Effects of psychogenic stress on oxidative stress and antioxidant capacity at different growth stages of rats: Experimental study.

This study examined the psychogenic stress (PS) effects on changes in oxidative stress and the antioxidant capacity of an organism at different growth stages. The experimental animals were male Wistar rats of five different ages from growth periods (GPs) to old age. The growth stages were randomly classified into control (C) and experimental (PS) groups. The PS was performed using restraint and water immersion once daily for 3 h for 4 weeks. Reactive oxygen metabolites (d-ROMs) and the biological antioxidant potential (BAP) were measured before and after the experiment. In addition, the liver and adrenal glands were removed, and the wet weight was measured. The d-ROM and BAP of all growth stages given PS increased significantly. The d-ROM in the C group without PS increased significantly in GPs while decreased significantly in old-aged rats. In addition, the BAP of the C group in GP and early adulthood were all significantly elevated. There were significant differences in organ weights between the C and PS groups at all growth stages. Oxidative stress and antioxidant capacity differed depending on the organism's developmental status and growth stage, and PS also showed different effects. In particular, the variability in oxidative stress was remarkable, suggesting that the effect of PS was more significant in the organism's immature organs.

[Estimating the prevalence of tuberculosis infection among healthcare workers in our hospital by repeat QFT-G testing].

OBJECTIVE: The QuantiFERON-TB (QFT) blood test is the major tool for the diagnosis of Mycobacterium tuberculosis (TB) infection among healthcare workers (HCWs). We used QFT tests to estimate the prevalence of TB infection among HCWs in our hospital. MATERIAL AND METHODS: Between 2003 and 2010, a total of 733 HCWs were enrolled in this study, and the prevalence of TB infection was analyzed according to the HCWs' jobs and work place. RESULTS: Among the 152 men and 581 women who were evaluated, 3 female HCWs had a history of TB. Fifty-eight HCWs (8 men and 50 women with a mean age of 56.3 years and 48.4 years, respectively) demonstrated positive QFT tests. The positive rate was 7.9% for all staff members throughout the study period. The QFT test was positive for 1 HCW who was treated for TB in 1998, and negative and inconclusive for 2 other HCWs treated for TB in 2002. The positive rate for QFT was 16.0% in the TB ward (12/75, 95% confidence interval [CI]: 7.7-24.3%), 9.9% in the other wards (22/222, 95% CI: 7.9-11.9), and 1.1% in the outpatient department (1/91, 95% CI: 0-2.2). According to the job category, the QFT positive rates were as follows: doctors, 4.3% (3/70, 95% CI: 1.9-6.7); nurses, 10.3 (4/35, 95% CI: 6.0-16.8). The positive rate among doctors working in the TB ward was 10.0%, and that for nurses was 24.3%. This indicates that the prevalence of infection among HCWs in the TB ward was significantly higher than that in other work places. A comparison of the results from 2003 through 2007 revealed that for a total of 307 workers, 90.6% and 5.2% remained negative and positive, respectively, while 1.6% converted from negative to positive, and 2.6% from positive to negative. CONCLUSION: The positive rate among HCWs in the TB ward was higher than that in other wards. This is especially remarkable for doctors and nurses working in the TB ward.

[Clinical factors of pulmonary tuberculosis in non-tuberculosis wards and its incidence among contact patients and healthcare workers in our hospital using QuantiFERON GOLD testing].

OBJECTIVE AND METHODS: In our hospital, we analyzed the clinical factors of pulmonary tuberculosis (TB) diagnosed in non-TB wards and the incidence of TB infection among contact patients and healthcare workers (HCWs) using QuantiFERON-TB GOLD (QFT) testing. MATERIAL: This study included 16 patients who were diagnosed with pulmonary TB in non-TB wards in our hospital from January 2008 to May 2011. Eight contact patients and 120 HCWs were also enrolled. RESULTS: The 16 TB patients comprised 11 men (77.7 years) and 5 women (74.4 years). Among them, only 9 patients exhibited positive results for Mycobacterium tuberculosis after the first acid-fast bacterial examination; the other 7 patients presented positive results only after the second or third examinations. Moreover, there were 3 cases of positive Mycobacterium avium samples in the first acid-fast bacterial examination. Among 16 pulmonary Mycobacterium tuberculosis cases, 8 were sputum smear and culture positive, 7 were sputum smear negative and culture positive, and 1 was sputum smear and culture negative. Moreover, 17 days had elapsed from the time of admission to the non-TB ward to diagnosis. TB contact examination revealed that QFT results for 2 HCWs changed from negative to positive. DISCUSSION: We suspected pulmonary aspergillosis or old TB when presented with cases with a history of TB. Moreover, we believe that the periods from admission to diagnosis were delayed when the first acid-fast bacterial sputum examination was negative or showed non-tuberculous mycobacteria.

