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Objectives: Laryngeal complications have been reported after endotracheal intubation and prone positioning in patients with critical coronavirus disease 2019 (COVID‐19), but their association is unclear. In this study, we investigated the rate of laryngeal complications in patients with COVID‐19 compared to an alternative condition (control group). Methods: We retrospectively analyzed the data of 40 patients who underwent endotracheal intubation for either COVID‐19 or an alternative condition (control group). Data on age, sex, body mass index (BMI), cardiovascular disease (CVD) risk factors, use of prone therapy, duration of endotracheal intubation, and duration from extubation/tracheostomy to laryngeal evaluation were collected from medical records. Results: There were no significant differences in BMI, frequency of CVD risk factors, duration of endotracheal intubation, or duration from extubation/tracheostomy to laryngeal evaluation between the two groups. In the COVID‐19 group, all patients adopted the prone position. In comparison, only one patient in the control group adopted the prone position. Significant differences were observed between the two groups regarding the incidence of vocal fold immobility and laryngeal granuloma. Conclusion: Laryngeal complications were more common in the COVID‐19 group than in the control group. Prone positioning may be a risk factor for these complications. Level of Evidence: 4. [ABSTRACT FROM AUTHOR]

The article presents an abstract of the medical research paper "The Damage of Spinal Cord Via Pathological Microglia Proliferation is Inhibited by Mild Hypothermia Treatment." This research examined the effect of mild hypothermia treatment on brain damage. It concluded that mild hypothermia is effective in treating secondary neuronal damage after spinal cord injury and inhibition of microglial proliferation may be one of the mechanisms of the mild hypothermia effect.

Background: Although multidisciplinary care (MDC) is necessary for controlling chronic kidney disease (CKD), its impact on compliance with management target values in the CKD guidelines remains unclear. This study was designed to clarify the relationship between compliance with management target values and renal prognosis in CKD outpatients who received MDC. Methods: There were 255 outpatients with pre-dialysis CKD who received MDC. Achievement rates of systolic, and diastolic blood pressure, hemoglobin, uric acid, low-density lipoprotein cholesterol, and hemoglobin A1c values determined according to CKD guidelines were compared before and 12 months after MDC. In addition, after dividing achievement rates of the target values at 12 months after MDC into four groups (A < 30% ≤ B < 60% ≤ C < 80% ≤ D), dialysis initiation and renal survival rates were compared. Results: There was a significant increase in the overall achievement rate from 62.8 to 69.1% (p < 0.001). The higher the achievement rate after MDC, the lower the dialysis initiation rate (A 72.7%, B 35.3%, C 20.5%, D 8.2%, p < 0.001). There was also a significantly higher renal survival rate (p < 0.001). These findings suggest that MDC for CKD raised awareness of health literacy, and improved the achievement rate of target values. Furthermore, the higher the achievement rate, the later the initiation of dialysis, which led to improvement of renal survival. Conclusions: MDC can improve compliance with management target values for CKD, suggesting that it may improve renal prognosis. [ABSTRACT FROM AUTHOR]

Background: In periacetabular osteotomy for the treatment of developmental dysplasia of the hip, impairments in ADL due to limitations in hip flexion can occur when anterior displacement is added to lateral displacement in order to obtain sufficient femoral head coverage. This study was conducted to determine, by the range of motion (ROM) simulation based on CT images, the minimum angle of hip flexion and internal rotation at 90° of flexion that is necessary to avoid ADL impairments after eccentric rotational acetabular osteotomy (ERAO) and to estimate the angles of anterior femoral head coverage on plain radiography that enable the above flexion. Methods: Of 47 hips treated with ERAO at our hospital from December 2007 to May 2012, 27 hips without progressive osteoarthritis which could be CT scanned were examined and included. The mean age at the time of surgery was 40.7 years (SD 1.8). The postoperative follow-up period was 30.2 months (SD 3.6). Two hips were in male patients and 25 hips were in female patients. The disease stage prior to surgery was pre-osteoarthritis in 5 hips, early in 11 hips, and progressive in 11 hips. We checked whether the patients were capable of activities that require deep hip flexion for the evaluation of postoperative ADL. Radiographic examination was performed before and one year after surgery to calculate LCE angle, Sharp angle, AHI, and VCA angle. The angle at which impingement of the displaced fragment of the bone and the femur appeared was measured using 3D CAD software, and the relationship between this angle and the physical findings, ADL impairment, or radiographic findings, were also examined. Results: 22 out of 27 hips that were capable of 116° or more of flexion or 42° or more of internal rotation at 90° of flexion in ROM simulation showed the absence of ADL impairment and a postoperative VCA angle ≤42°, whereas 5 hips with 110° or less of flexion or 40° or less of internal rotation at 90° of flexion in ROM simulation had ADL impairments associated with limitations in hip flexion and a postoperative VCA angle ≥46°. Conclusions: Anterior and lateral coverage requires a postoperative VCA angle of ≥20° to achieve anterior structural stability and an LCE angle of >25° to obtain adequate superior lateral coverage of the femoral head. A VCA angle ≤42° is required to avoid impingement during deep flexion. A VCA angle ≥46° is a probable risk factor for pincer FAI syndrome after ERAO. [ABSTRACT FROM AUTHOR]

