Effect of aluminium ion on bioavailability of levofloxacin following oral administration of cilexetil ester of levofloxacin as prodrug in rats.

A prodrug of levofloxacin (LVFX), cilexetil ester of LVFX (LVFX-CLX), was synthesized to examine whether the prodrug can avoid chelate formation with metal cations in the gastrointestinal tract. LVFX-CLX exhibited a 10-times higher partition coefficient than LVFX. In vitro, LVFX was precipitated by 76.1% in the presence of a 10-times higher concentration of aluminium chloride (Al3+), but LVFX-CLX was not. LVFX-CLX was rapidly hydrolyzed enzymatically by rat plasma, intestinal mucosal and liver homogenates at 37 °C, but not by pancreatic enzymes and luminal fluid. The minimum inhibitory concentration values of LVFX-CLX against S. aureus, E. coli and P. aeruginosa were far higher than that of LVFX. In rats, area under the plasma concentration-time curve from zero to 4 h (AUC0-4h) of LVFX after oral administration of LVFX-CLX was 1.34-fold higher than that after LVFX, though it did not reach significance level. Co-administration of Al3+ with LVFX and LVFX-CLX in rats decreased AUC0-4h of plasma LVFX by 75% and 60%, respectively, however, the AUC0-4h of plasma LVFX after co-administration of LVFX-CLX and Al3+ was 2.2-times higher than that after co-administration of LVFX and Al3+. These results suggested that the use of LVFX-CLX may reduce the modulation of intestinal microflora caused by LVFX and the suppressive effect of Al3+ on intestinal absorption of LVFX.

High efficiency penetration of antibody-immobilized nanoneedle thorough plasma membrane for in situ detection of cytoskeletal proteins in living cells.

BACKGROUND: The field of structural dynamics of cytoskeletons in living cells is gathering wide interest, since better understanding of cytoskeleton intracellular organization will provide us with not only insights into basic cell biology but may also enable development of new strategies in regenerative medicine and cancer therapy, fields in which cytoskeleton-dependent dynamics play a pivotal role. The nanoneedle technology is a powerful tool allowing for intracellular investigations, as it can be directly inserted into live cells by penetrating through the plasma membrane causing minimal damage to cells, under the precise manipulation using atomic force microscope. Modifications of the nanoneedles using antibodies have allowed for accurate mechanical detection of various cytoskeletal components, including actin, microtubules and intermediate filaments. However, successful penetration of the nanoneedle through the plasma membrane has been shown to vary greatly between different cell types and conditions. In an effort to overcome this problem and improve the success rate of nanoneedle insertion into the live cells, we have focused here on the fluidity of the membrane lipid bilayer, which may hinder nanoneedle penetration into the cytosolic environment. RESULTS: We aimed to reduce apparent fluidity of the membrane by either increasing the approach velocity or reducing experimental temperatures. Although changes in approach velocity did not have much effect, lowering the temperature was found to greatly improve the detection of unbinding forces, suggesting that alteration in the plasma membrane fluidity led to increase in nanoneedle penetration. CONCLUSIONS: Operation at a lower temperature of 4 °C greatly improved the success rate of nanoneedle insertion to live cells at an optimized approach velocity, while it did not affect the binding of antibodies immobilized on the nanoneedle to vimentins for mechanical detection. As these experimental parameters can be applied to various cell types, these results may improve the versatility of the nanoneedle technology to other cell lines and platforms.

Salivary Alpha-Amylase Activity and Mild Cognitive Impairment among Japanese Older Adults: The Toon Health Study.