Crystallization and preliminary X-ray diffraction analysis of orotate phosphoribosyltransferase from the human malaria parasite Plasmodium falciparum.

Orotate phosphoribosyltransferase (OPRT) catalyzes the Mg(2+)-dependent condensation of orotic acid (OA) with 5-α-D-phosphorylribose 1-diphosphate (PRPP) to yield diphosphate (PP(i)) and the nucleotide orotidine 5'-monophosphate. OPRT from Plasmodium falciparum produced in Escherichia coli was crystallized by the sitting-drop vapour-diffusion method in complex with OA and PRPP in the presence of Mg(2+). The crystal exhibited tetragonal symmetry, belonging to space group P4(1) or P4(3), with unit-cell parameters a = b = 49.15, c = 226.94 Å. X-ray diffraction data were collected to 2.5 Å resolution at 100 K using a synchrotron-radiation source.

Baseline Body Composition and Physical Activity Level Recommended for Optimal Bone Mineral Density in Young Women.

Aim: This study examined the influence of body composition and physical activity level (PAL) on bone mineral density (BMD) to determine the baseline values necessary for maintaining healthy trabecular bone and improving bone health in young women. Materials and Methods: The subjects, consisting of healthy young women, were assigned to either a BMD-low (BMD-L) or BMD-high (BMD-H) group using the osteosonic index. PAL was measured for 1 week and rated on a scale from PAL-1 to 8 according to intensity levels (metabolic equivalents [METs]). The Wilcoxon rank-sum test was performed for intergroup comparisons. Results: The BMD-H group had significantly higher fat-free muscle mass, skeletal muscle index, and basic metabolic rate than the BMD-L group (p < 0.001, each). Regarding the timing of physical activity in daily life according to intensity, PAL-6 (p < 0.01), PAL-7 (p < 0.001), and PAL-8 (p < 0.01) for the BMD-H group were significantly longer than those for the BMD-L group. Discussion and Conclusion: For young women in their early 20s, BMD may be associated with baseline physical fitness and strength, as determined by body composition, but it is not influenced by the duration of physical activity. A PAL ≥6.0 METs may improve or maintain the effect on BMD.

Toe Clearance Rehabilitative Slippers for Older Adults With Fall Risk: A Randomized Controlled Trial.

INTRODUCTION: To evaluate fall-prevention rehabilitative slippers for use by self-caring, independent older adults. MATERIALS AND METHODS: This assessor-blinded, randomized, and controlled 1-year study included 59 self-caring, independent participants (49 women) who attended day services. The mean age of participants was 84.0 ± 5.3 years. Participants were randomly selected from 8 nursing homes. We tested slippers top-weighted with a lead bead (200, 300, or 400 g). Intervention group participants walked while wearing the slippers for 10-20 min, 1-3 days/week at the day service center. Fall risk was measured using the Berg Balance Scale and the Tinetti Performance-Oriented Mobility Assessment (POMA) before and at 3-month intervals after the intervention/control phase. RESULTS: After 12 months, the intervention group demonstrated significant improvement. Berg Balance and POMA compared to the control group (p < .05 p < .01, respectively). Mobility scores improved significantly for both measurements in the intervention group before and after (p < .01), but the control group had significantly lower scores. DISCUSSION: Overall, falls decreased in the intervention group from 10 to 7, and control group falls increased from 9 to 16 (p = .02). No adverse events related to the intervention were reported. CONCLUSIONS: Rehabilitation training slippers may reduce falls in older adults.

Substrate-induced closing of the active site revealed by the crystal structure of pantothenate synthetase from Staphylococcus aureus.