Pex1p and Pex6p are required for the relocation of the import receptor Pex5p from the peroxisomal membrane to the cytosol. Weherein show that mammalian Pex26p directly binds to Pex14p, the initial docking receptor of Pex5p, and interacts with Pex5p via Pex14p. The binding affinity of Pex26p to Pex14p is altered by Pex5p. Further evidence suggests that the N-terminal region in Pex26p acts as a scaffold protein to recruit Pex14p.Pex5p complex together with Pex1p.Pex6p complexes on peroxisomes. Pex26p binding to Pex14p was suppressed by overexpression of Pex1p and Pex6p in an ATP-dependent manner, whereas Pex14p was not competed out by Pex1p and Pex6p from Pex26p mutant defective in peroxisomal matrix protein import. These results suggested that peroxisome biogenesis requires Pex1p- and Pex6p-regulated dissociation of Pex14p from Pex26p. Pex1p homo-oligomer directly binds to Pex5p as assessed by a surface plasmon resonance-based assay. Moreover, cytosolic Pex1p is likely to maintain the functional oligomer of Pex5p. Taken together, in the peroxisomal protein import, AAA peroxins modulate the interaction between Pex26p and Pex14p on peroxisome membrane as well as Pex5p oligomer in the cytosol. [ABSTRACT FROM AUTHOR]

The article presents the study "Rapid Determination of Nitroimidazole Residues in Honey by Liquid Chromatography/Tandem Mass Spectrometry," by Maki Kanda et al. The researchers developed a rapid and efficient means of determining residues of four nitroimidazoles and three hydrophilic metabolites in honey. The results showed overall recovery of the nitroimidazoles ranged from 76.1% to 98.5%.

Study Design:A basic study using a spinal cord injury (SCI) model in rats.Objectives:The effect of mild hypothermic treatment on histological changes and motor function after a rat spinal cord compression injury was assessed.Methods:Mild spinal cord compression was performed at the eleventh thoracic vertebral level by a 20 g weight for 20 min. Rats in the mild hypothermic model were kept at a body temperature of 33 °C and rats in the normothermic group were kept at 37 °C for 1 h from beginning of compression. Motor function was evaluated by measuring the frequency of standing. Microglia were stained by isolectin B4 and observed in the compressed portion of the spinal cord. The amount of tumor necrosis factor-α (TNF-α) in the compressed spinal cord was measured by the ELISA method.Results:In the normothermic rats, microglia proliferated up to 72 h after the compression. Proliferation was substantially inhibited at 48 and 72 h after compression in the hypothermic rats. The motor function of the hypothermic rats improved at 48 and 72 h after the compression, whereas no improvement was seen in the normothermic rats. The amount of TNF-α in the compressed portion of the spinal cord was lower in hypothermic rats compared with normothermic rats throughout the experiment.Conclusions:These results suggest that hypothermic treatment is effective for the amelioration of delayed motor dysfunction via inhibition of microglial inflammatory responses.Spinal Cord (2008) 46, 425–431; doi:10.1038/sj.sc.3102163; published online 4 March 2008 [ABSTRACT FROM AUTHOR]