BACKGROUND: There is growing interest in examining objective markers for early identification and behavioral intervention to prevent dementia and mild cognitive impairment in clinical and community settings. OBJECTIVE: To investigate the association between salivary alpha-amylase as an objective measure of psychological stress response and mild cognitive impairment for the implication of psychological stress in the development of mild cognitive impairment. DESIGN, SETTING, AND PARTICIPANTS: This cross-sectional study involved 865 participants aged ≥ 65 years. A saliva sample was collected in the morning, and the levels of salivary alpha-amylase were assayed. Mild cognitive impairment was evaluated using the Japanese version of the Montreal Cognitive Assessment; a score < 26 was indicative of mild cognitive impairment. A multivariable logistic regression model was used to examine the association of salivary alpha-amylase and mild cognitive impairment after adjusting for age, sex, current drinking status, current smoking status, body mass index, hypertension, diabetes mellitus, physical activity, education, social support, social network, and heart rate variability. RESULTS: Salivary alpha-amylase was associated with mild cognitive impairment (the multivariable-adjusted odds ratio [95% confidence interval] for the 1-standard deviation increment of log-transformed salivary alpha-amylase was 1.24 [1.07-1.44]). This significant association persisted after adjusting for various confounding factors. CONCLUSION: Elevation of salivary alpha-amylase was associated with mild cognitive impairment among Japanese community-dwelling older adults. This suggests that salivary alpha-amylase is a useful objective marker of psychological stress responses associated with mild cognitive impairment.

High-power laser experiment forming a supercritical collisionless shock in a magnetized uniform plasma at rest.

We present an experimental method to generate quasiperpendicular supercritical magnetized collisionless shocks. In our experiment, ambient nitrogen (N) plasma is at rest and well magnetized, and it has uniform mass density. The plasma is pushed by laser-driven ablation aluminum (Al) plasma. Streaked optical pyrometry and spatially resolved laser collective Thomson scattering clarify structures of plasma density and temperatures, which are compared with one-dimensional particle-in-cell simulations. It is indicated that just after the laser irradiation, the Al plasma is magnetized by a self-generated Biermann battery field, and the plasma slaps the incident N plasma. The compressed external field in the N plasma reflects N ions, leading to counterstreaming magnetized N flows. Namely, we identify the edge of the reflected N ions. Such interacting plasmas form a magnetized collisionless shock.

Evaluation of a high-speed but low-throughput RT-qPCR system for detection of SARS-CoV-2.

With the emergence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), a high-speed and convenient detection technology should be at the forefront of medical care worldwide. This study evaluated the usefulness of GeneSoC, a compact, high-speed reciprocal flow quantitative reverse transcription polymerase chain reaction system, for the detection of SARS-CoV-2. The results support the use of this system for the rapid identification of SARS-CoV-2. This approach can contribute to the strategic selection of initial management strategies for patients with COVID-19.

Bmf is a possible mediator in histone deacetylase inhibitors FK228 and CBHA-induced apoptosis.

Histone deacetylase (HDAC) inhibitors modify transcription of selected genes and eventually induce apoptosis. However, molecular mechanisms for their proapoptotic activity remain unclear. We here demonstrate that HDAC inhibitors FK228 and CBHA preferentially upregulated the BH3-only protein Bmf in a broad range of cancer cells. In contrast, HDAC1 overexpression distinctly reduced Bmf expression. FK228 induced histones H3 and H4 acetylation at Bmf promoter region, but not at its 3' region, suggesting that histone hyperacetylation causes Bmf transcriptional activation. Knockdown of Bmf transcripts rescued cells from FK228 or CBHA-induced cell death, disruption of mitochondrial membrane potential (DeltaPsim) and DNA fragmentation. Taken together, FK228 and CBHA activate Bmf transcription by histone hyperacetylation at its promoter region, and inhibition of this action decreased their proapoptotic activity, thereby highlighting a central role of Bmf in HDAC inhibitor-mediated apoptosis.

Interactions among four subunits of elongation factor 1 from rice embryo.

To establish the subunit construction of elongation factor EF-1, interactions among four non-identical subunits of rice embryo EF-1 (alpha, beta, beta', and gamma) were analyzed with polyacrylamide gel electrophoresis. Complexes beta beta', alpha beta, alpha beta', and beta gamma were formed by mixing the two respective subunits. However, no complex was formed between EF-1 beta' and EF-1 gamma. Complexes containing three subunits like alpha beta beta', alpha beta gamma, and beta beta' gamma, were formed by mixing the three respective subunits. EF-1 was reconstructed when each subunit was added in the following order, beta, beta', gamma, and alpha. The affinity of EF-1 alpha for other subunits was as follows, beta beta' gamma > beta beta' > beta not equal to beta'. Likewise, the affinity of EF-1 gamma for other subunits was: beta beta' gamma > beta >> beta'. Phe-tRNA binding activity of the reconstructed EF-1 was about 90% of that of the native EF-1. From these results, we concluded that rice embryo EF-1 is constructed of equimolar amount of four subunits, alpha, beta, beta' and gamma.

