Hypoxia inhibits L-arginine synthesis from L-citrulline in porcine pulmonary artery endothelial cells.
Both non-arginine-depleted and arginine-depleted pulmonary artery endothelial cells (PAEC) actively convert citrulline into arginine. Exposure to hypoxia for 4-24 h inhibited arginine synthesis from citrulline in intact cells and in cell homogenates. The conversion of L-citrulline to L-argininosuccinate by argininosuccinate synthetase (AS) was inhibited by exposure to hypoxia for 4, 12, or 24 h. The conversion of argininosuccinate to arginine by argininosuccinate lyase was inhibited by exposure to hypoxia for 24 h but not for 4-12 h. The decrease of L-arginine biosynthesis during hypoxia coincided with the increase of intracellular glutamine content and was abrogated by preventing an increase in intracellular glutamine. In addition, AS activity was inversely related to glutamine content in the medium. These results indicate that hypoxia inhibited the L-arginine biosynthetic pathway via decreased activity of AS. The latter is related to increased glutamine content. Hypoxic inhibition of arginine synthesis from citrulline did not result in a decrease of arginine content, suggesting that PAEC are able to maintain intracellular arginine for up to 24 h despite reduction in the L-arginine biosynthetic pathway.


Spectroscopic investigation of the binding interactions of a membrane potential molecule in various supramolecular confined environments: contrasting behavior of surfactant molecules in relocation or release of the probe between nanocarriers and DNA surface.
The fluorescence and optical properties of membrane potential probes are widely used to measure cellular transmembrane potentials. Hemicyanine dyes are also able to bind to membranes. The spectral properties of these molecules depend upon the charge shift from the donor moiety to the acceptor moiety. Changes in their spectral properties, i.e. absorption and emission maxima or intensities, are helpful in characterizing model membranes, microheterogeneous media, etc. In this article, we have demonstrated the binding interaction of a membrane potential probe, 1-ethyl-2-(4-(p-dimethylaminophenyl)-1,3-butadienyl)-pyridinium perchlorate (LDS 698), with various supramolecular confined environments. The larger dipole moment in the ground state compared to the excited state is a unique feature of hemicyanine dyes. Due to this unique feature, red shifts in the absorption maxima are observed in hydrophobic environments, compared with bulk solvent. On addition of surfactants and CT DNA to an aqueous solution containing LDS 698, significant increase in the emission intensity along with the quantum yield and lifetime indicate partition of the probe molecules into organized assemblies. In the case of the sodium dodecyl sulfate (SDS)-water system, due to interactions between the cationic LDS 698 and the anionic dodecyl sulfate moiety, the fluorescence intensity at ∼666 nm decreases and an additional peak at ∼590 nm appears at premicellar concentration (∼0.20 mM-4.50 mM). But at ∼5.50 mM SDS concentration, the absorbance in the higher wavelength region increases again, indicating encapsulation of the probe in micellar aggregates. This observation indicates that the premicellar aggregation behavior of SDS can also be judged by observing the changes in the UV-vis and fluorescence spectral patterns. The temperature dependent study also indicates that non-radiative deactivation of the dye molecules is highly restricted in the DNA micro-environment, compared with micelles. Besides, we have also investigated the specific interaction of surfactant micelles with DNA. Our observations reveal that, in the presence of CT DNA, LDS 698 interacts exclusively with SDS micelles, but that it preferentially releases from micelles and relocates to DNA surfaces in solutions containing TX-100 micelles.


Wound-healing defects in mice lacking fibrinogen.
In addition to its key role in the control of blood loss following injury, fibrin(ogen) has been proposed to play an important role in tissue repair by providing an initial matrix that can stabilize wound fields and support local cell proliferation and migration. To test directly these concepts, the effect of fibrinogen deficiency on cutaneous tissue repair in mice was investigated using incisional and excisional wounds. The time required to overtly heal wounds was similar in fibrinogen-deficient and control mice, but histologic evaluation revealed distinct differences in the repair process, including an altered pattern of epithelial cell migration and increased epithelial hyperplasia. Furthermore, granulation tissue in fibrinogen-deficient mice failed to adequately close the wound gap, resulting in persistent open wounds or partially covered sinus tracts. The tensile strength of these wounds was also reduced compared with control mice. The most profound defect in wound tissue organization was observed in fibrinogen-deficient mice following the subcutaneous implantation of a porous tubing chamber. Cells migrated into the wall of the implants at a similar rate as control mice, but cells from fibrinogen-deficient animals were unable to efficiently organize and migrate into wound fluid-filled dead space within the center of the implants. These studies show that re-epithelialization, granulation tissue formation, including the establishment of neovasculature, and the formation of fibrotic scar tissue can proceed in the absence of fibrin(ogen) and all of its proteolytic derivatives. However, fibrin (ogen) is important for appropriate cellular migration and organization within wound fields and in initially establishing wound strength and stability. (Blood. 2001;97:3691-3698)


HIV vaccine to induce cytotoxic T cells recognizing conserved HIV-1/2-epitopes derived from the most frequent HLA types of the human population.
Evaluation of: Pleguezuelos O, Stoloff GA, Caparros-Wanderley W. Synthetic immunotherapy induces HIV virus specific Th1 cytotoxic response and death of an HIV-1 infected human cell line through classic complement activation. Virol. J. 10(1), 107 (2013). AIDS vaccine development represents an unprecedented challenge in both immunogen design and delivery to induce potent and long-lasting HIV-specific immune responses, including neutralizing antibodies and cytotoxic T lymphocytes (CTL). Pleguezuelos and coworkers recognized that immunogen design must address both HIV and HLA diversity to make a global vaccine. The HIV-v synthetic polypeptide vaccine described here sets a new standard in antigen design by selecting conserved regions of global HIV-1 and HIV-2 isolates and epitopes from most frequent HLA types of the human population. The new vaccine induced both antibody and CTL responses. Importantly, the authors demonstrated vaccine-specific HLA restricted CD8(+) CTL responses for one HLA allele that was involved in the antigen design. HIV-v vaccine is a new candidate to be tested in human subjects carrying frequent HLA types.


[Comparison of the activities of silver nitrate with those of three antifungal agents against ocular pathogenic fungi in vitro].
To investigate antifungal activity of silver nitrate compared with fluconazole, ketoconazole and amphotericin B against ocular pathogenic fungi in vitro. It was an experimental study. Susceptibility tests were performed against 260 isolates (15 genera and 29 species) of ocular pathogenic fungi by broth dilution antifungal susceptibility testing of filamentous fungi (M38-A) approved by National Committee for Clinical Laboratory Standards (NCCLS). Final concentrations ranged from 0.031 to 16.000 mg/L for silver nitrate, ketoconazole and amphotericin B, from 0.5 - 256.0 mg/L for fluconazole. Minimum inhibitory concentration (MIC) was defined as the lowest drug concentration that showed absence of growth or complete growth inhibition (100%). The end points were determined as 100% growth inhibition for silver nitrate and amphotericin B, and > or = 75% growth inhibition for ketoconazole and fluconazole. The MICs at which 90% of isolates were inhibited (MIC(90)) of silver nitrate, ketoconazole, amphotericin B and fluconazole were 2.000, 512.000, 32.000 and 2.000 mg/L for Fusarium species, respectively; 1.000, 256.000, 2.000 and 2.000 mg/L for Aspergillus species, respectively; 2.000, 128.000, 4.000 and 2.000 mg/L for Alternaria alternate, respectively; 2.000, 4.000, 0.125 and 0.500 mg/L for Curvularia lunata, respectively; and 1.000, 256.000, 1.000 and 1.000 mg/L for unusual ocular pathogens, respectively. Silver nitrate was highly active against Aspergillus species (92.9% susceptible at a MIC of < or = 1.0 mg/L) and Fusarium species (96.3% susceptible at a MIC of < or = 2.0 mg/L). 95.6% of Fusarium species and 90.8% of Aspergillus species exhibited resistance to fluconazole, 44.1% of Fusarium species and 42.9% of Aspergillus species exhibited resistance to amphotericin B, 66.2% of Fusarium species exhibited resistance to ketoconazole. The activity of silver nitrate against the fluconazole-resistant, ketoconazole-resistant and amphotericin B-resistant strains was high. Silver nitrate has promising activity against a wide variety of ocular pathogenic fungi in vitro, and may have a role in future studies of antifungal eye drops and treating fungal keratitis.


Use of In Vitro Fertilisation Prediction Model in an Asian Population-Experience in Singapore.
This retrospective study was conducted to perform an external validation of the in vitro fertilisation (IVF) predict model developed by Scott Nelson et al in an Asian population. All IVF cycles registered in the study centre from January 2005 to December 2010 were included. Observed and predicted values of at least 1 live birth per cycle were compared by discrimination, calibration. Hosmer-Lemeshow test was used to assess the goodness-of-fit of the model calibration and Brier score was used to assess overall model performance. Among 634 IVF cycles, rate of at least 1 live birth was 30.6%. Causes of infertility were unexplained in 35.5% cases. Fifty-seven percent of women came for their first IVF treatment. First IVF cycle showed significantly higher success in comparison to subsequent cycles. The odds ratio of successful live birth was worse in women with endometriosis. Observed outcome was found to be more than the prediction of the model. The area under the curve (AUC) in this study was found to be 0.65 that was close to that of Nelson model (0.6335) done in internal validation. Brier score (average prediction error) of model was 0.2. Chi square goodness-of-fit test indicated that there was difference between the predicted and observed value (x² =18.28, df = 8, P = 0.019). Overall statistical findings indicated that the accuracy of the prediction model fitted poorly with the study population. Ovarian reserve, treatment centre and racial effect on predictability cannot be excluded. So it is important to make a good prediction model by considering the additional factors before using the model widely.


PACAP enhances mouse urinary bladder contractility and is upregulated in micturition reflex pathways after cystitis.
Pituitary adenylate cyclase-activating polypeptide (PACAP) elicits a transient contraction, sustained increase in the amplitude of spontaneous phasic contractions, and significantly increases the amplitude of nerve-mediated contractions in mouse urinary bladder smooth muscle (UBSM) strips. PACAP immunoreactivity (IR) is increased in micturition reflex pathways following cystitis. PACAP may contribute to altered sensation and bladder overactivity in the chronic bladder inflammatory syndrome, interstitial cystitis.


Behavior-constrained support vector machines for fMRI data analysis.
Statistical learning methods are emerging as a valuable tool for decoding information from neural imaging data. The noisy signal and the limited number of training patterns that are typically recorded from functional brain imaging experiments pose a challenge for the application of statistical learning methods in the analysis of brain data. To overcome this difficulty, we propose using prior knowledge based on the behavioral performance of human observers to enhance the training of support vector machines (SVMs). We collect behavioral responses from human observers performing a categorization task during functional magnetic resonance imaging scanning. We use the psychometric function generated based on the observers behavioral choices as a distance constraint for training an SVM. We call this method behavior-constrained SVM (BCSVM). Our findings confirm that BCSVM outperforms SVM consistently.