Recombination activating gene (RAG) re-expression and secondary Ig gene rearrangement in mature B lymphocytes have been reported. Here, we have studied RAG expression of peripheral blood B lymphocytes in humans. Normal B cells did not express RAG1 and RAG2 spontaneously. More than a half of circulating B cells expressed RAG proteins, when activated with Staphylococcus aureus Cowan I (SAC) + IL-2. DNA binding activity of the RAG complex has been verified by a gel shift assay employing the recombination signal sequence (RSS). Secondary Ig light chain rearrangement in the RAG-expressing B cells was confirmed by linker-mediated (LM)-PCR. Highly purified surface κ+ B cells activated by SAC + IL-2 became RAG+, and thereafter they started to express λ chain mRNA. 2 colour immunofluorescence analysis disclosed that a part of the RAG+ cells derived from the purified κ+ B cells activated by SAC + IL-2 turned to λ+ phenotype in vitro. Similarly, apoptosis induction was observed in a part of the RAG+ B cells. Our study suggests that a majority of peripheral blood B cells re-expresses RAG and the RAG+ B lymphocytes could be eliminated from the B cell repertoire either by changing Ag receptor specificity due to secondary rearrangement or by apoptosis induction. Thus, RAG expression of mature B cells in peripheral blood would contribute to not only receptor revision for further diversification of B cell repertoire but in some cases (or in some B cell subsets) to prevention or induction of autoAb responses at this differentiation stage in humans. [ABSTRACT FROM AUTHOR]

The potential risks to human health and reproduction from the xenoestrogen, epoxy-resins compounds, bisphenols have not been well established. This is due in part to the absence of accurate analytical methods to quantify bisphenols in food samples. In fact, the possibility of contamination of honey with epoxy-resins compounds arises from packing, contact materials, imported production, etc. First, an analytical method for the determination of bisphenol A (BPA) and bisphenol F (BPF) in honeys was developed. Reversed-phase liquid chromatography was used for measuring BPA and BPF in honey sample, prepared with solid-phase extraction. The recoveries of BPA and BPF spiked at 5.0 and 50 ng/g were above 90% with a relative standard deviation (RSD) of less than 10%. Second, this method was applied to various honey samples (n=107) from different countries; BPA was detected in only a few of them (detected levels: ND<2.0–33.3 ng/g), while BPF was not detected at all in the analyzed samples. Third, an assessment of BPA levels in honey was expressed by our analytical data and references. [Copyright &y& Elsevier]

SUMMARY We have reported previously that circulating anti-Fas ligand (FasL) autoantibodies able to inhibit Fas/FasL-mediated apoptosis were present in patients with systemic lupus erythematosus (SLE). In the present study, we describe the epitopes recognized by these anti-FasL autoantibodies. Rabbit antihuman antibody, raised against a FasL fragment consisting of amino acids (aa) 103–179 (fragment 2·0), inhibited Fas/FasL-mediated apoptosis, whereas an antibody against a FasL aa 103–146 fragment (fragment 1·0) did not. This suggested that an epitope around aa 146–179 was important for Fas/FasL interaction. Epitope mapping of anti-FasL autoantibodies using deletion mutants indicated that the epitope was located around aa 163–179. Three-dimensional molecular modelling of the Fas/FasL complex revealed that the aa 162–169 region was located on the outermost side of FasL, which suggested that the anti-FasL autoantibody would easily have access to the epitope. FasL point mutants involving aa positions 162–169 resulted in complete loss of apoptosis-inducing capability, which suggested that the aa 162–169 region was important for Fas/FasL interaction. A synthetic FasL peptide consisting of aa 161–170 blocked the binding of anti-FasL autoantibodies to FasL fragment 2·0 (aa 103–179). The FasL aa 161–170 sequence was found to be highly homologous with aa sequences from several infectious agents. Synthetic peptides derived from some of these microorganisms cross-reacted with the epitope recognized by the autoantibodies, suggesting that several foreign infectious agent-derived proteins may share an epitope with human FasL. As lymphocytes from SLE patients aberrartly expressed FasL, it is possible that infection by one of several infectious agents may trigger cross-reactive antibody responses, after which aberrantly expressed endogenous FasL might induce the shift from a cross-reactive... [ABSTRACT FROM AUTHOR]

Although the rate of preterm birth has increased in recent decades, a number of preterm infants have escaped death due to improvements in perinatal and neonatal care. Antenatal glucocorticoid (GC) therapy has significantly contributed to progression in lung maturation; however, its potential effects on other organs remain controversial. Furthermore, the effects of antenatal GC therapy on the fetal heart show both pros and cons. Translational research in animal models indicates that constant fetal exposure to antenatal GC administration is sufficient for lung maturation. We have established a premature fetal rat model to investigate immature cardiopulmonary functions in the lungs and heart, including the effects of antenatal GC administration. In this review, we explain the mechanisms of antenatal GC actions on the heart in the fetus compared to those in the neonate. Antenatal GCs may contribute to premature heart maturation by accelerating cardiomyocyte proliferation, angiogenesis, energy production, and sarcoplasmic reticulum function. Additionally, this review specifically focuses on fetal heart growth with antenatal GC administration in experimental animal models. Moreover, knowledge regarding antenatal GC administration in experimental animal models can be coupled with that from developmental biology, with the potential for the generation of functional cells and tissues that could be used for regenerative medical purposes in the future.