CXCL14 and MCP1 are potent trophic factors associated with cell migration and angiogenesis leading to higher regenerative potential of dental pulp side population cells.

INTRODUCTION: The release of trophic factors from mesenchymal stem cells (MSCs) is critical for tissue regeneration. A systematic investigation of the regenerative potential of trophic factors from different MSCs, however, has not been performed. Thus, in the present study, the regenerative potential of conditioned medium (CM) from dental pulp, bone marrow, and adipose tissue-derived CD31(-) side population (SP) cells from an individual source was compared in an ectopic tooth transplantation model. METHODS: The tooth root transplantation in an ectopic site model was used for investigation of the regenerative potential and trophic effects in vivo. Either pulp CD31(-) SP cell populations (1×10(6) cells) at the third to fourth passage or 5 µg/ml of CM from dental pulp, bone marrow, and adipose stem cells from four different individuals were injected into the root with collagen TE. Each root was transplanted subcutaneously in 5-week-old severe combined immunodeficiency mice. Each root with surrounding tissue was harvested for histology on days 7, 21, and 28 and for Western blot analysis and real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis on day 28. Furthermore, the trophic factors responsible for the regenerative potential were identified as the upregulated genes present in pulp CD31(-) SP cells when compared with the genes in both bone marrow and adipose CD31(-) SP cells by using microarray analysis, real-time RT-PCR, and Western blot analysis. RESULTS: Transplantation of pulp CM yielded increased volume of pulp regeneration, more bromodeoxyuridine (BrdU)-positive migrated cells, and fewer caspase 3-positive cells in the regenerated pulp compared with the others. Pulp CM also demonstrated significantly increased cell migration, anti-apoptosis, and angiogenesis in C2C12 cells. Higher expression of CXCL14 and MCP1 in pulp SP cells suggested candidate trophic factors. The stimulatory effects on both migration and angiogenesis of CXCL14 and MCP1 were demonstrated in vitro. In the regenerated tissue, BrdU-positive migrated cells expressed CXCR4 and CCR2, receptors for CXCL14 and MCP1, respectively. CONCLUSIONS: The higher regenerative potential of pulp SP cells may be due to potent trophic factors, including CXCL14 and MCP1, which promote migration and angiogenesis.

Occurrence of four subunits in high molecular weight forms of polypeptide chain elongation factor 1 from wheat embryo.

In contrast to high molecular weight forms of elongation factor 1 (EF-1H) from animal sources which contain three subunits, EF-1a, EF-1b, and EF-1c, EF-1H from wheat embryo consisted of four subunits, EF-1a, EF-1b, EF-1b', and EF-1c, in an equimolar ratio. The molecular weights of EF-1a, EF-1b, EF-1b', and EF-1c from wheat embryo were 52,000, 29,000, 28,000, and 48,000, respectively. In the animal system, EF-1a and EF-1b correspond functionally to EF-Tu and EF-Ts, respectively. In the wheat system, however, both EF-1b and EF-1b' had the EF-Ts-like activity to stimulate EF-1a-dependent binding of aminoacyl-tRNA to ribosomes. EF-1b and EF-1b' from wheat embryo gave 21 and 20 tryptic peptides, respectively. Twenty peptides were common.

Identification of the replication region of the Lactobacillus acidophilus plasmid pLA106.

The complete nucleotide sequence of plasmid pLA106 (2862 bp) from Lactobacillus acidophilus TK8912 was determined. Based on this sequence, the location of the genes for mobilization-plasmid recombination protein (mob), replication origin (ori), transcriptional repressor protein (repA) and replication initiation protein (repB) were predicted. Deletion analysis showed that the 1.4-kb PstI-EcoRV fragment carrying the ori, repA and repB genes is able to replicate in Lactobacillus and Escherichia coli cells. The plasmid pLA106 appears to have most features of the pLS1/pE194 plasmid family.