Pantothenate synthetase (PS, EC 6.3.2.1) is the last enzyme in the pantothenate biosynthesis pathway, a metabolic pathway identified as a potential target for new antimicrobials. PS catalyzes the ATP-dependent condensation of pantoate and beta-alanine to form pantothenate. Here we report the overexpression, purification, enzyme assay, and tertiary structure of PS from Staphylococcus aureus. PS activity was experimentally confirmed, indicating a k(cat) value comparable to those of enzymes from other organisms. The structures of the apoenzyme and the reaction intermediate (pantoyl adenylate; PA) complex were determined by X-ray crystallography to resolutions of 2.5 and 1.85 A, respectively. Structural analysis indicated that the apoenzyme adopts an open and relatively mobile structure, while the complex structure is closed and entirely rigid. Structural comparison of the apoenzyme and the complex revealed how S. aureus PS undergoes open/close conformational change, and also determined the key interactions with the adenine ring of PA for a hinge bending domain closure. In the complex structure, PA and acetate are bound in the active site. We suggest that the acetate mimics the substrate beta-alanine. Therefore, the complex structure seems to represent a catalytic state poised for in-line nucleophilic attack on PA. These data also offer an alternative strategy for designing novel compounds that selectively inhibit PS activity.

[Circumstances and comparisons of child rearing burnout scales between neonatal mothers with and without home visit nursing. Analysis with a child rearing burnout scale].

OBJECTIVE: The Japanese neonate home-visit nursing guidance program for mother-and-child health administration is unique on the world scale, because it makes support for every family possible. However, very few studies have been conducted concerning the efficacy of the neonate home-visiting guidance program. The purpose of this study was to examine the utility of the program and factor out necessary questionnaire entries to sreflect mother's child-rearing anxieties. METHOD: Questionnaire survey. The subjects were 169 mothers. RESULTS: Ninety-two (54.4%) out of 169 subjects had neonate home-visit nursing guidance. Five factors and twenty-two items were abstracted by the factor analysis method. The difference in factor scores between the groups with and without guidance was analyzed byt-test. There were significant differences in the first factor (feelings change), the second factor (body tiredness), and the fourth factor (brought up a child) between the group that had guidance and the group that had not, and the score for mother's child-care anxiety in the group that had guidance was high. Among subjects who answered "no" to the question whether their husbands gave them support in rearing their child, there were significant differences concerning child-rearing anxiety questions in the questionnaire. The attributes showing meaningful differences in child-rearing anxiety questions were: first delivery, having an extended family, unemployment, and delivery at over-30 years of age. CONCLUSION: The mother's idea that her husband is cooperative in raising their children showed a relationship with the mother's child-care anxiety, so it is important for home-visit guidance to find out whether the husband is a partner in raising children. The other necessary questionnaire items were: age, having birth experience, family structure, and being employed or unemployed. As the subjects who accepted neonate home-visit nursing guidance could be said to have some child-rearing anxiety subconsciously, it is concluded that the neonate home-visit nursing guidance program is effective for determination of mother's child rearing anxiety.

The Influence of the Type of Continuous Exercise Stress Applied during Growth Periods on Bone Metabolism and Osteogenesis.

BACKGROUND: In this study, we examined the influence of exercise loading characteristics on bone metabolic responses and bone morphology in the growth phase and adulthood. METHODS: Running exercise (RUN) and jumping exercise (JUM) were used for the exercise loading in 28-day-old male Wistar rats. Bone metabolism was measured by blood osteocalcin (OC) and tartrate-resistant acid phosphatase (TRACP) levels. For bone morphology, the maximum bone length, bone weight, and bone strength of the femur and tibia were measured. RESULTS: A pre- and post-exercise loading comparison in the growth phase showed significantly increased OC levels in the RUN and JUM groups and significantly decreased TRACP levels in the JUM group. On the other hand, a pre- and post-exercise loading comparison in adulthood showed significantly decreased TRACP levels in the RUN and JUM groups. Femur lengths were significantly shorter in the RUN and JUM groups than in the control (CON) group, while bone weight was significantly greater in the JUM group than in the CON group. CONCLUSIONS: Exercise loading activates OC levels in the growth phase and suppresses TRACP levels in adulthood. On the other hand, these results suggest that excessive exercise loading may suppress bone length.

The active site structure of quinohemoprotein amine dehydrogenase inhibited by p-nitrophenylhydrazine.

