Affinity-purified antigen-specific products produced by T cells share epitopes recognized by heterologous antisera raised against several different antigen-specific products from T cells.
Heterologous antisera to murine or rat T-cell antigen-binding molecules (T-ABM) were raised in rabbits or sheep. The T-ABM used for immunization were purified by affinity for antigen and did not bear known immunoglobulin isotypes. T-ABM and anti-T-ABM were raised in three separate laboratories. Antisera to T-ABM were exchanged and tested for binding to T-ABM in three separate laboratories. Thus antisera to at least three distinct T-ABM were tested directly for binding to T-ABM or by adsorption of biological activity. Rabbit antisera to murine trinitrophenol (TNP)-specific T-ABM or rat AgB-specific T-ABM bound both murine or rat T-ABM, indicating evolutionary conservation of T-ABM. Similar results were found with sheep antisera to murine T-ABM. In addition, all heterologous anti-T-ABM antisera used bound murine T-ABM specific for TNP, 4-hydroxy-3-nitrophenyl acetate (NP), SRBC, or T-cell membrane proteins with similar structure. Thus, there is a commonality of antigenic determinants between various T-ABM and T-cell membrane homologues which may be T-cell surface receptors for foreign antigen.


Functional intracellular glutaminase activity in intact astrocytes.
Numerous cellular metabolites such as glutamine, glutamate, phosphate, calcium, ammonia and acetyl derivatives are known to affect the phosphate-activated glutaminase activity in whole cell homogenates or extracts. Since measurements in extracts under non-physiological conditions may obscure the actual intracellular metabolic flux, the "functional" intracellular phosphate-activated glutaminase activity was measured by the formation of 3H2O from L-[2-3H]glutamine (Anal. Biochem. 127:134-142, 1982) in cultures of intact astrocytes, untreated and treated with dibutyryl c-AMP (DiBcAMP), in the presence of several potential effectors. These values were compared with enzyme levels determined in extracts from identical cells. The rate of 14CO2 release from L-[1-14C]glutamine was also measured in both untreated and DiBcAMP treated astrocytes. The intracellular activity of glutaminase for untreated cells assayed in MEM medium with 1 mM radioactive glutamine was 88 nmol/mg protein/h and in DiBcAMP treated cells the rate was 153 nmol/mg protein/h. However, the enzymatic activity measured under optimal conditions in extracts from both untreated and treated cells was much higher, but essentially the same, about 1,750 nmol/mg protein/h. The rate of 14CO2 release from L-[1-14C]glutamine was 74 and 133 nmol/mg protein/h in untreated and DiBcAMP treated cells, respectively. This represents approximately 85% of the intracellular glutaminase activity. Furthermore, increasing the concentration of glutamine in the medium from 1 to 6.4 mM increased glutaminase intracellular activity about 3 fold in both untreated and treated cells. Addition of 250 microM glutamate to the medium inhibited intracellular glutaminase activity by 70% under both treatment conditions. Deletion of glucose stimulated glutaminase activity. In contrast the removal of fetal bovine serum decreased activity by 35%.(ABSTRACT TRUNCATED AT 250 WORDS)


Development and Validation of an Interprofessional Collaboration Scale for Home Health Care for the Frail Elderly.
The purpose of this study was to develop an interprofessional collaboration (IPC) scale for home health care for frail elderly. The first items of the IPC scale included collaboration members' attitudes, awareness, motivation, team strength, communication, relationships, information, care recipients' interests, effects, development, utilization of social resources, contributions to the community, and crisis management. The subjects were 512 care managers who work in home care support offices across Japan. They manage interprofessional collaboration in home health care for frail elderly who need care at 65 years old and above. The scale's construct validity, internal consistency, the validity of known groups, concurrent validity, and test-retest reliability (193 subjects) were examined. The final IPC scale's items consisted of four factors (37 items): the strength of interprofessional teams (16), the management of collaborative systems (7), effects of collaboration (8), and communication (6). Four factors explained 58.6% of the total explained variance. The modified model fit of the scale achieved acceptable levels. The Cronbach's α coefficient for all items was .97. The sum of communication factor in the cities/wards group was lower than those in the towns/villages group. There were differences between the sum and each factor with different levels of ease to collaborate. The intraclass correlation coefficient for all items in the first and second assessments was .875. The validity and reliability of the IPC scale have been verified. This scale can be used to assess the IPC for home health care for frail elderly.


[Epidemiologic aspects of Hansen's disease at the 10th Regional Health Department (DRS/10) of Presidente Prudente, State of São Paulo, in December of 1984].
This paper relates the epidemiological situation of hanseniasis at the Health Region of Presidente Prudente, São Paulo State, Brazil. It was observed a high percentage from 1974 to nowadays, of diagnosed early forms of the disease (I). In 1984 were diagnosed 132 patients, V and D cases 36 (27.28%), I cases 62 (46.96%) and T cases 34 (25.15%). The rate of incidence in 1984 was 0.19% and the rate of prevalence was 1.83.


[Malignant mixed mesodermal tumor of the ovary. A review of the literature apropos of a case with immunohistochemical study].
A very large left ovarian tumor in a 76 year old woman, on microscopic study, consisted of an epithelial component: both glandular and squamous, and a sarcomatous component: both cartilage and striated muscle. Epithelial and sarcomatous patterns were intermingled in a spindle-cell stroma. Immunohistochemical study revealed, in these spindle cells, cytokeratin-positive anaplastic epithelial structures and vimentin-positive mesenchymal structures. A review of the literature concerning this rare ovarian tumor especially including cases with a immunohistochemical study is added. However, immunohistochemical study does not allow histogenetic interpretation.


Effects of elastase and cathepsin G on the levels of membrane and soluble TNFalpha.
Neutrophil elastase (NE) and cathepsin G (CG), the proteolytic enzymes localized in azurophil granules of neutrophils (PMN), are involved in PMN responses to various stimuli. When released at sites of inflammation, they participate in the degradation of numerous proteins involved in the regulation of the immune response. In this study, we employed ADAM17(-/-) fibroblasts stably transfected with cDNA of human pro-tumor necrosis factor alpha (proTNFalpha) (ADAM17(-/-)TNF(+)) to investigate the effects of NE and CG on shedding and degradation of TNFalpha. Both NE and CG were found to diminish the level of membrane TNFalpha (mTNFalpha) as measured by flow cytometry. This process was accompanied by the accumulation of biologically active soluble TNFalpha (sTNFalpha) in the culture medium, as determined by an increase in both the cytotoxic activity of TNFalpha and its ability to serve as a co-stimulator in the induction of inducible nitric oxide synthase (iNOS). However, in contrast to CG, NE at high concentrations was able to degrade sTNFalpha released from the cell surface. Using soluble recombinant human TNFalpha, we identified Val(93)-Ala(94) and Val(117)-Glu(118) as the NE cleavage sites within the sTNFalpha molecule. Taken together, the ability of NE and CG to modulate levels of membrane and soluble forms of TNFalpha may contribute to the proinflammatory activity of neutrophils.


The influence of ether lipid (AMG) on histamine release from isolated rat mast cells: synergistic interaction with TPA indicates protein kinase C activation.
We have previously shown that the ether lipid AMG (1-O-hexadecyl-2-O-methyl-sn-glycerol) has both synergistic and inhibitory effects on mast cell responses to the ionophore A23187. The present investigation showed only inhibitory effects of AMG in antigen- and compound 48/80-induced histamine release. Both enhancement and inhibition were noted in responses to the combination of A23187 and the phorbol ester TPA, at 2-5 microM and 10-20 microM and 10-20 microM AMG, respectively, as found earlier with A23187 alone. The synergistic response to AMG in combination with A23187 resembles that with TPA but required higher concentrations of A23187. The flavonoid phloretin was a potent inhibitor of the response to combinations of AMG and A23187. A pronounced synergistic interaction between AMG and TPA was found at very low concentrations of A23187. Our results do not provide much information about mechanisms involved in the inhibitory effect of AMG although some competition relating to protein kinase C activity might participate. The synergistic interactions indicate that AMG can activate protein kinase C but in a manner different from the phorbol ester.


Chemotherapy of schistosomal colonic polyposis with oxamniquine.
Schistosomal polyposis of the colon is a common complication of schistosomiasis. In severe infections it may be associated with considerable morbidity. Surgery is associated with high morbidity and mortality. Niridazole therapy is of limited therapeutic value because of its hepatotoxicity but oxaminiquine has been found to be effective and safe. Several dosage regimens have been tried but a total dose of 40 mg/kg over two days in two equally divided doses has been found to have an effective therapeutic action on the polypi, with correction of the anaemia, hypoalbuminaemia and serum iron.


CXCL3 contributes to CD133(+) CSCs maintenance and forms a positive feedback regulation loop with CD133 in HCC via Erk1/2 phosphorylation.
Although the chemotactic cytokine CXCL3 is thought to play an important role in tumor initiation and invasion, little is known about its function in hepatocellular carcinoma (HCC). In our previous study, we found that Ikaros inhibited CD133 expression via the MAPK pathway in HCC. Here, we showed that Ikaros may indirectly down-regulate CXCL3 expression in HCC cells, which leads to better outcomes in patients with CD133(+) cancer stem cell (CSC) populations. CD133 overexpression induced CXCL3 expression, and silencing of CD133 down-regulated CXCL3 in HCC cells. Knockdown of CXCL3 inhibited CD133(+) HCC CSCs' self-renewal and tumorigenesis. The serum CXCL3 level was higher in HCC patients' samples than that in healthy individual. HCC patients with higher CXCL3 expression displayed a poor prognosis, and a high level of CXCL3 was significantly associated with vascular invasion and tumor capsule formation. Exogenous CXCL3 induced Erk1/2 and ETS1 phosphorylation and promoted CD133 expression, indicating a positive feedback loop between CXCL3 and CD133 gene expression in HCC cells via Erk1/2 activation. Together, our findings indicated that CXCL3 might be a potent therapeutic target for HCC.


