT-PCR product suggested that active and inactive eNT arise from the same mRNA and are generated by a differential post-translational processing. Compared to the content in muscle, the amount of eNT mRNA was 12-fold higher in liver and kidney, eight-fold in heart and five-fold in lung. The relative content of eNT mRNA was unaffected by the deficiency of merosin.     

The organ distribution of selenium in German adults

The selenium concentrations were determined in liver, kidney, skeletal muscle, heart, brain, prostate, testis, bile, lung, and spleen of German traffic accident victims. In addition the nitrogen and phosphorus contents were determined in the same organs and tissues. On a per-weight unit basis, the highest selenium concentration was found in kidney. However, this corresponds to only 4% of the total body selenium. Most of the whole body selenium (50%) is present in skeletal muscle, which thus appears to act as a selenium storage organ. However, there is also evidence that selenium is required for muscle function. In plasma and interstitial fluid, .450 mg of Se, or 7.5% of the total body selenium is present. A comparison of the organ Se concentrations of the German traffic accident victims with the selenium concentrations of the same human organs as reported in different countries indicates that the organ concentrations of West Germans are comparable to that of the population of New Zealand, a low-Se country, and significantly lower than that observed in the organs of American, Canadian, and especially Japanese subjects. The international comparison of the organ selenium concentrations also revealed that the selenium uptake of kidney is higher at low- and adequate dietary Se intakes and lower if the dietary Se supply is high, as is the case for Japanese subjects. Estimates of the daily excretion of selenium with the bile indicate that the amounts are three times higher than the daily urinary losses and in the same order of magnitude as the daily dietary selenium intakes. Enterohepatic reabsorption of selenium from the bile appears to be a significant mechanism of conserving dietary selenium and to maintain Se balance at comparatively low dietary Se intakes.