Immunohistochemical Detection of Human Papillomavirus 16 E7 Oncoprotein in Cervical Lesions.
Almost all cervical cancer is associated with persistent infection of high-risk (hr) human papillomavirus (HPV) like HPV16. The E7 oncoprotein encoded by hrHPV plays a crucial role in carcinogenesis. The present study aimed to establish a reliable protocol of immunohistochemistry stains to detect HPV16 E7 protein in formalin-fixed and paraffin-embedded tissue specimens of various cervical lesions. Firstly, the HPV16 E7 gene was inserted into a prokaryotic expression vector pGEX-4T2. Then the recombinant plasmid pGEX-4T2-(HPV16-E7) was transformed into Escherichia coli JM109. The fusion protein containing a GST tag was purified, and New Zealand white rabbits were immunized to produce the HPV16 E7 polyclonal antibody. The anti-HPV16 E7 antibody was verified by western blotting and immunofluorescence stains, and applied in 182 HPV16 DNA-positive cervical tissue specimens and matched 36 HPV DNA-negative specimens by immunohistochemistry. Furthermore, A positive correlation between HPV16 E7 protein expression and malignancy grade was observed. But there is no relationship between HPV16 E7 protein expression and tumor sizes, tumor differentiation, lymph node metastasis, International Federation of Gynecology and Obstetrics (FIGO) stage, or lymphovascular space invasion in cervical cancer. These findings provide a basis for further research focusing on the role of HPV E7 protein in various HPV-related diseases.


Liver: single breath-hold dynamic subtraction CT with multi-detector row helical technology feasibility study.
Fifty-two patients with known or suspected hypervascular malignancy were examined to determine the technical feasibility of performing single-breath-hold dynamic subtraction computed tomography (CT) of the liver with multi-detector row helical CT. The precontrast and hepatic arterial CT scans, which were acquired during the same breath hold, were subtracted. The mean liver-to-muscle contrast ratio on the precontrast, hepatic arterial, and subtracted images was 1.3, 1.4, and 2.3, respectively. In 13 patients with lesions, the subtracted images showed a 2.5-fold increase in mean lesion contrast compared with the hepatic arterial CT scans.


The treatment of sickle cell anemia with induced hyponatremia.
Since the rate of polymerization of sickle hemoglobin is exquisitely dependent on its concentration, a small reduction in intracellular hemoglobin concentration should cause a significant inhibition of sickling. In three patients with homozygous sickle cell anemia, sustained hyponatremia was induced by a program consisting of a high fluid intake, a low salt diet and a vasopressin analog, DDAVP. During periods of hyponatremia, mean corpuscular hemoglobin concentration (MCHC) fell 13% and in vitro sickling was reduced as assessed by morphology and oxygen affinity. The frequency and duration of sickle cell crises appeared to be decreased during periods when patients were hyponatremic. These preliminary results indicate that reduction in intracellular hemoglobin concentration is an effective approach to the treatment of sickle cell anemia.


Purification and characterization of coenzyme F390 synthetase from Methanobacterium thermoautotrophicum (strain delta H).
Coenzyme F390 synthetase catalyzes the formation of 8-hydroxyadenylylated-coenzyme F420 (coenzyme F390-A) from coenzyme F420 and ATP in some methanogenic Archaea. The presence of coenzyme F390 was found when these organisms were exposed to oxygen. To get more insight into the defined function of coenzyme F390, the coenzyme F390 synthetase from Methanobacterium thermoautrophicum was purified from a cell-free extract and its catalytic properties were determined. The synthetase was purified 150-fold to a specific activity of 0.45 mumol.min-1.mg protein-1. The enzyme consisted of one polypeptide of approximately 51 kDa. The isolated enzyme showed a tendency to aggregate into dimers and tetramers upon concentration. Co-elution during purification of GTP-dependent coenzyme F390 synthetase activity suggested that the synthetase is also capable of 8-hydroxyguanylylated-coenzyme F420 (coenzyme F390-G) formation. Initial-velocity measurements of the two-substrate reaction showed that the enzyme kinetics for the coenzyme F390 synthetase reaction proceeded by a ternary-complex mechanism. The coenzyme F390 synthetase displayed a Km for coenzyme F420 of 39 microM and a Km for ATP of 1.7 mM. In contrast to the enzyme in the cell-free extract, the isolated enzyme was active under aerobic and anaerobic conditions. Treatment with air was not required to obtain the enzyme in an active form. However, 1,5-dihydro-coenzyme F420 (coenzyme F420H2) appeared to be a potent competitive inhibitor (Ki 3 microM) with respect to coenzyme F420. The latter findings may explain why the enzyme could only be detected in crude extracts that had been exposed to air, i.e. treatment with air causes the oxidation of reduced coenzyme F420 present in anaerobic extracts. The results of this study are discussed in view of the proposed role for coenzyme F390 in methanogenic metabolism.


Demonic possessions and mental illness: discussion of selected cases in late medieval hagiographical literature.
During the Middle Ages, demonic possession constituted an explanation for an erratic behavior in society. Exorcism was the treatment generally applied to demoniacs and seems to have caused some alleviation in the suffering of mentally distressed people. We have selected and analyzed some cases of demonic possession from thirteenth-century hagiographical literature. In the description of demoniacs we have been able to find traits of psychotic, mood, neurotic, personality disorders and epilepsy. The exorcisms analyzed in our article are the result of literary invention more than the description of a contemporary event. Nevertheless, the writers were witnesses of their time, transferred their knowledge about exorcism and possession in their narrative and presumably incorporated their actual experience with demoniacs.


Traditional Chinese medicines as immunosuppressive agents.
Traditional Chinese Medicines (TCM) have been used for centuries in China to treat various immune-mediated disorders. This review focuses on the clinical and experimental studies that have been performed with TCM as immunosuppressive agents for the treatment of systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), atopic eczema and solid organ transplantation. The "thunder god" vine, Tripterygium wilfordii Hook F (TWHf), has been extensively used in China to treat SLE and RA. TWHf not only inhibited mitogen-stimulated lymphoproliferation, but its active derivatives have also been shown to inhibit production of proinflammatory cytokines by monocytes and lymphocytes, as well as prostaglandin E2 production via the cyclooxygenase, COX-2, pathway, a potential mechanism of action in patients with RA. Demethylzelasteral (TZ-93), a triterpenoid isolated from the root cortex of TWHf, the plant alkaloid berbamine, and the hydrophobic extract of a Chinese herbal decoction, CMX-13, were all shown to be active in prolonging allograft survival in experimental animal models of heart, skin and single lung transplants, respectively. There are few well-designed randomised placebo-controlled clinical trials demonstrating the efficacy of TCM in various diseases. Zemaphyte, a decoction of 10 herbs, has been shown to be efficacious in the treatment of atopic dermatitis in both children and adults in two randomised double-blind placebo-controlled trials. There is both laboratory and clinical evidence that the derivatives of many of these herbs may have significant beneficial immunosuppressive effects, however, concerns of toxicity must also be addressed, as exact dosing of the active derivatives is difficult to achieve with the current prescriptions of TCM.


Growth of erythroid colonies in chronic myelogenous leukemia is independent of erythropoietin only in the presence of steel factor.
The mechanisms of the chronic myeloid leukemia (CML) clones proliferative advantage over normal clones are currently unknown. They may involve an insensitivity to a negative regulation of a growth factor-independent proliferation. Clonogenic progenitors from CML patient blood or marrow in chronic phase were grown either in the presence or absence of recombinant growth factors. No erythroid colonies were observed in the absence of any cytokine. In contrast, erythroid colonies composed of fully mature hemoglobinized erythroblasts (day 12 burst-forming units-erythroid) were obtained in the presence of Steel factor (SF) alone. Addition of erythropoietin (Epo) to SF either had no effect on the cloning efficiency or increased up to 50% the number of erythroid colonies. No erythroid growth was observed when cultures were stimulated by interleukin-3 or granulocyte-macrophage colony-stimulating factor alone. Similar erythroid growth in the presence of SF but without Epo was obtained in "serum-free" cultures when purified blood CML CD34+ cells were grown. This growth of erythroid colonies in the absence of Epo was not accounted for by an autocrine stimulation loop by Epo, because neutralizing antibodies against Epo did not inhibit it. This abnormal response to growth factor was specifically observed in the CML clone, as shown by the presence of the BCR-ABL transcript in all of these erythroid colonies. The direct implication of BCR-ABL was further documented (1) by studies of alpha-interferon-treated patients with a chimerism in which the abnormal growth correlates with the presence of the malignant clone and (2) by the use of antisense oligonucleotide against BCR-ABL transcript, which abrogated this abnormal growth. Finally, erythroid growth in the SF presence was greatly diminished by herbimycin A, whereas, at the same concentration, this tyrosine kinase inhibitor had no marked effect on erythroid colony formation in the presence of SF plus Epo on CML or normal marrow cells. This result suggests that the BCR-ABL kinase activity leads directly to this Epo-independent terminal differentiation requiring, however, the presence of SF.


Higher Coffee Consumption Is Associated With Reduced Cerebral Gray Matter Volume: A Mendelian Randomization Study.
Recently published two-sample Mendelian randomization (MR) studies showed that genetically predicted coffee consumption may be associated with increased risk of Alzheimer's disease and intracerebral hemorrhage but associated with a decreased risk of small vessel ischemic stroke. We aimed to investigate the effects of genetically predicted coffee consumption on magnetic resonance imaging (MRI) markers of cerebral small vessel disease and brain volume using the two-sample MR method. Twelve single nucleotide polymorphisms (SNPs) in up to 375,833 individuals were used as genetic instruments for cups consumed per day of coffee. Another four SNPs from an independent sample were used to perform the replication analysis. Three SNPs in up to 45,821 individuals were used as genetic instruments for high coffee consumption vs. low/no coffee consumption. Mendelian randomization analysis showed that coffee consumption (cups/day) was inversely associated with gray matter volume (beta = -0.371, 95% CI = -0.596 to -0.147, p = 0.001). Replication analysis and multivariable analyses after adjusting for other risk factors confirmed the effect. High coffee consumption was also suggestively associated with decreased gray matter volume (beta = -0.061, 95% CI = -0.109 to -0.013, p = 0.013) compared with low/no coffee consumption. All analyses did not find an effect of coffee consumption on other outcomes including white matter hyperintensity volume, mean diffusivity, fractional anisotropy, brain microbleed, total brain volume, white matter volume, and hippocampus volume. This two-sample MR study showed that genetically predicted higher coffee consumption is causally associated with reduced gray matter volume of the brain.