Sepsis is a leading cause of mortality in intensive care units due to multi-organ failure caused by dysregulated immune reactions. In this study, kinetic changes in the immune system were analyzed for 72 h in cecal ligation and puncture (CLP)-induced septic mice while preventing animal death by keeping body temperature. Increase of myeloid cells and decrease of B cells in circulation at 6 h after CLP were markedly observed. At the same time point, interleukin (IL)-10 expressing CD5+ regulatory B cells (Bregs) appeared. IL-10 and programmed death-ligand 1 (PD-L1) mRNA as well as IL-1β, IL-6 and interferon γ (IFNγ) mRNA was increased in the spleen at 6 h. A gradual decrease in Bcl-2 and abrupt increase of Bim expression in the spleen at the late phase were also found. These results showed that B lymphocytopenia with the appearance of Bregs is the earliest event, likely leading to immunoparalysis in sepsis. • Kinetic changes in the immune system were analyzed for 72 h in septic mice. • Animal death was prevented by keeping body temperature. • Decrease of total B cells in circulation at 6 h after the onset of sepsis were marked. • IL-10-expressing presumable regulatory B cells appeared at 6 h. • B lymphocytopenia and the IL-10-expressing Bregs may induce immunoparalysis. [ABSTRACT FROM AUTHOR]

Studies over the past decade have reported that the use of highly porous sockets in total hip arthroplasty (THA) results in osseointegration and long-term implant stability. However, some reports have raised concerns regarding radiographic evidence of poor osseointegration with features of fibrous tissue ingrowth. The purpose of this study was to compare clinical and radiographic assessments of highly porous sockets with those of hydroxyapatite (HA)-coated porous sockets in THA for hip dysplasia (DDH) at least 1 year after surgery. A total of 127 patients (136 hips) were recruited for the study. Of these, 94 patients (101 hips) received highly porous sockets with clustered screws, while 33 patients (35 hips) received HA-coated porous sockets with clustered screws. There was no difference in clinical outcomes between the two types of sockets. All HA-coated porous sockets were radiographically stable, without radiolucent lines. Fifteen hips had radiolucent lines in two or three DeLee and Charnley zones, accompanied by sclerotic lines along the circumferences of the highly porous sockets. A significant difference in the height of the preoperative osteophyte of the anterior acetabular wall was observed between 86 hips with one or no radiolucent lines and 15 hips with two or three radiolucent lines. In cases of DDH with atrophic bone remodeling pattern, highly porous sockets with multiple screws may be used, while HA-coated porous sockets with clustered screws result in better sealing of the bone-component interface. [ABSTRACT FROM AUTHOR]

Background: Comprehensive education about lifestyle, nutrition, medications and other types of treatment is important to prevent renal dysfunction in patients with chronic kidney disease (CKD). However, the effectiveness of multidisciplinary care on CKD progression has not been evaluated in detail. We aimed to determine whether multidisciplinary care at our hospital could help prevent worsening renal function associated with CKD. Methods: A total of 150 pre-dialysis CKD outpatients accompanied (n = 68) or not (n = 82) with diabetes mellitus (DM) were enrolled into this study. We assessed annual decreases in estimated glomerular filtration rates (ΔeGFR), and measured systolic blood pressure (SBP), diastolic blood pressure (DBP), hemoglobin (Hb), uric acid (UA), low-density lipoprotein cholesterol (LDL), hemoglobin A1c (HbA1c) values and urinary protein to creatinine ratios (UPCR) 12 months before and after multidisciplinary care. In addition, changes in the number of medications and prescription ratio before and after multidisciplinary care were assessed in 90 patients with CKD who could confirm their prescribed medications. Results: The ΔeGFR significantly improved between before and after multidisciplinary care from − 5.46 to − 0.56 mL/min/1.73 m2/year, respectively. The number of medications and prescription ratio showed no significant changes before and after multidisciplinary care. The ratios of improved ΔeGFR were found in 66.7% of all patients, comprising 63.1% of males and 76.9% of females, 64.8% without DM and 69.4% with DM. Values for UA, LDL, and HbA1c were significantly reduced among patients with improved ΔeGFR. Conclusion: Comprehensive multidisciplinary care of outpatients might help prevent worsening renal function among patients with CKD. [ABSTRACT FROM AUTHOR]