Clinical impact of low-volume lymph node metastases in early-stage cervical cancer: A comprehensive meta-analysis.
In order to define the clinical significance of low-volume metastasis, a comprehensive meta-analysis of published data and individual data obtained from articles mentioning micrometastases (MIC) and isolated tumor cells (ITC) in cervical cancer was performed, with a follow up of at least 3 years. We performed a systematic literature review and meta-analysis, following Cochrane's review methods guide and the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The primary outcome was the disease-free survival (DFS), and the secondary outcome was the overall survival (OS). The hazard ratio (HR) was taken as the measure of the association between the low-volume metastases (MIC+ITC and MIC alone) and DFS or OS; it quantified the hazard of an event in the MIC (+/- ITC) group compared to the hazard in node-negative (N0) patients. A random-effect meta-analysis model using the inverse variance method was selected for pooling. Forest plots were used to display the HRs and risk differences within individual trials and overall. Eleven articles were finally retained for the meta-analysis. In the analysis of DFS in patients with low-volume metastasis (MIC + ITC), the HR was increased to 2.60 (1.55-4.34) in the case of low-volume metastasis vs. N0. The presence of MICs had a negative prognostic impact, with an HR of 4.10 (2.71-6.20) compared to N0. Moreover, this impact was worse than that of MIC pooled with ITCs. Concerning OS, the meta-analysis shows an HR of 5.65 (2.81-11.39) in the case of low-volume metastases vs. N0. The presence of MICs alone had a negative effect, with an HR of 6.94 (2.56-18.81). In conclusion, the presence of MIC seems to be associated with a negative impact on both the DFS and OS and should be treated as MAC.


Voltage sensor current, SR Ca2+ release, and Ca2+ channel current during trains of action potential-like depolarizations of skeletal muscle fibers.
In skeletal muscle, CaV 1.1 serves as the voltage sensor for both excitation-contraction coupling (ECC) and L-type Ca2+ channel activation. We have recently adapted the technique of action potential (AP) voltage clamp (APVC) to monitor the current generated by the movement of intramembrane voltage sensors (IQ ) during single imposed transverse tubular AP-like depolarization waveforms (IQAP ). We now extend this procedure to monitoring IQAP , and Ca2+ currents during trains of tubular AP-like waveforms in adult murine skeletal muscle fibers, and compare them with the trajectories of APs and AP-induced Ca2+ release measured in other fibers using field stimulation and optical probes. The AP waveform remains relatively constant during brief trains (<1 sec) for propagating APs in non-V clamped fibers. Trains of 10 AP-like depolarizations at 10 Hz (900 ms), 50 Hz (180 ms), or 100 Hz (90 ms) did not alter IQAP amplitude or kinetics, consistent with previous findings in isolated muscle fibers where negligible charge immobilization occurred during 100 ms step depolarizations. Using field stimulation, Ca2+ release did exhibit a considerable decline from pulse to pulse during the train, also consistent with previous findings, indicating that the decline of Ca2+ release during a short train of APs is not correlated to modification of charge movement. Ca2+ currents during single or 10 Hz trains of AP-like depolarizations were hardly detectable, were minimal during 50 Hz trains, and became more evident during 100 Hz trains in some fibers. Our results verify predictions on the behavior of the ECC machinery in response to AP-like depolarizations and provide a direct demonstration that Ca2+ currents elicited by single AP-like waveforms are negligible, but can become more prominent in some fibers during short high-frequency train stimulation that elicits maximal isometric force.


Accuracy of convolutional neural networks-based automatic segmentation of pharyngeal airway sections according to craniofacial skeletal pattern.
This study aimed to evaluate a 3-dimensional (3D) U-Net-based convolutional neural networks model for the fully automatic segmentation of regional pharyngeal volume of interests (VOIs) in cone-beam computed tomography scans to compare the accuracy of the model performance across different skeletal patterns presenting with various pharyngeal dimensions. Two-hundred sixteen cone-beam computed tomography scans of adult patients were randomly divided into training (n = 100), validation (n = 16), and test (n = 100) datasets. We trained the 3D U-Net model for fully automatic segmentation of pharyngeal VOIs and their measurements: nasopharyngeal, velopharyngeal, glossopharyngeal, and hypopharyngeal sections as well as total pharyngeal airway space (PAS). The test datasets were subdivided according to the sagittal and vertical skeletal patterns. The segmentation performance was assessed by dice similarity coefficient, volumetric similarity, precision, and recall values, compared with the ground truth created by 1 expert's manual processing using semiautomatic software. The proposed model achieved highly accurate performance, showing a mean dice similarity coefficient of 0.928 ± 0.023, the volumetric similarity of 0.928 ± 0.023, precision of 0.925 ± 0.030, and recall of 0.921 ± 0.029 for total PAS segmentation. The performance showed region-specific differences, revealing lower accuracy in the glossopharyngeal and hypopharyngeal sections than in the upper sections (P <0.001). However, the accuracy of model performance at each pharyngeal VOI showed no significant difference according to sagittal or vertical skeletal patterns. The 3D-convolutional neural network performance for region-specific PAS analysis is promising to substitute for laborious and time-consuming manual analysis in every skeletal and pharyngeal pattern.


Radiorespirometry evidence for the discrimination between 13C-enriched glucose and unlabelled glucose molecules by Paracoccus denitrificans.
Paracoccus denitrificans was grown on either unlabelled glucose, [1-13C]glucose or [6-13C]glucose as the sole carbon source for growth. The cells were then incubated with a range of 14C-glucose substrates to compare the 14CO2-evolution rates between cells grown on the glucose and the 13C-labelled glucose. Cells grown on 13C-glucose had significantly faster rates of 14CO2-evolution than those grown on unlabelled glucose. The % yields of 14CO2, per [1-14C]-, [6-14C]- and [U-14C]glucose supplied were also substantially greater than those measured for cells grown on unlabelled glucose. The data indicated that growth of Paracoccus on 13C-enriched glucose substrates resulted in cells with notably different 14C-glucose oxidation metabolism compared to that observed in cells grown on unlabelled glucose.


Systems biology of the genomes' microsatellite signature of Orthopoxvirus including the Monkeypox virus.
This study is an attempt to extract and analyse the microsatellites or simple sequence repeats (SSRs) from the genomes of eight species of the genus Orthopoxvirus. The average size of genomes included in the study was 205 kb while the GC% was 33% for all but one. A total of 10,584 SSRs and 854 cSSRs were observed. POX2 with the largest genome of 224.499 kb had maximum of 1493 SSRs and 121 cSSRs (compound SSR) while POX7 with the smallest genome of 185.578 kb had minimum incident SSRs and cSSRs at 1181 and 96, respectively. There was significant correlation between genome size and SSR incidence. Di-nucleotide repeats were the most prevalent (57.47%) followed by mono- at 33% and tri- at 8.6%. Mono-nucleotide SSRs were predominantly T (51%) and A (48.4%). A majority of 80.32% SSRs were in the coding region. The three most similar genomes as per heat map POX1, POX7 and POX5 (93% similarity) are adjacent to one another in the phylogenetic tree. Ankyrin/Ankyrin like protein and Kelch protein which are associated with host determination and divergence have the highest SSR density in almost all studied viruses. Thus, SSRs are involved in genome evolution and host determination of viruses.


Acetylcholine-induced endothelium-dependent contractions in the SHR aorta: the Janus face of prostacyclin.
In the spontaneously hypertensive rat (SHR) and aging Wistar-Kyoto rats (WKY), acetylcholine releases an endothelium-derived contracting factor (EDCF) produced by endothelial cyclooxygenase-1, which stimulates thromboxane A2 receptors (TP receptors) on vascular smooth muscle. The purpose of the present study was to identify this EDCF by measuring changes in isometric tension and the release of various prostaglandins by acetylcholine. In isolated aortic rings of SHR, U 46619, prostaglandin (PG) H2, PGF2alpha, PGE2, PGD2, prostacyclin (PGI2) and 8-isoprostane, all activate TP receptors of the vascular smooth muscle to produce a contraction (U 46619>>8-isoprostane=PGF2alpha=PGH2>PGE2=PGD2>PGI2). The contractions produced by PGH2 and PGI2 were fast and transient, mimicking endothelium-dependent contractions. PGI2 did not relax isolated aortic rings of WKY and SHR. Acetylcholine evoked the endothelium-dependent release of thromboxane A2, PGF2alpha, PGE2, PGI2 and most likely PGH2 (PGI2>>PGF2alpha>or=PGE2>TXA2>8-isoprostane, PGD2). Dazoxiben abolished the production of thromboxane A2, but did not influence the endothelium-dependent contractions to acetylcholine. The release of PGI2 was significantly larger in the aorta of SHR than in WKY, and the former was more sensitive to the contractile effect of PGI2 than the latter. The inhibition of PGI-synthase was associated with an increase in PGH2 spillover and the enhancement of acetylcholine-induced endothelium-dependent contractions. Thus, in the aorta of SHR and aging WKY, the endothelium-dependent contractions elicited by acetylcholine most likely involve the release of PGI2 with a concomitant contribution of PGH2.


Prelabeled red nucleus and sensorimotor cortex neurons of the rat survive 10 and 20 weeks after spinal cord transection.
To demonstrate definitively the fate of the somata of rubrospinal and corticospinal neurons axotomized by a complete spinal cord transection at T-9, in young adult rats we prelabeled the neurons by injection into the lumbar enlargement of a retrogradely transported fluorescent dye, Fluoro-Gold, and four days later transected the cord. We found no loss in cell number ten or 20 weeks after axotomy. The average size of the neurons in each case is slightly but significantly reduced. These findings unequivocally demonstrate that the somata of long tract neurons of the rubrospinal and corticospinal systems persist in an atrophic and presumably inactive state for at least 20 weeks, and raise the possibility that treatment of spinal cord injury may normalize cell activity and allow long tract regeneration.


The effect of maternal multisensory stimulations on bath stress in premature infants: A randomized controlled clinical trial.
Bathing in Neonatal Intensive Care Unit (NICU) is a stressful experience to preterm infants. Reducing this stress is an important challenge in bathing preterm infants. The aim of present study was to evaluate the effects of maternal supportive interventions on the stress caused by swaddled bathing in preterm infants. In this randomized clinical trial, 48 preterm infants randomly allocated into control and intervention groups. In intervention group the mothers were asked to initiate multisensory supportive interventions 5 min before and through swaddled bathing. In controls the mothers were only present during bathing. To determine the level of stress, infant responses were recorded by camera 5 min before bathing and 5 min after bathing. Then infants' stress measured by using Newborn Stress Scale (NSS). Finding was shown that the level of stress in control group was especially increased after bathing that was statistically significant (p < 0.001). Although there was an increase in stress among interventions but it was not statistically significant (p > 0.05). Considering the positive effects of multisensory interventions in decreasing the stress of preterm infants, it can be recommended as cost-free and non-pharmacological care during infants bathing.


