Preoperative diabetes management protocol for adult outpatients.
The lack of evidence-based guidelines or nationally recognized protocols for the preoperative management of adult diabetics in the outpatient setting poses a challenge for the perianesthesia practitioner. The West Virginia University Hospital Perioperative Research Committee sought to develop an institutional protocol for management of this population. A retrospective chart review of diabetic outpatients who arrived the morning of surgery with elevated glucose levels was performed. The purpose of the chart review was to delineate indicators measured in the preoperative visit that could be predictive of patients arriving the morning of surgery with an elevated glucose level. Self-reported home fingerstick glucose values >200mg/dL were a statistically significant (p = .007) indicator of elevated arrival glucose in the preoperative holding area. Results of the chart review and best clinical evidence, as well as expert opinion, were used to develop an Adult Preoperative Diabetes Protocol and an accompanying Patient Instruction Tool.


Regulation of lysine metabolism and endosperm protein synthesis by the opaque-5 and opaque-7 maize mutations.
Two high lysine maize endosperm mutations, opaque-5 (o5) and opaque-7 (o7), were biochemically characterized for endosperm protein synthesis and lysine metabolism in immature seeds. Albumins, globulins, and glutelins, which have a high content of lysine, were shown to be increased in the mutants, whereas zeins, which contain trace concentrations of lysine, were reduced in relation to the wild-type lines B77xB79+ and B37+. These alterations in the storage protein fraction distribution possibly explain the increased concentration of lysine in the two mutants. Using two-dimensional polyacrylamide gel electrophoresis of proteins of mature grains, variable amounts of zein polypeptides were detected and considerable differences were noted between the four lines studied. The analysis of the enzymes involved in lysine metabolism indicated that both mutants have reduced lysine catabolism when compared to their respective wild types, thus allowing more lysine to be available for storage protein synthesis.


Expression of a rat liver microsomal cytochrome P-450 catalyzing testosterone 16 alpha-hydroxylation in Saccharomyces cerevisiae: vitamin D3 25-hydroxylase and testosterone 16 alpha-hydroxylase are distinct forms of cytochrome P-450.
Rat cytochrome P-450(M-1) cDNA was expressed in Saccharomyces cerevisiae TD1 cells by using a yeast-Escherichia coli shuttle vector consisting of P-450(M-1) cDNA, yeast alcohol dehydrogenase promoter and yeast cytochrome c terminator. The yeast cells synthesized up to 2 X 10(5) molecules of P-450(M-1) per cell. The microsomal fraction prepared from the transformed cells contained 0.1 nmol of cytochrome P-450 per mg of protein. The expressed cytochrome P-450 catalyzed 16 alpha- and 2 alpha-hydroxylations of testosterone in accordance with the catalytic activity of P-450(M-1), but did not hydroxylate vitamin D3 or 1 alpha-hydroxycholecalciferol at the 25 position. The expressed cytochrome P-450 also catalyzed the oxidation of several drugs and did not show 25-hydroxylation activity toward 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol. However, it cross-reacted with the polyclonal and monoclonal antibodies elicited against purified P-450cc25 which catalyzed the 25-hydroxylation of vitamin D3. These results indicated that P-450(M-1) cDNA coded the 2 alpha- and 16 alpha-hydroxylase of testosterone, and that these two positions of testosterone are hydroxylated by a single form of cytochrome P-450. Vitamin D3 25-hydroxylase and testosterone 16 alpha- and 2 alpha-hydroxylase are different gene products, although these two hydroxylase activities are immunochemically indistinguishable.