Introduction: Successful reconstruction during total hip arthroplasty for acetabular bone defects remains challenging. The purpose of our study was to evaluate clinical and radiographic results and performed a survivorship analysis for patients with acetabular bone loss undergoing total hip arthroplasty (THA) or revision THA using Kerboull-type acetabular reinforcement device(KT plate). Additionally, some independent prognostic factors for radiographic failure were identified.Methods: A retrospective cohort study was conducted. Thirty patients (36 hips) were available for follow-up examination between 1997 and 2012, with a mean follow-up period of 10 years. We evaluated clinical and radiographic results. The mean patient age at the time of the operation was 66 years.Results: Failure occurred in 6 cases, and further revisions were required for 2 hips. With failure for any reason as an endpoint, the cumulative survival at 10 years was 85%. Survival analysis at 10 years with failure as the endpoint revealed that the inclination angle of the KT plate ≤ 45° group showed higher survivorship than the inclination angle > 45° group (95.8 vs. 63.6%, P = 0.0047).Conclusions: With a mean follow-up of 10 years, the clinical and radiographic outcomes were satisfactory. While this study group was small, the results suggest that prosthesis longevity may be improved by setting the inclination angle of this reinforcement device at ≤ 45°. [ABSTRACT FROM AUTHOR]

Introduction: Midflexion stability can potentially improve the outcome of total knee arthroplasty (TKA). The purpose of this study was to evaluate the correlation between varus-valgus stability at 0° of extension and 90° of flexion and that at the midflexion range in posterior-stabilized (PS)-TKA.Materials and Methods: Forty-three knees that underwent PS-TKA were evaluated. Manual mild passive varus-valgus stress was applied to the knees, and the postoperative maximum varus-valgus stability was measured every 10° throughout range of motion, using a navigation system. Correlations between the stability at 0°, 90° of flexion, and that at each midflexion angle were evaluated using Spearman's correlation coefficients.Results: The stability of 0° modestly correlated with that of 10°-20°, but it did not significantly correlate with that of 30°-80°. However, the stability of 90° strongly correlated with that of 60°-80°, modestly correlated with that of 40°-50°, weakly correlated with that of 20°-30°, and did not correlate with that of 10°.Conclusions: The present study confirmed the importance of acquiring stability at 90° flexion to achieve midflexion stability in PS-TKA. However, initial flexion stability did not strongly correlate with the stability at either 0° or 90°. Our findings can provide useful information for understanding varus-valgus stability throughout the range of motion in PS-TKA. Attention to soft tissue balancing is necessary to stabilize a knee at the initial flexion range in PS-TKA. [ABSTRACT FROM AUTHOR]

Background aims. Adipose tissue has therapeutic potential for spinal cord injury (SCI) because it contains multipotent cells known as adipose-derived stem/stromal cells (ASCs). In this study, we attempted intravenous ASC transplantation in rats with SCI to examine the effect on functional recovery. Methods. ASCs (2.5 x 106) were intravenously infused into SCI rats, after which hindlimb motor function was evaluated. Distribution of transplanted ASCs was investigated and growth factor/cytokine levels were determined. Results. Intravenous transplantation of ASCs promoted the functional recovery in SCI rats and reduced the area of spinal cord cavitation. A distribution study revealed that ASCs gradually accumulated at the site of injury, but long-term survival of these cells was not achieved. Levels of growth factors increased only slightly in the spinal cord after ASC transplantation. Unexpectedly, cytokine-induced neutrophil chemoattractant (CINC)-l showed a transient but substantial increase in the spinal cord tissue and blood of the ASC group. CINC-1 was secreted by ASCs in vitro, and the sponge implantation assay showed that CINC-1 and ASCs induced angiogenesis. CINC-1 promoted functional recovery in SCI rats, which was similar to the ASCs. Expression of glial cell line-derived neurotrophic factor was greater in the ASC group than in the CINC-1 group, although both promoted extracellular signal-regulated kinase (ERK)l/2 phosphorylation, Akt phosphorylation was entranced in the spinal cord after ASC transplantation. Conclusions. Our findings indicated that intravenously transplanted ASCs gradually accumulated in the injured spinal cord, where cytokines such as CINC-1 activated ERK1/2 and Akt, leading to functional recovery. [ABSTRACT FROM AUTHOR]