Quinohemoprotein amine dehydrogenase (QH-AmDH) catalyzes the oxidative deamination of aliphatic and aromatic amines. The enzyme from Pseudomonas putida has an alpha beta gamma heterotrimeric structure with two heme c groups in the largest alpha subunit, and a novel quinone cofactor [cysteine tryptophylquinone (CTQ)] and hitherto unknown internal cross-bridges in the smallest gamma subunit. The crystal structure of the enzyme in the complex with the inhibitor [p-nitrophenylhydrazine (pNPH)] has been determined at a 2.0 A resolution.(1) The hydrazone of the cofactor with the inhibitor was nicely modeled into the omit electron density map, identifying the C6 carbonyl group as the reactive site of the cofactor. The Asp33 gamma is unambiguously determined as the catalytic base to abstract the alpha-proton from a substrate, because N beta atom of the inhibitor corresponding to the C alpha atom of the substrate amine is neighbored to Asp33 gamma. The bound inhibitor is completely enclosed in the active site pocket formed by the residues from the beta- and gamma-subunits. The cofactor-inhibitor adduct may be predominantly in the hydrazone with the azo form as a minor component. The binding of the inhibitor causes minor but important conformational changes in the residues surrounding the active site. The inhibitor may have access to the active site pocket through the water-filled crevice between the beta- and gamma-subunits.

Structure of the transition state analog of medium-chain acyl-CoA dehydrogenase. Crystallographic and molecular orbital studies on the charge-transfer complex of medium-chain acyl-CoA dehydrogenase with 3-thiaoctanoyl-CoA.

The flavoenzyme medium-chain acyl-CoA dehydrogenase (MCAD) eliminates the alpha-proton of the substrate analog, 3-thiaoctanoyl-CoA (3S-C8-CoA), to form a charge-transfer complex with deprotonated 3S-C8-CoA. This complex can simulate the metastable reaction intermediate immediately after the alpha-proton elimination of a substrate and before the beta-hydrogen transfer as a hydride, and is therefore regarded as a transition-state analog. The crystalline complex was obtained by co-crystallizing MCAD in the oxidized form with 3S-C8-CoA. The three-dimensional structure of the complex was solved by X-ray crystallography. The deprotonated 3S-C8-CoA was clearly located within the active-site cleft of the enzyme. The arrangement between the flavin ring and deprotonated 3S-C8-CoA is consistent with a charge transfer interaction with the negatively charged acyl-chain of 3S-C8-CoA as an electron donor stacking on the pyrimidine moiety of the flavin ring as an electron acceptor. The structure of the model complex between lumiflavin and the deprotonated ethylthioester of 3-thiabutanoic acid was optimized by molecular orbital calculations. The obtained theoretical structure was essentially the same as that of the corresponding region of the X-ray structure. A considerable amount of negative charge is transferred to the flavin ring system to stabilize the complex by 9.2 kcal/mol. The large stabilization energy by charge transfer probably plays an important role in determining the alignment of the flavin ring with 3S-C8-CoA. The structure of the highest occupied molecular orbital of the complex revealed the electron flow pathway from a substrate to the flavin ring.

Structure basis for the regulation of glyceraldehyde-3-phosphate dehydrogenase activity via the intrinsically disordered protein CP12.

The reversible formation of a glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-CP12-phosphoribulokinase (PRK) supramolecular complex, identified in oxygenic photosynthetic organisms, provides light-dependent Calvin cycle regulation in a coordinated manner. An intrinsically disordered protein (IDP) CP12 acts as a linker to sequentially bind GAPDH and PRK to downregulate both enzymes. Here, we report the crystal structures of the ternary GAPDH-CP12-NAD and binary GAPDH-NAD complexes from Synechococcus elongates. The GAPDH-CP12 complex structure reveals that the oxidized CP12 becomes partially structured upon GAPDH binding. The C-terminus of CP12 is inserted into the active-site region of GAPDH, resulting in competitive inhibition of GAPDH. This study also provides insight into how the GAPDH-CP12 complex is dissociated by a high NADP(H)/NAD(H) ratio. An unexpected increase in negative charge potential that emerged upon CP12 binding highlights the biological function of CP12 in the sequential assembly of the supramolecular complex.

Factors associated with late post-partum depression in Japan.