Application of a novel liquid biopsy in patients with hepatocellular carcinoma undergoing liver transplantation.
Circulating tumor cells (CTCs) serves a primary function in metastasis and recurrence of hepatocellular carcinoma (HCC). In the present study, in order to evaluate the analytical performance and clinical value of the liquid biopsy-based platform, a novel integrated subtraction enrichment and immunostaining-fluorescence in situ hybridization (iFISH®) platform was applied to analyze CTCs in patients with HCC undergoing liver transplantation (LT). In total, 30 patients with HCC undergoing LT and 10 healthy volunteers were enrolled. CTCs in peripheral blood that were obtained from each patient prior to LT and 3 months thereafter were detected using the iFISH® platform, and CellSearch® system was performed for each subject for comparison. Using iFISH® and CellSearch®, the percentage of CTCs in patients with pre-operative HCC were 70.00% and 26.67%, respectively. CTCs counted using iFISH® (iFISH-CTCs) were increased compared with CellSearch® (Cellsearch-CTCs) (P<0.01). A significant decrease in iFISH-CTCs was observed 3 months following LT (3.04±0.93/7.5 to 1.0±0.53/7.5 ml, P<0.05). Furthermore, patients with lower preoperative iFISH-CTCs level (<5/7.5 ml) had markedly increased recurrence-free survival compared with iFISH-CTCs (>5/7.5 ml, 15 vs. 5.5 months; P<0.01. iFISH® platform exhibits an increased analytical sensitivity, and may be used as a dynamic monitoring tool for CTCs, and CTCs may be a good prognostic indicator for patients with HCC undergoing LT.


A new Corononcodes Speiser from the Palaearctic Region, with a key to species (Diptera: Acroceridae).
Corononcodes ziegleri spec. nov. is described and illustrated based on material from Iran. The species represents the second Palaearctic representative of this panopine genus. In addition, C. siculus Bezzi is recorded from Fuerteventura (Canary Islands, Spain) for the first time. An identification key to the Palaearctic species of Corononcodes is presented. Additional faunistic records for the Iranian fauna comprise Acrocera orbiculus (Fabricius) and an apparently undescribed species of Ogcodes Latreille. DNA-barcodes for all Iranian taxa are provided.


Effect of a Quality Improvement Project to Reduce Noise in a Pediatric Unit.
Noise levels remain high in clinical settings, which may result in stress and sleep disruption, and can lead to immunosuppression, delayed healing, confusion, disorientation, delusions, and increased length of hospital stay. The purpose of this quality improvement project was to assess effects of a multidisciplinary noise reduction program on a pediatric unit in an acute care hospital in a developing country. A quality improvement project was carried out over 15 months in a pediatric unit. A three-phase study was conducted where the first phase included obtaining patient satisfaction ratings and recording sound levels, the second phase included implementing a noise reduction program and designing a noise detector machine, and the third phase included obtaining patient satisfaction data and recording noise levels over a 1-year period. There was a significant decrease in noise of 8 A-weighted decibels when comparing the values before and after implementing the quality improvement project at t = 6.44, p < 0.000. There was no significant difference in patient satisfaction ratings. Noise in the pediatric unit exceeded recommended guidelines; however, decreasing the levels was possible and sustainable, which can improve the psychological and physiological wellbeing of hospitalized children.


Developmental changes in visual line bisection in women throughout adulthood.
It has been suggested that some hemispheric asymmetries change in a systematic way from young adulthood to older age. However, little is known whether these changes are due to differential aging of a single hemisphere or based on age-related alterations of interhemispheric interactions. A sample of 281 right-handed neurologically healthy participants (151 women), ranging from age 20 to 79, was investigated with a line-bisection task. Previous studies indicate the midpoint estimation shows a consistent leftward bias from the veridical center, which is accentuated when the left hand is used to bisect lines. These findings support the view of a right hemispheric superiority in spatial attention. This study revealed this pattern to be stable in men throughout adulthood. However, women from 50 to 69 years of age showed a reduced leftward bias and a reduced hand effect compared to men and younger women. The results suggest that developmental changes in hemispheric asymmetry of spatial attention are more pronounced in women and support the view that neuromorphological changes during adulthood differ between sexes.


Vaccine development for potential bioterrorism agents.
Vaccines are considered to be one of the most effective ways of combating disease caused by bioterrorism agents. Such vaccines must be able to provide protection against pathogens which might enter the body by a number of routes, including the respiratory tract. They should also be able to induce protective immunity rapidly and would ideally be given non-invasively. There are few vaccines which currently meet these requirements. In part, this reflects the low level of research on many bioterrorism agents over the past few decades. Little is known about basic mechanisms of pathogenicity of many of these agents. However, by their very nature these agents cause serious disease, and must be handled in high containment laboratories. This requirement also limits the speed and ease with which research on these pathogens can now take place. Against this background, research on vaccines against potential bioterrorism agents is likely to proceed along two lines. Firstly because the genome sequences of most of the principal bioterrorism agents have either been completed or are close to completion, there is likely to be reliance on the exploitation of this information to devise improved vaccines. A number of groups are working on methodologies to identify vaccine antigens directly from genome sequences. Secondly, there will be a need to formulate such vaccines appropriately for the rapid induction of protective immunity after non-invasive delivery. The prospects for the development of a new generation of bioterrorism vaccines which exploit these technologies are reviewed in this manuscript.