The anti-emetic activity of GK-128 in Suncus murinus.

In Suncus murinus, various emetic responses and the anti-emetic activity of a new 5-hydroxytryptamine3 (5-HT3) receptor antagonist, GK-128 (2-[(2-methylimidazol-1-yl) methyl benzo[f]thiochromen-1-one monohydrochloride hemihydrate), were investigated. Cancer chemotherapeutic agents, cisplatin and cyclophosphamide, dose-dependently induced emesis of long-lasting duration. The 5-HT3 receptor agonist, 2-methyl-5-HT, and copper sulfate also induced emesis of short duration. However, another 5-HT3 receptor agonist, m-chlorophenylbiguanide, was not consistently emetic. GK-128 inhibited the emetic responses induced by chemotherapeutic agents and 2-methyl-5-HT with similar potency. The anti-emetic action of GK-128 was more potent than that of ondansetron, Y-25130, granisetron and metoclopramide. The order of potency of these drugs, except granisetron, was consistent with that of their 5-HT3 receptor binding affinity in rat cortex. GK-128 failed to inhibit copper sulfate-induced emesis. These data suggest that GK-128 has a potent inhibitory effect on emesis via the 5-HT3 receptor, and that the 5-HT3 receptor involved in emesis in Suncus murinus may be different from the classically defined 5-HT3 receptor in other animals such as rats, dogs and ferrets.

Inhibition of delayed hypersensitivity reaction in mice by cadmium.

The effect of a single exposure to Cd on DH was studied through exposure to Cd at different time intervals in relation to antigen administration. Exposure to Cd significantly suppressed the DH reaction, with a dose-response relationship, when administered 2 or 3 days after immunization of BALB/c and DBA/2 mice with sheep red blood cells. A strain difference in sensitivity to Cd was also demonstrated. These results suggest that a single exposure to Cd impairs the cellular immune response in mice.

Cadmium, copper and zinc-binding protein (metallothionein) in the liver of the water lizard Triturus pyrrhogaster.

A low-Mr copper (Cu) and zinc (Zn)-binding protein was present in the liver of a control water lizard Triturus pyrrhogaster. Cadmium (Cd) accumulated in the liver of the animals after intramuscular injections of Cd salt and was bound to Cu- and Zn-binding protein of a single isoform that had properties characteristic of metallothionein (MT). Effects of Cd loading on several essential elements in the liver and kidney were also determined.

Thymic atrophy in mice induced by cadmium administration.

The effects of cadmium (Cd) on the immune organs were examined histopathologically. On 2 or 3 days after a single i.p. injection of 1.8 mg Cd/kg body weight into mice, slight loss of body weight, significant decrease of thymus weight and marked increase of spleen weight were observed. Lymph node weight did not show any change. Histopathologically, cortical atrophy of the thymus was very marked. The white pulp of the spleen tended to diminish in size any many polymorphonuclear leukocytes and myeloid cells appeared in the red pulp.

Hydration of fibrinogen, fibrin, and fibrin degradation product (FDP) as estimated by nuclear magnetic resonance (NMR) spectroscopy.

The relaxation times (T1 and T2) of water proton in nuclear magnetic resonance (NMR) were measured with solutions containing bovine fibrinogen (Fbg), fibrin degradation products (FDP) and with fibrin-gel (Gel), at varying protein concentrations (0.7-70 mg/ml). Both T1 and T2 declined exponentially with increasing protein concentration. At a protein concentration of 35 mg/ml, the T1 of Fbg, Gel and FDP were 2.32, 2.12 and 2.82 s and the T2 values were 0.35, 0.17 and 0.70, respectively. The relaxation times for the control samples (0.2 M borate buffer) were 3.41 (T1) and 2.28 (T2). When the relaxation rates (the inverse of T1 and T2), R1 and R2 were plotted against the protein concentration, there were positive linear correlations between them. Using the slopes of the plots, the hydration value of each protein was calculated. The hydration value (g of H2O/g of protein) was 0.24 for Fbg, 0.34 for Gel and 0.14 for FDP.

Subclavian vein puncture under ultrasonic guidance.