Integrator is a genome-wide attenuator of non-productive transcription.
Termination of RNA polymerase II (RNAPII) transcription in metazoans relies largely on the cleavage and polyadenylation (CPA) and integrator (INT) complexes originally found to act at the ends of protein-coding and small nuclear RNA (snRNA) genes, respectively. Here, we monitor CPA- and INT-dependent termination activities genome-wide, including at thousands of previously unannotated transcription units (TUs), producing unstable RNA. We verify the global activity of CPA occurring at pA sites indiscriminately of their positioning relative to the TU promoter. We also identify a global activity of INT, which is largely sequence-independent and restricted to a ~3-kb promoter-proximal region. Our analyses suggest two functions of genome-wide INT activity: it dampens transcriptional output from weak promoters, and it provides quality control of RNAPII complexes that are unfavorably configured for transcriptional elongation. We suggest that the function of INT in stable snRNA production is an exception from its general cellular role, the attenuation of non-productive transcription.


Epidermal growth factor stimulation can substitute for c-Src overexpression in promoting breast carcinoma invasion.
Cancer progression is in large part dependent on the complex process of cell invasion, involving adhesion, motility, and enzymatic proteolysis. Overexpression of the Src proto-oncogene (c-Src), a nonreceptor tyrosine kinase, has been implicated in the progression of both colon and breast cancer. Our group has previously reported that overexpression of c-Src leads to a significant gain in invasive cell behavior in vitro. In this study, we sought to assess the relative importance of epidermal growth factor (EGF) stimulation and c-Src overexpression in conferring an invasive phenotype. Breast carcinoma cells and colon epithelial cells which naturally express low levels of c-Src were used for these studies. The cells were transfected so that they overexpressed c-Src; the mock-transfected parent lines were used as controls. Transfectants were tested for changes in invasion patterns after Src inhibition and EGF stimulation. Invasion assays in both cell systems confirmed the importance of c-Src in determining invasive potential. A significant correlation was shown between c-Src kinase protein and cell invasion. Furthermore, Src inhibition significantly inhibited invasion in a dose-dependent manner. To clarify the relative contribution of EGF and c-Src to cell invasion, the ability of cells to invade through growth-factor-reduced matrigel, with or without EGF added, was compared to invasion through intact matrigel. The breast and colon cell lines behave quite differently in this regard. In the colon model, overexpressed c-Src is critical for cell invasion and stimulation with EGF is synergistic with c-Src overexpression. Conversely, the breast carcinoma cells transfected with c-Src were unable to invade without EGF stimulation and did not demonstrate the same synergistic relationship between c-Src and EGF. Instead, our results indicate that in BT474 breast carcinoma cells, EGF can substitute for c-Src in promoting breast cancer cell invasion. Because most breast carcinomas overexpress c-Src, it behooves one to question the extent to which reducing the amount of EGF and consequent EGFR activity will decrease invasion. In this study, the effects of EGF on cell invasion were determined in light of a single alteration in c-Src expression. Our results show that EGF enhances the impact of c-Src overexpression on invasion. In breast cancer cells, EGF is capable of inducing invasion to the same extent as c-Src overexpression. This suggests that anti-EGFR therapies will be efficacious in retarding breast carcinoma invasion and metastasis.


The development of a fluorescence in situ hybridization assay for the detection of dysplasia and adenocarcinoma in Barrett's esophagus.
The goal of this study was to identify a set of fluorescence in situ hybridization probes for the detection of dysplasia and adenocarcinoma in patients with Barrett's esophagus. We examined 170 brushing specimens from 138 patients with Barrett's esophagus or a history of Barrett's esophagus using fluorescence in situ hybridization with probes to 5p15, 5q21-22, centromere 7, 7p12, 8q24.12-13, centromere 9, 9p21, centromere 17, 17p13.1, 17q11.2-12, 20q13.2, and centromere Y. Receiver-operator curves were used to determine the sensitivity and specificity of various four-probe combinations for detecting low-grade dysplasia, high-grade dysplasia, and esophageal adenocarcinoma. Endoscopic biopsy results were used as the gold standard. Numerous four-probe combinations provided a similarly high sensitivity and specificity. Of these, a set consisting of probes to 8q24, 9p21, 17q11.2, and 20q13.2 was found to have a sensitivity and specificity, respectively, of 70% and 89% for low-grade dysplasia, 84% and 93% for high-grade dysplasia, and 94% and 93% for esophageal adenocarcinoma. This probe set was chosen for future prospective clinical evaluations based on its high sensitivity and specificity, its ability to distinguish adenocarcinoma and high-grade or low-grade dysplasia from lesser diagnostic categories, and the favorable signal quality for each of the probes.


Supercontinuum generation in a photonic crystal fiber with two zero-dispersion wavelengths tapered to normal dispersion at all wavelengths.
We numerically study supercontinuum generation in photonic crystal fibers with two zero-dispersion wavelengths, weakly tapered to have normal dispersion at all wavelengths after a certain distance. We pump with 15 fs pulses with milliwatt average power and show that two distinct smooth spectral parts are generated, with improved stability due to the normal dispersion. We characterize the two spectral parts and show how the 3 dB bandwidth, the center wavelength, and the power of the two parts depend on the taper parameters and the pump power.


Pediatric Anesthesia Severe Adverse Events Leading to Anesthetic Morbidity and Mortality in a Tertiary Care Center in a Low- and Middle-Income Country: A 25-Year Audit.
The analysis of adverse events, including morbidity and mortality (M&M), helps to identify subgroups of children at risk and to modify clinical practice. There are scant data available from low- and middle-income countries. Our aim was to estimate the proportion of pediatric patients with various severe adverse events in the perioperative period extending to 48 hours and to describe the clinical situations and causes of those events. We reviewed the M&M database of the Department of Anesthesiology between 1992 and 2016. A data collection tool was developed, and the outcomes were standardized. Each case was reviewed independently and subsequently discussed between 2 reviewers to identify a major primary causative factor. The total number of pediatric cases during this period was 48,828. Seventy-six significant adverse events were identified in 39 patients (8 patients [95% confidence interval {CI}, 5.7-10.9] per 10,000). Thirteen patients had multisystem involvement, and hence the total number of events exceeded the number of patients. Respiratory events were the most common (33.5%). Thirteen patients had perioperative cardiac arrest within 48 hours of surgery (2.6 [95% CI, 1.3-4.3] per 10,000), 7 of these were infants (54%), 5 of whom had congenital heart disease (CHD). Eleven of these 39 patients died within 48 hours (2.0 [95% CI, 1.1-4.0] per 10,000).In 13 cases, anesthesia was assessed to be the predominant cause of morbidity (2.6 per 10,000), whereas in 26 cases, it contributed partially (5.32 per 10,000). There was only 1 death solely related to anesthesia (0.2 per 10,000), and this death occurred before the start of surgery. Adverse events were uncommon. Respiratory complications were the most frequent (33%). Infants, especially those with CHD, were identified as at a higher risk for perioperative cardiac arrest, but this association was not tested statistically. Twenty-eight percent of the patients who suffered events died within 48 hours. Increased access to anesthesia drugs and practice improvements resulted in a decline in perioperative cardiac arrests.


N-methyl-D-aspartate induces phase shifts in circadian rhythm of neuronal activity of rat SCN in vitro.
The suprachiasmatic nucleus (SCN) acts as a pacemaker for mammalian circadian rhythms. Receptors for excitatory amino acids like N-methyl-D-aspartate (NMDA) and non-NMDA receptors have both been found to play an important role in the transmission of photic information from the retina to the SCN. Therefore, we investigated whether the application of glutamate receptor agonists could reset the phase of the circadian rhythm of SCN firing activity in vitro. Treatment with NMDA (0.1-10 microM) for 15 min or 1 h during the early part of the subjective night produced phase delay, whereas treatment during the late subjective night caused an advance in phase. The phase-response curve for NMDA was similar to that previously obtained in response to light pulses in vivo. Application of DL-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid hydrobromide (AMPA) (1 or 10 microM), a non-NMDA-receptor agonist, also produced a dose-dependent phase delay of SCN activity. The NMDA-induced phase delay was antagonized by an NMDA-receptor antagonist MK-801. These findings suggest that both NMDA and non-NMDA receptors may be involved in the transmission of information to the SCN via the retinohypothalamic tract. In addition, both the advances and delays in phase caused by NMDA were potentiated by cotreatment with neuropeptide Y, whereas AMPA-induced phase delay was not potentiated by neuropeptide Y. This points to a functional link between NMDA and neuropeptide Y receptor-mediated mechanisms in the SCN.


Socio-economic factors, gender and smoking as determinants of COPD in a low-income country of sub-Saharan Africa: FRESH AIR Uganda.
In Uganda, biomass smoke seems to be the largest risk factor for the development of COPD, but socio-economic factors and gender may have a role. Therefore, more in-depth research is needed to understand the risk factors. The aim of this study was to investigate the impact of socio-economic factors and gender differences on the COPD prevalence in Uganda. The population comprised 588 randomly selected participants (>30 years) who previously completed the FRESH AIR Uganda study. In this post hoc analysis, the impact of several socio-economic characteristics, gender and smoking on the prevalence of COPD was assessed using a logistic regression model. The main risk factors associated with COPD were non-Bantu ethnicity (odds ratio (OR) 1.73, 95% confidence interval (CI) 1.06-2.82, P=0.030), biomass fuel use for heating (OR 1.76, 95% CI 1.03-3.00, P=0.038), former smoker (OR 1.87, 95% CI 0.97-3.60, P=0.063) and being unmarried (OR 0.087, 95% CI 0.93-2.95, P=0.087). A substantial difference in the prevalence of COPD was seen between the two ethnic groups: non-Bantu 20% and Bantu 12.9%. Additional analysis between these two groups showed significant differences in socio-economic circumstances: non-Bantu people smoked more (57.7% vs 10.7%), lived in tobacco-growing areas (72% vs 14.8%) and were less educated (28.5% vs 12.9% had no education). With regard to gender, men with COPD were unmarried (OR 3.09, 95% CI 1.25-7.61, P=0.015) and used more biomass fuel for heating (OR 2.15, 95% CI 1.02-4.54, P=0.045), and women with COPD were former smokers (OR 3.35, 95% CI 1.22-9.22, P=0.019). Only a few socio-economic factors (i.e., smoking, biomass fuel use for heating, marital status and non-Bantu ethnicity) have been found to be associated with COPD. This applied for gender differences as well (i.e., for men, marital status and biomass fuel for heating, and for women being a former smoker). More research is needed to clarify the complexity of the different risk factors.