The purpose of this study was to investigate whether postoperative combined anteversion (CA) can be kept within the safe zone while using cementless total hip arthroplasty (THA) using the operative technique which prepares the socket first for developmental dysplasia of the hip (DDH), by estimating the anteversion of the metaphyseal fit stem using preoperative three-dimensional (3D) computerized planning and by adjusting the anteversion of the socket using a navigation system that considers CA. Our subjects were 65 patients (65 hips) that had undergone cementless THA for DDH that could be observed for 1 year or more. Clinical assessments were made using the Japanese Orthopaedic Association's (JOA) hip score. For a radiological evaluation, we investigated 3D-planned stem versions, postoperative stem versions, preoperative and postoperative CA, and the relationship between CA and dislocation tendencies with temporary intraoperative reductions. JOA hip scores improved from 52.3 ± 11.4 points to 88.9 ± 8.6 points. CT evaluations revealed that 3D-planned stem versions were strongly correlated with postoperative stem versions ( r = 0.80; p < 0.01). Preoperative CA was 50.5° ± 7.2°, and postoperative CA was 41.3° ± 8.6°. Postoperative CA was kept within the safe zone in 61 hips. No intraoperative dislocation tendencies were observed in any hips. By estimating the anteversion of the cementless metaphyseal fit stem using 3D planning preoperatively and adjusting the angle of anteversion of the socket using a navigation system that considers CA intraoperatively, postoperative CA can very frequently be kept within the safe zone, even with cementless THA using the operative technique which prepares the socket first for DDH. [ABSTRACT FROM AUTHOR]

Abstract: Oxidative stress may contribute to the pathogenesis of diabetic nephropathy (DN), although the detailed mechanism of reactive oxygen species (ROS) regulation is still unclear. This study examined the effect of high-salt diet on ROS production and expression of antioxidant enzymes in control and experimentally diabetic rats. Wistar fatty rats (WFR) as a type 2 diabetes mellitus model and Wistar lean rats (WLR) as a control were fed a normal-salt diet (NS) and high-salt diet (HS) from the age of 6 to 14 weeks. We then examined the blood pressure, urinary albumin excretion (UAE), and urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels. The expression of antioxidant enzymes including α-catalase (CAT), Cu–Zn superoxide dismutase (SOD), Mn SOD, and glutathione peroxidase (GPx) were analyzed in the glomeruli of the rats using Western blotting. The expression of NAD(P)H oxidase p47phox and NFκB p65 was evaluated using immunohistochemical staining. By 14 weeks of age, the WFR-HS group exhibited hypertension and markedly increased UAE. The level of 8-OHdG, a marker of oxidative damage, in the WFR-HS group was also higher than that in the WLR groups or WFR-NS group. The expression of α-CAT and Mn SOD proteins was significantly decreased in isolated glomeruli in the WFR-HS group. GPx and Cu–Zn SOD expression did not differ between the WFR and WLR groups. High expression of ROS and decreases in antioxidants were seen in the glomeruli of diabetic rats with hypertension, suggesting that oxidative stress may be involved in the development of DN. [Copyright &y& Elsevier]

Eubacterial-type multi-subunit plastid RNA polymerase (PEP) is responsible for the principal transcription activity in chloroplasts. PEP is composed of plastid-encoded core subunits and one of multiple nuclear-encoded sigma factors that confer promoter specificity on PEP. Thus, the replacement of sigma factors associated with PEP has been assumed to be a major mechanism for the switching of transcription patterns during chloroplast development. The null mutant (sig6-1) of plastid sigma factor geneAtSIG6exhibited a cotyledon-specific pale green phenotype. Light-dependent chloroplast development was significantly delayed in thesig6-1mutant. Genetic complementation of the mutant phenotype by theAtSIG6cDNA demonstrated that AtSIG6 plays a key role in light-dependent chloroplast development. Northern and array-based global analyses for plastid transcripts revealed that the transcript levels of most PEP-dependent genes were greatly reduced in thesig6-1mutant, but that the accumulation of nuclear-encoded RNA polymerase (NEP)-dependent transcripts generally increased. As the PEPα subunit and PEP-dependenttrnVaccumulated at normal levels in thesig6-1mutant, theAtSIG6knockout mutant probably retained functional PEP, and the transcriptional defects are likely to have been directly caused byAtSIG6deficiency. Most of the AtSIG6-dependent genes are preceded byσ70-type promoters comprised of conserved−35/−10 elements. Thus, AtSIG6 may act as a major general sigma factor in chloroplasts during early plant development. On the other hand, the mutant phenotype was restored in older seedlings. Arabidopsis probably contains another late general sigma factor, the promoter specificity of which widely overlaps with that of AtSIG6. [ABSTRACT FROM AUTHOR]