Risk factors for acquisition of multidrug-resistant Acinetobacter baumannii among cancer patients.
Data regarding multidrug-resistant (MDR) Acinetobacter baumannii infections among cancer patients are limited. We conducted a case-control study to investigate the risk factors for acquisition of MDR A baumannii and the outcomes among cancer patients. Cases were inpatients with malignancy who had MDR A baumannii from any cultures between 2008 and 2011. Controls were inpatients with malignancy but no MDR A baumannii. A total of 31 case patients were matched with 62 control patients. Hematologic malignancy (P = .036), need for dialysis (P = .01), admission for other reasons except elective surgery (P = .03), transfer from other health care facilities (P = .02), prolonged intensive care unit stay (P = .004), mechanical ventilation (P < .001), pressor use (P = .001), tube feeding (P < .001), transfusion (P = .009), and prior antimicrobial use (P < .001) were identified as significant risk factors in univariate analysis. Need for dialysis (odds ratio [OR], 18.23; P = .04) and prolonged intensive care unit stay (OR, 19.28; P = .01) remained significant in multivariate analysis. Lengths of stay were 28 days for the case patients and 10 days for the control patients (P = .001). The 90-day mortality rates were 41.9% and 29.0%, respectively (P = .20). Acquisition of MDR A baumannii among cancer patients appears to be associated with general nosocomial infection risk factors rather than underlying malignancies.


V-type H+-ATPase in the human eccrine sweat duct: immunolocalization and functional demonstration.
We investigated for the presence of a vacuolar-type H+-ATPase (V-ATPase) in the human eccrine sweat duct (SD). With the use of immunocytochemistry, an anti-V- ATPase antibody showed a strong staining at the apical membrane and a weaker one in the cytoplasm. Cold preservation followed by rewarming did not alter this staining pattern. With the use of the pH-sensitive dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein on isolated and perfused straight SD under HCO-free conditions and in the absence of Na+, proton extrusion was determined from the recovery rate of intracellular pH (dpH(i)/dt) following an acid load. Oligomycin (25 microM), an inhibitor of F-type ATPases, decreased dpH(i)/dt by 88 +/- 6%, suggesting a role for an ATP-dependent process involved in pH(i) recovery. Moreover, dpH(i)/dt was inhibited at 95 +/- 3% by 100 nM luminal concanamycin A, a specific inhibitor of V-ATPases, whereas 10 microM bafilomycin A1, another specific inhibitor of V-ATPases, was required to decrease dpH(i)/dt by 73%. These results strongly suggest that a V-ATPase is involved in proton secretion in the human eccrine SD.


Micro magnetic tweezers for nanomanipulation inside live cells.
This study reports the design, realization, and characterization of a multi-pole magnetic tweezers that enables us to maneuver small magnetic probes inside living cells. So far, magnetic tweezers can be divided into two categories: I), tweezers that allow the exertion of high forces but consist of only one or two poles and therefore are capable of only exerting forces in one direction; and II), tweezers that consist of multiple poles and allow exertion of forces in multiple directions but at very low forces. The magnetic tweezers described here combines both aspects in a single apparatus: high forces in a controllable direction. To this end, micron scale magnetic structures are fabricated using cleanroom technologies. With these tweezers, magnetic flux gradients of nablaB = 8 x 10(3) T m(-1) can be achieved over the dimensions of a single cell. This allows exertion of forces up to 12 pN on paramagnetic probes with a diameter of 350 nm, enabling us to maneuver them through the cytoplasm of a living cell. It is expected that with the current tweezers, picoNewton forces can be exerted on beads as small as 100 nm.