International Commission for Protection Against Environmental Mutagens and Carcinogens. Cyclosporine A: review of genotoxicity and potential for adverse human reproductive and developmental effects. Report of a Working Group on the genotoxicity of cyclosporine A, August 18, 1993.
Cyclosporine is an important therapeutic agent for transplant recipients and for a growing number of autoimmune diseases. Experimental animal and human data has indicated that cyclosporine is unlikely to be genotoxic. In contrast, azathioprine, an agent often given with cyclosporine, is considered to be genotoxic making the assessment of the independent effects of cyclosporine difficult. Cyclosporine does appear to be related to the development of tumors, primarily lymphomas, in animals and humans, but the basis of its potential carcinogenicity is not completely understood. In terms of reproductive and developmental toxicity, cyclosporine produces some adverse effects in both experimental animals and humans. In animals, the effects are seen at high doses sufficient to cause maternal toxicity. In humans, outcomes such as growth retardation have been noted, but the confounding effects of renal toxicity and resultant pregnancy complications cloud the interpretation. An increase in congenital anomalies and genetic disease have not been found reported in human studies that are limited in sample size. Given that the present data indicate the lack of genotoxicity, a mutation epidemiology study of cyclosporine is not recommended. As indicated above, such a study is probably impractical for many genetic endpoints of interest. However, well-conducted, large multicenter studies of transplant patients and their offspring would allow for routine monitoring of an increased risk for some reproductive and developmental (possibly non-genetic) endpoints that are of public health importance.


Implementing evidence-based nursing practice.
A methodology for establishing and supporting evidence-based nursing practice is examined. Description of a clinical and administrative scenario serves as an example of a systematic appraisal of the relevant literature that had implications for clinical practice.


The characteristics and patterns of utilization of healthcare services among Omanis with substance use disorders attending therapy for cessation.
It is indicated that Oman is witnessing an increase in issues pertinent to alcohol and psychoactive substance use. The aim of this study was to identify the characteristics of Omanis with substance use disorder attending a specialized hospital in Oman and the pattern of their utilization of healthcare services. A related aim was to ascertain the age group most vulnerable to alcohol and substance use in Oman. A cross-sectional study was conducted in a tertiary care center specialized for treatment of those engaging in substance use in Oman. The participants in the study were selected from a convenience sample among patients seeking consultation at the center for alcohol and substance use. A six-part questionnaire was designed to obtain information regarding socio-demographic background, clinical history, healthcare utilization and perceived hurdles to access. Chi-square analyses were used to evaluate the significance of differences among categorical data. Logistic regression modelling was used to obtain measures of association after adjusting for confounding factors. Among the patients (n = 293) seeking cessation therapy, 99% were male and less than 30 years of age. Peer influences on the initiation of substance use were significant. Most patients had a history of polysubstance use, including intravenous substance use. Cannabis and alcohol were the first substances consumed by most patients and Hepatitis C and psychiatric disorders were found to be the most common co-morbidities. The participants that reported use of cannabis and benzodiazepines were more likely to perceive "improvement" upon receiving treatment. This study indicated that males below 30 years of age with a history of polysubstance use were likely to attend a hospital specialized in treating substance use disorder in Oman. This study identified information regarding socio-demographic background, risk factors and perceived hurdles to healthcare that could serve as groundwork for further studies conducted on newly emerging issues of substance use in Oman.


Multiple actions of 2,3-butanedione monoxime on contractile activation in frog twitch fibres.
1. The effects of 2,3-butanedione monoxime (BDM) on various steps in the excitation-contraction coupling sequence, including action potential, charge movement and twitch tension, were studied in twitch fibres of Rana temporaria. 2. The resting potential of intact fibres in whole muscle bathed in 20 mM-BDM was the same as control. The resting potential also remained stable after more than 100 min in 20 mM-BDM. 3. The action potential was measured in intact fibres of fibre bundles with an intracellular microelectrode. Applications of 5 and 7.5 mM-BDM had no effect on its amplitude, whereas 10 and 20 mM suppressed its amplitude by about 4 and 10%, respectively. Increasing concentrations of BDM prolonged the half-width and elevated the after-potential of the action potential progressively. The action potential was also measured in cut fibres mounted in a double Vaseline-gap chamber. Results were similar to those in intact fibres. 4. Charge movement was measured in intact fibres of halved muscles with the three-microelectrode voltage-clamp technique. The steady-state Q-V plot of the total charge measured in isotonic tetraethylammonium (TEA) Ringer solution with 20 mM-BDM appeared to be shifted about 10 mV in the depolarizing direction and to be slightly more shallow when compared with the control Q-V plot measured in hypertonic TEA Ringer solution with 350 mM-sucrose. After allowing for the voltage shift, 20 mM-BDM did not appear to affect the kinetics of both components of charge movement, but suppressed the maximum amount of total charge by about one-quarter. 5. Charge movement was also measured in cut fibres with the double Vaseline-gap voltage-clamp technique. In the presence of 20 mM-BDM, charge movement traces resembled those from intact fibres. Twenty millimolar BDM suppressed the maximum amount of total charge by about one-quarter, as in intact fibres. The steady-state Q-V plots from cut fibres were separated into Q beta (early current) and Q gamma (late hump current) components by least-squares fitting with a sum of two Boltzmann distribution functions. On average, 20 mM-BDM suppressed Q beta and Q gamma in roughly equal proportion, but did not affect the individual voltage distributions of Q beta and Q gamma. 6. Twitch tension was measured in single intact fibres stimulated extracellularly. BDM effectively reduced the peak amplitude, the time-to-peak and the half-width of twitch tension. The interaction of BDM with receptors appeared to follow more or less a simple 1:1 binding in fibres stretched to sarcomere lengths of about 3.6 microns.(ABSTRACT TRUNCATED AT 400 WORDS)


Coupled Dynamics of Stimulus-Evoked Gustatory Cortical and Basolateral Amygdalar Activity.
Gustatory cortical (GC) single-neuron taste responses reflect taste quality and palatability in successive epochs. Ensemble analyses reveal epoch-to-epoch firing-rate changes in these responses to be sudden, coherent transitions. Such nonlinear dynamics suggest that GC is part of a recurrent network, producing these dynamics in concert with other structures. Basolateral amygdala (BLA), which is reciprocally connected to GC and central to hedonic processing, is a strong candidate partner for GC, in that BLA taste responses evolve on the same general clock as GC and because inhibition of activity in the BLA→GC pathway degrades the sharpness of GC transitions. These facts motivate, but do not test, our overarching hypothesis that BLA and GC act as a single, comodulated network during taste processing. Here, we provide just this test of simultaneous (BLA and GC) extracellular taste responses in female rats, probing the multiregional dynamics of activity to directly test whether BLA and GC responses contain coupled dynamics. We show that BLA and GC response magnitudes covary across trials and within single responses, and that changes in BLA-GC local field potential phase coherence are epoch specific. Such classic coherence analyses, however, obscure the most salient facet of BLA-GC coupling: sudden transitions in and out of the epoch known to be involved in driving gaping behavior happen near simultaneously in the two regions, despite huge trial-to-trial variability in transition latencies. This novel form of inter-regional coupling, which we show is easily replicated in model networks, suggests collective processing in a distributed neural network.SIGNIFICANCE STATEMENT There has been little investigation into real-time communication between brain regions during taste processing, a fact reflecting the dominant belief that taste circuitry is largely feedforward. Here, we perform an in-depth analysis of real-time interactions between GC and BLA in response to passive taste deliveries, using both conventional coherence metrics and a novel methodology that explicitly considers trial-to-trial variability and fast single-trial dynamics in evoked responses. Our results demonstrate that BLA-GC coherence changes as the taste response unfolds, and that BLA and GC specifically couple for the sudden transition into (and out of) the behaviorally relevant neural response epoch, suggesting (although not proving) that: (1) recurrent interactions subserve the function of the dyad as (2) a putative attractor network.


Body weight loss as an indicator of breast cancer recurrence.
Body weight loss (BWL), a major prognostic factor in breast cancer, was included as a parameter to be monitored in the recent breast cancer surveillance guidelines of the American Society of Clinical Oncology. The aim of this work was prospectively to evaluate BWL as an indicator of breast cancer recurrence. Body weight was measured every 2 months for 10.4+/-3.7 (SD) months in 109 disease-free breast cancer patients in stage II node-positive and stage III disease. The correlation between unexplained BWL and recurrence was studied. Attempts were made to define the limits in weight variations among disease-free patients beyond which recurrence could be suspected. Unexplained BWL was observed in 16/19 (84%) patients developing recurrence, versus 9/90 (10%) patients remaining disease-free. There was a significant (p < 0.001) correlation between BWL and recurrence. BWL anticipated the diagnosis of recurrence by 6 (range 4-12) months. Based on the average percentage weight variation +/- 2 SD (95% confidence interval) of the disease-free group, the limits for BWL beyond which recurrence could be suspected were a 5.8% decrease in the last 6 months, 3.6% in the last 2 months or 3.0% of the patient's mean weight. However, because of the large variation in the amplitude of individual weight oscillations among disease-free patients (from < 0.5% to > 5.9% of the mean weight), individual limits derived from the patient's own body weight curve seemed more reliable. The results suggest that unexplained BWL is a valuable indicator of incipient breast cancer recurrence. Careful monitoring of body weight in breast cancer patients during follow-up is encouraged.


Contrast-induced nephrotoxicity: clinical landscape.
Over 80 million doses of iodinated intravascular contrast media (CM) were administered in the most recent tabulations of 2003, corresponding to approximately 8 million liters, making it one of the highest volume medical drugs used compared to any other pharmaceutical. The evolution of CM has focused on minimizing adverse events by eliminating ionicity, increasing hydrophilicity, lowering osmolality and increasing the number of iodine atoms per molecule. Contrast media are classified into three general categories based on their osmolality relative to blood: high osmolar (5 times or greater than blood), low osmolar (2-3 times blood) and iso-osmolar (the same as blood). All imaging modalities that employ CM, especially computerized tomography (CT), have shown rapid growth. In the last two decades, the use of CT scanning has increased by 800%. From 1979 to 2002, the number of cardiac catheterization procedures in the USA increased by 390% and in Europe from 1992 to 1999 by 112%. There is a general consensus that renal insufficiency and diabetes are major risk factors for contrast-induced nephropathy (CIN), particularly when co-existing. The US Renal Data System documents a 'relentless' increase in kidney failure, projecting a 90% increase by 2010. Diabetes affects 194 million people worldwide and the number is anticipated to increase by 75% by 2025. The unavoidable conclusion is that patient exposure and prevalence of risk factors for CIN will continue to increase.


