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Major depression （MD） is the most prevalent mood disorder worldwide. Electroconvulsive treatment （ECT） is a highly effective treatment in psychiatry and shows efficacy in patients who were resistant to pharmacotherapy. Recent retrospective cohort studies have shown that ECT is highly effective in ameliorating depressive symptoms. Similar ECT efficacy has been established also in various rodent models of MD. To establish further efficacy of ECT in rodent depression models, we investigated the effect of ECT on depressive-like behaviour induced by lipopolysaccharide （LPS） in rats. Methods: Adult male Sprague-Dawley （SD） rats were randomly divided into four groups, namely, control, LPS, sham ECT, and LPS + ECT. The SD rats received intraperitoneal injection of LPS or sterile saline, followed by ECT or sham treatment for 7 consecutive days. Subsequently, the forced swimming test （FST） and Y-maze test were performed. Results: The FST showed that the immobility time in the LPS group （169.73 ± 15.46 s） was significantly longer than that in the control group （63.16 ± 4.48 s）. However, ECT administration to LPS-injected rats significantly shortened the prolonged immobility. The Y-maze test showed a significant decrease in % spontaneous alternation behaviour （SAB） in the LPS group compared to the control group. ECT administration to LPS-injected rats significantly restored such a decrease in % SAB. Conclusions: Our results suggest that repeated ECT ameliorated LPS-induced depressive-like behaviour in SD rats. Further studies are warranted to elucidate the molecular mechanism of such therapeutic effects of ECT.

Type I collagen, which consists of two α1 and one α2 chains, is an abundant extracellular matrix ( ECM ) protein. Changes in type I collagen are often associated with various diseases including fibrosis, osteogenesis imperfecta ( OI ), and Ehlers-Danlos syndrome ( EDS ). In the present study, we developed a method for quantification of type I collagen α1 ( COL1A1 ) in a culture medium of LX-2 human hepatic stellate cells by using nano-liquid chromatography tandem mass spectrometry ( nano-LC/MS/MS ). After selecting a specific peptide of COL1A1, unlabeled and stable isotope-labeled peptides were used for quantitative analysis of COL1A1 by nano-LC/MS/MS. The concentration of secreted COL1A1 in the LX-2 cell culture medium was 38.78 ng/mL. The results indicate that this method is useful for quantifying COL1A1 in a cell culture medium.

Osteoarthritis (OA) is a degenerative disorder that is characterized by articular cartilage destruction. The level of tenascin-C (TNC), an extracellular matrix protein, is increased in synovial fluid (SF) from OA patients, and TNC concentration in SF correlates with the severity of OA in the knee. Tenascin-X (TNX), another member of the tenascin family, has been identified as a causative protein of classical-like Ehlers-Danlos syndrome (clEDS). In this study, we investigated the correlation between TNX concentration and the degree of cartilage denaturation in OA patients. TNX concentrations in SF from patients with meniscus tears and from OA patients were lower than those of control sera. However, significant difference in TNX concentration in SF was not observed between patients with meniscus tears and OA patients. Furthermore, a significant correlation was not found between TNX concentration in SF and severity of OA in the knees of patients. These results suggest that TNX might not be as useful as TNC as a biomarker.

A caffeine overdose is most likely to result from the consumption of nutritional supplements or caffeine pills. Herein, we describe two fatal cases of caffeine intoxication. Case 1: A Japanese man in his 30s was found dead, with two bottles of water and unopened tablets for motion sickness near his body. Caffeine concentrations were 99.6 μg / mL in the cardiac blood and 169.1 μg / mL in stomach contents. Case 2: A Japanese man in his 40s was found dead, with an empty bottle of caffeine and six empty boxes of Travelmin ( a motion sickness medication ) in a trash can. Caffeine concentrations were 79.6 μg / mL in the cardiac blood and 876 μg / mL in the stomach contents. Fatal caffeine poisoning is comparatively uncommon, but it has recently been increasing due to the easy availability. For the prevention of caffeine overdoses, there should be heightened public awareness of the potential dangers of a caffeine overdose.