A multiresidue analytical method was developed for the quantification of benzylpenicillin (PCG), phenoxymethylpenicillin (PCV), oxacillin (MPIPC), cloxacillin (MCIPC), nafcillin (NFPC) and dicloxacillin (MDIPC) in bovine tissues using liquid chromatography– electrospray ionization tandem mass spectrometry (LC–ESI MS/MS) with a multiple reaction monitoring technique. Using the deuterated PCG and NFPC as internal standard was effective for improvement of repeatability and accuracy. We chose [M - H-141]- as a monitor ion of MRM analysis and [M - H]- as a precursor ion for each penicillin. Combination of an ion-exchange cartridge clean-up and ion-pair LC enable us to determine the residual penicillins using the standard curves made from standard solutions without the influence of sample matrix on the MS. The average recoveries of PCG, PCV, MPIPC, MCIPC, NFPC and MDIPC from bovine liver, kidney and muscle at the same concentrations as the tolerance levels of PCG (50 μg/kg) ranged from 77 to 101% with the coefficients of variation ranging from 0.7 to 4.2% (n=5). The limits of quantification for the six penicillins were 2–10 μg/kg in bovine muscle, liver and kidney (S/N ratio >10). [Copyright &y& Elsevier]

Th1 and Th2 cells, resulting from antigenic stimulation in the presence of IL-12 and IL-4. respectively, are implicated in the pathology of various diseases including allergic and autoimmune diseases. Txk Rlk is a member of Tee family tyrosine kinases. We reported that Txk acts as a Th1 specific transcription fritter in the T lymphocytes. In this study we have asked whether administration of txk expression plasmid brings about a Th1 Th2 shift in vivo of the mice, and subsequent reduction of circulating IgE. Mice were administered a txk expression plasmid with hemagglutinating virus of Japan (HVJ) envelope vector. Txk expressions in spleen cells were assessed by immunoblotting and immunocytochemical staining. Cytokine productions By the spleen cells and serum Ig content rations were studied by ELISA. Administration of a txk expression plasmid with HVJ vector induced expression of Txk in the spleen cells. The spleen cells showed enhanced Th1 specific cytokine production: spleen ceils from the txk administered mice produced more IFN-γ as compared with those from control plasmid administered mice in an antigen specific manner. IL-2 and IL-4 secretions of the spleen cells were comparable between the two mouse groups. Txk administration did not reduce serum IgG concentration. It markedly reduced total IgE level and an IgGI IgG2a ratio, reflection of Th1 Th2 balance, in sera. Furthermore, txk administration reduced ovalbumin (OVA)-specific IgE levels in sera of the OVA sensitized mice. Thus, Txk enhances IFN-γ secretion and thus modulates Th1 Th2 cytokine balance. leading to reduction of serum IgE. [ABSTRACT FROM AUTHOR]

Since the rat is an atherosclerosis-resistant species, the study of atherosclerosis using rats is limited. The present study was undertaken to develop an atherosclerotic model in rats, to investigate the effect of nitric oxide (NO) inactivation and hyperlipidemia, and to evaluate the effect of pitavastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) inhibitor, on NO inactivation and on hyperlipidemia-induced changes in the cardiovascular system. Four-month-old male spontaneously hypertensive hyperlipidemic rats (SHHR) and Sprague-Dawley (SD) rats were used to study 1) the effect of the period of treatment with NG-nitro-L-arginine methyl ester (L-NAME, 100 mg/L) on high fat diet (HFD)-treated SHHR and SD rats, and 2) the effect of pitavastatin (Pit, 0.3 mg/kg/day) on the changes in the aorta of L-NAME- and HFD-treated SHHR and SD rats. L-NAME administration for 1 month then HFD feeding for 2 months markedly increased the deposition of lipids and the thickness of the endothelium in SHHR. Continuous L-NAME treatment with HFD produced severe injury and stripped of endothelium in both strains. The plasma total cholesterol of L-NAME + HFD-treated and L-NAME + HFD + Pit-treated SHHR was significantly higher than that of control SHHR. Lipid deposition, however, was comparatively less in the aorta of L-NAME + HFD + Pit-treated SHHR. The concentration of cholesterol in the aorta of control SHHR was significantly lower than that in the aorta of L-NAME + HFD-treated SHHR, whereas that of L-NAME + HFD + Pit-treated SHHR was the same as that in control SHHR. These data indicated that Pit blocked lipid deposition in the aorta of L-NAME + HFD treated SHHR without changing plasma lipid profiles. In conclusion, NO inactivation and HFD induce lipid deposition in the endothelium, and the HMG-CoA reductase inhibitor blocks the deposition in SHHR. [Copyright &y& Elsevier]