Characterization of Three Species of Sordariomycetes Isolated from Freshwater and Soil Samples in Korea.
During a survey of fungal diversity in the class Sordariomycetes, 3 fungal strains, CNUFC-KMHY6-1, CNUFC-MSW24-2-11, and CNUFC-GW2S-4 were isolated from soil and freshwater samples, respectively in Korea. The strains were analyzed both morphologically and phylogenetically on the basis of internal transcribed spacer and RNA polymerase II second largest subunit gene sequences. On the basis of their morphology and phylogeny, CNUFC-KMHY6-1, CNUFC-MSW24-2-11, and CNUFC-GW2S-4 isolates were identified as Arcopilus aureus, Memnoniella echinata, and Stachybotrys sansevieriae, respectively. To the best of our knowledge, Ar. aureus and M. echinata have not been previously recorded in Korea, and this is the first report of S. sansevieriae from freshwater niche.


Identification of Bordetella pertussis in nasopharyngeal swabs by PCR amplification of a region of the adenylate cyclase gene.
The polymerase chain reaction (PCR) was used to amplify a 522-bp region of the adenylate cyclase toxin (cyaA) gene of Bordetella pertussis. As few as 100 cfu from a suspension of B. pertussis could be detected by this procedure when the amplified PCR product was detected by ethidium bromide staining of agarose gels. However, simulated clinical specimens, prepared from swabs impregnated with known numbers of B. pertussis cells, only yielded a positive reaction with > or = 10(4) cfu. Hybridisation of a Southern blot of the PCR products from the swab samples with a cya-specific probe gave a positive reaction with as few as 8 cfu, but the hybridisation signal was uniformly weak with fewer than 10(4) cfu. Nevertheless, three of 13 nasopharyngeal swabs, taken from suspected clinically defined cases of whooping cough and stored frozen for up to 18 months, gave a positive PCR reaction.


Prenatal blockade of vasoactive intestinal peptide alters cell death and synaptic equipment in the murine neocortex.
Vasoactive intestinal peptide (VIP) is a potent growth factor that stimulates murine neocortical astrocyte genesis during the period of ontogenesis corresponding to premature delivery in humans. In rodents, part of the VIP supplied to the fetal brain is maternal VIP that crosses the placenta. If these data also apply to human brain development, premature newborns may be partly VIP-deficient because of loss of the maternal supply, and this may adversely affect their brain development. The goal of the present study was to determine the effects of VIP blockade during mouse neocortical astrocyte genesis on neuritic survival and maturation. VIP blockade by a specific VIP antagonist on embryonic d 17 and 18 induced transient, postnatal depletion of astrocytes in the upper neocortical layers. Combined use of in situ DNA fragmentation analysis (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method, a marker of cell death); immunohistochemical detection of synaptophysin, microtubule-associated proteins, and neurofilaments; and quantification of mRNA for synaptophysin and N-methyl-D-aspartate R1 receptor subunit revealed that early VIP blockade significantly altered programmed neuritic death and impaired neuritic differentiation. VIP inhibition induced 1) exaggerated postnatal terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling of cortical neurons, 2) long-term overexpression of synaptophysin and N-methyl-D-aspartate R1 receptor subunit, and 3) long-term overexpression of microtubule-associated protein-5 and neurofilament 160 kD. Although the functional consequences of this deviant pattern of murine neocortical development remain to be determined, these data open up new avenues for investigating some of the cognitive deficits observed in human premature infants.


Neuropsychiatric symptoms and syndromes in a large cohort of newly diagnosed, untreated patients with Alzheimer disease.
Neuropsychiatric symptoms are common in patients with Alzheimer disease (AD). Treatment for both AD and psychiatric disturbances may affect the clinical observed pattern and comorbidity. The authors aimed to identify whether particular neuropsychiatric syndromes occur in untreated patients with AD, establish the severity of syndromes, and investigate the relationship between specific neuropsychiatric syndromes and AD disease severity. Cross-sectional, multicenter, clinical study. A total of 1,015 newly diagnosed, untreated outpatients with AD from five Italian memory clinics were consecutively enrolled in the study from January 2003 to December 2005. All patients underwent thorough examination by clinical neurologists/geriatricians, including neuropsychiatric symptom evaluation with the Neuropsychiatric Inventory. Factor analysis revealed five distinct neuropsychiatric syndromes: the apathetic syndrome (as unique syndrome) was the most frequent, followed by affective syndrome (anxiety and depression), psychomotor (agitation, irritability, and aberrant motor behavior), psychotic (delusions and hallucinations), and manic (disinhibition and euphoria) syndromes. More than three quarters of patients with AD presented with one or more of the syndromes (N = 790, 77.8%), and more than half exhibited clinically significant severity of symptoms (N = 603, 59.4%). With the exception of the affective one, all syndromes showed an increased occurrence with increasing severity of dementia. The authors' study supports the use of a syndrome approach for neuropsychiatric evaluation in patients with AD. Individual neuropsychiatric symptoms can be reclassified into five distinct psychiatric syndromes. Clinicians should incorporate a thorough psychiatric and neurologic examination of patients with AD and consider therapeutic strategies that focus on psychiatric syndromes, rather than specific individual symptoms.


Complete Genome Sequence of Porcine Epidemic Diarrhea Virus Strain COL/Cundinamarca/2014 from Colombia.
Porcine epidemic diarrhea virus (PEDV) has been found throughout Europe and Asia, and has emerged in North and South America. A whole genome sequence was obtained from a paraffin-embedded tissue sample from the Instituto Colombiano Agropecuario (ICA), Colombia through Next Generation Sequencing techniques to further understand the evolution of PEDV.


Spontaneous cardiomyopathy and exophthalmos in cotton rats (Sigmodon hispidus).
Exophthalmos and clinical signs of heart failure occurred sporadically in 3- to 12-month-old cotton rats (Sigmodon hispidus) in a colony originally derived from three male and four female littermates. Macroscopic lesions in severely affected animals included subcutaneous edema, hydrothorax, right ventricular dilatation, unilateral or bilateral atrial thrombosis, and exophthalmos. Hearts from 17 cotton rats that were found dead or were euthanatized because of exophthalmos or dyspnea and 33 control cotton rats were examined microscopically. Myocardial lesions were present in 46 of 46 cotton rats > or = 1 month of age and consisted of multifocal cardiac myocyte necrosis, mineralization, and mononuclear inflammatory cell infiltration. Cotton rats > 5 months of age also had foci of interstitial fibrosis and myocyte atrophy. Twelve of 24 (50%) necropsied cotton rats had chronic pulmonary congestion, and livers from eight of 24 (33%) had chronic periacinar congestion and atrophy. Thrombi were present in one or both cardiac atria in nine of 50 (18%) hearts, and in at least one orbital venous sinus in 14 of 24 (58%) necropsied cotton rats and in 12 of 14 (86%) with exophthalmos. Exophthalmos in this colony of cotton rats appears to have resulted predominantly from orbital venous sinus thrombosis caused by stasis of venous blood secondary to right heart failure associated with a heritable cardiomyopathy.


Two Kinds of Protein Glycosylation in a Cell-Free Preparation from Developing Cotyledons of Phaseolus vulgaris.
Membrane preparations from developing cotyledons of red kidney bean (Phaseolus vulgaris L.) transferred radioactive mannose from GDP-mannose (U-[(14)C]mannose) to endogenous acceptor proteins. The transfer was inhibited by the antibiotic tunicamycin, suggesting the involvement of lipidoligosaccharide intermediates typical of the pathway for glycosylation of asparagine residues. This was supported by the similarity of the linkage types of radioactive mannose in lipid-oligosaccharide and glycoprotein products; both contained labeled 2-linked mannose, 3,6-linked and terminal mannose typical of glycoprotein "core" oligosaccharides. As expected for "core" glycosylation, the transfer of labeled N-acetylglucosamine (GlcNAc) from UDP-GlcNAc (6-[(3)H]GLcNAc) to 4-linkage in endogenous glycoproteins could also be demonstrated. However, most of the radioactive GlcNAc was incorporated into terminal linkage, in a reaction insensitive to tunicamycin. The proteins receiving "core" oligosaccharide in vitro were heterogeneous in size, in contrast to those receiving most of the GlcNAc (which chiefly comprised the seed reserve-proteins phaseolin and phytohemagglutinin). It is suggested that following "core" glycosylation, single GlcNAc residues are attached terminally to the oligosaccharides of these seed proteins, without the involvement of lipid-linked intermediates. Phaseolin from mature seeds does not possess a significant amount of terminal GlcNAc and so it is possible that these residues are subsequently removed in a processing event.


Greater systemic lipolysis in women compared with men during moderate-dose infusion of epinephrine and/or norepinephrine.
Women have lower circulating catecholamine levels during metabolic perturbations, such as exercise or hypoglycemia, but similar rates of systemic lipolysis. This suggests women may be more sensitive to the lipolytic action of catecholamines, while maintaining similar glucoregulatory effects. The aim of the present study, therefore, was to determine whether women have higher rates of systemic lipolysis compared with men in response to matched peripheral infusion of catecholamines, but similar rates of glucose turnover. Healthy, nonobese women (n = 11) and men (n = 10) were recruited and studied on 3 separate days with the following infusions: epinephrine (Epi), norepinephrine (NE), or the two combined. Tracer infusions of glycerol and glucose were used to determine systemic lipolysis and glucose turnover, respectively. Following basal measurements of substrate kinetics, the catecholamine infusion commenced, and measures of substrate kinetics continued for 60 min. Catecholamine concentrations were similarly elevated in women and men during each infusion: Epi, 182-197 pg/ml and NE, 417-507 pg/ml. There was a significant sex difference in glycerol rate of appearance and rate of disappearance with the catecholamine infusions (P < 0.0001), mainly due to a significantly greater glycerol turnover during the first 30 min of each infusion: glycerol rate of appearance during Epi was only 268 +/- 18 vs. 206 +/- 21 micromol/min in women and men, respectively; during NE, only 173 +/- 13 vs. 153 +/- 17 micromol/min, and during Epi+NE, 303 +/- 24 vs. 257 +/- 21 micromol/min. No sex differences were observed in glucose kinetics under any condition. In conclusion, these data suggest that women are more sensitive to the lipolytic action of catecholamines, but have no difference in their glucoregulatory response. Thus the lower catcholamine levels observed in women vs. men during exercise and other metabolic perturbations may allow women to maintain a similar or greater level of lipid mobilization while minimizing changes in glucose turnover.