Synthesis and characterization of new ion-imprinted polymer for separation and preconcentration of uranyl (UO2(2+)) ions.
UO(2)(2+) ion-imprinted polymer materials used for solid-phase extraction were prepared by copolymerization of a ternary complex of uranyl ions with styrene and divinyl benzene in the presence of 2,2'-azobisisobutyronitrile. The imprinted particles were leached by HCl 6M. Various parameters in polymerization steps such as DVB/STY ratio, time of polymerization and temperature of polymerization were varied to achieve the most efficient uranyl-imprinted polymer. X-ray diffraction (XRD), infra-red spectroscopy (IR), thermo gravimetric analysis (TGA), UV-vis and nitrogen sorption were used to characterize the polymer particles. The XRD results showed that uranyl ions were completely removed from the polymer after leaching process. IR Analysis indicated that the N,N'-ethylenebis(pyridoxylideneiminato) remained intact in the polymer even after leaching. Some parameters such as pH, weight of the polymer, elution time, eluent volume and aqueous phase volume which affects the efficiency of the polymer were studied.


Paediatric presentation and outcome of congenital protein C deficiency in Japan.
Severe heritable protein C (PC) deficiency is quite rare, although heterozygous PROC mutation is the second leading cause of genetic predisposition to thrombosis in Japanese adults. The aim of the study was to search the optimal management, the paediatric onset and outcomes of PC deficiency were characterized in Japan. The genetic study, postmarketing survey of activated PC(aPC) concentrate (Anact(®)C) and intensive review in Japan for 20 years enabled the analysis of the disease onset, genotype, treatment and prognosis. Symptomatic PC deficiency was determined in 27 Japanese children. All but two patients presented within 16 days after birth (three prenatal and six neonatal onsets). Postnatal-onset cases had normal growth at full-term delivery. Of the 27 patients, 19 suffered intracranial thrombosis or haemorrhage (ICTH) (three foetal hydrocephalies), 16 developed purpura fulminans (PF) and 10 had both at the first presentation. ICTH preceded PF in both affected cases. Low PC activities of 18 mothers and/or 12 fathers indicated 20 inherited PC deficiencies (2 homozygotes, 11 compound heterozygotes and 7 heterozygotes) and seven unidentified causes of PC deficiency. Nine of 11 patients studied had PROC mutations. Four unrelated patients (50%) carried PC nagoya (1362delG). No PC-deficient parents had experienced thromboembolism. Of the 18 patients with aPC therapy, two died and eight evaluable survivors had neurological sequelae. This first comprehensive study of paediatric PC deficiency suggested that perinatal ICTH was the major presentation, occurring earlier than neonatal PF. PC nagoya was prevalent in paediatric, but not adult, patients in Japan. Early maternal screening and optimal PC therapy are required for newborns at risk of PC deficiency.


Desensitization of alpha 2-adrenergic receptors in NG 108 15 cells by (-)-adrenaline and phorbol 12-myristate 13-acetate.
alpha 2-Adrenergic receptors on NG 108 15 cell membranes were identified by [3H]rauwolscine binding: Bmax. = 661 +/- 81 fmol/mg of protein, Kd = 6.9 +/- 2.5 nM (mean +/- S.E.M., n = 6). On intact cells, stimulation of these receptors by (-)-adrenaline inhibited the prostaglandin-E1-stimulated adenylate cyclase activity by about 60%. The effect of (-)-adrenaline was pertussis-toxin-sensitive, indicating the involvement of an inhibitory G protein. (-)-Adrenaline/[3H]rauwolscine competition-binding experiments revealed that only 50% of the alpha 2 receptors were coupled to G proteins (i.e. displayed high agonist affinity). Pre-treatment of the cells with 20 microM-(-)-adrenaline provoked homologous desensitization of the alpha 2 receptors. The alpha 2-adrenergic response decreased after a time lag of about 2 h, to reach a minimum after 12 h. The bradykinin and muscarinic responses were not affected. The alpha 2-receptor concentration decreased without time lag. The high-agonist-affinity sites disappeared more rapidly (t1/2 = 42 min) than did the low-affinity uncoupled sites (t1/2 approx. 20 h). In contrast, pertussis-toxin-mediated [32P]ADP-ribosylation of inhibitory G proteins was unaffected by the pre-treatment. Pretreatment of intact NG 108 15 cells with 1 microM-phorbol 12-myristate 13-acetate (PMA) provoked a rapid decrease of the alpha 2-adrenergic response. The effect was nearly complete after 40 min. PMA also decreased the bradykinin response, suggesting a heterologous desensitization process. The alpha 2-receptor concentration, the (-)-adrenaline competition-binding curves and the pertussis- and cholera-toxin-mediated [32P]ADP-ribosylation of their respective G proteins were not affected.


CRISPR/Cas9-based knockout pipeline for reverse genetics in mammalian cell culture.
We present a straightforward protocol for reverse genetics in cultured mammalian cells, using CRISPR/Cas9-mediated homology-dependent repair (HDR) based insertion of a protein trap cassette, resulting in a termination of the endogenous gene expression. Complete loss of function can be achieved with monoallelic trap cassette insertion, as the second allele is frequently disrupted by an error-prone non-homologous end joining (NHEJ) mechanism. The method should be applicable to any expressed gene in most cell lines, including those with low HDR efficiency, as the knockout alleles can be directly selected for.


Evaluating health promotion: a longitudinal quasi-experimental design.
A quasi-experimental research design is used to evaluate Carolina Healthstyle, a health promotion project for South Carolina state employees. A 10% stratified random sample of employees was surveyed in the Spring of 1983 and again in 1984. Eighteen agencies were intervention agencies and the rest comparison that year. This article reports changes with simple before-after comparisons in the intervention agencies and matched pair analysis and randomized block designs to compare intervention and comparison agencies. Results are reported for smoking, exercise safety, nutrition, stress, and alcohol. Significant increases in exercise were found in both intervention and comparison agencies. The number of smokers decreased in intervention from 30%-26% with no change in comparison agencies. Safety practices changed at similar rates in both intervention and comparison groups. Consumption of chicken increased significantly only in the intervention agencies. Few other diet or stress changes were found. There were changes in alcohol consumption in intervention agencies only. The presence of the comparison group helps to separate the program effects from secular trends.


Impact of embolization during coronary artery bypass grafting on outcome and length of stay.
Transcranial Doppler ultrasonography detects emboli in most patients during coronary artery bypass grafting. However, the significance of these emboli has not yet been established. We monitored 82 patients during coronary artery bypass grafting with this technique and related the numbers of emboli to the outcomes and length of hospital stay. We detected cerebral emboli in all patients. Patients with stroke (n = 4; 4.9%) had a mean of 449 emboli, as compared with 169 emboli in patients without stroke (n = 78) (p = 0.005). Patients with major cardiac complications (n = 7) had a mean of 392 emboli, as compared with 163 in patients without such complications (n = 75) (p = 0.003). The mean hospital stay of survivors was 8.6 days in patients with fewer than 100 emboli (n = 40), 13.5 days in patients with 101 to 300 emboli (n = 23), 16.3 days in those with 301 to 500 emboli (n = 16), and 55.8 days in patients with more than 500 emboli (n = 6) (p = 0.0007). This relation was unchanged when patients with complications were excluded. The correlation between embolization and outcome was independent of the extent of aortic atheroma or age. Emboli detected during coronary artery bypass grafting are significantly related to major cardiac and neurologic complications and affect length of stay in all patients, even in the absence of such specific complications.


Cost-effectiveness of HMG-CoA reductase inhibition for primary and secondary prevention of coronary heart disease.
To determine the cost-effectiveness of HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase inhibitors (such as lovastatin) for the primary and secondary prevention of coronary heart disease, we used the Coronary Heart Disease Policy Model, a computer-stimulated model that estimates the risk factor-specific annual incidence of coronary heart disease and the risk of recurrent coronary events in persons with prevalent coronary heart disease. When used for secondary prevention, 20 mg/d of lovastatin was estimated to save lives and save costs in younger men with cholesterol levels above 250 mg/dL (6.47 mmol/L) and to have a favorable cost-effectiveness ratio regardless of the cholesterol level except in young women with cholesterol levels below 250 mg/dL (6.47 mmol/L). Doses of 40 mg/d of lovastatin had favorable incremental cost-effectiveness ratios in men with cholesterol levels above 250 mg/dL (6.47 mmol/L). By comparison, primary prevention had favorable cost-effectiveness ratios only in selected subgroups based on cholesterol levels and other established risk factors. We conclude that current national recommendations regarding medication for secondary prevention are not as aggressive as our projections would suggest, while recommendations regarding the use of medications for primary prevention should consider the cost of medication as well as the risk factor profile of the individual patient.


Overall Survival with Ribociclib plus Letrozole in Advanced Breast Cancer.
In a previous analysis of this phase 3 trial, first-line ribociclib plus letrozole resulted in significantly longer progression-free survival than letrozole alone among postmenopausal patients with hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced breast cancer. Whether overall survival would also be longer with ribociclib was not known. Here we report the results of the protocol-specified final analysis of overall survival, a key secondary end point. Patients were randomly assigned in a 1:1 ratio to receive either ribociclib or placebo in combination with letrozole. Overall survival was assessed with the use of a stratified log-rank test and summarized with the use of Kaplan-Meier methods after 400 deaths had occurred. A hierarchical testing strategy was used for the analysis of progression-free survival and overall survival to ensure the validity of the findings. After a median follow-up of 6.6 years, 181 deaths had occurred among 334 patients (54.2%) in the ribociclib group and 219 among 334 (65.6%) in the placebo group. Ribociclib plus letrozole showed a significant overall survival benefit as compared with placebo plus letrozole. Median overall survival was 63.9 months (95% confidence interval [CI], 52.4 to 71.0) with ribociclib plus letrozole and 51.4 months (95% CI, 47.2 to 59.7) with placebo plus letrozole (hazard ratio for death, 0.76; 95% CI, 0.63 to 0.93; two-sided P = 0.008). No new safety signals were observed. First-line therapy with ribociclib plus letrozole showed a significant overall survival benefit as compared with placebo plus letrozole in patients with HR-positive, HER2-negative advanced breast cancer. Median overall survival was more than 12 months longer with ribociclib than with placebo. (Funded by Novartis; MONALEESA-2 ClinicalTrials.gov number, NCT01958021.).