Background: We have found the association of single nucleotide polymorphism （SNP） rs6180 in the growth hormone receptor （GHR） gene with plural tissue weight. On the other hand, the DNase I gene（ DNASE1） is considered to be one of the susceptibility genes for liver disease, gastric and colorectal carcinoma, and myocardial infarction, based on the polymorphic study of rs1053874. Therefore, we investigated the association of rs1053874 in DNASE1 with plural tissue weight in the present study. Materials and methods: Blood samples （n = 198; 99 female and 99 male） were collected from Japanese subjects autopsied in Shimane Prefecture, and genomic DNA was extracted. SNP （rs1053874, A > G substitution） was analyzed using a polymerase chain reaction, followed by a mismatched-restriction fragment length polymorphism analysis. Results: Except for the pancreatic weight and appendix, all parameters were significantly related to gender. All parameters were significantly related to age except for the left and right renal weight. In the present study, we focused on analyzing the potential effect of a variation in DNASE1 and investigated the effect of SNP rs1053874 on human organ weight using autopsied samples. Conclusions: In the present study, the rs1053874 polymorphism does not contribute to organ weight, cardiac hypertrophy index, or body surface areas, and these associations were not observed in Japanese subjects. This study is the first to investigate the association of SNP rs1053874 （A > G substitution） in DNASE1 and data routinely measured at autopsy, such as organ weight.

Caffeine is commonly used as a stimulant, and fatal caffeine overdoses are increasing. In the present study, two case reports showing a fatal blood concentration of caffeine were reported and compared to previously published case reports. Case 1: A 19-year-old female was found dead in her room. It was speculated that she ingested 84 tablets （100 mg） of a sleep-inhibitor product. The caffeine levels in her cardiac blood and stomach were 116 μg/ml and 619 μg/ml, respectively. There was no macroscopic indication of any natural or unnatural cause of death. Case 2: The condition of a 47-year-old male deteriorated suddenly. Hypothermia and excess sweating were observed. After vomiting several times, he lost consciousness. At autopsy, severe pulmonary edema was observed. His cardiac blood caffeine level was determined to be 171 μg/ml, which exceeds fatal blood caffeine levels. However, there is no evidence of his ingesting massive amounts of caffeine. The blood caffeine concentration in previously reported fatal caffeine ingestions ranged from 33 to 350 μg/ml. The two present cases are within this range. The accumulation of data on blood caffeine concentrations and ingestion amounts is helpful to predict the intake amount of caffeine.

X-ray repair cross-complementing group 1 (XRCC1) repairs single-strand breaks in DNA. Several reports have shown the association of single nucleotide polymorphisms (SNPs) (Arg194Trp, Pro206Pro, Arg280His, Arg399Gln) in XRCC1 to diseases. Limited population data are available regarding SNPs in XRCC1, especially in African populations. In this study, genotype distributions of four SNPs in worldwide populations were examined and compared with those reported previously.Four SNPs (Arg194Trp, Pro206Pro, Arg280His, Arg399Gln) in XRCC1 from genomic DNA samples of 10 populations were evaluated by using polymerase chain reaction followed by restriction fragment length polymorphism analysis.The frequency of the minor allele corresponding to the Trp allele of XRCC1Arg194Trp was higher in Asian populations than in African and Caucasian populations. As for XRCC1Pro206Pro, Africans showed higher minor allele frequencies than did Asian populations, except for Tamils and Sinhalese. XRCC1 Arg280His frequencies were similar among Africans and Caucasians but differed among Asian populations. Similarly, lower mutant XRCC1 Arg399Gln frequencies were observed in Africans.This study is the first to show the existence of a certain genetic heterogeneity in the worldwide distribution of four SNPs in XRCC1.