Role of the FtsA C terminus as a switch for polymerization and membrane association.
Together with ATP, the C-terminal region of the essential streptococcal FtsA protein acts as an intramolecular switch to promote its polymerization and attachment to the membrane. During septation, FtsA is known to anchor the constricting FtsZ ring and, subsequently, the divisome to the membrane. Truncation of the C terminus of the streptococcal FtsA (FtsAΔCt) facilitates a more rapid ATP-dependent polymerization in solution than is seen with the full-length protein (FtsA(+)). The FtsAΔCt polymers are more organized and compact than those formed in solution by FtsA(+), resembling the shape of the membrane-associated FtsA(+) polymers. We find that ATP, besides being needed for polymerization, is required for the attachment of FtsA(+) to lipid monolayers and to vesicle membranes. We propose a model in which the binding of ATP activates a switch favoring the polymerization of FtsA and at the same time driving the amphipathic helix at its C terminus to become attached to the membrane. Conversely, when FtsA is in the cytoplasm, the C terminus is not engaged in the attachment to the membrane, and it obstructs polymerization. ATP-dependent polymerization of FtsA inside membrane vesicles causes vesicle shrinkage, suggesting that, besides providing a membrane attachment for FtsZ, the FtsA C terminus may also introduce local alterations in the membrane to facilitate septation. FtsA is a protein needed in many bacteria to construct a septum that divides one fully grown cell, producing two daughters. We show that the region located at the C-terminal end of the Streptococcus pneumoniae FtsA protein works as a switch triggered by ATP, a molecule that stores energy. This region contains an amphipathic helix that obstructs the assembly of FtsA into polymers in the cytoplasm. In the presence of ATP, the obstruction is removed by switching the position of the helix. The switch directs the helix to the membrane and simultaneously facilitates the polymerization of the protein. The accumulation of FtsA molecules at the membrane causes distortions, an effect produced also by proteins such as MinD, MreB, and SepF that also contain amphipathic helixes as membrane attachment devices. In the case of FtsA, these distortions may also facilitate the initial events that lead to the division of bacteria.


Subacute normobaric oxygen and hyperbaric oxygen therapy in drowning, reversal of brain volume loss: a case report.
A 2-year-old girl experienced cardiac arrest after cold water drowning. Magnetic resonance imaging (MRI) showed deep gray matter injury on day 4 and cerebral atrophy with gray and white matter loss on day 32. Patient had no speech, gait, or responsiveness to commands on day 48 at hospital discharge. She received normobaric 100% oxygen treatment (2 L/minute for 45 minutes by nasal cannula, twice/day) since day 56 and then hyperbaric oxygen treatment (HBOT) at 1.3 atmosphere absolute (131.7 kPa) air/45 minutes, 5 days/week for 40 sessions since day 79; visually apparent and/or physical examination-documented neurological improvement occurred upon initiating each therapy. After HBOT, the patient had normal speech and cognition, assisted gait, residual fine motor and temperament deficits. MRI at 5 months after injury and 27 days after HBOT showed near-normalization of ventricles and reversal of atrophy. Subacute normobaric oxygen and HBOT were able to restore drowning-induced cortical gray matter and white matter loss, as documented by sequential MRI, and simultaneous neurological function, as documented by video and physical examinations.


Histone modification in the TGFbetaRII gene promoter and its significance for responsiveness to HDAC inhibitor in lung cancer cell lines.
We previously reported silencing of the TGF-beta type II receptor gene (TGFbetaRII), involving histone deacetylation, instead of DNA methylation (DNA-Me). Because different histone modifications may play crucial roles in the epigenetic alterations, we further studied links with silencing of the TGFbetaRII gene promoter in six lung cancer cell lines. ChIP assays demonstrated three chromatin patterns for this gene silencing (Pattern I: histone H3 acetylation (H3-Ac)(+/-)/histone H3 lysine 4 methylation (H3K4-Me)(+)/DNA-Me(-), Pattern II; H3-Ac(-)/H3K4-Me(+/-)/DNA-Me(-), and Pattern III; H3-Ac(-)/H3K4-Me(-)/DNA-Me(+)), indicating possible progressive alterations with H3K4-Me alteration. With exposure to a histone deacetylase inhibitor (HDAC-I), trichostatin A, cell lines with the pattern II demonstrated strong and persistent induction of TGFbetaRII expression, while those with the pattern III showed only weak or no induction. ACC-LC-91 cell line, one of the pattern II examples demonstrated strong and continuous induction of H3K4-Me similar to TGFbetaRII expression. In contrast, ACC-LC-176 with the pattern III showed only weak and transient induction of H3K4-Me, similar to TGFbetaRII expression. Treatment with 5-aza-2'-deoxycytidine (5aza-dC) in addition to HDAC-I resulted in strong and continuous induction of TGFbetaRII expression and H3K4-Me in ACC-LC-176, although 5aza-dC alone was without such effects. In ACC-LC-91, both H3-Ac and H3K4-Me were promptly and simultaneously induced by HDAC-I, and similarly inhibited by wortmannin, a PI3K family inhibitor, together with TGFbetaRII induction. These findings suggested progressive alterations of chromatin configuration including H3K4-Me alteration in TGFbetaRII gene silencing. A possible involvement of a wortmannin-sensitive kinase in histone modification was also suggested.