In order to investigate the mechanism of delayed progressive or secondary neuronal damage after the spinal cord injury, we developed a mild-compression injury model in the rat thoracic spinal cord. Our compression device consists of a soft silicone point of contact to the dura, in order to prevent violent injury that may cause axonal tears or hemorrhages in the spinal cord. Since rats often assume a ‘standing’ posture, i.e. raising head with lifting their fore-limbs, damage to the thoracic spinal cord was evaluated by measuring the frequency of ‘standing’, which effectively indicates hind limb function. Twenty-four hours after compression by a 20 g weight for 10 or 20 min, the standing frequency of the injured rat was almost the same as that of sham animals that underwent laminectomy without compression. However, the standing frequency decreased with time; the frequency of standing at 72 h was approximately 30–50% that of sham animals. In the compressed spinal cord tissue, microglial cells, detected by lectin staining, proliferated with time. An enormous amount of microglia was observed at 48 and 72 h after compression, although only a small amount of cells were positive to lectin staining at 24 h after the compression. These results suggest that our mild-compression spinal cord injury model showed late-onset or delayed neuronal damage that may be related to pathological microglia proliferation. [Copyright &y& Elsevier]

Under hypergravity conditions, the cell wall of stem organs becomes mechanically rigid and elongation growth is suppressed, which can be recognized as the mechanism for plants to resist gravitational force. The changes in gene expression by hypergravity treatment were analyzed in Arabidopsis hypocotyls by the differential display method, for identifying genes involved in hypergravity-induced growth suppression. Sixty-two cDNA clones were expressed differentially between the control and 300 g conditions: the expression levels of 39 clones increased, whereas those of 23 clones decreased under hypergravity conditions. Sequence analysis and database searching revealed that 12 clones, 9 up-regulated and 3 down-regulated, have homology to known proteins. The expression of these genes was further analyzed using RT-PCR. Finally, six genes were confirmed to be up-regulated by hypergravity. One of such genes encoded 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGR), which catalyzes a reaction producing mevalonic acid, a key precursor ofterpenoids such as membrane sterols and several types of hormones. The expression of HMGR gene increased within several hours after hypergravity treatment. Also, compactin, an inhibitor of HMGR, prevented hypergravity-induced growth suppression, suggesting that HMGR is involved in suppression of Arabidopsis hypocotyl growth by hypergravity. In addition, hypergravity increased the expression levels of genes encoding CCR1 and ERD15, which were shown to take part in the signaling pathway of environmental stimuli such as temperature and water, and those of the α-tubulin gene. These genes may be involved in a series of cellular events leading to growth suppression of stem organs under hypergravity conditions. [Copyright &y& Elsevier]

Although dysesthesia is one of the most serious problems in patients with spinal cord injury, most of them being unresponsive to conventional treatments. In this study, we established a rat thoracic spinal cord mild-compression model that revealed thermal hyperalgesia in the hind limb. The thoracic spinal cord was compressed gently, using a 20 g weight for 20 min. The withdrawal latency of the thermal stimulation of the bilateral hind-limb was monitored using Hargreaves’ Plantar test apparatus. In this model, thermal-hyperalgesia was observed for 1 week after the injury. The spinal cord injury-induced thermal-hyperalgesia was mimicked by the intrathecal application of metergoline, a non-selective 5-HT antagonist, 1-(2-methoxyphenyl)-4-[4-(2-phthalimido) butyl]-piperazine hydrobromide (NAN190), a selective 5-HT1 antagonist, and 3-tropanyl-3,5-dichlorobenzoate (MDL72222), a selective 5-HT3 antagonist. Intraperitoneal application of fluvoxamine maleate, a selective serotonin reuptake inhibitor, reduced the intensity of hyperalgesia induced by spinal cord injury. The inhibitory effect of fluvoxamine maleate on thermal hyperalgesia was prevented by the application of the aforementioned nonselective or selective 5-HT receptor antagonists. Intrathecal application of fluvoxamine maleate and selective 5-HT receptor agonists, i.e., 8-hydroxy-2-(di-n-proplyamino)-tetralin hydrobromide (8-OH-DPAT: 5HT-1 selective) and 2-methyl-5-hydroxytryptamine maleate (2-m-5-HT: 5HT-3 selective), inhibited the spinal cord injury-induced hyperalgesia. These results suggest that the change in the descending serotonergic signal plays an important role in hyperalgesia after the spinal cord injury, and that the application of selective serotonin reuptake inhibitors will be one of the candidates for new therapeutic methods against post-spinal cord injury dysesthesia. [Copyright &y& Elsevier]