A "bottle-around-ship" like method synthesized yolk-shell Ag3PO4@MIL-53(Fe) Z-scheme photocatalysts for enhanced tetracycline removal.
A novel yolk-shell Ag3PO4@MIL-53(Fe) Z-scheme photocatalyst was fabricated via a "bottle-around-ship" like method. Experiments on the treatment of tetracycline upon visible light irradiation showed that the as-prepared photocatalyst possessed excellent photocatalytic performance. Experimental results showed that tetracycline removal efficiency of the yolk-shell Ag3PO4@MIL-53(Fe) Z-scheme photocatalyst was almost 3 times higher than that of MIL-53(Fe). The enhanced photocatalytic performance of Ag3PO4@MIL-53(Fe) nanocomposite could be contributed to its higher surface area, better absorption capability, and greater charge separation efficiency. In addition, the H2O2 concentration detection results for Ag3PO4 (154 μmol/L) and Ag3PO4@MIL-53(Fe) (52 μmol/L) indicated that a big part of generated H2O2 on the Ag3PO4 core would be quickly decomposed by the MIL-53(Fe) shell and generated more reactive species through the photo-Fenton-like reaction, which is beneficial for the improvement of photocatalytic performance. This is a promising approach to fabricate yolk-shell structure photocatalyst and a different aspect to design multiple semiconductor composites heterojunction for environmental remediation.


Coupling of early response gene expression to distinct regulatory pathways during alpha-interferon and phorbol ester-induced plasmacytoid differentiation of B chronic lymphocytic leukaemia cells.
Phorbol esters (phorbol 12-myristate 13-acetate; PMA) and alpha-interferon (alpha-IFN) act through divergent signal transduction pathways to induce B chronic lymphocytic leukaemia cells (B-CLL) to undergo plasmacytoid differentiation in vitro. By using a panel of PMA-inducible early response gene probes we show that these two different effectors are coupled to second messenger pathways that do not converge on a common gene regulatory programme in differentiation of B-CLL cells. Moreover, using the calcium ionophore, A23187, four distinct regulatory classes of early response gene could be defined implying that multiple regulatory pathways may mediate the process of terminal differentiation in B lymphocytes.


How do normal subjects learn a simple adaptative task: how and why do paranoid schizophrenic patients fail?
The authors studied the behavior of normal subjects and paranoid schizophrenic patients in a simple problem-solving situation. The schizophrenics were divided into two sample groups, one of individuals under treatment and the other of individuals not under treatment. The learning process involved in this problem-solving situation is very similar to an instrumental conditioning, and can be understood by means of the following assumptions: (1) the subjects use decision functions in reacting to the stimuli, although they may be not fully aware of this; (2) learning is the result of successive transformations of these decisions in the course of time; (3) the changes have specific probabilities and are related to (a) those responses which are made to the latest stimuli, and (b) a differential probability for decision functions which were effective, or only interrupted painful reinforcement, or were completely ineffective. In schizophrenics further factors of importance were (1) an 'inertia' factor and (2) the rigidly continued use of unsuccessfuly or only partially successful decision criteria. The authors used a systems theory based on Galois field theory and a calculus of operators specifying three groups of subjects. A computer program based on thses hypotheses was tested in a stimulation experiment. The statistical evaluation of the results showed a congruence between the theoretical approach and the experimental data.


Comparison of pulsed light inactivation kinetics and modeling of Escherichia coli (ATCC-29055), Clostridium sporogenes (ATCC-7955) and Geobacillus stearothermophilus (ATCC-10149).
Pulsed light (PL) inactivation kinetics of Escherichia coli K-12, Clostridium sporogenes and Geobacillus stearothermophilus were evaluated under different treatment conditions. The PL system was factory set to operate at three pulses per second with a pulse width of 360 μs exposing samples placed on one of the 9 trays on a rack. Two PL parameters were evaluated in the study: number of pulses (a time factor) and the tray position (a spatial distance factor) both influencing the amount of light energy absorbed. As expected, the level of microbial inactivation increased with an increase in the number of pulses (from 1 to 15) and decreased with an increase in the Spatial distance (Tray # 1 to 9) away from the light source. Both the number of pulses and spatial distance as well as their interactions were found to have a significant effect (P < 0.05) on the extent of microbial inactivation. Vegetative cells of E. coli were most sensitive to PL treatment with a maximum 5 logarithmic reductions on Tray 1 after a 12-pulse treatment (4 s). G. stearothermophilus was more resistant to PL than C. sporogenes. Overall, the PL treatments (12-15 pulses) achieved a minimum four logarithmic reductions in the populations of all three microorganisms on the top tray at doses still below 12 J/cm2, the FDA-approved limit.


Hypothermia differentially modulates the formation and decay of NBS1, γH2AX and 53BP1 foci in U2OS cells exposed to gamma radiation.
In studies on the mechanism of DNA damage response where ionizing radiation is used as the DNA damaging agent, cells are often exposed to ionizing radiation on melting ice (corresponding to 0.8 °C). The purpose of this procedure is to inhibit cellular processes i.e. DNA repair. Low temperature at exposure has been shown to act in a radioprotective manner at the level of cytogenetic damage, but its mechanisms of action are poorly understood. The aim of the study was to analyze the effect of hypothermia at the level of formation and decay of NBS1, γH2AX, and 53BP1 foci, micronuclei, survival, cell cycle progression and oxidative stress in U2OS cells. The results show that hypothermia alone induced oxidative stress and foci. When applied in combination with radiation but only during the exposure time, it potentiated the formation of γH2AX and 53BP1 but not of NBS1 foci. When applied during irradiation and subsequent repair time, 53BP1 and NBS1 foci formed and decayed, but the levels were markedly lower than when repair was carried out at 37 °C. The frequency of micronuclei was elevated in cells irradiated at 0.8 °C, but only when analysed 20 h after irradiation which is likely due to a reduced G2 cell cycle block. Hypothermia reduced cell survival, both with and without radiation exposure. The temperature effect should be considered when cooling cells on melting ice to inhibit DNA repair in the induction of DNA damage.


Temperature-mediated heteroduplex analysis for the detection of drug-resistant gene mutations in clinical isolates of Mycobacterium tuberculosis by denaturing HPLC, SURVEYOR nuclease.
Denaturing high-performance liquid chromatography (DHPLC) is a relatively new technique, which utilizes heteroduplex formation between wild-type and mutated DNA strands to identify point mutations. Heteroduplex molecules are separated from homoduplex molecules by ion-pair, reverse-phase liquid chromatography on a special column matrix with partial heat denaturation of the DNA strands. In order to investigate the application of this method for point mutation detection in drug-resistant genes of Mycobacterium tuberculosis, katG, rpoB, embB, gyrA, pncA and rpsL genes, which are responsible for isoniazid, rifampicin, ethambutol, fluoroquinolone, pyrazinamide and streptomycin resistance, respectively, were detected by temperature-mediated DHPLC in 10 multidrug-resistant and 10 drug-susceptible clinical isolates. The DHPLC data were compared with those from a conventional MIC test. The results show that DHPLC is cost-effective with high capacity and accuracy, and is potentially useful for genotypic screening for mutations associated with anti-tuberculosis drug resistance.


Reference directions and reference objects in spatial memory of a briefly viewed layout.
Two experiments investigated participants' spatial memory of a briefly viewed layout. Participants saw an array of five objects on a table and, after a short delay, indicated whether the target object indicated by the experimenter had been moved. Experiment 1 showed that change detection was more accurate when non-target objects were stationary than when non-target objects were moved. This context effect was observed when participants were tested both at the original learning perspective and at a novel perspective. In Experiment 2, the arrays of five objects were presented on a rectangular table and two of the non-target objects were aligned with the longer axis of the table. Change detection was more accurate when the target object was presented with the two objects that were aligned with the longer axis of the table during learning than when the target object was presented with the two objects that were not aligned with the longer axis of the table during learning. These results indicated that the spatial memory of a briefly viewed layout has interobject spatial relations represented and utilizes an allocentric reference direction.


The non-anticoagulant heparin-like K5 polysaccharide derivative K5-N,OSepi attenuates myocardial ischaemia/reperfusion injury.
Heparin and low molecular weight heparins have been demonstrated to reduce myocardial ischaemia/reperfusion (I/R) injury, although their use is hampered by the risk of haemorrhagic and thrombotic complications. Chemical and enzymatic modifications of K5 polysaccharide have shown the possibility of producing heparin-like compounds with low anticoagulant activity and strong anti-inflammatory effects. Using a rat model of regional myocardial I/R, we investigated the effects of an epimerized N-,O-sulphated K5 polysaccharide derivative, K5-N,OSepi, on infarct size and histological signs of myocardial injury caused by 30 min. ligature of the left anterior descending coronary artery followed by 1 or 24 h reperfusion. K5-N,OSepi (0.1-1 mg/kg given i.v. 15 min. before reperfusion) significantly reduced the extent of myocardial damage in a dose-dependent manner. Furthermore, we investigated the potential mechanism(s) of the cardioprotective effect(s) afforded by K5-N,OSepi. In left ventricular samples, I/R induced mast cell degranulation and a robust increase in lipid peroxidation, free radical-induced DNA damage and calcium overload. Markers of neutrophil infiltration and activation were also induced by I/R in rat hearts, specifically myeloperoxidase activity, intercellular-adhesion-molecule-1 expression, prostaglandin-E(2) and tumour-necrosis-factor-α production. The robust increase in oxidative stress and inflammatory markers was blunted by K5-N,OSepi, in a dose-dependent manner, with maximum at 1 mg/kg. Furthermore, K5-N,OSepi administration attenuated the increase in caspase 3 activity, Bid and Bax activation and ameliorated the decrease in expression of Bcl-2 within the ischaemic myocardium. In conclusion, we demonstrate that the cardioprotective effect of the non-anticoagulant K5 derivative K5-N,OSepi is secondary to a combination of anti-apoptotic and anti-inflammatory effects.


The use of the Dienes test in the epidemiology of proteus urinary tract infections in patients with spinal cord disease.
The incidence of urinary tract infections in 47 patients with spinal cord injuries (11 women and 36 men) was studied and the aetiological agents identified. A high incidence of Proteus mirabilis infection was found and both auto- and cross-infection were identified by the Dienes test.


An electrophysiological analysis of the anticonvulsant action of cannabidiol on limbic seizures in conscious rats.
The effects of cannabidiol (CBD) on electrically evoked kindled seizures were studied in conscious, unrestrained rats with chronically implanted cortical and limbic electrodes, and the results were compared with those of delta 9-tetrahydrocannabinol (delta 9-THC), phenytoin (PHT), and ethosuximide (ESM). All drugs were anticonvulsant, but there were marked differences in their effects on afterdischarge (AD) threshold, duration, and amplitude. CBD, like PHT and delta 9-THC, elevated the AD threshold; in contrast, ESM decreased the threshold but suppressed AD spread. CBD, however, also resembled ESM inasmuch as both drugs decreased AD duration and amplitude. Electrophysiologically, the antiseizure effects of CBD were a combination of those of PHT and ESM. The combination of effects may account for the observation that CBD was the most efficacious of the drugs tested against limbic ADs and convulsions. Other properties of CBD were also noted: For example, compared with delta 9-THC, it is a much more selective anticonvulsant vis-à-vis motor toxicity. CBD also lacks the CNS excitatory effects produced by delta 9-THC, PHT, and ESM. These characteristics, combined with its apparently unique set of electrophysiological properties, support the suggestion that CBD has therapeutic potential as an antiepileptic.