Membrane-bound Na,K-ATPase: target size and radiation inactivation size of some of its enzymatic reactions.
Frozen samples of membrane-bound pig kidney Na,K-ATPase were subjected to target size analysis by radiation inactivation with 10-MeV electrons at -15 degrees C. The various properties investigated decreased monoexponentially with radiation dose, and the decay constants, gamma, were independent of the presence of other proteins and of sucrose concentrations above 0.25 M. The temperature factor was the same as described by others. Irradiation of four proteins of known molecular mass, m, showed that gamma for protein integrity was proportional to m with a proportionality factor about 20% higher than that conventionally used. By this standard curve, glucose-6-phosphate dehydrogenase activity used as internal standard gave a radiation inactivation size of 110 +/- 5 kDa, very close to m = 104-108 kDa for the dimer, as expected. For Na+/K+-transporting ATPase the following target sizes and radiation inactivation size values were very close to m = 112 kDa for the alpha-peptide: peptide integrity of alpha, 115 kDa; unmodified binding sites for ATP and vanadate, 108 kDa; K+-activated p-nitrophenylphosphatase activity, 106 kDa. There was thus no sign of dimerization of the alpha-peptide or involvement of the beta-peptide. In contrast, optimal Na+/K+-transporting ATPase activity had a radiation inactivation size = 189 +/- 7 kDa, and total nucleotide binding capacity corresponded to 72 +/- 3 kDa. These latter results will be extended and discussed in a forthcoming paper.


A possible mechanism of insulin resistance in the rat adipose cell in streptozotocin-induced diabetes mellitus. Depletion of intracellular glucose transport systems.
The effects of insulin-dependent diabetes mellitus on glucose transport activity and on the concentrations of glucose transport systems in the plasma and low density microsomal membranes in adipose cells isolated from streptozotocin-induced diabetic rats have been examined. Glucose transport activity was assessed by measuring 3-O-methylglucose transport and the concentration of glucose transport systems estimated by measuring specific D-glucose-inhibitable cytochalasin B-binding. Basal glucose transport activity decreases from 0.19 to 0.12 fmol/cell per min with the induction of diabetes, but remains constant per unit cellular surface area and is accompanied by a constant 6 pmol of glucose transport systems/mg of membrane protein in the plasma membrane fraction. Maximally insulin-stimulated glucose transport activity decreases from 3.16 to 1.05 fmol/cell per min and from 0.26 to 0.12 amol/micrometers 2 per min, and is accompanied by a decrease from 25 to 15 pmol of glucose transport systems/mg of plasma membrane protein. These diminished effects of insulin on glucose transport activity and the concentration of glucose transport systems in the plasma membrane fraction are paralleled by a 45% decrease in the basal number of glucose transport systems per milligram of membrane protein in the low density microsomal membrane fraction, the source of those glucose transport systems appearing in the plasma membrane in response to insulin. Thus, the "insulin resistant" glucose transport of the adipose cell in the streptozotocin-induced diabetic rat appears to be the consequence of a depletion of glucose transport systems in the intracellular pool.


The impact of anxiety and depressive symptoms on chronic pain in conservatively and operatively treated hand surgery patients.
The aim of this prospective study was to examine to what extent anxiety and depressive symptoms predict the level of pain at 4-month follow-up in hand surgery patients. A total of 132 consecutive patients (mean age: 51.5±17.1 years, 51.9% female) of a tertiary center for hand surgery participated in this study. The patients underwent conservative or operative treatment, depending on the nature of their hand problem. The initial pain assessment included psychometric testing with the hospital anxiety and depression scale. Ninety-nine patients underwent operative treatment and 33 patients were conservatively treated. At 4-month follow-up, the amount of pain was measured with a visual analog scale (0-10). After controlling for age, sex, and pre-surgical pain intensity, depressive symptoms were a significant predictor for increased pain levels at follow-up in conservatively treated patients. In operatively treated patients, anxiety symptoms showed a trend for being a predictor of pain level at follow-up. The findings support the assumption that psychological factors may have an impact on pain outcome in patients presenting to hand surgery clinics.


Effects of tetrandrine on cardiac and vascular remodeling.
Tetrandrine (Tet) is an alkaloid isolated from the Chinese herb Radix of Stephaniae tetrandrae S Moore. Cardiac and vascular remodeling confers a very definite risk of increased cardiovascular morbidity and mortality. Remodeling reversal has been achieved in human and experimental animals treated with some antihypertensive drugs but not all. This review will focus on cardiovascular remodeling and therapeutic effects of Tet. Three models, SHR, RHR (high renin), and DOCA-Salt HR (low renin) were used. Left ventricular and vascular remodeling had been developed in rats with 8-week untreated hypertension. Tet was administrated by ig 50 mg/kg/d for 9 weeks. Tet lowered SBP, left ventricular weight to body weight ratio, vascular media thickness, media to lumen ratio, cardiac and vascular wet weight, and collagen content. Tet decreased markedly the density and total number of dihydropyridine binding sites and also decreased Ca2+ overload in myocardium and vessels. Tet improved haemodynamic changes during remodeling special diastolic function such as LV compliance and stiffness, increased cardiac myosin ATPase activity and Na+-K+, Ca2+ ATPase activity, and normalized vascular reactivity. Tet inhibited proliferation of vascular smooth muscle cells, induced and sensitized VSMCs to pro-apoptosis stimulation, improved the endothelial function, and increased NO production. These results suggest that Tet was not only an anti-hypertensive drug but also an excellent drug to reverse cardiac and vascular remodeling.


Using State Transition Models to Explore How the Prevalence of Subtherapeutic Posaconazole Exposures Impacts the Clinical Utility of Therapeutic Drug Monitoring for Posaconazole Tablets and Oral Suspension.
Therapeutic drug monitoring (TDM) has been recommended in guidelines for patients receiving posaconazole oral suspension, but its utility in patients receiving posaconazole tablet, which has an improved bioavailability, remains unclear. We used state transition models with first-order Monte Carlo microsimulation to re-examine the posaconazole exposure-response relationships reported in two Phase III clinical trials (prophylaxis with posaconazole oral suspension - Models 1 & 2) and a third multicenter observational TDM study (Model 3). We simulated the impact of TDM-guided interventions to improve initial average posaconazole concentrations (Cavg) to reduce clinical failure (in Models 1 & 2) and breakthrough invasive fungal disease (bIFD) in Model 3. Simulations were then repeated using posaconazole tablet Cavg distributions in place of the oral suspension formulation. In all three models with posaconazole oral suspension, TDM interventions associated with maximal improvement in posaconazole Cavg reduced absolute rates of subtherapeutic exposures (Cavg < 700 ng/mL) by 25-49%. Predicted reductions in absolute clinical failure rates were 11% in Model 1 and 6.5% in Model 2, and a 12.6% reduction in bIFD in Model 3. With the tablet formulation, maximally-effective TDM interventions reduced subtherapeutic exposures by approximately 5% in all three models and absolute clinical failure rates by 3.9% in Model 1, and 1.6% in Model 2; and a 1.6% reduction in bIFD in Model 3. Our modeling suggests that routine TDM during prophylaxis with posaconazole tablets may have limited clinical utility unless populations with higher prevalence (>10%) of subtherapeutic exposures can be identified based on clinical risk factors.


Assessment of endotoxin levels in the home and current asthma and wheeze in school-age children.
The relationship between household endotoxin and asthma in children is not clear. To further investigate the relationship between sources of endotoxin and childhood asthma, we conducted a case-control study of children with and without asthma and examined their more frequent household exposures in the home. Children ages 6-13 years with current asthma (n = 70) or wheeze only (n = 19) were sex and age matched (+/-1 year) to 107 controls. Play area and mattress dust were collected for endotoxin analysis. Atopic status was determined by skin prick testing for allergies. A family size of >4 per household was associated with higher endotoxin levels (EU/mg) in the bed dust (P < 0.05). Passive smoking (P < 0.05) and the presence of a cat were associated with higher levels of endotoxin in mattress dust. Endotoxin levels in either the play dust or the bed dust did not differ between cases and controls. Within atopic cases, those with higher endotoxin loads (EU/m2) in bed or play areas were more likely to miss school for chest illness (P < 0.05). In this study, household endotoxin is not a risk factor for current asthma overall but may be associated with increased severity in children with atopic asthma. This study did not find that household sources of endotoxin were associated with asthma. However, within atopic asthmatics, asthma severity (as measured by a history of being kept home from school because of a chest illness in the past year) was associated with higher levels of endotoxin in dust from the child's bed. There is a need to further investigate the nature of the relationship between household endotoxin and asthma severity in children which could lead to better management of childhood asthma.


Acupressure therapy and Liu Zi Jue Qigong for pulmonary function and quality of life in patients with severe novel coronavirus pneumonia (COVID-19): a study protocol for a randomized controlled trial.
In December 2019, pneumonia associated with the 2019 novel coronavirus (COVID-19) emerged in Wuhan, China. The number of cases has increased rapidly. Patients with severe disease have a poor prognosis, and there are no effective therapies for COVID-19. Only rapid advice guidelines for symptomatic supportive care have been used. A traditional Chinese medicine rehabilitation (TCMR) program consisting of acupressure therapy and Liu Zi Jue Qigong can be used as a complementary therapy for COVID-19. Hence, we designed a randomized trial to evaluate the efficacy and advantages of TCMR for treating patients with severe COVID-19. This is a parallel-design, two-arm, analyst assessor-blinded, randomized controlled trial. A total of 128 patients with COVID-19 aged from 20 to 80 years will be recruited and assigned randomly into a guideline therapy group and a guideline therapy plus TCMR group at a 1:1 ratio. Patients in both groups will receive guideline therapy. The patients in the intervention group will perform acupressure therapy and Liu Zi Jue Qigong exercises in addition to conventional treatments twice a day and will be persistent from admission to discharge. The primary outcome will be measured with the Modified Medical Research Council Dyspnea Scale, and the secondary outcomes will include the Activities of Daily Living Barthel Index Scale, Patient Health Questionnaire-9 Scale, and the Respiratory Symptoms Scale. The assessments of the clinical scales will be performed at three points (before treatment, the 7th day during hospitalization, and the discharge day). Adverse events will be noted and recorded for the safety evaluation. This trial will provide high-quality evidence of the value of TCMR, which consists of acupressure therapy and Liu Zi Jue Qigong exercises, for treating patients with severe COVID-19. Chinese Clinical Trial Registry ChiCTR2000029994 . Registered on 18 February 2020.


Moderate evidence for a Lombard effect in a phylogenetically basal primate.
When exposed to enhanced background noise, humans avoid signal masking by increasing the amplitude of the voice, a phenomenon termed the Lombard effect. This auditory feedback-mediated voice control has also been found in monkeys, bats, cetaceans, fish and some frogs and birds. We studied the Lombard effect for the first time in a phylogenetically basal primate, the grey mouse lemur, Microcebus murinus. When background noise was increased, mouse lemurs were able to raise the amplitude of the voice, comparable to monkeys, but they did not show this effect consistently across context/individuals. The Lombard effect, even if representing a generic vocal communication system property of mammals, may thus be affected by more complex mechanisms. The present findings emphasize an effect of context, and individual, and the need for further standardized approaches to disentangle the multiple system properties of mammalian vocal communication, important for understanding the evolution of the unique human faculty of speech and language.