Flow resistance characteristics of the stem and root from conifer (Sabina chinensis) xylem tracheid.
Xylem tracheids are the channels for water transport in conifer. Tracheid flow resistance is composed of tracheid lumen resistance and pit resistance. The single tracheid structure parameters in the stem and root of Sabina chinensis were obtained by dissociation and slicing, combined with numerical simulation to analyze the tracheid flow resistance characteristics. The results showed that the tracheid lumen resistance was determined by the tracheid width and tracheid length. The pit resistance was determined by the number of pits and single pit resistance. The single pit resistance was composed of four elements: the secondary cell wall, the border, the margo and the torus. The margo contributed a relatively large fraction of flow resistance, while the torus, the border and the secondary cell wall formed a small fraction. The size and position of the pores in the margo had a significant effect on the fluid velocity. The number of pits were proportional to tracheid length. The power curve, S-curve and inverse curve were fitted the scatter plot of total pit resistance, total resistance, total resistivity, which was found that there were the negative correlation between them. The three scatter plot values were larger in the stem than in the root, indicating that the tracheid structure in the root was more conducive to water transport than the stem. The ratio of tracheid lumen resistance to pit resistance mainly was less than 0.6 in the stem and less than 1 in the root, indicating that the pit resistance was dominant in the total resistance of the stem and root.


Glucosamine attenuates drug resistance in Mitoxantrone-resistance breast cancer cells.
This study was aimed at investigating the cytotoxicity and multi-drug resistance (MDR) reversal effect of Glucosamine (GlcN) on resistant BCRP-overexpressing breast cancer MCF-7/MX cells. After confirming the overexpression of BCRP, the cytotoxicity and MDR reversing potential of GlcN on MCF-7/MX mitoxantrone-resistant and MCF-7 sensitive breast cancer cells were assessed via MTT assay. The effects of GlcN on mitoxantrone accumulation were analyzed through flow cytometry. Finally, the expression of BCRP and Epithelial-Mesenchymal Transition (EMT)-related markers following the exposure to GlcN were assessed by real-time RT-PCR. This study showed that glucosamine had an inhibitory effect on the proliferation of human breast cancer cells. The respective IC50 values for MCF-7/MX cells following exposure to mitoxantrone (MX) in the presence of GlcN (0, 0.5 and 1 mm) for 72 h were 3.61 ± 0.21, 0.598 ± 0.041 and 0.284 ± 0.016 μm, respectively. Furthermore, GlcN reduced the expression of BCRP mRNA without any significant effect on EMT-related markers in breast cancer cells. These results proposed that glucosamine as a natural sugar could down regulate the BCRP expression and increased MX cytotoxicity in breast cancer cells.


Opposite CD4/CD8 lineage decisions of CD4+8+ mouse and rat thymocytes to equivalent triggering signals: correlation with thymic expression of a truncated CD8 alpha chain in mice but not rats.
Unselected CD4+8+ rat thymocytes, generated in vitro from their direct precursors, are readily converted to functional TCRhigh T cells by stimulation with immobilized TCR-specific mAb plus IL-2. Lineage decision invariably occurs toward CD4-8+, regardless of the timing of TCR stimulation after entry into the CD4+8+ compartment or the concentration of TCR-specific mAb used for stimulation. CD4-specific mAb synergizes with suboptimal TCR-specific mAb in inducing T cell maturation, but lineage decision remains exclusively CD4-8+. These results contrast with those obtained in mice, in which Abs to the TCR complex were shown to promote CD4+8- T cell maturation from CD4+8+ thymocytes. Surprisingly, when rat and mouse CD4+8+ thymocytes were stimulated with PMA/ionomycin under identical conditions, the opposite lineage commitment was observed, i.e., mouse thymocytes responded with the generation of CD4+8- and rat thymocytes with the generation of CD4-8+ cells. It thus seems that CD4+8+ thymocytes of the two species respond with opposite lineage decisions to strong activating signals such as given by TCR-specific mAb or PMA/ionomycin. A possible key to this difference lies in the availability of p56lck for coreceptor. supported signaling. We show that in contrast to mouse CD4+8+ thymocytes, which express both a complete and a truncated CD8 alpha-chain (CD8 alpha') unable to bind p56lck, rat thymocytes only express full-length CD8 alpha molecules. Mice, but not rats, therefore may use CD8 alpha' as a "dominant negative" coreceptor chain to attenuate the CD8 signal, thereby facilitating MHC class II recognition through the higher amount of p56lck delivered, and rats may use a different mechanism for MHC class distinction during positive selection.


Six-year outcomes of carotid artery stenting performed with multidisciplinary management in a single center.
This study aimed to evaluate the first 30-day results of clinical, periprocedural asymptomatic cranial embolism, and long-term restenosis of the multidisciplinary conducted and evaluated carotid artery stenting (CAS) procedure in our patient group with real-life data. A total of 610 patients who were subjected to consecutive CAS procedures in our center between December 2010 and February 2019 were clinically and radiologically followed up for a mean duration of 6 years. Of the 610 patients, 274 (45%) were symptomatic for carotid artery stenosis, whereas 336 (55%) were identified as asymptomatic. As embolism protection methods, distal protection, proximal protection, and double (distal + proximal) protection was used in 52%, 43%, and 0.3% of patients, respectively. The success rate of the CAS procedure was 96%. Procedure-related death was reported in 4 (0.6%) patients who successfully underwent the CAS procedure. Moreover, acute carotid artery stent thrombosis, hyperperfusion syndrome, periprocedural major stroke, and periprocedural minor stroke was observed in 4 (0.6%), 2 (0.3%), 2 (0.3%), and 12 (1.9%) patients, respectively. The total clinical complication rates during the first 30 periprocedural days were 1.6% (10 patients) and 3.1% (19 patients) in the asymptomatic and symptomatic groups, respectively. On cranial magnetic resonance imaging performed, asymptomatic ipsilateral cranial microembolism, asymptomatic contralateral cranial microembolism, and bilateral asymptomatic cranial microembolism was detected in 61 (11.6%), 20 (3.8%), 23 (4.4%) patients, respectively. Asymptomatic restenosis was observed in 24 (3.9%) patients. The CAS procedure is a reliable treatment option applicable with acceptable complication and success rates as outlined in the guidelines, when performed following a multidisciplinary evaluation, in the treatment of symptomatic and asymptomatic carotid artery stenosis, including high-risk patient groups.


Granzyme B deficiency exacerbates lung inflammation in mice after acute lung injury.
Granzyme B (GzmB) is a serine protease with intracellular and extracellular activities capable of regulating inflammation through cytokine processing and the apoptosis of effector cells. We tested the hypothesis that GzmB expression in T regulatory cells (Tregs) is required for the control of inflammatory responses and pathology during acute lung injury. To substantiate the clinical relevance of GzmB during lung injury, we performed GzmB immunohistochemistry on lung tissue from patients with acute respiratory distress syndrome (ARDS) and healthy control subjects. We also performed in vivo experiments with wild-type (WT) C57BL/6 and GzmB(-/-) mice exposed to a single intranasal instillation of bleomycin to model lung injury. Our results demonstrate that the expression of GzmB was elevated in ARDS lung sections, relative to healthy control samples. Bleomycin-exposed GzmB(-/-) mice exhibited greater morbidity and mortality, which was associated with increased numbers of lung lymphocytes. Bleomycin induced an equal increase in CD4(+)/CD25(+)/FoxP3(+) Treg populations in WT and GzmB(-/-) mice. GzmB expression was not significant in Tregs, with the majority of the expression localized to natural killer (NK)-1.1(+) cells. The expression of GzmB in NK cells of bleomycin-exposed WT mice was associated with greater lymphocyte apoptosis, reduced total lymphocyte numbers, and reduced pathology relative to GzmB(-/-) mice. Our data demonstrate that GzmB deficiency results in the exacerbation of lymphocytic inflammation during bleomycin-induced acute lung injury, which is associated with pathology, morbidity, and mortality.


Pivotal role of the AREB/ABF-SnRK2 pathway in ABRE-mediated transcription in response to osmotic stress in plants.
Water availability is one of the main limiting factors for plant growth and development. The phytohormone abscisic acid (ABA) fulfills a critical role in coordinating the responses to reduced water availability as well as in multiple developmental processes. Endogenous ABA levels increase in response to osmotic stresses such as drought and high salinity, and ABA activates the expression of many genes via ABA-responsive elements (ABREs) in their promoter regions. ABRE-binding protein/ABRE-binding factor (AREB/ABF) transcription factors (TFs) regulate the ABRE-mediated transcription of downstream target genes. Three subclass III sucrose non-fermenting-1 related protein kinase 2 (SnRK2) protein kinases (SRK2D/SnRK2.2, SRK2E/SnRK2.6/OST1 and SRK2I/SnRK2.3) phosphorylate and positively control the AREB/ABF TFs. Substantial progress has been made in our understanding of the ABA-sensing system mediated by Pyrabactin resistance1/PYR1-like/regulatory components of ABA receptor (PYR/PYL/RCAR)-protein phosphatase 2C complexes. In addition to PP2C-PYR/PYL/RCAR ABAreceptor complex, the AREB/ABF-SnRK2 pathway, which is well conserved in land plants, was recently shown to play a major role as a positive regulator of ABA/stress signaling through ABRE-mediated transcription of target genes implicated in the osmotic stress response. This review focuses on current progress in the study of the AREB/ABF-SnRK2 positive regulatory pathway in plants and describes additional signaling factors implicated in the AREB/ABF-SnRK2 pathway. Moreover, to help promote the link between basic and applied studies, the nomenclature and phylogenetic relationships between the AREB/ABFs and SnRK2s are summarized and discussed.


Analysis of instantaneous amplitude and frequency of intracranial EEG signal to characterize epileptic seizure stages.
A graphical and analytical description of epileptic seizures based on amplitude modulation and frequency modulation components of intracranial EEG (iEEG) is proposed. This representation allows the characterization of seizures and their different stages from the iEEG by means of triangles whose vertexes are the centroids (cm) of the signal during preictal, ictal and postictal periods. The centroid is the point defined by the average values of instantaneous amplitude and frequency, ai and fi respectively. Data were obtained from 8 patients with recurrent epilepsy, 170 records were processed, 62 of which were seizures and 108 interictal signals. Results show that the centroids of the ictal periods are located in a region of the space ai-fi distant from the centroids corresponding to interictal and postictal periods. This original representation of epileptic seizures can facilitate the visualization of stage-transitions and discrimination between the different stages of the iEEG signal. An additional advantage of the method is that the information contained in the signal is synthesized significantly.


Structural investigations of the neutral polysaccharide of Linum usitatissimum L. seeds mucilage.
The heterogeneity of the purified water-soluble neutral fraction coming from the mucilage extract of the yellow flaxseed was investigated. After fractionation by size-exclusion chromatography, the analyse of the neutral monosaccharides composition showed a mixture of three major families of polymers. They were all identified as arabinoxylans with a constant A/X ratio of 0.24, but varying in their galactose and fucose residues in the side chains. Furthermore, the molecular weight (M(w)) analysis acquired by multi-angle laser light scattered, revealed the association of two high M(w) polymers [5.7x10(6) (11.1%) and 9.3x10(5) (42.4%) g mol(-1)] with a smaller one [3.2x10(5) g mol(-1) (45.3%)].