Immune-adjuvant effect of vitamin A and probiotics supplementation on humoral response to cell culture rabies vaccine in rabbits.
This study was carried out to evaluate the effects of vitamin A (Vit A) and probiotic co-supplementation with rabies vaccine on humoral immune response in New Zealand white (NZW) rabbits. For this experiment, 54 rabbits were randomized into six experimental and three control groups. Mixed cultures of commercial probiotics supplements and a dose of Vit A were administered to each animal. Results were compared with the control group fed with only basal diet. Animals in different treatment groups showed significantly higher sero-conversions against rabies vaccine. There was a significant increase (p < 0.001) in the titers of rabies antibodies in all treatment groups on 14th and 35th days than control C3 group. Both commercial probiotics irrespective of brand increase the humoral immune response of rabbits against rabies vaccine. The mean titer values of all groups G1-G6 and sub-controls (C1, C2) were generally above 3.6 EU/ml on day 14th and between 3.7 and 3.9 EU/ml, showing highest sero-conversion on 35th day than mean titer of C3 control = 3.091 and 3.505 EU/ml respectively on both days. The maximum titer values were obtained with the addition of organic carrots to the daily diet. These results suggest that simple dietary interventions using probiotics and Vit A in natural form may enhance the efficacy of rabies vaccine in the host. These cost-effective strategies can be applied for getting higher yields of polyclonal antibody production in animal models, thus providing promising means of improving the final product yield and can be adopted easily by the manufacturers.


Long-Term Evaluation of Endoscopic and Pretrichial Open Forehead Lifts: A Morphometric Analysis.
The aim of this study was to evaluate and compare the long-term (average, 6 years; range, 3 to 11 years) follow-up results of both the endoscopic and pretrichial open forehead lift. Both the amount of eyebrow elevation and patient satisfaction (FACE-Q questionnaires) were analyzed. Preoperative and postoperative eyebrow positions of 65 patients were measured. Both eyebrow sides were measured at three different points. Thirty-two patients completed the FACE-Q questionnaires (10 domains, Dutch translation). Both the endoscopic and the pretrichial open forehead lift raised the eyebrow significantly at all measured points for each eye. This elevation effect was maintained significantly after long-term evaluation (average, 6 years; range 3 to 11 years), with no significant difference reported between techniques. After both procedures, patients were satisfied or strongly satisfied according to the FACE-Q questionnaires. Both the endoscopic forehead lift and the pretrichial open forehead lift raise the eyebrow significantly, and both have a long-term effect with either satisfied or strongly satisfied patients, as evaluated according to the FACE-Q questionnaire. Therapeutic, III.


Identification of epigenetic histone modifications and analysis of histone lysine methyltransferases in Alexandrium pacificum.
Alexandrium pacificum is a toxic dinoflagellate that can cause harmful algal blooms (HABs). The molecular mechanisms of HABs are still poorly understood, especially at the epigenetics level. Organism growth and metabolic processes are affected by histone modifications, an important mode of epigenetic regulation. In this study, various types of modifications, including methylation, acetylation, ubiquitination, and phosphorylation in A. pacificum cells were identified by using pan-antibodies, mass spectrometry, and an H3 modification multiplex assay kit. The modification abundance of H3K4me2 and H3K27me3 of A. pacificum varied under different growth conditions detected by Western blots. A class of SET domain genes (SDGs) encoding histone lysine methyltransferase was analyzed. A total of 179 SDG members were identified in A. pacificum, of which 53 sequences encoding complete proteins were classified into three categories by phylogenetic analysis, conserved domains and motifs analysis. Expression analysis and real-time polymerase chain reaction validation showed that the expressions of some SDGs were significantly influenced by light, nitrogen, phosphorus and manganese supplements. The results revealed that histone lysine methylation played an important role in responding to HABs inducing conditions. This study provided useful information for the further exploration of the role and regulatory mechanism of SDGs in the rapid growth of A. pacificum.


Selective extraction of desmosomal proteins by low ionic strength media.
Desmosomes, isolated using an acidic buffer, have been subjected to extraction at low ionic strength. This treatment removes more than 35% of their protein in the form of two polypeptide chains of molecular weight 210 000 and 230 000, but the desmosomes show only subtle changes in ultrastructure. It is concluded that the use of low ionic strength media for desmosome isolation yields residual structures specifically depleted in high molecular weight proteins.


Risk factors of early recurrence within 6 months after esophagectomy following neoadjuvant chemotherapy for resectable advanced esophageal squamous cell carcinoma.
Esophagectomy following neoadjuvant chemotherapy (NAC) is a standard treatment for resectable advanced esophageal cancer in Japan. However, approximately 10 % of patients with resectable advanced esophageal cancer experience recurrence within 6 months. One hundred twenty-eight patients with resectable advanced esophageal cancer underwent NAC between October 2008 and July 2015 in Kumamoto University Hospital. Among them, 82 patients with esophageal squamous cell carcinoma (SCC), who underwent curative esophagectomy without adjuvant treatment, were eligible. Clinicopathological factors correlated with early recurrence were retrospectively analyzed. Of 82 patients, 14 (17 %) recurred within 6 months after surgery. The logistic regression analysis suggested that CRP before NAC ≥ 0.5 mg/dl [hazard ratio (HR) 33.8, 95 % confidence interval (CI) 2.767-413.9; p = 0.006), presence of poorly differentiated SCC component (HR 138, 95 % CI 5.339-3576; p = 0.003), and pathological vessel invasion (HR 16.3, 95 % CI 1.960-136.1; p = 0.010) were candidates for independent risk factors of early recurrence. Patients with at least two factors frequently recurred (82 %). Of 14 patients with early recurrence, 13 (93 %) had a distant metastasis. Patients with resectable advanced esophageal cancer with at least two factors of CRP before NAC ≥ 0.5 mg/dl, presence of poorly differentiated SCC component, and pathological vessel invasion might be at high risk for early recurrence after esophagectomy following NAC. These patients might be considered for additional treatment and should be meticulously followed up after treatment.


Changes of the DNA packaging mode during boar sperm maturation.
Changes in the mode of DNA packaging in nuclei during spermatogenesis were studied by measuring of the fluorescence anisotropy decay of an ethidium dye intercalated in the DNA in whole nuclei. The nuclei were isolated from boar spermatid or sperm cells at three distinct stages of spermatogenesis: just before the completion of a maturation process in the testis (late spermatid), immediately after a subsequent transformation into spermatozoa (caput spermatozoon), and after full maturation (cauda spermatozoon). Although these three kinds of nucleus were morphologically indistinguishable from each other, the anisotropy decay detected a clear difference. In the late spermatid nuclei, in which the replacement of histones by protamine was still in progress, the anisotropy decayed extensively. The decay suggested that the DNA in the spermatid nuclei contained very flexible regions, in which the interaction of the DNA and proteins may be weak. The rapid and extensive anisotropy decay was absent in the caput and cauda nuclei. The flexible portions must have turned into very rigid structure during transformation from the late spermatid into the caput spermatozoon.


[Conservative treatment of congenital patellar dislocation].
This article presents the rare case of a boy who was born in our hospital with valgus deformity and external rotation of the right lower leg because of congenital patellar dislocation. In the case presented a stable repositioning of the patella could be achieved by redressment with a plaster cast and leg brace. During a 4-year follow-up there were no tendencies towards dislocation during the clinical examination and no dislocation events were documented. In selected cases an attempt at conservative repositioning and retention treatment appears to be worthwhile before surgical treatment is indicated.


Ganoderic acid A/DM-induced NDRG2 over-expression suppresses high-grade meningioma growth.
N-myc downstream-regulated gene 2 (NDRG2) is down-regulated in grade-III meningioma [anaplastic meningioma (AM)] and associated with clinically aggressive behavior. Current therapies in the treatment of high-grade meningioma are lacking with limited success. This study aims to validate the effect of NDRG2-targeted therapy using structurally related bioactive triterpene compounds derived from the edible mushroom Ganoderma lucidum (ganoderic acid A:GA-A/ganoderic acid DM:GA-DM) in human AM in relevant pre-clinical models. Tissue samples from the AM tumor regions of three human patients and control non-tumor samples were used to analyze the expression pattern of NDRG2. In vitro cell culture and in vivo cell-line-derived orthotopic xenograft animal models of AM were utilized to assess efficacy of treatment with GA-A/DM. Downregulation of NDRG2 expression was observed in surgically resected high-grade meningiomas compared to normal brain. These results prompt us to use NDRG2-targeting agents GA-A/DM. In vitro results showed that 72-h treatments of 25 µM GA-A/DM induced AM cell death, upregulate NDRG2 protein expression, downregulate NDRG2 promoter methylation in meningioma cells as compared to azacitidine and decitabine, the most commonly used demethylating agents. Our results also demonstrated that GA-A/DM does not have any detrimental effect on normal human neurons and arachnoid cells. GA-A/DM promoted apoptotic factors (Bax) while suppressing MMP-9, p-P13K, p-AKT, p-mTOR, and Wnt-2 protein expression. RNAi-mediated knockdown of NDRG2 protein expression increased tumor proliferation, while forced expression of wt-NDRG2 decreased proliferation in an in vitro model. Magnetic resonance (MR) imaging and Hematoxylin (H&E) staining demonstrated gross reduction of tumor volume in GA-A/DM treated mice at 5 weeks when compared with saline-treated orthotopic AM xenografted controls. There was an overall decrease in tumor cell proliferation with increased survival in GA-A/DM-treated animals. Enzyme assays showed that GA-A/DM did not negatively impact hepatic function. GA-A/DM may be a promising natural therapeutic reagent in the treatment of AM by suppressing growth via NDRG2 modulation and altering of intracellular signal pathways. We have shown it could potentially be an effective treatment for AM with decreased cellular proliferation in vitro, decreased tumor volume and increased survival in vivo.


Scaling-up antiretroviral treatment in Southern African countries with human resource shortage: how will health systems adapt?
Scaling-up antiretroviral treatment (ART) to socially meaningful levels in low-income countries with a high AIDS burden is constrained by (1) the continuously growing caseload of people to be maintained on long-term ART; (2) evident problems of shortage and skewed distribution in the health workforce; and (3) the heavy workload inherent to presently used ART delivery models. If we want to imagine how health systems can react to such challenges, we need to understand better what needs to be done regarding the different types of functions ART requires, and how these can be distributed through the care supply system, knowing that different functions rely on different rationales (professional, bureaucratic, social) for which the human input need not necessarily be found in formal healthcare supply systems. Given the present realities of an increasingly pluralistic healthcare supply and highly eclectic demand, we advance three main generic requirements for ART interventions to be successful: trustworthiness, affordability and exclusiveness--and their constituting elements. We then apply this analytic model to the baseline situation (no fundamental changes) and different scenarios. In Scenario A there are no fundamental changes, but ART gets priority status and increased resources. In Scenario B the ART scale-up strengthens the overall health system: we detail a B1 technocratic variant scenario, with profoundly re-engineered ART service production, including significant task shifting, away from classical delivery models and aimed at maximum standardisation and control of all operations; while in the B2 community-based variant scenario the typology of ART functions is maximally exploited to distribute the tasks over a human potential pool that is as wide as possible, including patients and possible communities. The latter two scenarios would entail a high degree of de-medicalisation of ART.