Shear bond strength of veneering composite to high performance polymers.
High performance polymers like PEEK (polyetheretherketone) and FRC (fiberreinforced composite) could substitute metallic alloys for removable partial dentures. However, these polymers require aesthetic veneering. This study was to determine the bond strength to direct composite. Specimens made of PEEK and FRC were produced and air-abraded (50 μm aluminum-oxide). Specimens were allocated to four experimental groups: Luxatemp Glaze & Bond, Scotchbond Universal, SR Nexco Connect and iBond Universal. Specimens were divided into three subgroups for short-term, long-term and no artificial aging and shear bond strength (SBS) was evaluated. SBS of specimens made of PEEK with no artificial aging showed values between 10.79-14.00 MPa, short-term artificial aging resulted in values between 3.78-13.85 MPa and after long-term artificial aging SBS decreased to 0-8.75 MPa. SBS measurement of FRC specimens resulted in values between 9.83-12.1 MPa without aging, after short-term artificial aging values decreased to 8.36-11.98 MPa and after long-term aging SBS showed a degradation to 4.52-7.82 MPa.


Trafficking: a perspective from Asia.
The main theme of this article is market development and trafficking as a business. It touches upon most of the aspects of the phenomenon, which have been encountered elsewhere, and translates them into the relatively unfamiliar context of many of the Asian and South-East Asian economies. Equally, the literature cited is also probably unfamiliar. Themes touched upon include democratization, inter-state relations, human rights, and scale and perspectives, together with the problems of definitions, theory, and the reliability of data. The directions and characteristics of trafficking flows together with routes and border control are also considered. Coordinated official responses to criminality and criminal organizations, as well as to trafficked individuals, are beginning to emerge. There is a note of caution sounded that contextual and cultural perspectives, particularly on sex workers, must be viewed somewhat differently to those in Western societies. The article concludes that as long as countries in Asia maintain their policies of restrictive immigration, trafficking can be expected to continue and almost certainly increase. This is because accelerating development creates demand for labor at various skill levels and because even in times of recession migrants and brokers will seek to side-step attempts to expel immigrants and restrict access to labor markets. The elimination of trafficking is unlikely to be realistically achieved through legislation and declarations of intent but by improvements in the socioeconomic status of the population.


H. pylori, the most common bacterial infection in Africa: a random serological study.
This study was carried out to determine the prevalence of antibodies to Helicobacter pylori in northern Nigeria, a region with a low incidence of peptic ulceration. In a random, serological survey of 268 subjects, 228 (85%) of the population studied had IgG antibodies to H. pylori. Fifty-eight of these subjects had experienced dyspepsia in the preceding 6 months. The majority of the population (82%) is infected between the ages of 5 and 10. Despite the high prevalence of antibodies to H. pylori, peptic ulcer is uncommon, suggesting that H. pylori is not important in the etiology of peptic ulcer in this population. Indeed, most patients infected by H. pylori are asymptomatic. The possible reasons for this are discussed.


Synthesis and biological activity of a series of potent fluoromethyl ketone inhibitors of recombinant human calpain I.
Calpain I, an intracellular cysteine protease, has been implicated in the neurodegeneration following an episode of stroke. In this paper, we report on a series of potent dipeptide fluoromethyl ketone inhibitors of recombinant human calpain I (rh calpain I). SAR studies revealed that while calpain I tolerates a variety of hydrophobic groups at the P1 site, Leu at P2 is preferred. However, the nature of the N-terminal capping group has a significant effect on the inhibitory activity of this series of compounds. Compound 4e [(1,2,3,4-tetrahydroisoquinolin-2-yl)carbonyl-Leu-D,L-Phe-CH2F+ ++], having a tetrahydroisoquinoline containing urea as the N-terminal capping group, is the most potent dipeptide fluoromethyl ketone inhibitor of calpain I (with a second-order rate constant for inactivation of 276,000 M-1 s-1) yet reported; tripeptide 4k (Cbz-Leu-Leu-D,L-Phe-CH2F) is equipotent. A number of compounds presented in this study displayed excellent selectivity for calpain I over cathepsins B and L, two related cysteine proteases. Compounds which exhibited good inhibitory activity in the assay against isolated rh calpain I also inhibited intracellular calpain I in a human cell line. Thus, in an intact cell assay, compounds 4e and 4k inhibited calpain I with IC50 values of 0.2 and 0.1 microM, respectively. Finally, we also disclose the first example of fluorination of a dipeptide enol silyl ether to generate the corresponding dipeptide fluoromethyl ketone.


Properties of Barley Seed Chitinases and Release of Embryo-Associated Isoforms during Early Stages of Imbibition.
Barley (Hordeum vulgare L.) seeds contain at least five proteins with chitinase (CH) activity. Two of these (CH1 and CH2) are found primarily in the aleurone and endosperm tissues, and the other three (CH3, CH4, and CH5) are enriched in the embryo. From the bran fraction, three of these CHs (CH1, CH2, and CH3) were purified to apparent homogeneity. These three CHs have apparent molecular masses of 27, 34, and 35 kilodaltons and isoelectric points of 9.3, 9.2, and 8.7, respectively. CH2 and CH3 have amino terminal sequences resembling a portion of the chitin-binding domain of lectins and other plant defense proteins. CH1 lacks this domain. All three CHs exhibit antifungal activity and inhibit the mycelial growth of some species of trichoderma and Fusarium in vitro. During the early period of imbibition by seeds, two of the embryo-associated CHs are selectively released into the surrounding aqueous medium.


Versatile Construction of Single-Tailed Giant Surfactants with Hydrophobic Poly(ε-caprolactone) Tail and Hydrophilic POSS Head.
Giant surfactants refer to a new kind of amphiphile by incorporating functional molecular nanoparticles with polymer tails. As a size-amplified counterpart of small-molecule surfactants, they serve to bridge the gap between small-molecule surfactants and amphiphilic block copolymers. This work reports the design and synthesis of single-tailed giant surfactants carrying a hydrophobic poly(ε-caprolactone) (PCL) as the tail and a hydrophilic cage-like polyhedral oligomeric silsesquioxane (POSS) nanoparticle as the head. The modular synthetic strategy features an efficient "growing-from" and "click-modification" approach. Starting from a monohydroxyl and heptavinyl substituted POSS (VPOSS-OH), a PCL chain with controlled molecular weight and narrow polydispersity was first grown by the ring-opening polymerization (ROP) of ε-CL under the catalysis of stannous octoate, leading to a PCL chain end-capped with heptavinyl substituted POSS (VPOSS-PCL). To endow the POSS head with adjustable polarity and functionality, three kinds of hydrophilic groups, including hydroxyl groups, carboxylic acids, and amine groups, were installed to the periphery of POSS molecule by a high-efficiency thiol-ene "click" reaction. The compounds were fully characterized by NMR, gel permeation chromatography (GPC), MALDI-TOF mass spectrometry, and TGA analysis. In addition, the preliminary self-assembly study of these giant surfactants was also investigated by TEM and dynamic laser light scattering (DLS), which indicated that they can form spherical nanoparticles with different diameters in aqueous solution. This work affords a straightforward and versatile way for synthesizing single-tailed giant surfactants with diverse head surface functionalities.


Effects of dioxane on cytochrome P450 enzymes in liver, kidney, lung and nasal mucosa of rat.
The effect of acute and chronic dioxane administration on hepatic, renal, pulmonary and nasal mucosa P450 enzymes and liver toxicity were investigated in male rats. The acute treatment consisted of two doses (2 g/kg) of dioxane given for 2 days by gavage, whereas the chronic treatment consisted of 1.5% of dioxane in drinking water for 10 days. Both the acute and chronic dioxane treatments induced cytochrome P450 2B1/2- and P450 2E1-dependent microsomal monooxygenase activities (pentoxyresorufin O-depentylase and p-nitrophenol hydroxylase) in the liver, whereas in the kidney and nasal mucosa, only the 2E1 marker activities were enhanced. In addition in the liver, an induction of 2alpha-testosterone hydroxylase (associated with the constitutive and hormone-dependent P450 2C11) was also revealed, whereas the hepatic P450 4A-dependent omega-lauric acid hydroxylase was not enhanced by any dioxane treatment. These inductions were mostly confirmed by western blot analysis of liver, kidney and nasal mucosa microsomes. In the lung, no alteration of P450 activities was observed. To assess the mechanism of 2E1 induction, the hepatic, renal and nasal mucosa 2E1 mRNA levels were also examined. Following two kinds of dioxane administration, in the liver the 2E1 induction was not accompanied by a significant alteration of 2E1 mRNA levels, while both in the kidney and nasal mucosa the 2E1 mRNA increased about 2- to 3-fold, indicating an organ-specific regulation of this P450 isoform. Furthermore, dioxane was unable to alter the plasma alanine aminotransferase activity and hepatic glutathione (GSH) content, examined as an index of toxicity, when it was administered into rats with P450 2B1/2 and 2E1 preinduced by phenobarbital or fasting pretreatment. These results support the lack of or a poor formation of reactive and toxic intermediates during the biotrasformation of this solvent, even when its metabolism was enhanced by P450 inducers. The chronic administration of dioxane was also unable to induce the palmitoyl CoA oxidase, a marker of peroxisome proliferation, excluding this as a way to explain its toxicity. Thus, although the mechanism of dioxane carcinogenicity remains unclear, the present results suggest that the induction of 2E1 following a prolonged administration of dioxane might provide oxygen radical species, and thereby contribute to its organ-specific toxicity.


MiRNA-99a alleviates inflammation and oxidative stress in lipopolysaccharide-stimulated PC-12 cells and rats post spinal cord injury.
Spinal cord injury (SCI) is caused by spinal fracture after the displacement of the spine or broken bone fragments protruding into the spinal canal, resulting in different degrees of injury to the spinal cord or spinal nerves. Expression levels of miR-99a and nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in cerebrospinal fluid of SCI patients were analyzed. Rat adrenal gland pheochromocytoma cell line PC-12 were stimulated with lipopolysaccharide (LPS) to mimic the in vitro environment of SCI. A rat mode of SCI was established by laminectomy. Reactive oxygen species (ROS) levels were measured by 2',7'-Dichlorodihydrofluorescein diacetate staining assay. Western blot was conducted to evaluate the expression levels of apoptotic indexes and proinflammatory cytokines. The interaction between miR-99a and NOX4 was verified by dual-luciferase reporter assay. The expression level of miR-99a was reduced while NOX4 expression was upregulated in cerebrospinal fluid of SCI patients and LPS-treated PC-12 cells. LPS impeded cell viability and promoted inflammation, apoptosis and ROS levels of PC-12 cells. Overexpression of miR-99a significantly promoted cell viability and reduced inflammation, apoptosis and oxidative stress in LPS-stimulated PC-12 cells. Dual-luciferase reporter assays verified that NOX4 was a target of miR-99a. Moreover, the expression of NOX4 was reduced in PC-12 cells after transfection with miR-99a mimic. Overexpression of NOX4 partly abolished the protective effect of miR-99a in LPS-treated PC-12 cells. To sum up, miR-99a suppresses NOX4 expression to relieve the LPS-induced inflammation, apoptosis and the progression of oxidative stress in SCI.