In the present study, acute toxicity of ZnO NPs and silica-coated ZnO NPs following intravenous injection using mice were evaluated. ZnO dispersion （average size: 148 nm） at a dose of 10 mg/kg and 30 mg/kg, and a Silica-coated ZnO （ZnOSiO2）at a dose of 10 mg/kg, 30 mg/kg and 130 mg/kg were injected intravenously via tail vein of mouse. All mice （n = 6） died within 5 min following intravenous ZnO dose of 10 mg/kg and 30 mg/kg. Impaired motility and convulsion were observed before the death. On the other hand, silica-coating can reduce the toxicity. All mice survived and impaired motility was not observed following intravenous ZnO-SiO2 （average size: 81 nm） treatment at the same dose of ZnO NPs. At a dose of 130 mg/kg, impaired motility and convulsion was observedin all mice, one mouse out of 5 mice died after 5 min. Blood coagulation was observed in all died mice,which may be related to the toxicity of ZnO NPs.

An elderly woman in her late 80s was living with her husband and her son in his mid-60s. The woman had urinary incontinence. Her son took her to the bathroom, showered her in the bathtub with her clothes on, and then left her inside the bathtub. Five days after on a hot summer day, the woman was found dead inside the waterless bathtub. A forensic autopsy was performed 2 days after the discovery of her death on suspicion of the son’s negligence as a guardian. The woman was underweight, and the low BMI value in addition to the lack of sufficient subcutaneous fat and musculature, had an effect on the woman’s vulnerability to hypothermia. Furthermore, the woman’s clothes were drenched. We believe that these factors contributed greatly to her susceptibility to hypothermia. Examining all of the environmental factors and autopsy results, the cause of death was concluded to be hypothermia.

Elevated urinary albumin excretion rate（UAER) is a modifiable risk factor for renal and cardiovascular diseases in type 2 diabetes. Blockade of the renin-angiotensin system（RAS）lowers UAER. In the present study, 46 patients（male: 25, female: 21, mean age 62.8±5.9 years old）with CKD and hypertension accompanied with type 2 diabetes were enrolled, among whom there were wide ranges of blood-pressure values. Patients had been used amlodipine or nifedipine to control blood pressure between 2007 and 2009 at Nagami Clinic, Shimane Japan. However, UAER did not decrease and estimated glomerular filtration rate（eGFR）did not increase during the first 24 month follow up period. To those 46 patients, losartan which is one of angiotensin receptor blockers（ARBs）and azelnidipine which is one of calcium channel blockers（CCBs）were administered in order to examine whether combined administration of losartan and azelnidipine（Los/Azl treatment）provides superior reduction in proteinuria compared to nifedipine or amlodipine in hypertensive type 2 diabetes patients with CKD. All 46 patients received either 25 mg per day losartan, which was increased up to 50 mg per day, and 8 mg per day azelnidipine, which was increased up to 16mg per day, over 24 months. Mean systolic and diastolic pressure was controlled less than 120/80 mmHg. This present study was a 2 years’prospective cohort study. During the first 24 month follow up period, there was not a significant difference of the mean eGFR and UAER between amlodipine group and nifedipine group. However, the present changes in eGFR and UAER by Los/Azl treatment at 6, 12, 18 and 24 month showed 5.4, 7.0, 9.0, and 13.2 % for eGFR and -15.2, -24.8, -37.3, and -48.7% for UAER, respectively. In conclusion, it was demonstrated that Los/Azl treatment should be considered for hypertensive diabetic patients with CKD.

Adult height is a highly heritable trait involving multiple genes. Recent genome-wide association studies have identified that SNP rs12338076 in the LHX3-QSOX2 locus, and rs1457595 and rs17032362 in the IGF1 locus are associated with human height in the Japanese population (Okada et al. (2010) [9]). We performed a replication study to examine the associations between these three SNPs and adult height in the Japanese population based on autopsy cases. However, it was not possible to confirm that all these SNPs influenced adult height in the study population. We first conducted a wide-ranging survey of these three SNPs in the above genes using nine different populations including Asians, Africans and Caucasians, and demonstrated that the genotypes of rs12338076 and rs17032362 were distributed in an ethnicity-dependent manner; even within Asian populations, the genotype distributions of the SNPs differed widely. Although there are differences in height distribution between different populations, possibly due to genetic factors and/or gene–environmental interactions, the contradictory results of the association study and ethnic differences in genotype distribution allow us to assume that these height-related SNPs in the genes may contribute to adult height to a slight extent, at least in the Japanese population. It is anticipated that the present information will be useful for developing a reliable tool for personal identification through elucidation of the genetic basis of human height.