[Reverse total shoulder arthroplasty for the treatment of comminuted fracture of proximal humerus in elderly patients with rotator cuff injury].
To investigate clinical results of reverse total shoulder arthroplasty in treating comminuted fracture of proximal humerus in elderly patients with rotator cuff injury. From January 2017 to December 12, 12 comminuted fracture or dislocation of proximal humerus elderly patients were diagnosed as rotator cuff injury by preoperative MRI and operative exploration, and treated by reverse total shoulder arthroplasty. Among them, including 7 males and 5 females aged from 65 to 86 years old; 5 patients injured on the left side and 7 patients injured on the right side. Range of motion, postoperative complication were observed, VAS score was used to evaluate pain release and UCLA score was used to assess recovery of shoulder joint. All patients were followed up from 8 to 18 months. At the latest follow-up, shoulder range of motion conditions were as following:forward bend and lifts ranged from 90° to 150°, external rotation ranged from 10°to 30°, internal rotation could reached L₃ level(S₁-L₁), VAS was for 0 to 6 points, UCLA score ranged from 18 to 32 points, 5 patients were good and 7 patients were poor. No infection, prothesis loosening, shoulder stress fracture, injury of vessel and nerve occurred. Reverse total shoulder arthroplasty in treating comminuted fracture of proximal humerus in elderly patients with rotator cuff injury has advantages of early recovery of shoulder joint range of motion, less pain and high patients' satisfactory.


Age differences in the safety and effectiveness of the HEART Pathway accelerated diagnostic protocol for acute chest pain.
The HEART Pathway is a validated protocol for risk stratifying emergency department (ED) patients with possible acute coronary syndrome (ACS). Its performance in different age groups is unknown. The objective of this study is to evaluate its safety and effectiveness among older adults. A pre-planned subgroup analysis of the HEART Pathway implementation study was conducted. This prospective interrupted time series accrued adult ED patients with possible ACS who were without ST-elevation across three US sites from 11/2013-01/2016. After implementation, providers prospectively used the HEART Pathway to stratify patients as low-risk or non-low-risk. Patients were classified as older adults (≥65 years), middle-aged (46-64 years), and young (21-45 years). Primary safety and effectiveness outcomes were 30-day death or MI and hospitalization at 30 days, determined from health records, insurance claims, and death index data. Fisher's exact test compared low-risk proportions between groups. Sensitivity for 30-day death or MI and adjusted odds ratios (aORs) for hospitalization and objective cardiac testing were calculated. The HEART Pathway implementation study accrued 8474 patients, of which 26.9% (2281/8474) were older adults, 45.5% (3862/8474) middle-aged, and 27.5% (2331/8474) were young. The HEART Pathway identified 7.4% (97/1303) of older adults, 32.0% (683/2131) of middle-aged, and 51.4% (681/1326) of young patients as low-risk (p < 0.001). The HEART Pathway was 98.8% (95% CI 97.1-100) sensitive for 30-day death or MI among older adults. Following implementation, the rate of 30-day hospitalization was similar among older adults (aOR 1.25, 95% CI 1.00-1.55) and cardiac testing increased (aOR 1.25, 95% CI 1.04-1.51). The HEART Pathway identified fewer older adults as low-risk and did not decrease hospitalizations in this age group.


Successful Treatment of Refractory Hypoxemia Secondary to Disseminated Histoplasmosis Using Extracorporeal Membrane Oxygenation Support.
Refractory hypoxemia secondary to acute respiratory distress syndrome (ARDS) is associated with high mortality. Extracorporeal membraneoxygenation (ECMO) is an accepted strategy for treating refractory hypoxemia in patients with ARDS but is relatively contraindicated in the setting of systemic infections. We present a case of successful ECMO use in a host with refractory hypoxemia secondary to disseminated histoplasmosis with fungemia and discuss our management approach to this difficult patient.


Vitamin A concentration in amniotic fluid and maternal serum related to neural-tube defects.
The vitamin A concentration of amniotic fluid and maternal serum was measured during the second trimester of pregnancy in 106 women, 12 of whom had a baby with a neural-tube defect. In these 12 pregnancies the amniotic fluid vitamin A concentration was significantly higher than in 94 normal pregnancies. There was a highly significant correlation between amniotic fluid vitamin A and both zinc and alpha-fetoprotein (AFP) levels. The maternal serum vitamin A levels were also significantly related to serum zinc levels. Women with a raised serum AFP level, but a normal baby, had significantly higher amniotic fluid vitamin A levels and significantly lower serum vitamin A levels compared with those in women with normal serum AFP levels.


Utilization of cyanobacteria in photobioreactors for orthophosphate removal from water.
The effectiveness of photosynthetic free-living and polyurethane foam (PU) immobilized Anabaena variabilis cells for removal of orthophosphate (P) from water in batch cultures and in a photobioreactor was studied. Immobilization in PU foams was found to have a positive effect on P uptake by cyanobacteria in batch cultures. The efficiency of P uptake by immobilized cells was higher than by free-living cells. A laboratory scale photobioreactor was constructed for removal of P from water by the immobilized cyanobacteria. The photobioreactor was designed so that the growth medium (water) from a reservoir was pumped through a photobioreactor column with immobilized cyanobacteria and back to the reservoir. This created a closed system in which it was possible to measure P uptake. No leakage of cells into the photobioreactor medium reservoir was observed during the operation. The immobilized cells incorporated into a photobioreactor column removed P continuously for about 15 d. No measurable uptake was demonstrated after this period. Orthophosphate uptake efficiency of 88-92% was achieved by the photobioreactor.


Sequencing batch-reactor control using Gaussian-process models.
This paper presents a Gaussian-process (GP) model for the design of sequencing batch-reactor (SBR) control for wastewater treatment. The GP model is a probabilistic, nonparametric model with uncertainty predictions. In the case of SBR control, it is used for the on-line optimisation of the batch-phases duration. The control algorithm follows the course of the indirect process variables (pH, redox potential and dissolved oxygen concentration) and recognises the characteristic patterns in their time profile. The control algorithm uses GP-based regression to smooth the signals and GP-based classification for the pattern recognition. When tested on the signals from an SBR laboratory pilot plant, the control algorithm provided a satisfactory agreement between the proposed completion times and the actual termination times of the biodegradation processes. In a set of tested batches the final ammonia and nitrate concentrations were below 1 and 0.5 mg L(-1), respectively, while the aeration time was shortened considerably.


Genome-wide identification of seven superoxide dismutase genes in the marine rotifer Brachionus rotundiformis and modulated expression and enzymatic activity in response to microplastics and nutritional status.
Microplastics (MPs) pollution has attracted worldwide attention. Superoxide dismutase (SOD) is a sensitive indicator for assessing the toxic effects of MPs in aquatic organisms. However, few studies have been performed to identify all genes encoding SOD in aquatic invertebrates. Especially, effects of MPs on SOD activity and expression in aquatic organisms under starvation or a subsequent refeeding status are unclear. In the present study, all full-length genes encoding SOD were cloned and characterized from the marine rotifer Brachionus rotundiformis, including CuZnSOD1, CuZnSOD2, CuZnSOD3, CuZnSOD4, CuZnSOD5, MnSOD1, and MnSOD2. The CuZnSOD1, CuZnSOD2 and MnSOD2 are homologous to SODs from vertebrates and the other SOD proteins are rotifer-specific according to the results from the phylogenetic tree. The conserved signature sequences and binding sites of Cu2+, Zn2+and Mn2+ were also identified in the seven SOD proteins. Compared with feeding, starvation down-regulated SOD activity and mRNA expression of CuZnSOD2, CuZnSOD4, CuZnSOD5, MnSOD1 and MnSOD2 while refeeding maintained SOD activity comparable to the feeding level and up-regulated CuZnSOD5 and MnSOD2. Intake of MPs by B. rotundiformis was observed by examining fluorescence signals from the fluorescently-labeled microplastics under different nutritional status. Exposure to MPs reduced rotifer density and increased malondialdehyde (MDA) content and SOD activity in the rotifers under the refeeding condition, but did not affect these indicators under the feeding and starvation conditions. However, mRNA expression of some tested genes was responsive to MPs in the fed, starved and refed rotifers. The present study for the first time demonstrated a nutritional status-dependent effect of MPs on oxidative stress response, and provided more sensitive molecular biomarkers for assessing the toxicity of MPs using B. rotundiformis as a model animal.


A phenomenological base semi-physical thermodynamic model for the cylinder and exhaust manifold of a natural gas 2-megawatt four-stroke internal combustion engine.
This paper presents the application of a systematic methodology to obtain a semi-physical model of phenomenological base for a 2 MW internal combustion engine to generate electric power operating with natural gas, as a function of the average thermodynamic value normally measured in industrial applications. Specifically, the application of the methodology is focused on the cylinders, exhaust manifold, and turbocharger turbine sections. The proposed model was validated with actual operating data, obtaining an error rate not exceeding 5%, which allow a thermal characterization of the Jenbacher JMS 612 GS-N based on the model. A parametric analysis is conducted; considering the volumetric efficiency, the output electric power, the effective efficiency, the exhaust gas temperature, the turbine mass flow, the specific fuel consumption under the nominal operation conditions, which is 1.16 bar in the gas pressure, 65 °C in the cooling water temperature, 35 °C in the average ambient temperature, and 1500 rpm. The results of this model can be used to evaluate the thermodynamic performance parameters of waste heat recovery systems. On the other hand, new control strategies and the implementation of state observers for the detection and diagnosis of failures can be developed based on the proposed model.


Sensory substance P innervation of the stomach and pancreas. Demonstration of capsaicin-sensitive sensory neurons in the rat by combined immunohistochemistry and retrograde tracing.
The sensory innervation of the stomach and pancreas has been identified by retrograde tracing using the fluorescent dye True Blue (Illing, Gross-Umstadt, Federal Republic of Germany), coupled with the immunohistochemical localization of substance P. Labeled cells were visualized in spinal ganglia, nodose ganglia, celiac ganglion, and dorsal motor nucleus of the vagus after injection of True Blue into both stomach and pancreas. Substance P immunoreactivity was found in 35%-50% of gastric spinal afferent neurons and in approximately 15% of pancreatic spinal afferents. In rats treated at birth with the sensory neurotoxin capsaicin there was a reduction of about 70% in True Blue-labeled cells in the spinal and nodose ganglia, and virtually complete loss of substance P in these ganglia. There was also a marked depletion of substance P-immunoreactive fibers in the pancreas, and in the submucosa of the stomach. The results suggest that the substance P-containing spinal afferents that project to the gastric submucosa are an important component of the gastric sensory innervation.