Research methodology in NSAID monitoring: plasma concentrations of chiral drugs.
The poor concentration-efficacy correlation generally observed with chiral nonsteroidal antiinflammatory drugs administered as racemates may, in part, be the result of conclusions based on the correlation of total drug concentration (R- plus S-enantiomers) with efficacy. The disposition kinetics of the enantiomers may be stereoselective. The extent of this stereoselectivity may range from substantial for fenoprofen and ibuprofen to small or negligible for flurbiprofen, ketoprofen and tiaprofenic acid. Pathophysiological factors such as gastrointestinal residence time and age may influence the stereoselectivity. Thus basic and clinical pharmacological data generated without considering stereoselectivity may be misleading.


Herba Gelsemii elegantis is detoxified by ramulus et folium Mussaendae pubescentis extract by modulating hepatic cytochrome P450 and glutathione S-transferase enzymes in rats.
Herba Gelsemii elegantis (GE) has been frequently used as a Chinese folk medicine but has high acute toxicity. In Traditional Chinese Medicine, it may be detoxified by Ramulus et Folium Mussaendae pubescentis (MP), but the detoxification mechanism has remained elusive. The present study aimed to evaluate the detoxification mechanisms by which MP modulates the effect of GE in rats, including the inhibition of hepatic cytochrome P (CYP)450 and glutathione S-transferase (GST) enzymes. Male Sprague Dawley rats were orally administered GE at three doses (0.36, 0.43 or 0.54 g/kg) alone and, at the highest dose, in combination with MP (21.6 g/kg) every day for 7 consecutive days. The control group of animals received the same volume of saline. The mRNA and protein expression of hepatic CYPs representative of two subfamilies (CYP2E1 and CYP1A2) were separately assessed by reverse-transcription quantitative polymerase chain reaction (RT-qPCR), western blot and immunohistochemistry assays. The mRNA and protein expression as well as enzyme activity of hepatic GST were assessed by RT-qPCR, western blot and colorimetric assays, respectively. The results indicated that GE significantly inhibited CYP2E1 mRNA and protein expression in a dose-dependent manner. Co-administration of MP increased CYP2E1 mRNA and protein expression as compared with the high GE dose alone. Cells expressing CYP2E1, located around the hepatic vascular plexus under a clear background, were identified by immunohistochemical staining. The results for CYP1A2 were similar to those for CYP2E1. At all concentrations used, GE significantly inhibited GST mu 1 (GSTm1) mRNA and protein expression in a dose-dependent manner, as compared with the control. Combination of GE and MP increased the mRNA and protein expression of GSTm1 as compared with the high dose of GE. However, the differences in GST-pi mRNA and protein expression between the GE and GE + MP groups were not significant. Of note, rats co-treated with MP were significantly protected from the decrease in GST activity produced by GE. The present study indicated that co-administration of GE and MP upregulated the activities of CYP450 and GST enzymes when compared with GE alone. This modulation may explain for the effect of MP in reducing the acute toxicity of GE.