Prothrombin, alone or in complex concentrates or plasma, reduces bleeding in a mouse model of blood exchange-induced coagulopathy.
Prothrombin complex concentrates (PCC) are fractionated plasma protein drugs that reverse warfarin anticoagulation. PCC may control more general bleeding. We sought to identify the dominant procoagulant factor in PCC in vivo. We tested PCC or coagulation factor (F) treatment in CD1 mice made coagulopathic by exchange of whole blood for washed red cells. Anesthetized mice were transfused with murine fresh-frozen plasma (mFFP), PCC, mixtures of human vitamin K-dependent proteins (VKDP) (prothrombin, FVII, FIX, or FX), or purified single human VKDP, immediately prior to tail transection (TT), liver laceration (LL), or intravascular laser injury (ILI). Plasma donor mice were treated with vehicle or control antisense oligonucleotide (ASO-CON) or ASO specific for prothrombin (FII) (ASO-FII) to yield mFFP or ASO-CON mFFP or ASO-FII mFFP. Blood losses were determined spectrophotometrically (TT) or gravimetrically (LL). Thrombus formation was quantified by intravital microscopy of laser-injured arterioles. PCC or four factor- (4F-) VKDP or prothrombin significantly reduced bleeding from TT or LL. Omission of prothrombin from 4F-VKDP significantly reduced its ability to limit bleeding. Mice transfused with ASO-FII mFFP demonstrated inferior haemostasis versus those transfused with ASO-FII following TT, LL, or ILI. Prothrombin is the dominant procoagulant component of PCC and could limit bleeding in trauma.


Recommending healthy meal plans by optimising nature-inspired many-objective diet problem.
Healthy eating is an important issue affecting a large part of the world population, so human diets are becoming increasingly popular, especially with the devastating consequences of Coronavirus Disease (Covid-19). A realistic and sustainable diet plan can help us to have a healthy eating habit since it considers most of the expectations from a diet without any restriction. In this study, the classical diet problem has been extended in terms of modelling, data sets and solution approach. Inspired by animals' hunting strategies, it was re-modelled as a many-objective optimisation problem. In order to have realistic and applicable diet plans, cooked dishes are used. A well-known many-objective evolutionary algorithm is used to solve the diet problem. Results show that our approach can optimise specialised daily menus for different user types, depending on their preferences, age, gender and body index. Our approach can be easily adapted for users with health issues by adding new constraints and objectives. Our approach can be used individually or by dietitians as a decision support mechanism.


Micropenis.
Micropenis is part of a larger group of conditions broadly known as inconspicuous penis; however, it is fundamentally different from the other diagnoses in this group, such as webbed penis and buried penis, in that the underlying problem is the size of the penis itself, not with the surrounding and overlying skin. This condition is usually the result of a defect in the hypothalamic-pituitary-gonadal axis, although iatrogenic causes are identified infrequently. Management revolves around testosterone (direct administration or encouraging the patient's body to make its own), and long-term results with respect to increase in penile length are promising. Reconstructive surgery is based on the use of a vascular pedicle free flap and is reserved for patients who fail to respond to hormonal treatment. Although substantial long-term data are lacking, adult patients with micropenis appear to report dissatisfaction with penile appearance, but the majority appear to have adequate sexual function.


A role for Dehalobacter spp. in the reductive dehalogenation of dichlorobenzenes and monochlorobenzene.
Previously, we demonstrated the reductive dehalogenation of dichlorobenzene (DCB) isomers to monochlorobenzene (MCB), and MCB to benzene in sediment microcosms derived from a chlorobenzene-contaminated site. In this study, enrichment cultures were established for each DCB isomer and each produced MCB and trace amounts of benzene as end products. MCB dehalogenation activity could only be transferred in sediment microcosms. The 1,2-DCB-dehalogenating culture was studied the most intensively. Whereas Dehalococcoides spp. were not detected in any of the microcosms or cultures, Dehalobacter spp. were detected in 16S rRNA gene clone libraries from 1,2-DCB enrichment cultures, and by PCR using Dehalobacter-specific primers in 1,3-DCB and 1,4-DCB enrichments and MCB-dehalogenating microcosms. Quantitative PCR showed Dehalobacter 16S rRNA gene copies increased up to 3 orders of magnitude upon dehalogenation of DCBs or MCB, and that nearly all of bacterial 16S rRNA genes in a 1,2-DCB-dehalogenating culture belonged to Dehalobacter spp. Dehalobacter 16S rRNA genes from DCB enrichment cultures and MCB-dehalogenating microcosms showed considerable diversity, implying that 16S rRNA sequences do not predict dehalogenation-spectra of Dehalobacter spp. These studies support a role for Dehalobacter spp. in the reductive dehalogenation of DCBs and MCB, and this genus should be considered for its potential impact on chlorobenzene fate at contaminated sites.


A New, High-Efficacy, Noninvasive Transcranial Electric Stimulation Tuned to Local Neurodynamics.
In this paper, we pose the following working hypothesis: in humans, transcranial electric stimulation (tES) with a time course that mimics the endogenous activity of its target is capable of altering the target's excitability. In our case, the target was the primary motor cortex (M1). We identified the endogenous neurodynamics of hand M1's subgroups of pyramidal neuronal pools in each of our subjects by applying Functional Source Separation (FSS) to their EEG recordings. We then tested whether the corticospinal excitability of the hand representation under the above described stimulation, which we named transcranial individual neurodynamics stimulation (tIDS), was higher than in the absence of stimulation (baseline). As a check, we compared tIDS with the most efficient noninvasive facilitatory corticospinal tES known so far, which is 20 Hz transcranial alternating current stimulation (tACS). The control conditions were as follows: (1) sham, (2) transcranial random noise stimulation (tRNS) in the same frequency range as tIDS (1-250 Hz), and (3) a low current tIDS (tIDSlow). Corticospinal excitability was measured with motor-evoked potentials under transcranial magnetic stimulation. The mean motor-evoked potential amplitude increase was 31% of the baseline during tIDS (p < 0.001), and it was 15% during tACS (p = 0.096). tRNS, tIDSlow, and sham induced no effects. Whereas tACS did not produce an enhancement in any subject at the individual level, tIDS was successful in producing an enhancement in 8 of the 16 subjects. The results of the present proof-of-principle study showed that proper exploitation of local neurodynamics can enhance the efficacy of personalized tES.SIGNIFICANCE STATEMENT This study demonstrated that, in humans, transcranial individual neurodynamics stimulation (tIDS), which mimics the endogenous dynamics of the target neuronal pools, effectively changes the excitability of these pools. tIDS holds promise for high-efficacy personalized neuromodulations based on individual local neurodynamics.


Effects of differentiation inducing chemicals on in vivo malignancy and NK susceptibility of metastatic lymphoma cells.
In the present investigation we have determined the effects of the differentiation-inducing chemicals dimethyl sulfoxide (DMSO) and sodium butyrate on the growth and tumorigenicity of a highly malignant/metastatic cell line (RAW117-H10) and its less tumorigenic parental lymphoma cell line (RAW117-P). Both of these agents at doses shown to be nontoxic slowed the growth of these cells in suspension culture and significantly lengthened the doubling time while reducing colony formation in the agar tumor stem cell assay. Corresponding to these observations, the in vivo tumorigenicity of the highly malignant RAW117-H10 line was reduced by both chemicals but particularly by butyrate treatment. In addition, both agents increased the expression of some glycoproteins and the glycolipid asialo GM1. There was also a corresponding increase in the NK susceptibility of the normally NK resistant RAW117-H10 cells. To determine if the decreased malignancy of the highly malignant cells following treatment with the chemical agents was primarily due to the alteration in cell surface glycoconjugates or merely due to concomitant decreased growth potential, we transplanted highly glycosylated membrane fragments from normal syngeneic thymocytes to both RAW117-P and RAW117-H10 cells using a Sendai virus mediated membrane fusion technique. The in vivo tumorigenicity of the membrane altered RAW117-H10 cells was significantly decreased. These results strongly suggest that the decreased in vivo malignancy of RAW117-H10 cells resulting from treatment with chemical differentiation agents is caused by their increased susceptibility to NK cell mediated lysis which in turn results from cell surface changes involving altered, primarily increased, expression of certain glycosylated surface molecules. The cell surface glycoconjugates, such as receptors for certain lectins and glycolipid asialo GM1, can be used as markers for malignant potential and NK sensitivity of malignant lymphoid cells.


Ultrasonic standing wave manipulation technology integrated into a dielectrophoretic chip.
Several cell-based biological applications in microfluidic systems require simultaneous high-throughput and individual handling of cells or other bioparticles. Available chip-based tools for contactless manipulation are designed for either high-precision handling of individual particles, or high-throughput handling of ensembles of particles. In order to simultaneously perform both, we have combined two manipulation technologies based on ultrasonic standing waves (USWs) and dielectrophoresis (DEP) in a microfluidic chip. The principle is based on the competition between long-range ultrasonic forces, short-range dielectrophoretic forces and viscous drag forces from the fluid flow. The ultrasound is coupled into the microchannel resonator by an external transducer with a refractive element placed on top of the chip, thereby allowing transmission light microscopy to continuously monitor the biological process. The DEP manipulation is generated by an electric field between co-planar microelectrodes placed on the bottom surface of the fluid channel. We demonstrate flexible and gentle elementary manipulation functions by the use of USWs and linear or curved DEP deflector elements that can be used in high-throughput biotechnology applications of individual cells.


The use of machine learning for the identification of peripheral artery disease and future mortality risk.
A key aspect of the precision medicine effort is the development of informatics tools that can analyze and interpret "big data" sets in an automated and adaptive fashion while providing accurate and actionable clinical information. The aims of this study were to develop machine learning algorithms for the identification of disease and the prognostication of mortality risk and to determine whether such models perform better than classical statistical analyses. Focusing on peripheral artery disease (PAD), patient data were derived from a prospective, observational study of 1755 patients who presented for elective coronary angiography. We employed multiple supervised machine learning algorithms and used diverse clinical, demographic, imaging, and genomic information in a hypothesis-free manner to build models that could identify patients with PAD and predict future mortality. Comparison was made to standard stepwise linear regression models. Our machine-learned models outperformed stepwise logistic regression models both for the identification of patients with PAD (area under the curve, 0.87 vs 0.76, respectively; P = .03) and for the prediction of future mortality (area under the curve, 0.76 vs 0.65, respectively; P = .10). Both machine-learned models were markedly better calibrated than the stepwise logistic regression models, thus providing more accurate disease and mortality risk estimates. Machine learning approaches can produce more accurate disease classification and prediction models. These tools may prove clinically useful for the automated identification of patients with highly morbid diseases for which aggressive risk factor management can improve outcomes.