AIM: Postnatal depression is one of the life-threatening events faced by women. As the factors associated with postnatal depression have not been investigated fully in Japan, we studied the factors associated with postnatal depression. METHODS: One-hundred-and-sixty-nine women who visited the health center of a city in Aomori Prefecture, Japan, at 4 months after childbirth for regular examination fulfilled the selection criteria and completed self-reporting questionnaires on postnatal depression using the Edinburgh Postnatal Depression Scale (EPDS) and a life and social events scale. RESULTS: The primiparas showed a significantly higher EPDS score than the multiparas. The EPDS score decreased with the frequency of delivery in the groups of mothers in their twenties and thirties. For the multiparas, the number of participants who suffered obstetric events was lower, the number of participants who went back to their home was higher, and the number of participants who were taken care of by their mother was lower than for the primiparas. General health abnormalities, sociability, and worries about baby care were significantly associated with the EPDS for both the primiparas and multiparas. The cooperation of the husband was associated with a decreased EPDS score, both for the primiparas and multiparas, irrespective of the family structure. CONCLUSION: The EPDS score decreased with an increased frequency of delivery, suggesting that the experience of delivery would impact on postnatal depression, partly because of decreased obstetric events. However, a social assistance system is needed for women with general health abnormalities, less sociability, worries about baby care, and limited cooperation of the husband for both primiparas and multiparas.

Production of completely flavinylated histamine dehydrogenase, unique covalently bound flavin, and iron-sulfur cluster-containing enzyme of nocardioides simplex in Escherichia coli, and its properties.

The hmd gene of histamine dehydrogenase from Nocardioides simplex was overexpressed in Escherichia coli, and the resulting enzyme was purified to homogeneity. The purified recombinant enzyme is almost identical with the native enzyme in view of molecular weight and specific activity, and is stoichiometrically assembled with the three cofactors 6-S-cysteinyl FMN, 4Fe-4S cluster, and ADP.

6-S-cysteinyl flavin mononucleotide-containing histamine dehydrogenase from Nocardioides simplex: molecular cloning, sequencing, overexpression, and characterization of redox centers of enzyme.

Histamine dehydrogenase from Nocardioides simplex is a homodimeric enzyme and catalyzes oxidative deamination of histamine. The gene encoding this enzyme has been sequenced and cloned by polymerase chain reactions and overexpressed in Escherichia coli. The sequence of the complete open reading frame, 2073 bp coding for a protein of 690 amino acids, was determined on both strands. The amino acid sequence of histamine dehydrogenase is closely related to those of trimethylamine dehydrogenase and dimethylamine dehydrogenase containing an unusual covalently bound flavin mononucleotide, 6-S-cysteinyl-flavin mononucleotide, and one 4Fe-4S cluster as redox active cofactors in each subunit of the homodimer. The presence of the identical redox cofactors in histamine dehydrogenase has been confirmed by sequence alignment analysis, mass spectral analysis, UV-vis and EPR spectroscopy, and chemical analysis of iron and acid-labile sulfur. These results suggest that the structure of histamine dehydrogenase in the vicinity of the two redox centers is almost identical to that of trimethylamine dehydrogenase as a whole. The structure modeling study, however, demonstrated that a putative substrate-binding cavity in histamine dehydrogenase is quite distinct from that of trimethylamine dehydrogenase.

Crystal structure of quinohemoprotein amine dehydrogenase from Pseudomonas putida. Identification of a novel quinone cofactor encaged by multiple thioether cross-bridges.

The crystal structure of a quinohemoprotein amine dehydrogenase from Pseudomonas putida has been determined at 1.9-A resolution. The enzyme comprises three non-identical subunits: a four-domain alpha-subunit that harbors a di-heme cytochrome c, a seven-bladed beta-propeller beta-subunit that provides part of the active site, and a small gamma-subunit that contains a novel cross-linked, proteinous quinone cofactor, cysteine tryptophylquinone. More surprisingly, the catalytic gamma-subunit contains three additional chemical cross-links that encage the cysteine tryptophylquinone cofactor, involving a cysteine side chain bridged to either an Asp or Glu residue all in a hitherto unknown thioether bonding with a methylene carbon atom of acidic amino acid side chains. Thus, the structure of the 79-residue gamma-subunit is quite unusual, containing four internal cross-links in such a short polypeptide chain that would otherwise be difficult to fold into a globular structure.