Effect of gonadotropin-inhibitory hormone on luteinizing hormone secretion in humans.
Gonadotropin-inhibitory hormone (GnIH, human homologue of RFRP-3) suppresses gonadotropin secretion in animal models, but its effects have not been studied in the human. We tested the hypotheses that exogenous GnIH inhibits LH secretion (i) in postmenopausal women and (ii) in men concurrently administered exogenous kisspeptin. Following in vitro and in vivo preclinical studies to functionally characterize the GnIH peptide, a dose-finding study (human GnIH: 1·5-150 μg/kg/h, iv for 3 h) was undertaken, and 50 μg/kg/h selected for further evaluation. Five postmenopausal women were administered 50 μg/kg/h iv infusion for 3 h or vehicle on two separate days. Four men were administered kisspeptin-10 (0·3 μg/kg iv bolus) with simultaneous infusion of GnIH (50 μg/kg/h, iv for 3 h) or vehicle. Healthy postmenopausal women (mean age 58 ± 2 years, LH: 30·8 ± 2·9 IU/l, FSH: 78·7 ± 6·4 IU/l, oestradiol: <50 pmol/l) and men (39·8 ± 2·1 years, mean total testosterone 12·1 ± 1·8 nmol/l, LH 2·2 ± 0·2 IU/l). Change in area under curve (AUC) of LH during GnIHvs vehicle. During GnIH administration in postmenopausal women, LH secretion decreased (ΔAUC: -9·9 ± 1·8 IU/3 h) vs vehicle (ΔAUC: -0·5 ± 1·7 IU/3 h; P = 0·02). Kisspeptin-10-stimulated LH responses in men were not affected by GnIH co-administration (60-min AUC of LH 6·2 ± 0·8 IU/h with kisspeptin-10 alone, 6·3 ± 1·0 IU/h, kisspeptin-10 with GnIH, P = 0·72). Exogenous GnIH was well tolerated, with no adverse events reported. Gonadotropin-inhibitory hormone decreased LH secretion in postmenopausal women in this first-in-human study. Kisspeptin-stimulated LH secretion in men was not inhibited during concomitant administration of GnIH.


Refeeding enhances intestinal repair during an acute enteritis in infant rabbits subjected to protein-energy malnutrition.
We examined the effects of refeeding during an acute bacterial enteritis on small intestinal repair in infant rabbits subjected to protein-energy malnutrition and in noninfected and infected dietary controls. Malnutrition was induced by litter expansion at 7 d of age. Randomly selected litters from both dietary groups were infected on d 17 with Yersinia enterocolotica. Inflammation and intestinal damage were observed in the jejunum and ileum at the "acute stage" of infection in 23-d-old animals from both dietary groups, as evidenced by an inflammatory infiltrate, blunted villi, and reduced disaccharidase activities. In addition, ileal glucose-stimulated Na+ absorption was depressed. On d 24, a 7-d period of ad libitum refeeding of breast milk and rabbit feed was initiated in randomly selected litters of infected-malnourished animals and all dietary controls. Mucosal repair was nearly complete at 31 d of age in infected dietary controls and in the infected-malnourished animals that were refed, as demonstrated by the recovery of segmental mucosal mass and ileal glucose-stimulated Na+ transport in association with the resolution of inflammation and diarrhea. Only mucosal disaccharidase activities remain depressed. In contrast, in 31-d-old infected-malnourished animals subjected to ongoing nutrient deprivation, severe intestinal damage persisted as evidenced by increased mortality, ongoing intestinal inflammation, mucosal hypoplasia, depressed disaccharidase activities, and reduced glucose-stimulated Na+ transport. We conclude that a refeeding regimen introduced during an acute bacterial enteritis is well tolerated and promotes recovery of intestinal mass, structure, and function in malnourished infant rabbits and dietary controls.


Response of blood pressure after percutaneous transluminal renal artery angioplasty and stenting.
To evaluate the short and intermediate term outcome of percutaneous transluminal renal artery angioplasty (PTRA) and stenting particularly on blood pressure (BP) control and renal function and to evaluate predictors of poor BP response after successful PTRA and stenting. We conducted a prospective analysis of all patients who underwent PTRA and stenting in our institute between August 2010 to September 2012. A total number of 86 patients were underwent PTRA and renal stenting. Selective angiography was done to confirm at least 70% angiographic stenosis. The predilatation done except few cases with critical stenosis, direct stenting was done in the rest of cases. All patients received aspirin 325 mg orally, and clopidogrel 300 mg orally within 24 h before the procedure. Heparin was used as the procedural anticoagulant agent. Optimal results with TIMI-III flow obtained in all cases. Following stent placement, aspirin 150 mg orally once daily was continued for a minimum of 12 mo and clopidogrel 75 mg orally once daily for at least 4 wk. The clinical, radiological, electrocardiography, echocardiography and treatment data of all patients were recorded. The BP measurement, serum creatinine and glomerular filtration rate (GFR) were recorded before the procedure and 1 and 6 mo after PTRA. A total of 86 patients were included in the study. The mean age of study population was 55.87 ± 11.85 years old and 67 (77.9%) of patients were male. There was a significant reduction in both systolic and diastolic BP at 1 mo after the procedure: 170.15 ± 20.10 mmHg vs 146.60 ± 17.32 mmHg and 98.38 ± 10.55 mmHg vs 89.88 ± 9.22 mmHg respectively (P = 0.0000). The reduction in BP was constant throughout the follow-up period and was evident 6 mo after the procedure: 144.23 ± 18.19 and 88.26 ± 9.79 mmHg respectively (P = 0.0000). However, no improvement in renal function was observed at any time during the follow-up period. After multivariate analysis, we found male sex, low GFR (< 60 mL/min) and higher baseline mean BP as a poor predictors of successful outcome on BP response after PTRA and stenting. The PTRA and stenting can be considered as an effective therapeutic intervention for improving BP control with minimal effect on renal function. The male sex, higher baseline BP and low GFR are associated with poor BP response after successful PTRA and stenting.


Frequency-Comb Spectrum of Periodic-Patterned Signals.
Using arbitrary periodic pulse patterns we show the enhancement of specific frequencies in a frequency comb. The envelope of a regular frequency comb originates from equally spaced, identical pulses and mimics the single pulse spectrum. We investigated spectra originating from the periodic emission of pulse trains with gaps and individual pulse heights, which are commonly observed, for example, at high-repetition-rate free electron lasers, high power lasers, and synchrotrons. The ANKA synchrotron light source was filled with defined patterns of short electron bunches generating coherent synchrotron radiation in the terahertz range. We resolved the intensities of the frequency comb around 0.258 THz using the heterodyne mixing spectroscopy with a resolution of down to 1 Hz and provide a comprehensive theoretical description. Adjusting the electron's revolution frequency, a gapless spectrum can be recorded, improving the resolution by up to 7 and 5 orders of magnitude compared to FTIR and recent heterodyne measurements, respectively. The results imply avenues to optimize and increase the signal-to-noise ratio of specific frequencies in the emitted synchrotron radiation spectrum to enable novel ultrahigh resolution spectroscopy and metrology applications from the terahertz to the x-ray region.


Effects of wheat variety and specific weight on dietary apparent metabolisable energy concentration and performance of broiler chicks.
1. This study used 4 wheat cultivars (Brigadier, Chaucer, Consort, Reaper) from three locations (Crossnacreevy, C; Downpatrick, D; Limavady, L), which had given rise to differences in wheat specific weight (SW), to examine the relationships between apparent metabolisable energy (AME) concentration, broiler performance and wheat SW. 2. The diets contained (g/kg): wheat 744, casein 142, blended vegetable fat 50, dicalcium phosphate 22, potassium bicarbonate 10.8, sodium bicarbonate 7.5, arginine 5, methionine 2, binder 8, trace minerals/vitamins 7.2, titanium dioxide 1.5. The diets were heat-treated (80 degrees C for 2 min) prior to pelleting (3 mm die). 3. SW ranged from 63 to 77 kg/hectolitre (hl), averaging 66, 69 and 76 kg/hl at D, C and L, respectively. In vitro viscosity of the wheat samples ranged from 5.2 to 17.5 cps and thousand grain weight (TGW) from 33.4 to 47.3 g. Mean TGW was similar at C and D (38.7, 37.0 g) but higher at L (43.1 g). In vitro viscosity was similar at C and L (11.2, 10.2 cps) but somewhat higher at D (14.4 cps). Crude protein (6.25 N) ranged from 116 to 147 g/kg and tended to be higher at D. Starch, which ranged from 612 to 656 g/kg, was least at D (617 g/kg) and greatest at L (641 g/kg). 4. Crude protein, crude fibre and total non-starch polysaccharide (NSP) were negatively correlated with SW, the R2 being respectively 0.38 (P<0.05), 0.16 (NS) and 0.45 (P<0.05). TGW and starch concentration were positively correlated with SW (R2=0.70, 0.44, respectively). There was a weak (NS) negative relationship (R2=0.19) between in vitro viscosity and SW. For both TGW and in vitro viscosity, correlations improved when variety was taken into account (R2=0.95, 0.92, respectively). 5. There were no significant effects of variety on dry matter (DM) intake or live weight gain (LWG). Gain: food was significantly higher (P<0.05) for Consort than for the other three varieties and the metabolisable energy ratio (ME:GE) just failed to attain significance (P=0.062). Calculated wheat AME (MJ/kg DM) was significantly (P<0.05) higher for Consort than for the other three wheats. There was a good correlation (R2=0.49) for the total data set between gain:food and ME:GE. In vivo viscosity varied from 13.6 to 28.6 cps for individual treatments and was significantly affected by variety (P<0001). 6. Although there were no significant differences in DM intake or LWG due to site the values for L (SW 76) were 6 and 5% lower, respectively, than for D (SW 66). Gain:food was lower (P<0.05) for C (SW 69) than for D. ME:GE, wheat AME and ME:gain were not significantly different between sites. 7. There was a weak (R2=0.18) positive relationship between ME:GE and SW corresponding to a 2.5% increase in energy value for a 10 kg/hl increase in SW and no relationship between gain:food and SW. When variety was taken into the regression the slope was similar but R2 increased to 0.82. 8. ME:GE and wheat AME concentration were negatively correlated with wheat in vitro viscosity (R2=0.64, 0.55, respectively). 9. It was concluded that in vitro viscosity appears to provide a better basis than SW for prediction of the nutritive value of wheats of unknown variety. If the variety is known then SW could be used to predict energy value. However, the effect of quite a large change in SW (10 kg/hl) was relatively small.