Tetrachlorodibenzo-p-dioxin induction of renal microsomal enzyme systems: ultrastructural effects on pars recta (S3) proximal tubule cells on the rat kidney.
A single oral administration of 2,3,7,8-tetrachlorodibenzo-p-dioxin to rats at a dose level of 25 microgram/kg produced marked proliferation of smooth endoplasmic reticulum in renal proximal tubule cells of the straight (S3) segments. This ultrastructural effect was associated with a pronounced induction of the microsomal enzymes glucuronyl transferase and benzopyrene hydroxylase which persisted for at least 16 days after treatment. Renal dissection studies disclosed that the activities of these enzymes in 2,3,7,8-tetrachlorodibenzo-p-dioxin-treated animals were not uniformly distributed within the kidney but followed the general known distribution of S3 cells. Enzyme activities were highest in the outer stripe of the medulla and cortex and lowest in the medulla. These studies demonstrate that the kidney does possess inducible microsomal enzyme systems, capabilities of which have not been appreciated previously due to the lack of a suitable inducing agent and their apparent concentration in a relatively small population of proximal tubule cells.


Bilateral congenital lacrimal fistula in a Brown Swiss bull.
A five-year-old Brown Swiss bull was referred to the Department of Farm Animals, University of Zurich, because of bilateral epiphora that was unresponsive to treatment. Clinical examination revealed a fistulous opening medial to the medial canthus of both eyes and mucopurulent discharge from both openings. Attempts to flush the nasolacrimal duct via the lacrimal points resulted in the fluid exiting via the fistulous opening. Retrograde flushing of the nasolacrimal duct from the nasolacrimal opening resulted in the flush fluid flowing back out the nasolacrimal opening. Bilateral lacrimal fistula medial to the medial canthus of the eye was diagnosed based on the findings. The same anomaly was diagnosed a year later in 4 related female animals referred to our Department for other reasons. Three of the cases were sired by the bull described above and one was sired by his half-brother. Therefore, an autosomal recessive mode of inheritance of this anomaly was assumed. Clinical, epidemiological and molecular studies of the offspring of both bulls are underway to further investigate this anomaly.


Perioperative anaesthetic management of patients undergoing thoracic cytoreductive surgery and HITHOC.
Historically, patients with pleural malignant mesothelioma have had a poor prognosis and survival rate. Recently, new surgical approaches and chemotherapy delivery techniques have been developed. One of this treatment options is thoracic cytoreductive surgery and HITHOC (hyperthermic intrathoracic chemotherapy perfusion), a promising strategy in selected patients, offering significantly longer median survival length and tumour-free survival rate. However, there is little experience and little is known regarding the optimal perioperative management of this patients. Given that they usually present with poor preoperative status and the surgery is aggressive, prolonged and associated with significant hemodynamic repercussions, this procedure poses a true challenge to the anaesthesiologist. We will discuss optimal patient selection and optimization, as well as premedication, recommended monitoring aspects on top of the usual for any anaesthetic procedure, induction and anaesthetic agents, blood management and one lung ventilation. Also, we expose the importance of adequate pain control during the surgery and postoperatively, the hemodynamic disturbances that occur during the procedure and the potential complications that could occur afterwards. In a few words, this review intends to offer recommendations for the management of patients undergoing cytoreductive surgery and HITHOC for the perioperative care, based on the scarce evidence and our clinical experience.


Pregnant women carrying microcephaly foetuses and Zika virus contain potentially pathogenic microbes and parasites in their amniotic fluid.
Microcephaly has become a major public health problem in Brazil. The total number of newborns with microcephaly was reported to be >4000 in June 2016. Studies suggest that Zika Virus is a major cause of new microcephaly cases in Brazil. Inside the uterus, the foetus is surrounded by the Amniotic Fluid, a proximal fluid that contains foetal and maternal cells as well as microorganisms and where Zika Virus was already found. A previous study reported the presence of the Zika Virus in the amniotic fluid (collected in the 28th gestational week) of two pregnant women carrying microcephaly foetuses in Brazil. The virus was detected by means of real-time PCR and metatranscriptomic analysis. We compared the microbiome of these two cases with metatranscriptomic sequences from 16 pregnant women collected at various times in their pregnancies CONCLUSION: Several strains of bacteria (e.g., Streptococcus and Propionibacterium) found in Amniotic Fluid may be involved in neurological diseases. When the foetus is infected by the Zika Virus, due to neurological damage, they do not move inside the uterus, thus changing the Amniotic Fluid environment, potentially leading to secondary problems. Zika infection could also lead to an immunodeficient state, making bacterial colonization of the foetuses easier. An altered microbial composition during pregnancy may also result in harmful secondary metabolite production from certain microbes that further impair foetal brain development. However, these observations of potentially harmful microbial species are correlations and thus cannot be assumed to be causative agents of (microcephaly) disease. In our study, microbial and parasitic diversity of the Amniotic Fluid was lower in patients infected by ZIKV, compared to that of Prenatal and Preterm controls. The present study was a first attempt to shed light on the microbial and parasitic diversity associated with ZIKV-infected pregnant women bearing microcephaly foetuses, and the presence of diverse microbial and parasite communities in the Amniotic Fluid suggests a poor health status of both the pregnant women and the foetuses they carry.


Direct evidence for tautomerization of the uracil moiety within the Pb2+/uridine-5'-monophosphate complex: a combined tandem mass spectrometry and IRMPD study.
The structure of the [Pb(UMP)-H](+) (UMP = uridine-5'-monophosphate) complex was studied in the gas phase by combining electrospray ionization (ESI), tandem mass spectrometry, and mid-infrared multiple photon dissociation (IRMPD) spectroscopy. The results obtained show that Pb(2+) ions interact not only with the deprotonated phosphate group but also with a carbonyl group of the nucleobase moiety by folding of the mononucleotide, resulting in macrochelates that are not likely to be present in solution. Comparison between the IRMPD and DFT-computed spectra suggests that the ESI-generated complex likely corresponds to a mixture of several structures, and establishes the enolic tautomers as the most abundant species for the [Pb(UMP)-H](+) ion, while the very weak IRMPD signal observed at ∼1763 cm(-1) points to a minor population of oxo forms. Our data also suggest that losing the nucleobase residue under CID conditions does not necessarily mean a lack of interaction between the metal and the nucleobase moiety, as commonly reported in the literature for large oligonucleotides.


Modified sublimation to isolate phenanthrene-degrading bacteria of the genera Sphingomonas and Burkholderia from Xiamen oil port.
Sublimation was developed by Alley and Brown (2000) in order to isolate bacterial strains that were capable of degrading water insoluble compounds. In this study, sublimation was modified by the use of nutritional agar plates, instead of mineral salt agar, to isolate phenanthrene-degrading bacteria from a mixed culture that had been enriched under the selective pressure of high phenanthrene content. Five strains were obtained with different morphology and degradation ability. Based on the 16S rDNA sequence, two of them were classified as species of the genus Sphingomonas; the others as species of the genus Burkholderia. Denaturing gradient gel electrophoresis (DGGE) was introduced to detect dynamic changes in the bacterial community during enrichment batch culture, and to determine any correlation between the five isolates and the phenanthrene-degrading consortium. The DGGE profile indicated that these five isolates corresponded to four dominant bands of the consortium. Compared to traditional means of isolation, we concluded that modified sublimation is effective and more convenient.


Adalimumab-induced sensory vasculitic neuropathy.
Vasculitis and other autoimmune conditions are known complications of tumour necrosis factor alpha (TNF-α) inhibitor use. By definition, TNF-α inhibitor induced vasculitis is a secondary systemic vasculitis. However, its phenotype is varied and can present as an isolated vasculitic neuropathy. This presents a diagnostic challenge as the gold standard for diagnosis of a vasculitic neuropathy is a peripheral nerve biopsy that meets predefined histopathological criteria. Given the poor sensitivity of the peripheral nerve biopsy, it is important that clinicians take a good history and maintain a high index of suspicion, as this is a treatable iatrogenic condition. Here we present a case of adalimumab-induced sensory vasculitic neuropathy, treated according to the Peripheral Nerve Society guideline for non-systemic vasculitic neuropathy, given her disease phenotype.


Sociodemographic predictors of response to antidepressant treatment.
Our goal was to assess whether sociodemographic variables such as gender, marital status, level of education, and employment status are related to the changes in social functioning that have been reported after drug treatment in outpatients with major depressive disorder. Eligible subjects were 166 depressed outpatients participating in a study involving open treatment with fluoxetine 20 mg/day for eight weeks. Diagnosis of major depressive disorder was made with the use of the Structured Clinical Interview for DSM-III-R-Patient Edition (SCID-P), and patients were required to have a seventeen-item Hamilton Rating Scale for Depression (HAM-D-17) score > or = at study entry. All subjects were administered the HAM-d-17 and the Social Adjustment Scale-Self-Report (SAS-SR) before and after treatment with fluoxetine. We found that SAS-SR scores decreased significantly following treatment with fluoxetine from a mean score at baseline of 2.6 +/- 0.7 to a mean score at endpoint of 2.3 +/- 0.6. After adjusting for the degree of change in HAM-D-17 scores, we found a significant relationship between degree of change in SAS-SR and level of education. No statistically significant relationships were observed between SAS-SR change and age, gender, marital status, and employment status. The degree of improvement in psychosocial functioning observed in depressed outpatients following antidepressant treatment appears to be related to the level of education at study entry, but not to other sociodemographic variables. Further studies need to investigate the nature of this relationship.


Publication output in telemedicine in Spain.
We searched the MEDLINE database for publications about telemedicine from 1966 to 2003. Only publications with a first institutional address in Spain and from journals listed in the JCR Science Edition 2002 were included in the study. A total of 118 publications were found. The first publication appeared in 1988 and publication output showed a sudden increase starting in 1995, reaching a maximum in 2002 with 21 publications. The number of authors per publication in non-multicentre studies ranged from 1 to 14 (median 6.5). Most authors (79%) contributed to only a single publication; 10% of authors participated in three or more publications. Most publications were full papers (88%). The most active Autonomous Community in the number of publications relative to inhabitants was Galicia (0.91 publications per 100,000 inhabitants). The Community that produced the highest absolute number of publications was Madrid (29% of the total). The mean impact factor of the journals in which the publications appeared was 0.961. Production of telemedicine publications in Spain has followed the same temporal course as has been observed worldwide. There is an unequal geographical and institutional distribution of publications. The highest production is concentrated in a few institutions and only a small number of authors show steady research activity.