N-linked glycosylation of protease-activated receptor-1 second extracellular loop: a critical determinant for ligand-induced receptor activation and internalization.
Protease-activated receptor-1 (PAR1) contains five N-linked glycosylation consensus sites as follows: three residing in the N terminus and two localized on the surface of the second extracellular loop (ECL2). To study the effect of N-linked glycosylation in the regulation of PAR1 signaling and trafficking, we generated mutants in which the critical asparagines of the consensus sites were mutated. Here, we report that both the PAR1 N terminus and ECL2 serve as sites for N-linked glycosylation but have different functions in the regulation of receptor signaling and trafficking. N-Linked glycosylation of the PAR1 N terminus is important for transport to the cell surface, whereas the PAR1 mutant lacking glycosylation at ECL2 (NA ECL2) trafficked to the cell surface like the wild-type receptor. However, activated PAR1 NA ECL2 mutant internalization was impaired compared with wild-type receptor, whereas constitutive internalization of unactivated receptor remained intact. Remarkably, thrombin-activated PAR1 NA ECL2 mutant displayed an enhanced maximal signaling response compared with wild-type receptor. The increased PAR1 NA ECL2 mutant signaling was not due to defects in the ability of thrombin to cleave the receptor or signal termination mechanisms. Rather, the PAR1 NA ECL2 mutant displayed a greater efficacy in thrombin-stimulated G protein signaling. Thus, N-linked glycosylation of the PAR1 extracellular surface likely influences ligand docking interactions and the stability of the active receptor conformation. Together, these studies strongly suggest that N-linked glycosylation of PAR1 at the N terminus versus the surface of ECL2 serves distinct functions critical for proper regulation of receptor trafficking and the fidelity of thrombin signaling.


[Localized fibrous tumor of the pleura: radiological findings].
Localized fibrous tumors of the pleura are rare tumors that represent less than 5% of the pleural tumors, although they have also been described in extrathoracic locations such as the abdomen, head and neck or central nervous system. A total of 80% begin in the visceral pleura and are not related with environmental risk factors. They generally occur in patients over 50 with a mild predominance in the woman. Up to 50% of the patients are asymptomatic, so that it appears as an incidental finding on the chest X-ray. These are slow growing tumors. They are seen on the X-ray as well-defined rounded lesions dependent on the pleura and 50% are pediculated. It is important to know its radiological characteristics for its diagnosis of suspicion and correct treatment since surgical resection is generally curative. Local recurrence after the surgery is very common if it is not completely excised.


Is the presence of transient evoked otoacoustic emmissions in ears with acoustic neuroma significant?
One hundred sixty-eight patients admitted for acoustic neuroma removal were involved in this study. In all cases, the size of the tumor and the presence or absence of tinnitus and vertigo or dizziness were evaluated. Investigated functional procedures included bilateral measures of pure-tone audiometry (PTA), auditory brainstem response (ABR), electronystagmography (ENG), and transient evoked otoacoustic emissions (TEOAEs). Thirty-five patients (21%) had normal preoperative TEOAEs in the affected ear, whereas 133 patients (79%) failed to show reproducible responses. The aims of this study were 1. to evaluate what distinguishes patients with preoperative TEOAEs in the pathological ear (group A) from those who had no TEOAE (group B); and 2. to determine in cases of attempted hearing preservation whether preoperative TEOAE presence in the neuroma ear (group C) was predictive of postoperative hearing preservation compared with the group of patients without TEOAEs (group D). The presence of vertigo or dizziness was significantly less frequent, the age was lower, and preoperative mean PTA loss in both ears was lower in group A compared with group B. Frequency of the other studied parameters and ABR threshold were similar in both groups. When hearing preservation was attempted, the mean preoperative PTA loss of group C patients was lower in both ears compared with group D. However, postoperative mean PTA loss did not significantly differ in the two groups. In group C, the percentage of hearing preservation (66.6%) was significantly higher than the percentage of deafness (33.4%), whereas in group D the percentage of postoperative preserved audition and deafness did not significantly differ (respectively 44.4% and 55.6%). The findings suggest that 1. TEOAEs in ears with acoustic neuromas are found in younger patients with a lower preoperative mean PTA loss and are accompanied by fewer functional complaints, perhaps because preserved TEOAEs indicate a better preservation of inner ear vasculature; and, 2. along with radiological and electrophysiological investigations, TEOAE presence in the pathological ear could provide an additional criterion or predictive factor for the successful outcome of attempted hearing-conservation surgery in ears with acoustic neuromas.