Detection of mycoplasma in leukemia-lymphoma cell lines using polymerase chain reaction.
The specific, sensitive and reliable detection of mycoplasma contamination in cell cultures is an important part of mycoplasma control. We have sought to develop and validate a method for mycoplasma detection which is sensitive and accurate, but also practical in the sense of time spent, costs, and applicability in the standard laboratory; finally, the method should be suitable for screening large numbers of test specimens. To that end, we adapted a previously developed polymerase chain reaction (PCR) method for daily routine application. This single-step PCR uses a mixture of primers annealing to gene sequences coding for evolutionarily conserved 16S rRNA of different mycoplasma species, including the ones most commonly found in cell cultures. An internal control was introduced to exclude any false-negative tests resulting from technical PCR problems. This mycoplasma detection by PCR has been validated prospectively on 201 consecutive leukemia-lymphoma cell lines received at the institute over a 3-year period and on 118 initially positive cell lines after anti-mycoplasma treatment with antibiotics. The sensitivity (detection of true positives) of this PCR detection assay was 96% and the specificity (detection of true negatives) was also 96%, with positive and negative predictive values (probability of correct result) of 86% and 99%, respectively. PCR defined the mycoplasma status with 96% accuracy (detection of true positives and true negatives). Besides the high sensitivity and specificity, further attractive features of the PCR approach are the ease and speed with which large numbers of specimens can be tested. PCR mycoplasma analysis provides a readily available, quick and reliable test system with which to manage the important issue of mycoplasma contamination of cell lines.


[The pharmacological effects of hippocampus capsule on enhancing sexual functions of rats].
In this paper, the effects of marine health food Hippocampus Capsule on the intercoursing ability of sexually-mature rats were studied. The results showed that Hippocampus Capsule could enhance the intercoursing ability of sexually-mature male rats, shorten the latency of males catching females and ejeculation after their combination. The catching rate, intercoursing rate and the ejeculation status was markedly different from the blank control group (P < 0.05). Therefore, Hippocampus Capsule can markedly enhance the intercoursing ability of the male rats and the sexual function.


Prediction of Vitamin A Stores in Young Children Provides Insights into the Adequacy of Current Dietary Reference Intakes.
Limited data were available in infants and children when vitamin A (VA) DRIs were established; recommendations were developed based on average breast milk VA intake and extrapolation of data from adults. Our objective was to evaluate whether DRIs and reported intakes, with and without VA from intervention programs, would be sufficient to develop adequate VA stores from birth to age 5 y in Bangladeshi, Filipino, Guatemalan, and Mexican children. A mathematical relationship was established, defined by a series of equations, to predict VA total body stores (TBS) as a function of age based on VA intake and utilization. TBS calculated using reported VA intakes, with and without additional VA from intervention programs, were compared to those predicted using DRIs (specifically, Adequate Intake and RDA). Liver VA concentrations were also estimated. Our predictions showed that for these 4 groups, DRIs were sufficient to attain liver VA concentrations >0.07 μmol/g by 1 wk of age and sustain positive VA balance for 5 y. Using reported intakes, which were lowest in Bangladeshis from 1 y on and highest in Guatemalans, predicted VA stores in Bangladeshi and Filipino children increased until ∼2-3 y, then TBS stabilized and liver VA concentrations decreased with age. When VA interventions were included, stores exceeded those predicted using DRIs by 12-18 mo. In contrast, reported intakes alone in Guatemalan and Mexican children resulted in VA stores that surpassed those calculated using DRIs. For all populations, reported intakes were sufficient to build liver concentrations >0.07 μmol/g by 3 mo. Although more information is needed to better define dietary VA requirements in children, our results suggest that for an average, generally healthy child in a low- or middle-income country, current DRIs are sufficient to maintain positive VA balance during the first 5 y of life.


Activation of anti-Trypanosoma cruzi drugs to genotoxic metabolites promoted by mammalian microsomal enzymes.
Salmonella typhimurium TA100 and its nitroreductase-deficient derivative, TA100 NR, were used to reevaluate the mutagenic activities of benznidazole and nifurtimox. Mutagenicity and toxicity of nifurtimox were abolished in the TA100 NR tester strain under aerobic or anaerobic conditions and addition of rat liver extracts did not alter the results. However, benznidazole showed a significant mutagenicity and toxicity to the nitroreductase-deficient strain TA100 NR under hypoxic conditions. Addition of rat liver extracts enhanced the observed mutagenicity and toxicity of benznidazole even more. In the presence of O2 the genotoxic activities of benznidazole to the TA100 NR tester strain were eliminated. These results lead us to conclude that bacterial enzymes were responsible for the previously observed genotoxic effects of nifurtimox and benznidazole on S. typhimurium TA100. Moreover, under anaerobic conditions, only benznidazole could be metabolized by mammalian nitroreductases into a mutagenic derivative.


Surgical approach to l-dopa-induced dyskinesias.
Many patients treated chronically with l-dopa for Parkinson disease (PD) become functionally disabled by l-dopa-induced dyskinesias (LID). Evolved from early empirical procedures, modern stereotactic surgical lesioning techniques and deep brain stimulation (DBS) can effectively treat LID while simultaneously improving the cardinal motor symptoms of PD. Here we review the common surgical targets used to treat LID, and compare their relative efficacy. We explain the anti-dyskinetic action of surgery at each of these targets based on evolving models of basal ganglia function. Finally, we discuss the appropriate selection of patients with LID for surgery and address relevant technical and management issues in these patients.


Heavy-chain antibodies in Camelidae; a case of evolutionary innovation.
The emergence in Camelidae species of functional antibodies devoid of light chains (referred to as heavy-chain antibodies or HCAbs) is an intriguing evolutionary event. Homodimeric HCAbs have also been documented in spotted ratfish ( Cos5-Abs) and nurse shark (NAR). To reveal the evolutionary history of HCAbs, we evaluated the phylogenetic and phenotypic relationships among HCAbs and conventional antibodies across taxa and confirmed the current viewpoint that different groups of HCAbs have evolved independently in the three lineages. At least, in the camelids, HCAbs are not the result of resuscitation of dormant genes. They are derived from the conventional antibodies within the Camelidae lineage, and are apparently the outcome of more recent adaptive changes occurring in the compartment of heteromeric antibodies. The shared structural properties of HCAbs across taxa are therefore explained by convergent evolution due to similar constraints related to the absence of pairing to the light chain. It appears that innovative evolutionary changes in Camelidae have led to a new level of antigen binding repertoire diversification and have allowed acquisition of novel antigen-receptor properties.


Uptake characteristics of galactosylated emulsion by HepG2 hepatoma cells.
Galactosylated (Gal) emulsions containing various molar ratios of cholesten-5-yloxy-N-(4-((1-imino-2-D-thiogalactosylethyl)amino)butyl)formamide (Gal-C4-Chol) as a ligand for asialoglycoprotein receptors were prepared to study the effect of the galactose content of Gal-emulsions labeled with [3H]cholesteryl hexadecyl ether on their targeted delivery to hepatocytes. The uptake characteristics of Gal-emulsions having Gal-C4-Chol of 1, 3, 4, 6, and 9 mol% were evaluated in HepG2 cells which possess asialoglycoprotein receptors and NIH3T3 cells which are lack of asialoglycoprotein receptors. The uptake and internalization by HepG2 cells was enhanced by the addition of Gal-C4-Chol to the Gal-emulsions whereas the uptake of Gal-emulsions by NIH3T3 cells was not much and was comparable with that of bare-emulsions. In the presence of excess Gal-BSA, the uptake of Gal-emulsions having Gal-C4-Chol of 4, 6, and 9% was inhibited suggesting asialoglycoprotein receptor mediated uptake. Moreover, Gal-emulsions having Gal-C4-Chol of 4, 6, and 9% showed a slight increase in surface binding and exhibited extensive uptake and internalization into HepG2 cells. The present study strongly suggested that the Gal-emulsions are taken up by the asialoglycoprotein receptor-mediated endocytosis and galactose density of Gal-emulsions is important for effective recognition and cell internalization.


Patient-provider language concordance and colorectal cancer screening.
Patient-provider language barriers may play a role in health-care disparities, including obtaining colorectal cancer (CRC) screening. Professional interpreters and language-concordant providers may mitigate these disparities. DESIGN, SUBJECTS, AND MAIN MEASURES: We performed a retrospective cohort study of individuals age 50 years and older who were categorized as English-Concordant (spoke English at home, n = 21,594); Other Language-Concordant (did not speak English at home but someone at their provider's office spoke their language, n = 1,463); or Other Language-Discordant (did not speak English at home and no one at their provider's spoke their language, n = 240). Multivariate logistic regression assessed the association of language concordance with colorectal cancer screening. Compared to English speakers, non-English speakers had lower use of colorectal cancer screening (30.7% vs 50.8%; OR, 0.63; 95% CI, 0.51-0.76). Compared to the English-Concordant group, the Language-Discordant group had similar screening (adjusted OR, 0.84; 95% CI, 0.58-1.21), while the Language-Concordant group had lower screening (adjusted OR, 0.57; 95% CI, 0.46-0.71). Rates of CRC screening are lower in individuals who do not speak English at home compared to those who do. However, the Language-Discordant cohort had similar rates to those with English concordance, while the Language-Concordant cohort had lower rates of CRC screening. This may be due to unmeasured differences among the cohorts in patient, provider, and health care system characteristics. These results suggest that providers should especially promote the importance of CRC screening to non-English speaking patients, but that language barriers do not fully account for CRC screening rate disparities in these populations.


[Urinary NAG activity and diabetic microangiopathy].
The behaviour of urinary NAG was monitored in diabetic patients some of whom presented more or less severe vascular alterations. The results showed a relationship between urinary NAG levels and the severity of the vascular condition (in fact the greatest NAG activity was found in patients with overt kidney damage). NAG may therefore reveal the existence of a vascular nephropathy that is still too slight to be identified by the standard non-invasive clinical examinations.