Conformational rearrangements in the pro-apoptotic protein, Bax, as it inserts into mitochondria: a cellular death switch.
The B-cell lymphoma 2 (Bcl-2) family of proteins regulates the activation of apoptosis through the mitochondria pathway. Pro- and anti-apoptotic members of this family keep each other in check until the correct time to commit to apoptosis. The point of no return for this commitment is the permeabilization of the outer mitochondrial membrane. Translocation of the pro-apoptotic member, Bax, from the cytosol to the mitochondria is the molecular signature of this event. We employed a novel method to reliably detect Förster resonance energy transfer (FRET) between pairs of fluorophores to identify intra-molecular conformational changes and inter-molecular contacts in Bax as this translocation occurs in live cells. In the cytosol, our FRET measurement indicated that the C-terminal helix is exposed instead of tucked away in the core of the protein. In addition fluorescence correlation spectroscopy (FCS) showed that cytosolic Bax diffuses much slower than expected, suggesting possible complex formation or transient membrane interaction. Cross-linking the C-terminal helix (α9) to helix α4 reduced the potential of those interactions to occur. After translocation, our FRET measurements showed that Bax molecules form homo-oligomers in the mitochondria through two distinct interfaces involving the BH3 domain (helix α2) and the C-terminal helix. These findings have implications for possible contacts with other Bcl-2 proteins necessary for the regulation of apoptosis.


Direct Attenuation Correction Using Deep Learning for Cardiac SPECT: A Feasibility Study.
Dedicated cardiac SPECT scanners with cadmium-zinc-telluride cameras have shown capabilities for shortened scan times or reduced radiation doses, as well as improved image quality. Since most dedicated scanners do not have integrated CT, image quantification with attenuation correction (AC) is challenging and artifacts are routinely encountered in daily clinical practice. In this work, we demonstrated a direct AC technique using deep learning (DL) for myocardial perfusion imaging (MPI). Methods: In an institutional review board-approved retrospective study, 100 cardiac SPECT/CT datasets with 99mTc-tetrofosmin, obtained using a scanner specifically with a small field of view, were collected at the Yale New Haven Hospital. A convolutional neural network was used for generating DL-based attenuation-corrected SPECT (SPECTDL) directly from noncorrected SPECT (SPECTNC) without undergoing an additional image reconstruction step. The accuracy of SPECTDL was evaluated by voxelwise and segmentwise analyses against the reference, CT-based AC (SPECTCTAC), using the 17-segment myocardial model of the American Heart Association. Polar maps of representative (best, median, and worst) cases were visually compared to illustrate potential benefits and pitfalls of the DL approach. Results: The voxelwise correlations with SPECTCTAC were 92.2% ± 3.7% (slope, 0.87; R2 = 0.81) and 97.7% ± 1.8% (slope, 0.94; R2 = 0.91) for SPECTNC and SPECTDL, respectively. The segmental errors of SPECTNC scattered from -35% to 21% (P < 0.001), whereas the errors of SPECTDL stayed mostly within ±10% (P < 0.001). The average segmental errors (mean ± SD) were -6.11% ± 8.06% and 0.49% ± 4.35% for SPECTNC and SPECTDL, respectively. The average absolute segmental errors were 7.96% ± 6.23% and 3.31% ± 2.87% for SPECTNC and SPECTDL, respectively. Review of polar maps revealed successful reduction of attenuation artifacts; however, the performance of SPECTDL was not consistent for all subjects, likely because of different amounts of attenuation and different uptake patterns. Conclusion: We demonstrated the feasibility of direct AC using DL for SPECT MPI. Overall, our DL approach reduced attenuation artifacts substantially compared with SPECTNC, justifying further studies to establish safety and consistency for clinical applications in stand-alone SPECT systems suffering from attenuation artifacts.


Note: Is it symmetric or not?
The reasons by which a molecule might distort from an idealized high symmetry configuration (for example, D3h for the nitrate radical) in a quantum-chemical computation are well-known, but briefly reviewed here in light of considerable recent debate on the BNB molecule. The role of the pseudo-Jahn-Teller effect in such cases is emphasized, as is the ultimate relevance and proper interpretation of the title question in cases where the adiabatic potential energy surface is extremely flat.


Prognostic significance of CTCs and CSCs of tumor drainage vein blood in Dukes' stage B and C colorectal cancer patients.
The clinical significance of circulating tumor cells (CTCs) including cancer stem cells (CSCs) (CTC/CSC) in the tumor drainage vein blood of patients with colorectal cancer (CRC) is unclear. In this study, we investigated the prognostic value of CTC/CSC that express carcinoembryonic antigen (CEA) cytokeratin 19 (CK19), CK20 and/or CD133 (CEA/CK/CD133) mRNA in the tumor drainage blood of CRC patients with Dukes' stage B and C. We examined tumor drainage blood from 197 patients with Dukes' stage B and C CRC. CTCs that expressed CEA, CK19, CK20 and CD133 mRNA were detected using the quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay. Each mRNA level was normalized with GAPDH mRNA levels. In the relationship between the expression of CEA/CK/CD133 in the tumor drainage blood and clinicopathological factors, a significant correlation was observed between CEA/CK/CD133 expression and Dukes' stage (p<0.041). In CRC patients with Dukes' stage B and C, disease-free (DFS) and overall survival (OS) of patients with CEA/CK/CD133 positive in the tumor drainage blood were significantly worse than that of marker gene negative patients. In contrast, in patients with Dukes' stage A, no significant differences were shown between these groups. By Cox progression analysis, it was shown that CEA/CK/CD133 mRNA in tumor drainage blood was an independent prognostic factor for DFS and OS in patients with Dukes' stage B and C. These results suggest that detecting CEA/CK/CD133 mRNA in tumor drainage blood by the real-time RT-PCR method would have a prognostic value in CRC patients with Dukes' stage B and C.


Regulation of beta-amyloid precursor protein isoform mRNAs by transforming growth factor-beta 1 and interleukin-1 beta in astrocytes.
In cultured astrocytes, all three major transcripts of beta-amyloid precursor protein (APP) were expressed with the ratio for APP695, APP751 and APP770 isoform mRNAs being 1:4:2. In comparison with controls, treatment of astrocytes with transforming growth factor-beta 1 (TGF-beta 1) produced about 6 fold increase in total APP mRNA, while elevation in the interleukin-1 beta (IL-1 beta) treated group was small and may relate to the mitogenic effect of IL-1 beta on astrocytes. Treatment of astrocytes with cytokines also produced marked changes in the upregulation in expression of different APP isoforms. The net increase in mRNAs of KPI-containing isoforms APP751 and APP770 was relatively more than for the APP695 isoform. This phenomenon was mainly related to the differences in the expression of KPI-containing APP isoforms and APP695 isoform in the controls. The present findings provide further evidence for the involvement of astrocytes in a cascade of events leading to the development of senile plaques in Alzheimer's disease and Down's syndrome.


Retrieving Spectra of Pure Components from the DOSY-NMR Experiment via a Comprehensive Approach Involving the 2D Asynchronous Spectrum, 2D Quotient Spectrum, and Genetic Algorithm Refinement.
When diffusion coefficients of different components in a mixture are similar, NMR spectra of pure individual components are difficult to be obtained via a diffusion-ordered spectroscopy (DOSY) experiment. Two-dimensional correlation spectroscopy (2D-COS) is used to analyze the data from the DOSY experiment. Through the properties of the systematic absence of cross-peak (SACP) in the 2D asynchronous spectra, spectra of pure components can be obtained even if their diffusion coefficients are similar. However, fluctuations in peak-position and peak-width are often unavoidable in NMR spectra, which makes SACPs unrecognizable. To address the problem, a 2D quotient spectrum is used to identify the masked SACPs. However, undesirable interference peaks due to the fluctuations in peak-position and peak-width are still present when we extract a spectrum of a component by slicing the 2D asynchronous spectrum across the SACP. A genetic algorithm (GA) is used to select a suitable subset of spectra where the diversities of peak-position and peak-width are significantly reduced. Then, we used the selected spectra to construct a refined 2D asynchronous spectrum so that the spectra of pure components with significant attenuated interference can be obtained. The above approach has been proven to be effective on a model system and a real-world example, demonstrating that 2D-COS possesses a bright perspective in the analysis of the bilinear data from DOSY experiments.


A framework for considering the risk-benefit trade-off in designing noninferiority trials using composite outcome approaches.
When a new treatment regimen is expected to have comparable or slightly worse efficacy to that of the control regimen but has benefits in other domains such as safety and tolerability, a noninferiority (NI) trial may be appropriate but is fraught with difficulty in justifying an acceptable NI margin that is based on both clinical and statistical input. To overcome this, we propose to utilize composite risk-benefit outcomes that combine elements from domains of importance (eg, efficacy, safety, and tolerability). The composite outcome itself may be analyzed using a superiority framework, or it can be used as a tool at the design stage of a NI trial for selecting an NI margin for efficacy that balances changes in risks and benefits. In the latter case, the choice of NI margin may be based on a novel quantity called the maximum allowable decrease in efficacy (MADE), defined as the marginal difference in efficacy between arms that would yield a null treatment effect for the composite outcome given an assumed distribution for the composite outcome. We observe that MADE: (1) is larger when the safety improvement for the experimental arm is larger, (2) depends on the association between the efficacy and safety outcomes, and (3) depends on the control arm efficacy rate. We use a numerical example for power comparisons between a superiority test for the composite outcome vs a noninferiority test for efficacy using the MADE as the NI margin, and apply the methods to a TB treatment trial.


Neurotoxicity with single dose intrathecal midazolam administration.
The aim of study was to investigate the electron microscopic changes in the medulla of the spinal cord that occur with intrathecal midazolam administration. Twenty-eight albino rabbits of New Zealand type were randomized into two groups. Following anaesthesia, 16 rabbits were given 300 microg of midazolam (Group M) and 12 rabbits were given 0.3 mL of normal saline solution (Group C) intrathecally. Eight rabbits from Group M (Group M1) and 6 rabbits from Group C (Group C1) were sacrificed 24 h after the anaesthesia and 8 rabbits from Group M (Group M2) and 6 rabbits from Group C (Group C2) were sacrificed 6 days after the anaesthesia. The lumbosacral portion was removed by laminectomy and thin sections were examined microscopically. Severe separation in myelin lamella of the large axons, honeycomb appearance, slight separation in myelin lamella of small to moderately large axons, degenerate vacuoles in the cytoplasm and nuclear membrane irregularity were observed in neurons of Groups M1 and M2. Myelin lamella and nuclear membranes were found to be regular, vacuoles and oedema were observed in the neurons in the Groups C1 and C2. Midazolam administered at single dose by the intrathecal route may have neurotoxic effects on the neurons and myelinated axons at 24 h and 6 days following administration.