Prolactin (PRL) is associated with lactation, and also plays an important role in the development and progression of breast cancer (BC). PRL exerts its effects via its receptor, the PRL-receptor (PRLR), so it is beneficial to study the distribution and alteration in expression of PRLR in order to elucidate the biologic effects of PRL on BC. In 84 BCs, PRLR expression was examined by an immunohistochemical technique. PRLR immunoreactivity was detected mainly in the cytoplasm. And the overall positive rate (cut-off > 10%) was 71% (60/84), while invasive BC showed a significantly higher positive rate (75%, 57/76) than non-invasive BC (38%, 3/8) (p=0.0264). The PRLR expression showed no correlation with age, primary tumor, nodal involvement, distant metastasis, or clinical stage; however, it was significantly correlated with estrogen receptor (ER) expression (p=0.0361), especially in invasive BC (p=0.0098), but not with the progesterone receptor (PgR) expression or HER-2 expression. PRLR was highly expressed in IDC ( invasive ductal carcinoma) and correlated with ER expression, suggesting that PRL and PRLR may play important roles in the development and progression of ER (+) BC.

Recently, SNPs and hap10types in the DNase 11 gene were shown to be associated with renal disorder in SLE patients. In this study, DNASE2*Ser145Ser and DNASE2 *Va1206Ile polymorphisms were determined by using the PCR-RFLP technique in healthy subjects of Japanese, Koreans, Ovambos, Ghanaians, and Xhosans (n=871). For DNASE2*Ser145Ser, the Japanese and Korean populations exhibited higher 2630T frequencies than the Ovambos, Ghananians, and Xhosans, whose allele frequencies were higher for 2630C. Population data from the HapMap Project showed similar allele frequencies. For DNASE2 * Va1206Ile, Koreans were the only population with the DNASE2*VaI206Ile polymorphism. A previous study revealed an association with the risk of renal disorders in Korean SLE patients and the DNASE2 * Ser145Ser polymorphism. In this study, we verified significant ethnic diversity in the DNASE2*Ser145Ser polymorphisms. It is questionable whether a correlation between the risk of renal disorder in SLE patients and the DNASE2*Ser145Ser polymorphism exists in populations other than the Korean population.

Deoxyribonuclease I (DNase I) from equine parotid gland was purified by reformation with a yield of 48 % to electrophoretic homogeneity using three-step conventional column chromatography. The purified enzyme showed a molecular mass of about 37kDa and maximum activity at pH 7.0. It required divalent cations Mg2+ and Ca2+ for its activity and was inhibited by EDTA and EGTA. The purified enzyme preparation was found to contain no other detectable nucleases. An antibody against the purified enzyme was found to be monospecific against the equine parotid gland and the pure antigen, and completely blocked the activity of the purified enzyme. Inhibition by globular actin (G-actin) was investigated in seven mammalian enzymes. Activity inhibition by G-actin was very strong in an equine enzyme. In G-actin-inhibited human DNases I, four amino acid residues, Tyr-65, Val-66, Val-67 and Ala-114, were involved in actin binding. In equine, Tyr-65 is substituted with Phe-65. Considering that the enzymatic activity of a rabbit enzyme with Phe-65 and Ala-67 was less efficiently inhibited by G-actin, the structural involvement of Phe-65 and Val-67may result in a strong DNase I and G-actin complex.