Epigenetic regulation and measurement of epigenetic changes.
Epigenetic mechanisms provide an adaptive layer of control in the regulation of gene expression that enables an organism to adjust to a changing environment. Epigenetic regulation increases the functional complexity of deoxyribonucleic acid (DNA) by altering chromatin structure, nuclear organization, and transcript stability. These changes may additively or synergistically influence gene expression and result in long-term molecular and functional consequences independent of the DNA sequence that may ultimately define an individual's phenotype. This article (1) describes histone modification, DNA methylation, and expression of small noncoding RNA species; (2) reviews the most common methods used to measure these epigenetic changes; and (3) presents factors that need to be considered when choosing a specific tissue to evaluate for epigenetic changes.


Generating ultrabroadband terahertz radiation based on the under-compression mode of velocity bunching.
We propose and analyze a scheme to generate enhanced ultrabroadband terahertz (THz) radiation through coherent transition radiation emitted by ultrashort electron beams based on a 10.5 m beamline at Tsinghua University. The proposed scheme involves the initial compression of the electron beam with a few hundred pC charges using a velocity bunching scheme (i.e., RF compression) in an under-compression mode instead of the usual critical-compression mode in order to maintain a positive energy chirp at the exit of the traveling wave accelerator. After a long drift segment, the particles in the tail catch up with the bunch head. More than 80% of the particles are distributed in a spike with an rms length less than 20 fs. Such beams correspond to an ultrabroadband coherent transition radiation (CTR) spectrum of 0.1 THz to 25 THz, with the single-pulse THz radiation energy of up to 50 μJ. The principle of CTR and under-compression mode of velocity bunching are introduced in this paper. And the ASTRA simulation parameters and the stability of the system are also discussed.


Low levels of ADAMTS-13 with high anti-ADAMTS-13 antibodies during remission of immune-mediated thrombotic thrombocytopenic purpura highly predict for disease relapse: A multi-institutional study.
Immune-mediated thrombotic thrombocytopenic purpura (iTTP) is a life-threatening immune-mediated thrombotic microangiopathy. Daily therapeutic plasma exchange (TPE) and the optimized use of rituximab have strikingly improved the outcome of this disease, however the rate of disease recurrence remains high. Specific predictors of relapse in patients in remission can be relevant for an optimal patient management. In this study, we aimed to identify predictive variables of disease relapse in a multicenter cohort of 74 out of 153 iTTP patients. They were tested at different time points during remission for the levels of ADAMTS-13 activity and autoantibody, and did not receive pre-emptive treatment for ADAMTS-13 activity deficiency during remission. The results showed that the association of ADAMTS13 activity ≤20% with a high anti-ADAMTS-13 titer at remission, and the time to response to first line treatment ≥13 days, were independent predictive factors of disease relapse. In addition, the use of rituximab in patients with exacerbation or refractoriness to TPE was significantly associated with reduced relapse rate. By Cox regression analysis, patients with ADAMTS-13 activity ≤20% plus anti-ADAMTS13 antibody titer ≥15 U/mL at remission had an increased risk of relapse (HR 1.98, CI 95% 1.087-3.614; P < .02). These findings may help to outline more personalized therapeutic strategies in order to provide faster and sustained responses to first-line iTTP treatment and prevent relapses in these patients.


In-plane aromaticity in cycloparaphenylene dications: a magnetic circular dichroism and theoretical study.
The electronic structures of [8]cycloparaphenylene dication ([8]CPP(2+)) and radical cation ([8]CPP(•+)) have been investigated by magnetic circular dichroism (MCD) spectroscopy, which enabled unambiguous discrimination between previously conflicting assignments of the UV-vis-NIR absorption spectral bands. Molecular orbital and nucleus-independent chemical shift (NICS) analysis revealed that [8]CPP(2+) shows in-plane aromaticity with a (4n + 2) π-electron system (n = 7). This aromaticity appears to be the origin of the unusual stability of the dication. Theoretical calculations further suggested that not only [8]CPP(2+) but also all [n]CPP (n = 5-10) dications and dianions exhibit in-plane aromaticity.