Ultrasonically guided subclavian venipuncture is described. Since this method permits direct tapping of the subclavian vein and control of the insertion to the innominate vein under ultrasonic guidance, complications such as pneumothorax, accidental subclavian artery puncture, and malposition of the catheter, which often accompany the conventional method, can be avoided. As a result, this technique produces no radiation damage.

Th1/Th2 balance in childhood idiopathic nephrotic syndrome.

AIMS: In view of the conflicting evidence of helper T cell type 1 (Th1) or type 2 (Th2) pattern of cytokine synthesis in childhood idiopathic nephrotic syndrome (INS) this study examined the balance of Th1 and Th2 which are characterized by intracellular cytokine production of interferon-gamma (IFNgamma) and interleukin-4 (IL-4), respectively. SUBJECTS AND METHODS: Sixteen children with steroid-sensitive INS (mean age 9.0 years) were included in this study, together with 15 healthy normal children (mean age 7.9 years) for the control group. Intracellular production of both IFNgamma and IL-4 in helper T cell (CD4+ cell) was investigated by a 3-color flow cytometry. RESULTS: The cross-sectional data showed no significant differences of percentages of Th0 (IFNgamma+ IL-4+ CD4+ cell), Th1 (IFNgamma+ lL-4- CD4+ cell) and Th2 (IFNgamma- IL-4+ CD4+ cell) in CD4+ cells (p > 0.05). The Th1/Th2 ratio during nephrotic relapse did not differ from those during nephrotic remission and in normal healthy children (p > 0.05). CONCLUSION: We conclude that there is no significant skew of Th1/Th2 balance in childhood INS and that the cardinal immunological abnormality does not lie in helper T cells but in other cells, such as suppressor/cytotoxic T cells, natural killer cells or monocytes/macrophage. To clarify the pathogenesis of INS, comprehensive studies for these cells would be worthwhile.

[Clinical evaluation of cefroxadine in the field of obstetrics and gynecology].

Cefroxadine (CXD), an oral cephalosporin antibiotic was studied in the field of obstetrics and gynecology and the following results were obtained. CXD was orally given to 22 cases at daily dose of 1,500 mg 3 times a day. CXD administration was given to 22 cases in all; 4 with cervicitis, 6 with endometritis, 2 with puerperal fever, 4 with bartholinitis, 5 with adnexitis and 1 with vulvitis, respectively. Overall efficacy rate was 77.3% (17/22) (excellent 4, good 13, fair 5). As for side effects, a slight diarrhea was observed. CXD was considered to be a useful antibiotic in the field of obstetrics and gynecology by above the results.

[Circulating negative inotropic substance produced by the lung, affected by prostaglandin and thromboxane].

The Lung is capable of the synthesis and the release of prostaglandins responded to a number of pathophysiological stimuli, which may impair myocardial performance. Present study was undertaken to determine the release of circulating negative inotropic substance(s) from the lung and whether or not blockage of prostanoid synthesis, or infusion of prostaglandin are related to the production of this substance(s). Sixteen isolated left lower lobes were divided into 3 groups: (1) untreated, isolated blood-perfused lungs (n = 5), (2) prostaglandin I2, 1 microgram/min, continuously added to the perfusate (n = 6), and (3) five lobes pretreated with ibuprofen (12.5 mg/kg) in addition to 1 microgram/min prostaglandin I2. Perfusion of an isolated lobe with heparinized whole blood caused thrombocytopenia. The plasma 4 hours after perfusion led to the generation of a humoral agent(s) that reduced developed tension (Tpd) of a papillary muscle (p less than 0.05), and Ca++ -and Mg++-ATPase of sarcoplasmic reticulum (SR) (p less than 0.01). With prostaglandin I2, thromboxane B2 rose from 0.076 to 0.362 ng/ml at a level higher than control (p less than 0.05). The plasma significantly lowered SR-ATPase and myofibril-Mg++-ATPase activities and further reduced papillary muscle Tpd. By pretreatment with ibuprofen, adverse prostaglandin I2 effects were eliminated. The observation suggests that the lung produces prostaglandin related negative inotropic substance(s) that may reduced contractility, affected by energy availability.