Oxyallyl Cation Capture via Electrophilic Deborylation of Organoboronates: Access to α,α'-Substituted Cyclic Ketones.
An umpolung strategy to synthesize α,α'-substituted cyclic ketones through the nucleophilic addition of organoboronates to α-hydroxyl silyl enol ethers is described. The reaction proceeds via the trapping of in situ generated oxyallyl cations via the electrophilic deborylation of C(sp2) and C(sp) borates. This efficient and straightforward method provides direct access to α-substituted silyl enol ethers in high yield with complete regioselectivity. Desilylation in a one-pot procedure provides the corresponding α,α'-disubstituted ketones with high diastereoselectivity.


Technology to Produce High-Purity Anhydrous Rubidium Perrhenate on an Industrial Scale.
Technology used to produce high purity anhydrous rubidium perrhenate on an industrial scale from high purity perrhenic acid and rubidium nitrate by the ion-exchange method is described in this paper. This material is dedicated to catalyst preparation, therefore, strict purity requirements have to be fulfilled. These are satisfied by combining rubidium ion sorption on an ion exchange column and the subsequent elution of the high purity perrhenic acid solution, followed by crystallization, evaporation, purification, and drying. In the current study, rubidium and rhenium contents were found to be 22.5 wt.% and 55.4 wt.%, respectively, while contaminations were as follows: <2 ppm As, <2 ppm Bi, <5 ppm Ca, <5 ppm Cu, <3 ppm Fe, <10 ppm K, <3 ppm Mg, <5 ppm Mo, <2 ppm Na, <5 ppm Pb, and <3 ppm Zn.


A single-stranded DNA binding protein binds the origin of replication of the duplex kinetoplast DNA.
Replication of the kinetoplast DNA (kDNA) minicircle of trypanosomatids initiates at a conserved 12-nt sequence, 5'-GGGGTTGGTGTA-3', termed the universal minicircle sequence (UMS). A sequence-specific single-stranded DNA-binding protein from Crithidia fasciculata binds the heavy strand of the 12-mer UMS. Whereas this UMS-binding protein (UMSBP) does not bind a duplex UMS dodecamer, it binds the double-stranded kDNA minicircle as well as a duplex minicircle fragment containing the origin-associated UMS. Binding of the minicircle origin region by the single-stranded DNA binding protein suggested the local unwinding of the DNA double helix at this site. Modification of thymine residues at this site by KMnO4 revealed that the UMS resides within an unwound or otherwise sharply distorted DNA at the minicircle origin region. Computer analysis predicts the sequence-directed curving of the minicircle origin region. Electrophoresis of a minicircle fragment containing the origin region in polyacrylamide gels revealed a significantly lower electrophoretic mobility than expected from its length. The fragment anomalous electrophoretic mobility is displayed only in its native conformation and is dependent on temperature and gel porosity, indicating the local curving of the DNA double helix. We suggest that binding of UMSBP at the minicircle origin of replication is possible through local unwinding of the DNA double helix at the UMS site. It is hypothesized here that this local melting is initiated through the untwisting of unstacked dinucleotide sequences at the bent origin site.


Soluble suppressor supernatants elaborated by concanavalin A-activated human mononuclear cells. II. Characterization of a soluble suppressor of B cell immunoglobulin production.
Human peripheral blood mononuclear cells (PBMC) activated with the mitogenic lectin concanavalin A (Con A) elaborate a soluble immune suppressor supernatant (SISS) that contains at least 2 distinct suppressor factors. One of these, SISS-B, inhibits polyclonal B cell immunoglobulin production whereas the other, SISS-T, suppresses T cell proliferation to both mitogens and antigens. The latter mediator is discussed in the companion paper. Characteristics of the human soluble suppressor of B cell immunoglobulin production (SISS-B) include: 1) inhibition by a noncytotoxic mechanism, 2) loss of activity in the presence of the monosaccharide L-rhamnose, 3) appearance within 8 to 16 hr after the addition of Con A, 4) elaboration by cells irradiated with 500 or 2000 rads, 5) production by highly purified T cells, 6) stability at pH 2.5 but instability at 56 degrees C, and 7) m.w. of 60 to 80,000. These data indicate that after Con A activation, selected T cells not only become potent suppressor cells, but also generate a soluble saccharide-specific factor(s) that inhibits polyclonal immunoglobulin production by human B cells.


The effect of psychological distress on health outcomes: A systematic review and meta-analysis of prospective studies.
Distress effects are widely examined in cross-sectional studies with less known about effects on future health. This review summarizes distress impacts on health among adults in prospective studies and describes available distress measurement tools. Four inter-disciplinary databases were searched. Effects of distress on mortality and other outcomes were reviewed and estimated in a meta-analysis. A total of 19 studies were assessed which incorporated 10 distress tools. Distress had a detrimental effect on health regardless of the population studied, distress tool used, and health outcome examined. There was an increased mortality risk among those reporting high versus low distress (pooled hazard ratio (95% confidence interval) = 1.29 (1.15-1.46)).


A calendar savant with episodic memory impairments.
Patients with memory disorders have severely restricted learning and memory. For instance, patients with anterograde amnesia can learn motor procedures and retain some restricted ability to learn new words and factual information. However, such learning is inflexible and frequently inaccessible to conscious awareness. Here we present a case of patient AC596, a 25-year-old male with severe episodic memory impairments, presumably due to anoxia during a preterm birth. In contrast to his poor episodic memory, he exhibits savant-like memory for calendar information that can be flexibly accessed by day, month, and year cues. He also has the ability to recollect the exact date of a wide range of personal experiences over the past 20 years. The patient appears to supplement his generally poor episodic memory by using memorized calendar information as a retrieval cue for autobiographical events. These findings indicate that islands of preserved memory functioning, such as a highly developed semantic memory system, can exist in individuals with severely impaired episodic memory systems. In this particular case, our patient's memory for dates far outstripped that of normal individuals and served as a keen retrieval cue, allowing him to access information that was otherwise unavailable.


Comparative antibacterial activity of an arylglycyl oral cephalosporin, LY164846.
LY164846 is a semisynthetic arylglycyl cephalosporin which can be absorbed orally. It had in vitro activity comparable to that of cefaclor against beta-hemolytic streptococcal species and was two- to fourfold more active than cephalexin. Enterococci and Listeria species were resistant, and its activity against staphylococci was similar to that of other oral cephalosporins. Although some Bacteroides species were inhibited, the MICs for 25% were greater than or equal to 16 micrograms/ml. LY164846 was hydrolyzed by Staphylococcus aureus beta-lactamase and by cephalosporinases, but it was more stable than cefaclor.


Molecular Fingerprint of BMD Patients Lacking a Portion in the Rod Domain of Dystrophin.
BMD is characterized by a marked heterogeneity of gene mutations resulting in many abnormal dystrophin proteins with different expression and residual functions. The smaller dystrophin molecules lacking a portion around exon 48 of the rod domain, named the D8 region, are related to milder phenotypes. The study aimed to determine which proteins might contribute to preserving muscle function in these patients. Patients were subdivided, based on the absence or presence of deletions in the D8 region, into two groups, BMD1 and BMD2. Muscle extracts were analyzed by 2-D DIGE, label-free LC-ESI-MS/MS, and Ingenuity pathway analysis (IPA). Increased levels of proteins typical of fast fibers and of proteins involved in the sarcomere reorganization characterize BMD2. IPA of proteomics datasets indicated in BMD2 prevalence of glycolysis and gluconeogenesis and a correct flux through the TCA cycle enabling them to maintain both metabolism and epithelial adherens junction. A 2-D DIGE analysis revealed an increase of acetylated proteoforms of moonlighting proteins aldolase, enolase, and glyceraldehyde-3-phosphate dehydrogenase that can target the nucleus promoting stem cell recruitment and muscle regeneration. In BMD2, immunoblotting indicated higher levels of myogenin and lower levels of PAX7 and SIRT1/2 associated with a set of proteins identified by proteomics as involved in muscle homeostasis maintenance.


FXYD3 protein involved in tumor cell proliferation is overproduced in human breast cancer tissues.
FXYD3, also known as Mat-8 (Mammary tumor 8 kDa), is one of mRNAs highly expressed in mouse and human breast cancers. Here, we newly found that FXYD3 protein was also overexpressed in human breast cancer specimens; invasive ductal carcinomas and intra-ductal carcinomas, whereas its expression was low in benign lesion specimens; mastopathy, fibroadenoma and phyllodes tumors. Although human FXYD3 has two isoforms (shorter FXYD3a and longer FXYD3b), there have been no reports in which these two mRNAs were separately quantified. Here, we found that FXYD3a mRNA is a major transcript product expressed in human normal tissues as well as in breast, colon, stomach and pancreas cancer cell lines. In addition, expression level of FXYD3b was much lower than that of FXYD3a in these cancer cell lines. Cell proliferation rate of MCF-7 breast cancer cell line was drastically decreased when FXYD3a and 3b mRNAs were suppressed by the small interfering RNA. These results suggest that FXYD3a protein is highly expressed in breast cancers, and responsible for cancer cell proliferation.


Dopamine in vivo inhibits VEGF-induced phosphorylation of VEGFR-2, MAPK, and focal adhesion kinase in endothelial cells.
Vascular permeability factor (VPF)/VEGF is a potent multifunctional cytokine and growth factor that has critical roles in vasculogenesis and in both physiological and pathological angiogenesis. Because it has been recently shown that the neurotransmitter dopamine at pharmacological dose can inhibit VEGF/VPF-mediated microvascular permeability, proliferation, and migration of endothelial cells in vitro, we therefore hypothesized that endogenous dopamine may regulate the actions of VPF/VEGF in vivo. We report that VPF/VEGF-induced phosphorylation of VEGF receptor 2, focal adhesion kinase, and MAPK in the endothelial cells is strikingly increased in both dopamine-depleted and dopamine D(2) receptor knockout mice compared with normal controls, thereby indicating that endogenous dopamine regulate these critical signaling cascades required for the in vivo endothelial functions of VPF/VEGF. Together, these observations provide new mechanistic insight into the dopamine-mediated inhibition of the activities of VPF/VEGF and suggest that endogenous neurotransmitter dopamine might be an important physiological regulator of VPF/VEGF activities in vivo.


Peptide-mediated allo-recognition of HLA-B27 by cytotoxic T lymphocytes.
The molecular basis of the recognition of class-I HLA antigens by allo-reactive cytotoxic T lymphocytes (CTL) remains obscure. This article reviews our work in which HLA-B27-specific allo-reactive CTL clones were obtained and their fine specificity was analyzed with a panel of structurally defined HLA-B27 natural variants and site-directed mutants expressed on human and mouse cells. The results have implications for the involvement of endogenous peptides in determining the clonal diversity of HLA-B27 allogeneic responses and the fine specificity of T-cell recognition when HLA-B27 is expressed on different cell types.