Prevalence and incidence of multiple sclerosis in Ardabil, Northwest of Iran.
Multiple sclerosis (MS) epidemiology is studies in many populations; however, studying populations with unique characteristics could provide opportunities to deepen the understanding of the underlying reason of the disease. In this regard, we aimed to study the epidemiology of MS in Ardabil, a province in northwest Iran, where the majority are of Iranian Azerbaijanis. A retrospective population-based study was conducted from 2008 to 2018 in Ardabil, based on the data of Iran's Ministry Of Health. Collected information includes sex, age, age at disease onset, education and type of MS. We used t-test to compare means and chi-square test to analyze the association among variables. The total number of patients was 760 with 533 (70.13%) females and 227 (29.87%) males. The crude prevalence was 59.37 per 100,000 in 2018 (95% CI: 55.31, 63.73). The crude incidence rate was 7.65 per 100,000 in 2018 (95% CI: 6.28, 9.32). The most frequent educational level was high school diploma (38.36%). The relapsing-remitting (RR) form was the most frequent type of MS (48.16%). The F/M ratio was 2.92:1 and the mean onset age was 33.14 (95% CI: 31.56, 34.72) in 2018, significantly higher in males (P value= 0.01). Ardabil is a medium risk zone of MS. The different ethnicity of its population, climate and environmental features, may explain the factors modulating the risk of MS in similar areas and present windows of opportunity to understand the causes of MS.


Indonesia free from pasung: a policy analysis.
Many people with mental illnesses remain isolated, chained, and inside cages, called Pasung in Indonesia. Despite numerous policies introduced to eradicate Pasung, Indonesia has made slow progress in decreasing this practice. This policy analysis examined existing policies, plans and initiatives in Indonesia targeted at eradicating Pasung. Policy gaps and contextual constraints are identified in order to propose stronger policy solutions. Eighteen policy documents were examined, including government news releases and organisational archives. A content analysis was undertaken of national-level policies that address Pasung within the context of the health system, social system and human rights since the establishment of Indonesia. This was followed by a case study analysis of policy and program responses particularly in West Java Province. While policy to address Pasung exists at a national level, implementation at national and local levels is complicated. Pasung policy has generated a sense of awareness but the different directions and ambiguous messaging across all stakeholders, including policy actors, has created a lack of clarity about institutions' roles and responsibilities in the implementation process, as well as accountability for outcomes. This situation is exacerbated by an incomplete decentralisation of healthcare policymaking and service delivery, particularly at the primary level. It is possible that policymakers have overlooked international obligations and lessons learned from successful policymaking in comparable regional countries, resulting in disparities in target-setting, implementation mechanisms, and evaluation. While the public has become more informed of the need to eradicate Pasung, ongoing communication with the various clusters of policy actors on the aforementioned issues will be critical. Addressing the various segments of the policy actors and their challenges in response to policy will be critical as part of building the evidence base to establish a feasible and effective policy to combat Pasung in Indonesia.


Effectiveness of complementary and self-help treatments for anxiety in children and adolescents.
To review the evidence for the effectiveness of complementary and self-help treatments for anxiety disorders and situational anxiety in children and adolescents. Systematic literature search using PubMed, PsycINFO and the Cochrane Library for 111 treatments up to February 2006. There were 11 treatments for which intervention studies had been undertaken and reported. Studies on each treatment were reviewed by one author and checked by a second. A consensus was reached for level of evidence. Relevant evidence was available for bibliotherapy, dance and movement therapy, distraction techniques, humour, massage, melatonin, relaxation training, autogenic training, avoiding marijuana, a mineral-vitamin supplement (EMPower +) and music therapy. Findings from case-control studies, individual cohort studies or low quality randomised controlled trials indicated that several treatments may have potential to reduce anxiety, including bibliotherapy, massage, melatonin, and relaxation training. Although some complementary and self-help treatments might be useful for children and adolescents with anxiety, they need to be tested adequately through randomised controlled trials before they could be recommended.


Detection and analysis of graduate students' academic emotions in the online academic forum based on text mining with a deep learning approach.
The possibility of mental illness caused by the academic emotions and academic pressure of graduate students has received widespread attention. Discovering hidden academic emotions by mining graduate students' speeches in social networks has strong practical significance for the mental state discovery of graduate students. Through data collected from online academic forum, a text based BiGRU-Attention model was conducted to achieve academic emotion recognition and classification, and a keyword statistics and topic analysis was performed for topic discussion among graduate posts. Female graduate students post more than male students, and graduates majoring in chemistry post the most. Using the BiGRU-Attention model to identify and classify academic emotions has a performance with precision, recall and F1 score of more than 95%, the category of PA (Positive Activating) has the best classification performance. Through the analysis of post topics and keywords, the academic emotions of graduates mainly come from academic pressure, interpersonal relationships and career related. A BiGRU-Attention model based on deep learning method is proposed to combine classical academic emotion classification and categories to achieve a text academic emotion recognition method based on user generated content.


Dietary hawthorn-leaves flavonoids improves ovarian function and liver lipid metabolism in aged breeder hens.
Hawthorn-leaves flavonoids (HF), extracted from hawthorn leaves, were reported to exert antioxidant, anti-inflammatory and hypolipidemic properties. The aim of our study was to investigate the effects of dietary HF on the reproduction performance and liver lipid metabolism of aged breeder hens. A total of 270 aged Qiling breeder hens (60-wk-old) were randomly divided into 3 treatments: 1) basic corn-soybean diet (CON); 2) basic corn-soybean diet supplemented with 30 mg/kg HF (LHF); 3) basic corn-soybean diet supplemented with 60 mg/kg HF (HHF). The results showed that supplemented HF significantly improved the egg-laying rate and hatching rate of aged breeder hens (P < 0.05). HF treatment reduced the serum TG, T-CHO and L-LDL levels (P < 0.05), and upregulated the mRNA expressions of ESR1, ESR2, VTGⅡ, ApoB, and ApoVI in the liver (P < 0.05). Serum estrogen levels in HF treated groups were elevated compared with the CON group (P < 0.05). In the HHF group, the number of the primordial follicles was higher in comparison with the CON group (P < 0.05). Furthermore, dietary supplementation with HF improved the activity of antioxidant enzymes (T-AOC, GSH-Pχ) (P < 0.05), following with the reversed ovarian apoptosis and morphological damage. In addition, 60 mg/kg dietary HF upregulated the protein expression of PCNA and Nrf2 in the ovary (P < 0.05). In summary, dietary supplementation with HF could improve the reproduction performance through regulating liver lipid metabolism and improving ovarian function in aged breeder hens.


Raman and infrared spectroscopic characterization of the phosphate mineral paravauxite Fe2+Al2(PO4)2(OH)2.8H2O.
We have undertaken a vibrational spectroscopic study of paravauxite the Siglo XX mine, Bustillo Province, northern of Potosí department, Bolivia. This mine is important source for rare and unusual secondary phosphate minerals and is the type locality for a number of rare phosphates such as vauxite, sigloite, metavauxite and for jeanbandyite. The chemical formula of the studied sample was determined as Fe(2+)(0.9)5, Al(0.07)Σ1.02 (Al)2.09 (PO4)1:97 (OH)1.98 · 7.90(H2O). The Raman spectrum is dominated by an intense Raman band at 1020 cm(-1) assigned to the PO4(3-) ν1 symmetric stretching mode. Low intensity Raman bands found at 1058, 1115 and 1148 cm(-1) are assigned to the PO4(3-) ν3 antisymmetric stretching vibrations. Raman bands of paravauxite at 537, 570, 609 and 643 cm(-1) are assigned to the ν4 PO4(3-) bending modes whilst the Raman bands at 393 and 420 cm(-1) are due to the ν2 PO4(3-) bending modes. The Raman spectral profile of paravauxite in the hydroxyl stretching region is broad with component bands resolved at 3086, 3215, 3315, 3421, 3505 and 3648 cm(-1). Vibrational spectroscopy enables the assessment of the molecular structure of paravauxite to be undertaken.


Dissolving microneedles for alopecia treatment.
Alopecia is a treatable benign disease, however, approximately 15-30% of women and 50% of men suffer from alopecia, which greatly affects patient's self-esteem and quality of life. Currently, commercial products for alopecia treatment include topical minoxidil solution, oral finasteride tablets and oral baricitinib tablets. However, the barrier of stratum corneum, systemic adverse effects and poor cure rate limit the application of commercial products. Therefore, researchers investigated the mechanism of alopecia, and developed new drugs that could target lactate dehydrogenase-related pathways, remove excessive reactive oxygen in hair follicles, and reduce the escape of hair follicle stem cells, thus injecting new strength into the treatment of alopecia. Moreover, starting from improving drug stratum corneum penetration and reducing side effects, researchers have developed hair loss treatment strategies based on dissolved microneedles (MNs), such as drug powders/microparticles, nanoparticles, biomimetic cell membranes, phototherapy and magnetically responsive soluble microneedles, which show exciting alopecia treatment effects. However, there are still some challenges in the practical application of the current alopecia treatment strategy with soluble microneedles, and further studies are needed to accelerate its clinical translation.


Studies on the properties of triphosphoinositide phosphomonoesterase and phosphodiesterase of rabbit iris smooth muscle.
The rabbit iris smooth muscle has been shown to contain triphosphoinositide phosphomonoesterase (phosphatidyl-myo-inositol-4,5-bisphosphate phosphohydrolase, EC 188.8.131.52) and phosphodiesterase (triphosphoinositide inositoltrisphosphohydrolase, EC 22.214.171.124) activities. Under our experimental conditions about 77% of the phosphomonoesterase and 61% of the phosphodiesterase activities were localized in the particulate fraction. The kinetic properties of the enzymes in the microsomal fraction were examined. The enzyme preparation was specific to polyphosphoinositides; it did not attack phosphatidylinositol under the present assay condition. The effects of Ca2+ and Mg2+ were also studied. Although the microsomal enzymes did not require added divalent cations for their activities, both the phosphomonoesterase and phosphodiesterase were appreciably inhibited by 1 mM EDTA. Phosphodiesterase and phosphomonoesterase were stimulated by Ca2+ and Mg2+, respectively. The demonstration of triphosphoinositide phosphodiesterase in the iris muscle, coupled with the findings that this enzyme is activated by Ca2+ and is not influenced by acetylcholine add further support to our previous conclusion (J. Pharmacol. Exp. Ther. (1978) 204, 655--668; J. Neurochem. (1978) 30, 517--525) that an increased Ca2+ influx, following the interaction between the neurotransmitter and its receptor, could act to stimulate the phosphodiesterase, thus leading to increased triphosphoinositide breakdown and increased phosphatidic acid via increased diacylglycerol.