Deoxyribonuclease I (DNase I) polymorphism n exon 8 (A2317G) designated as DNASEI is considered to be one of the susceptibility genes for gastric and colorectal carcinoma, and myocardial infarction. Moreover, the presence of a variable number of tandem repeat (VNTR) polymorphism, designated as HumDN1, in intron 4. was found. In this study, the simultaneous genotyping of DNASEI and HumDN1 polymorphisms within the DNase I gene was performed in Ovambo, Turk, Mongolian, Korean and Japanese populations. Genotype distributions of DNASEI and HumDNI Ioci differed among these populations. While there was a general uniformity for the two polymorphisms in the three Asian populatlons, a significant difference in genotype distribution were found between the Ovambo and Turk populations. The DNASE1*1 and HumDN1*3 alleles were found to be the most predominant in the Ovambos, with the Turks showing the highest allele frequency for DNASE1*2, HumDN1*4 and HumDN1*5. From this study, there was the existence of a certain genetic heterogeneity in the distribution of these two DNase l polymorphisms. Furthermore, linkage disequilibrium between these two polymorphisms was revealed in all of the populations except the Ovambos. The combination of the two polymorphisms within a DNase I gene may be potentially useful for clinical purposes and in population genetic studies.

A deoxyribonuclease I (DNase I)-genotyping method using polymerase chain reaction (PCR) has been developed, underlying the molecular basis of the genetic polymorphism controlled by four codominant alleles, DNASE1*1, *2, *3 and *4. Two new alleles, DNASE1*5 and DNASE1*6, were recently discovered. In this study, we have added a new DNase I-genotyping method based upon both allele-specific amplification and mismatched PCR followed by Bam HI digestion. Using this method, the genotype distribution of DNase I in unrelated individuals from Shimane Prefecture in Japan was examined and compared with the results of other Japanese and/or ethnic population studies. The DNASE1 allele frequencies for the Japanese individuals from Shimane Prefecture were determined to be DNASE1*1 0.5763, DNASE1*2 0.4153 and DNASE1*4 0.0084 (X2=1.8304, 0.75> p >0.60). The existence of a decreasing north-to-south gradient in the DNASE1*2 allele was revealed from the study of 10 Japanese populations containing Shimane Prefecture. The allele frequency of DNASE1*2 of the Shimane Prefecture population was relatively lower than those of the German and Turkish populations but higher than that of the Ovambos population. These findings suggest the presence of a different mechanism in the chance fluctuaions of DNASE1 alleles among different populations.

We have confirmed for the first time, both biochemically and immunologically, the existence of deoxyribonuclease I (DNase I) in the human tears. Isoelectric focusing of tear samples on polyacrylamide gels (pH 3.5-5), followed by dried agarose film overlay detection, was used to determine the phenotypes of tear DNase I. Because of its high degree of sensitivity and its high band resolution, this detection method allowed determination of DNase I types from tear samples of about 5 μL. Pretreatment of tear samples with the enzyme sialidase was essential for typing in order to markedly enhance the sensitivity and simplify the isozyme pattern. DNase I types in all tear samples were consistently related to types found in corresponding blood and urine samples. DNase I typing may, therefore, provide a novel discriminating characteristic in the genetic, clinical and forensic examination of tears.

Because of their sensitivity and high level of discrimination, short tandem repeat (STR) maker systems are currently the method of choice in routine forensic casework and data banking, usually in multiplexes up to 1517 loci. Constraints related to sample amount and quality, frequently encountered in forensic casework, will not allow to change this picture in the near future, notwithstanding the technological developments. In this study, we present a free online calculator named PopAffiliator (http://cracs.fc.up.pt/popaffiliator) for individual population affiliation in the three main population groups, Eurasian, East Asian and sub-Saharan African, based on genotype profiles for the common set of STRs used in forensics. This calculator performs affiliation based on a model constructed using machine learning techniques. The model was constructed using a data set of approximately fifteen thousand individuals collected for this work. The accuracy of individual population affiliation is approximately 86%, showing that the common set of STRs routinely used in forensics provide a considerable amount of information for population assignment, in addition to being excellent for individual identification.