Holter Monitoring and Loop Recorders: From Research to Clinical Practice.
Holter monitors are tools of proven efficacy in diagnosing and monitoring cardiac arrhythmias. Despite the fact their use is widely prescribed by general practitioners, little is known about their evolving role in the management of patients with cryptogenic stroke, paroxysmal atrial fibrillation, unexplained recurrent syncope and risk stratification in implantable cardioverter defibrillator or pacemaker candidates. New Holter monitoring technologies and loop recorders allow prolonged monitoring of heart rhythm for periods from a few days to several months, making it possible to detect infrequent arrhythmias in patients of all ages. This review discusses the advances in this area of arrhythmology and how Holter monitors have improved the clinical management of patients with suspected cardiac rhythm diseases.


The efficacy of routine outpatient management without antibiotics of fever in selected infants.
A previous study produced a protocol for outpatient management of febrile infants (FIs) judged to be at low risk for serious bacterial illness (SBI). This Philadelphia protocol demonstrated that 40% of FIs seen in the emergency department could be safely managed without antibiotics at home; and it was established by the emergency department staff as the standard of care at our institution. To determine 1) the actual practices of management of FIs 18 months after establishment of the Philadelphia protocol as the standard of care, and 2) the continued efficacy of noninvasive outpatient management of fever in FIs who, using the Philadelphia protocol, were identified as low risk for SBI. Thirty-six-month consecutive cohort study. Urban pediatric emergency department. Four hundred twenty-two infants, 29 to 60 days of age, with rectal temperatures >/=38.0 degrees C. Interventions. After a complete history taking, physical examination, and workup for SBI, infants were managed at the discretion of the attending physician in the emergency department. Subsequently, those management practices were reviewed and compliance with the Philadelphia protocol was evaluated. In addition, the overall efficacy and safety of that standard during 8 years of use was assessed. Of the 422 FIs enrolled, 101 (24%) were prospectively identified as low risk for SBI, and safe for management without antibiotics. Twenty-eight (6%) FIs were managed out of accordance with the Philadelphia protocol. Seven were admitted out of accordance, 10 (2 with UTI) were discharged out of accordance, and 11 inpatients (1 with bacterial gastroenteritis) initially received no antibiotics out of accordance with the protocol. Physician failure to consider the results of the complete blood count or urinalysis accounted for errors involving FIs with SBI. None of the 43 FIs with SBI were identified by the Philadelphia protocol to be at low risk for SBI. The Philadelphia protocol for outpatient management without antibiotics of FIs at low risk for SBI remains practical, reliable, and safe. Because breaches do occur, physicians must carefully scrutinize protocol compliance, especially with regard to the complete blood count and urinalysis.


Early prediction of long-term cognitive impairment after cardiac arrest.
This prospective study evaluated the prognostic value of early neurobiochemical markers, neuron-specific enolase and astroglial protein S-100B, for long-term cognitive outcome after cardiac arrest. Six months after admission of a cohort of 80 consecutive patients, 26 survivors were able to undergo a neuropsychological test battery. Survivors showed low test performances in attention, learning/memory, and executive functioning. Neuropsychological bedside screening during the first month significantly differentiated between patients with and without long-term cognitive impairment. The neurobiochemical marker S-100B at day 3 after admission was found to predict significant proportions of variance in specific cognitive domains (learning/memory and executive functioning). The results indicate that early neuropsychological assessment might help identify patients who run at risk of long-term neuropsychological dysfunction. This study also suggests that especially the protein S-100B provides valuable information on long-term cognitive outcomes. To understand the exact relationship, results have to be replicated in larger trials.


Physicians' attitudes and behaviors regarding hepatitis B immunization.
Hepatitis B virus (HBV) infections are a serious health problem in the United States, where approximately 18,000 cases are reported each year to the Centers for Disease Control. Even among health care providers, reported vaccination rates have ranged from 17% to 68%. The purpose of this study was to determine the important factors influencing physicians in an academic medical center to receive the HBV vaccination since the introduction of recombinant yeast-derived vaccines. The 1282 house staff and attending physicians in a university medical center were surveyed regarding their HBV vaccination history. The characteristics of vaccinated and nonvaccinated physicians and their attitudes and concerns regarding vaccination were compared. Of the 813 physicians who responded, 54.0% had been vaccinated. Vaccination rates varied with level of training, from 91.9% among first- and second-year residents to 32.2% among attending physicians. Although physicians in specialties at higher risk for infection were more likely to have been vaccinated, only 40.0% of pathologists and 51.9% of obstetrician-gynecologists reported having been vaccinated. Using multivariate analysis, we found important demographic predictors of HBV vaccination included physician sex and years since graduation, as well as level of training and specialty. Physicians who had been offered the vaccine were more likely to have been vaccinated. These results show that many physicians in an academic medical center, particularly those at an early stage of their training, have received HBV vaccination. Our results suggest that programs offering hepatitis B vaccinations to physicians can be effective in reducing this group's risk of hepatitis B infections. Special efforts may be necessary to reach physicians who have completed their training.


Feedback-stabilized fractional fringe laser interferometer for plasma density measurements.
A feedback stabilization technique is described for a fractional fringe interferometer measuring plasma electron densities. Using this technique, a CO2 laser Michelson interferometer with a pyroelectric detector exhibited a sensitivity of 3.4 x 10(-4) fringe on a 1-ms time scale and, due to acoustic pickup, 1.8 x 10(-2) fringe on a 10-ms time scale. The rise time is 45 micros. Stabilization against slow drifts in mirror distances is achieved by an electromechanically translated mirror driven by a servo system having a 0.2-s response time. A mechanical chopper in one of the two beam paths generates the signal which drives the servo system.


Thymine methyls and DNA-protein interactions.
Evidence is summarized showing that thymine methyls are as important in the recognition of specific sequences by proteins as are the more widely recognized hydrogen bonding sites of bases in the major groove (1). Strongest evidence has come from experiments using functional group mutagenesis (2) in which thymines in a specific recognition sequence (e.g., promoters, operators and restriction sites) are replaced by oligonucleotide synthesis with methyl-free uracil or cytosine and 5-methylcytosine. Such experiments have shown that thymine methyls can provide contact points via van der Waals interactions with amino acid side chains of specific DNA binding proteins. Actual contact between a thymine methyl and carbons of a glutamine side chain has been observed in a cocrystal of the phage 434 repressor and its operator by X-ray analysis. The issue of why thymine occurs in DNA is discussed in light of these findings.


IgG-class insulin autoantibodies in autoimmune thyroid disease.
We studied the incidence of insulin autoantibodies (IAA) in 97 patients with autoimmune thyroid disease with an enzyme-linked immunosorbent assay. The sera were also tested for islet cell antibodies (ICA) and thyrotropin receptor antibodies (TRAb). IAA as assessed by a standard deviation score were present in 7 patients (7.2%). None of the patients were seropositive for ICA. Data from this random sampling and a retrospective study of 12 patients on antithyroid drug treatment indicated that the presence of IAA was not associated with TRAb.


Rh(II)-catalyzed reactions of differentially substituted bis(diazo) functionalities.
The chemoselective reaction of donor/acceptor (D/A) and acceptor/acceptor (A/A) diazo moieties in the same molecule was examined using 3-diazo-1-(ethyl 2-diazomalonyl)indolin-2-one under rhodium(II) catalysis. The D/A diazo group undergoes selective cyclopropanation as well as XH-insertion, leaving behind the second diazo group for a further intramolecular dipolar cycloaddition reaction.


Liver venous deprivation for resection of advanced hilar cholangiocarcinoma-a case report and review of the literature.
Hilar cholangiocarcinoma is a rare primary malignancy associated with a dismal prognosis. Currently, complete extended right or left-sided hepatectomy is the primary curative therapy. Achieving a negative resection margin is associated with long-term survival and better quality of life, while post-hepatectomy liver failure (PHLF) due to insufficient liver remnant remains the most dreaded complication with a negative effect on overall survival. Precise preoperative management with sufficient future remnant liver (FRL) volume is the key to achieving good results in the treatment of bile duct carcinoma. To present a case report and a literature review for preoperative FRL optimization prior to major hepatectomies for hilar cholangiocarcinoma. Improvement of postoperative outcomes after extended liver resections in the case of hilar cholangiocarcinoma. A 62-year-old Caucasian woman with Lynch syndrome presented to our department with a hilar cholangiocarcinoma Bismuth type IIIa. The patient had an insufficient future liver volume for extended liver resection. She underwent preoperative preconditioning using a liver venous deprivation (LVD) and underwent two weeks later a right trisectorectomy without any interventional complications. Liver function remained stable postoperatively. The patient was discharged on the 20th postoperative day without major surgical post-operative complications or the need for readmission. LVD is a technically feasible, safe, and effective procedure to increase the FRL in a short period of time with low intra and post-operative complications and therefore improving the survival of patients with hilar cholangiocarcinoma.


Effects of advanced glycation end products on dental pulp calcification.
The main aim of this study was to elucidate the effects of advanced glycation end products (AGEs) on the calcification of cultured rat dental pulp cells (RDPCs) and to investigate the crystallisation ability of glycated collagen. AGEs were prepared via non-enzymatic glycation of a dish coated with type I collagen using dl-glyceraldehyde. To investigate the effects of AGEs on RDPCs, we performed WST-1 and lactate dehydrogenase assays; alkaline phosphatase, Alizarin Red S and immunohistochemical staining; and real-time quantitative reverse transcription PCR. In addition, we performed crystallisation experiments on glycated collagen. All microstructures were analysed using scanning electron microscopy/energy-dispersive X-ray spectroscopy and transmission electron microscopy/diffraction pattern analysis. AGEs did not affect the proliferation or differentiation of RDPCs, but enhanced the calcification rate and cytotoxicity. No major calcification-related genes or proteins were involved in these calcifications, and glycated collagen was found to exhibit a negative polarity and form calcium phosphate crystals. Cytotoxicity due to drastic changes in the concentration of pericellular ions led to dystrophic calcification, assumed to represent an aspect of diabetic pulp calcifications. Glycated collagen-containing AGEs provide a nurturing environment for crystallisation and have a significant effect on the early calcification of RDPCs.


[The value of lymphography in testicular, bladder and kidney tumors].
On the basis of 143 lymphographical findings of testicular tumours, 95 of vesical tumours and 55 of renal tumours, this method of examination for urological tumours is evaluated. Lymphography has great diagnostic value for testicular tumours. It is very accurate, and the examination is essential for staging, the planning and checking of therapy and for prognosis. If possible it should always be carried out in combination with computerized tomography. With vesical and renal tumours the value of this examination is often limited by the low lymphogenic rate of metastasis, the low accuracy with vesical tumours and often by therapeutic procedures which are independent of lymph node findings. Nevertheless, even with these two types of tumour lymphography is justified by the information it gives on further course, possible inoperability and the help it gives in field delimitation and checking of therapy. Its use in connection with vesical and testicular tumours will depend on the therapeutic preference of the particular clinician.