Every child, every day, back to play: the PICUstars protocol - implementation of a nurse-led PICU liberation program.
As admissions to paediatric intensive care units (PICU) rise and mortality rates decline, the focus is shifting from survival to quality of survivorship. There is paucity of internationally accepted guidelines to manage complications like over-sedation, delirium, and immobility in the paediatric setting. These have a strong adverse impact on PICU recovery including healthcare costs and long-term functional disability. The A2F bundle (ABCDEF), or ICU Liberation, was developed to operationalise the multiple evidence-based guidelines addressing ICU-related complications and has been shown to improve clinical outcomes and health-care related costs in adult studies. However, there is little data on the effect of ICU Liberation bundle implementation in PICU. PICU-STARS will be a single centre before-and-after after trial and implementation study. It is designed to evaluate if the multidimensional, nurse-led ICU Liberation model of care can be applied to the PICU and if it is successful in minimising PICU-related problems in a mixed quaternary PICU. In a prospective baseline measurement, the present practises of care in the PICU will be assessed in order to inform the adaptation and implementation of the PICU Liberation bundle. To assess feasibility, implementation outcomes, and intervention effectiveness, the implementation team will use the Consolidated Framework for Implementation Research (CIFR) and process assessment (mixed methods). The implementation process will be evaluated over time, with focus groups, interviews, questionnaires, and observations used to provide formative feedback. Over time, the barriers and enablers for successful implementation will be analysed, with recommendations based on "lessons learned." All outcomes will be reported using standard descriptive statistics and analytical techniques, with appropriate allowance for patient differentials in severity and relevant characteristics. The results will inform the fine-tune of the Liberation bundle adaptation and implementation process. The expected primary output is a detailed adaptation and implementation guideline, including clinical resources (and investment) required, to adopt PICU-STARS in other children's hospitals. The authors thank the PICU education and Liberation Implementation team, and our patients and families for their inspiration and valuable comments on protocol drafts. Results will be made available to critical care survivors, their caregivers, relevant societies, and other researchers. ACTRN, ACTRN382863 . Registered 19/10/2021 - Retrospectively registered. recruiting.


Prevention of chronic allograft nephropathy with vitamin D.
Chronic allograft nephropathy (CAN) is the leading cause of late allograft loss in kidney transplantation. Interstitial fibrosis and glomerulosclerosis are characteristic of CAN. Transforming growth factor beta-1 (TGFbeta-1) is associated with both of these histologic findings in the transplant setting. Recent studies have suggested that vitamin D signaling pathways may interact with and regulate TGFbeta-1 mediated events. We examined the efficacy of 1,25-dihydroxyvitamin D(3), the active metabolite of vitamin D [1,25-(OH)(2)D(3)], the active metabolite of vitamin D, as monotherapy to prolong allograft survival and preserve renal function in a rat model of CAN, the Fisher 344 to Lewis model. Recipients went without treatment or were treated with cyclosporine A (CSA; 10 days) or 1,25(OH)(2)D(3) (1000, 500 or 250 ng/kg/day). Grafts were harvested at the time of rejection or at 24 weeks post-transplant. A portion of the graft was processed for histology and immunohistochemistry and a second portion was analyzed for protein expression by western blotting. Not only did 1,25-(OH)(2)D(3) treatment significantly prolong graft survival, but it also prevented histological changes associated with CAN. 1,25-(OH)(2)D(3) treatment significantly decreased Smad 2 expression. This TGFbeta signaling molecule is likely involved in fibrosis. Moreover, 1,25-(OH)(2)D(3) treatment increased Smad 7 expression, an important feedback molecule in the TGFbeta-1 signaling pathway. This suggests that 1,25-(OH)(2)D(3) interacts with TGFbeta-1 in limiting histological injury in this model of CAN. Furthermore, 1,25-(OH)(2)D(3), treatment increased expression of matrix metalloproteinase 2 (MMP-2), thus directly affecting levels of another important matrix molecule. Taken together our data suggests that 1,25-(OH)(2)D(3) mitigates CAN in this model by altering TGFbeta-1 and matrix-regulating molecules.


Evaluation of stability of cefuroxime axetil in solid state.
The effect of temperature and relative atmospheric humidity on the stability of the crystalline form of cefuroxime axetil (CFA) in solid state was investigated. CFA is a mixture of diastereomers A and B. Changes in the concentration of the two diastereomers (A and B) of CFA were recorded by means of HPLC with UV detection. The degradation of diastereomers of CFA occurring at 0% relative humidity (RH) of the ambient air is a reversible first order reaction, while that occurring in humid air (RH>50%) is an autocatalytic first order reaction relative to substrate concentration. Although it has been found, that diastereomer B is the more stable isomer, humidity has a stronger effect on this very diastereomer.


Automatic multispectral MRI segmentation of human hippocampal subfields: an evaluation of multicentric test-retest reproducibility.
Accurate and reproducible automated segmentation of human hippocampal subfields is of interest to study their roles in cognitive functions and disease processes. Multispectral structural MRI methods have been proposed to improve automated hippocampal subfield segmentation accuracy, but the reproducibility in a multicentric setting is, to date, not well characterized. Here, we assessed test-retest reproducibility of FreeSurfer 6.0 hippocampal subfield segmentations using multispectral MRI analysis pipelines (22 healthy subjects scanned twice, a week apart, at four 3T MRI sites). The harmonized MRI protocol included two 3D-T1, a 3D-FLAIR, and a high-resolution 2D-T2. After within-session T1 averaging, subfield volumes were segmented using three pipelines with different multispectral data: two longitudinal ("long_T1s" and "long_T1s_FLAIR") and one cross-sectional ("long_T1s_FLAIR_crossT2"). Volume reproducibility was quantified in magnitude (reproducibility error-RE) and space (DICE coefficient). RE was lower in all hippocampal subfields, except for hippocampal fissure, using the longitudinal pipelines compared to long_T1s_FLAIR_crossT2 (average RE reduction of 0.4-3.6%). Similarly, the longitudinal pipelines showed a higher spatial reproducibility (1.1-7.8% of DICE improvement) in all hippocampal structures compared to long_T1s_FLAIR_crossT2. Moreover, long_T1s_FLAIR provided a small but significant RE improvement in comparison to long_T1s (p = 0.015), whereas no significant DICE differences were found. In addition, structures with volumes larger than 200 mm3 had better RE (1-2%) and DICE (0.7-0.95) than smaller structures. In summary, our study suggests that the most reproducible hippocampal subfield FreeSurfer segmentations are derived from a longitudinal pipeline using 3D-T1s and 3D-FLAIR. Adapting a longitudinal pipeline to include high-resolution 2D-T2 may lead to further improvements.


Threat learning impairs subsequent associative inference.
Despite it being widely acknowledged that the most important function of memory is to facilitate the prediction of significant events in a complex world, no studies to date have investigated how our ability to infer associations across distinct but overlapping experiences is affected by the inclusion of threat memories. To address this question, participants (n = 35) encoded neutral predictive associations (A → B). The following day these memories were reactivated by pairing B with a new aversive or neutral outcome (B → CTHREAT/NEUTRAL) while pupil dilation was measured as an index of emotional arousal. Then, again 1 day later, the accuracy of indirect associations (A → C?) was tested. Associative inferences involving a threat learning memory were impaired whereas the initial memories were retroactively strengthened, but these effects were not moderated by pupil dilation at encoding. These results imply that a healthy memory system may compartmentalize episodic information of threat, and so hinders its recall when cued only indirectly. Malfunctioning of this process may cause maladaptive linkage of negative events to distant and benign memories, and thereby contribute to the development of clinical intrusions and anxiety.


NGF regulates sertoli cell growth and prevents LPS-induced junction protein damage via PI3K/AKT/NFκB signaling.
Damage to Sertoli cell junction proteins caused by inflammation can lead to male infertility. Nerve growth factor (NGF) plays an important role in reproductive and inflammatory disease; however, whether and how NGF regulates Sertoli cell function remains unclear. Here, we aimed to assess the effect of NGF on the growth of Sertoli cells isolated from the testes of dairy goats and evaluate if NGF has a protective effect on these cells. We confirmed that Sertoli cell viability, proliferation, and ATP content increased following NGF treatment. In addition, qPCR results suggested that Sertoli cell apoptosis was inhibited after NGF treatment. To investigate the protective effect of NGF on Sertoli cells under pathological inflammatory conditions, LPS was used to induce inflammatory response in Sertoli cells. Post-treatment, the entangled filamentous pseudopodia of the cells loosened and no longer spanned adjacent cells. The expression of several junction proteins (ZO-1, occludin, CX-43, β-catenin, and N-cadherin), which was down-regulated after inflammatory response induction, was restored following NGF treatment. LPS-induced changes in cytotoxicity and transepithelial electrical resistance were reversed and the intercellular connections became tighter after NGF treatment. We further demonstrated that NGF prevented the inflammatory response of Sertoli cells via the PI3K/AKT/NFκB signaling pathway, similar to the effect of the PI3K-inhibitor, LY294002, which is modified by the PI3K activator, 740Y-P. These results provide insights for devising strategies for protecting the male reproductive system and curing or preventing associated pathological conditions.


Pathways of insulin degradation in isolated adipocytes: evaluation by gel filtration and differential precipitation.
Insulin degradation by isolated rat adipocytes was evaluated using gel filtration and a new technique of differential precipitation to fractionate the sample by molecular size using polyethylene glycol and trichloracetic acid. At 37 degrees C, 125I-insulin bound to adipocytes was rapidly degraded into small fragments or iodotyrosine. 125I-insulin in the medium was also degraded into iodotyrosine, as well as fragments intermediate in molecular weight between insulin and iodotyrosine. Lowering the temperature to 15 degrees C or adding bacitracin to the medium inhibited degradation in the medium but had little effect on cell-associated degradation. Methylamine, on the other hand, inhibited cell-associated degradation, but had little effect on the insulin degradation in the medium. Addition of methylamine or bacitracin or lowering of the temperature increased the amount of 125I-insulin bound to the cell and prolonged the steady-state of binding. Bacitracin also produced a slight shift to the left in the dose response curve for insulin-stimulated glucose oxidation. Methylamine increased basal glucose oxidation, but had no effect on insulin sensitivity as measured in the glucose oxidation bioassay. These data suggest that isolated adipocytes in vitro exhibit at least two distinct pathways of insulin degradation, a cell-associated pathway which can be inhibited by methylamine and a medium pathway which can be inhibited by bacitracin. Neither pathway, however, appears to be closely linked to insulin's ability to stimulate glucose metabolism in these cells.


Single coronary artery with bicuspid aortic valve stenosis and aneurysm of the ascending aorta: report of a case.
A 73-year-old man with a severely stenosed bicuspid valve and an aneurysm of the ascending aorta underwent valve and aortic surgery. Preoperative imaging revealed a single coronary artery arising from the right side of the sinus of Valsalva and a branch that perfused into the left side of the heart to pass through the front of the pulmonary artery. We replaced the aortic valve and ascending aorta, painstakingly avoiding damage to the coronary artery and obstruction of the sole coronary ostium.