An Occurrence of Metastable Cristobalite in High-Pressure Garnet Granulite
High-pressure (0.8 gigapascals) granulite facies garnet from Gore Mountain, New York, hosts multiple solid inclusions containing the low-pressure silica polymorph cristobalite along with albite and minor ilmenite. Identification of cristobalite is based on Raman spectra, electron microprobe analysis, and microthermometric measurements on the alpha/beta phase transformation. The cristobalite plus albite inclusions may have originated as small, trapped samples of hydrous sodium-aluminum-siliceous melt. Diffusive loss of water from these inclusions under isothermal, isochoric conditions may have resulted in a large enough internal pressure decrease to promote the metastable crystallization of cristobalite.


Evidence of cell-mediated immune response and specific local mucosal immunoglobulin (Ig) A production against Lawsonia intracellularis in experimentally infected swine.
The purpose of this study was to detect cell-mediated and local humoral immune responses to Lawsonia intracellularis in pigs inoculated with a pure culture of the pathogenic isolate or with an intestinal mucosa homogenate. Twenty-four 5-week-old pigs were inoculated with a pure culture of L. intracellularis (n = 10), an intestinal mucosa homogenate from proliferative enteropathy diseased pigs (n = 10), or a control solution (n = 4). All animals were bled 0, 7, 14, and 20 d post-inoculation (pi). Serum was tested for immunoglobulin (Ig) G against L. intracellularis and for the production of interferon (IFN)-gamma by peripheral blood mononuclear cells (PBMC) after inoculation with L. intracellularis total proteins. Delayed-type hypersensitivity (DTH) reactions were evaluated 24 and 48 h after intra-dermal injection of different concentrations of L. intracellularis antigen 20 d pi. All animals were euthanized on day 22, intestinal lavages of ileum and IgA titrations were done. Weak IFN-gamma production was detected in 1 pig from the pure culture group and 2 pigs from the mucosal homogenate group 14 d pi, and in 2 animals from both groups 20 d pi. All pigs, in both inoculated groups, were seropositive for IgG on day 20. Inoculated pigs from both groups showed very weak dose-dependent DTH reactions, which were more evident at 24 h than 48 h pi. Eight pigs from the pure culture group and 7 from the mucosa homogenate group had detectable IgA titers in the intestinal lavage 22 d pi. In conclusion, specific local intestinal humoral and weak cell-mediated immune responses can be detected in pigs experimentally infected with L. intracellularis.


Reconstruction of Flight Parameters of a Bat for Flapping Robots.
The flight of bats is comparatively less documented and understood than birds and insects and may provide novel inspiration for the design of flapping flight robots. This study captured the natural flight of short-nosed fruit bats (Cynopterus sphinx) by an optical motion capture system, "OptiTrack", with pasted markers on the wings and body to reconstruct the flight parameters. Due to the self-occlusion at some moments, points on the membrane wings cannot be captured by any cameras. To draw a smooth trajectory, it is desired to reconstruct all missing data. Therefore, an algorithm is proposed by using numerical techniques, accompanied by modern mathematical and computational tools, to envisage the missing data from the captured flight. The least-square fitted polynomial engendered the parameter equations for x-, y-, and z-coordinates of marked points which were used to reconstruct the trajectory of the flight. The parameter equations of position coordinates were also used to compute the morphological and aerodynamic characteristics of the flight. The most outstanding contribution of the work is that not only the trajectory, velocity, and velocity field but also the morphing areas of the membrane wings were recreated using the reconstructed data. These data and reconstructed curves of trajectory and velocity field will be used for the further aerodynamic analysis and mechanism design of the flapping robot. This method can also be generalized to reconstruct the performance parameters of any other animals for bionic design.


Vaccine-Specific Immune Responses against Mycobacterium ulcerans Infection in a Low-Dose Murine Challenge Model.
The neglected tropical disease Buruli ulcer (BU) is an infection of subcutaneous tissue with Mycobacterium ulcerans There is no effective vaccine. Here, we assessed an experimental prime-boost vaccine in a low-dose murine tail infection model. We used the enoyl reductase (ER) domain of the M. ulcerans mycolactone polyketide synthases electrostatically coupled with a previously described Toll-like receptor 2 (TLR-2) agonist-based lipopeptide adjuvant, R4Pam2Cys. Mice were vaccinated and then challenged via tail inoculation with 14 to 20 CFU of a bioluminescent strain of M. ulcerans Mice receiving either the experimental ER vaccine or Mycobacterium bovis bacillus Calmette-Guérin (BCG) were equally protected, with both groups faring significantly better than nonvaccinated animals (P < 0.05). To explore potential correlates of protection, a suite of 29 immune parameters were assessed in the mice at the end of the experimental period. Multivariate statistical approaches were used to interrogate the immune response data to develop disease-prognostic models. High levels of interleukin 2 (IL-2) and low gamma interferon (IFN-γ) produced in the spleen best predicted control of infection across all vaccine groups. Univariate logistic regression revealed vaccine-specific profiles of protection. High titers of ER-specific IgG serum antibodies together with IL-2 and IL-4 in the draining lymph node (DLN) were associated with protection induced by the ER vaccine. In contrast, high titers of IL-6, tumor necrosis factor alpha (TNF-α), IFN-γ, and IL-10 in the DLN and low IFN-γ titers in the spleen were associated with protection following BCG vaccination. This study suggests that an effective BU vaccine must induce localized, tissue-specific immune profiles with controlled inflammatory responses at the site of infection.


Referral success among HIV-infected women and HIV-exposed children referred for monitoring and treatment in Tanga, Tanzania.
Long-term retention of patients in the rapidly expanding antiretroviral programs against HIV/AIDS is essential for the success of these programs. Unfortunately, high percentages of patients in need of treatment are lost to follow up in resource-poor settings. From June 2006 to March 2007 269 HIV-infected pregnant women were screened for participation in a Prevention of Mother to Child Transmission (PMTCT) study in Tanga, Tanzania. Ninety-one women were referred to Care and Treatment Center (CTC) for monitoring and antiretroviral therapy regardless of inclusion in the PMTCT study. Both women and HIV-exposed children were traced at CTC from mid-February to mid-March 2008. Follow-up visits for all patients were examined. Only 10% (9/91) of the referred women were still attending CTC by February/March 2008 and 64% (58/91) never attended CTC. Not a single child could be traced at CTC. The largest loss of patients occurs between referral and the first attendance at CTC. Every effort must be made to clarify reasons for loss of patients at this point. Further, measures to improve patient retention must be increased as must initiatives to improve the ability to trace and retain children within the clinics.


Patient-reported outcome after primary and aseptic revision hip arthroplasty: 1-year follow-up of 3,559 primary and 406 revision THAs in an institutional registry.
Background and purpose - Patient-reported outcomes (PROMs) after primary total hip arthroplasty (THA) and revision THA are important information in the preoperative shared decision-making process. We present 1-year results on pain, function, and quality of life following primary and revision THA. Patients and methods - From 2010 to 2018, 3,559 primary THA and 406 revision THAs were included in our institutional quality registry. PROMs were registered preoperatively, 3 months, and 1 year after surgery, numeric rating scale (0-10) for pain during mobilization and at rest, healthrelated quality of life (EQ-5D), and a hip-specific physical function score (HOOS-PS). 2 anchor questions were asked 1 year after surgery concerning joint function and willingness to go through surgery again. Results - There were statistically significant improvements in all PROMs at the 3-month follow-up in both groups. All PROMs improved more in the primary group relative to the revision group. 1 year after surgery, pain during mobilization was reduced with a mean change of 5.1 (SD 2.6) for primary THA and 2.9 (SD 3.0) for revision THA. 93% of primary THA patients reported both better function 1 year after surgery and that they would have gone through surgery again, compared with 78% and 79% in the revision THA group. Interpretation - Primary THA patients reported better function and more pain relief than the revision THA group 1 year after surgery. Pain during mobilization shows the most marked improvement in both groups, which is important preoperative information for patients.


CYP3A4 genetic polymorphism influences repaglinide's pharmacokinetics.
To investigate the effects of CYP3A4 and CYP2C8 enzymes on repaglinide's pharmacokinetics in healthy Malaysian subjects. Subjects (n = 121) received oral repaglinide (4 mg). Blood samples were taken at 0, 30, 60, 120, 180 and 240 min and serum concentrations of repaglinide were determined using high-performance liquid chromatography. Subjects were also genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for CYP3A4*4, *5 and*18 and by an allele-specific multiplex PCR for CYP2C8*2, *3, *4 and *5 alleles. The allele frequencies of CYP2C8*1, *2, *3, *4 and *5 were 95.04, 0.40, 0.40, 0 and 4.13%, respectively. The frequencies of the CYP3A4*1, *4, *5 and *18 alleles were 97.93, 0, 0 and 2.07%, respectively. CYP2C8 and CYP3A4 genotypes were not significantly associated with repaglinide's blood glucose-lowering effect. However, the CYP3A4 genotype significantly influenced some of repaglinide's pharmacokinetics, where the mean elimination rate constant was 44.0% lower (p = 0.04) and the mean half-life was 33.8% higher (p = 0.04) in subjects with the CYP3A4*1/*18 genotype as compared to those with the normal CYP3A4*1/*1 genotype. This result confirms that CYP3A4 plays a large role in metabolizing repaglinide. Genetic polymorphisms of CYP3A4, specifically CYP3A4*18, play a major role in contributing to the interindividual variability in repaglinide's pharmacokinetics.


Ultrasound in treatment of back pain resulting from prolapsed intervertebral disc.
The efficacy of ultrasound therapy in the management of back pain resulting from rupture of the intervertebral disc was evaluated in 3 groups: treatment, placebo, and control. Range of motion of total flexion and extension, total side flexion and total rotation of the lumbar spine, and subjective assessment of pain were used as criteria to determine the efficacy of treatment. Group comparisons showed statistical significance in favor of the treatment group (p 0.01), leading to the conclusion that ultrasound therapy is significantly effective in the treatment of back pain resulting from prolapse of the lumbar intervertebral disc.


A quantitative evaluation of confidence measures for stereo vision.
We present an extensive evaluation of 17 confidence measures for stereo matching that compares the most widely used measures as well as several novel techniques proposed here. We begin by categorizing these methods according to which aspects of stereo cost estimation they take into account and then assess their strengths and weaknesses. The evaluation is conducted using a winner-take-all framework on binocular and multibaseline datasets with ground truth. It measures the capability of each confidence method to rank depth estimates according to their likelihood for being correct, to detect occluded pixels, and to generate low-error depth maps by selecting among multiple hypotheses for each pixel. Our work was motivated by the observation that such an evaluation is missing from the rapidly maturing stereo literature and that our findings would be helpful to researchers in binocular and multiview stereo.