Focal EEG slowing and chorea: electroclinical clues to the diagnosis of anti-NMDAR encephalitis.
Variations in clinical presentation can lead to delays in the diagnosis and initiation of treatment of anti-N-methyl-D-aspartate receptor encephalitis. Most patients have an EEG study performed early in the course of their illness. Although not specific, there may be clues in the electroclinical features that should alert clinicians and electroencephalographers to the possibility of this diagnosis. This case is a reminder that anti- anti-N-methyl-D-aspartate receptor encephalitis may present initially with a movement disorder as the sole symptom, without features of an encephalopathy. In addition, it adds to the growing body of evidence that recognition of certain electroclinical clues may shorten the time to diagnosis. [Published with video sequence].


Detection of invA, sivH, and agfA Virulence Genes in Salmonella spp. Isolated from Broiler Breeder Farms in Alborz Province, Iran.
Salmonellosis, among poultry infectious diseases, not only imposes economic losses in the field of poultry breeding but also is considered a zoonotic disease. This study aimed to investigate the presence of invA, sivH, and agfA virulence genes in Salmonella species. The present study was conducted on 30 Salmonella strains. Samples were cultured on selective and differential media, and afterward, the isolates were serotyped using specific antisera based on the Kauffman-White table. Subsequently, the samples were analyzed to detect invA, sivH, and agfA genes by polymerase chain reaction technique. The results indicated that 30 (100%) isolates had invA and agfA virulence genes and 28 (93.33%) isolates had a sivH virulence gene. The highest frequency of serotypes was related to Salmonella infantis. Among the studied serotypes, Salmonella uno and Salmonella O35 lacked the sivH virulence gene, unlike other serotypes. The findings of this study could pave the way for Salmonella monitoring and be used as a pattern to detect Salmonella bacteria-bearing genes encoding invasion and fimbria.


Does Gender of Administrator Matter? National Study Explores U.S. University Administrators' Attitudes About Retaining Women Professors in STEM.
Omnipresent calls for more women in university administration presume women will prioritize using resources and power to increase female representation, especially in STEM fields where women are most underrepresented. However, empirical evidence is lacking for systematic differences in female vs. male administratorsŠ attitudes. Do female administrators agree on which strategies are best, and do men see things differently? We explored United States college and university administratorsŠ opinions regarding strategies, policies, and structural changes in their organizations designed to increase women professorsŠ representation and retention in STEM fields. A comprehensive review of past research yielded a database of potentially-effective, recommended policies. A survey based on these policies was sent to provosts, deans, associate deans, and department chairs of STEM fields at 96 public and private research universities across the U.S. These administrators were asked to rate the quality and feasibility of each strategy; 474 provided data, of which 334 contained complete numerical data used in the analyses. Our data revealed that female (vs. male) administrators believed the 44 strategies were higher in quality overall-but not higher in feasibility-with 9 strategies perceived differently by women and men, after imposing conservative statistical controls. There was broad general agreement on the relative-quality rankings of the 44 strategies. Women (vs. men) gave higher quality ratings to increasing the value of teaching, service, and administrative experience in tenure/promotion decisions, increasing flexibility of federal-grant funding to accommodate mothers, conducting gender-equity research, and supporting shared tenure lines enabling work-life balance. Women (vs. men) believed it was more feasible for men to stop the tenure clock for 1 year for childrearing and for universities to support requests for shared tenure lines, but less feasible for women to chair search committees. Our national survey thus supported the belief that placing women into administration creates greater endorsement of strategies to attract and retain women in STEM, although the effectiveness of these strategies was outside the scope of this research. Topics of disagreement between women and men are potentially important focuses of future policy, because female administrators may have insights into how to retain women that male administrators do not share.


Spororrminone A and 2-epi-spororrminone A, two new chromones from an endolichenic fungus Sporormiella irregularis.
Two new chromones, spororrminone A (1, (S)-5-hydroxy-2-methyl-4-oxo-2-((S)-5-oxotetrahydrofuran-2-yl)chroman-7-carboxylic acid) and 2-epi-spororrminone A (2, (R)-5-hydroxy-2-methyl-4-oxo-2-((S)-5-oxotetrahydrofuran-2-yl)chroman-7-carboxylic acid), were isolated from an EtOAc extract of an endolichenic fungal strain Sporormiella irregularis (No. 71-11-4-1). The structures of these compounds were identified by spectroscopic analyses. The absolute configuration of 1 was established by single-crystal X-ray diffraction. Spororrminone A (1) and 2-epi-spororrminone A (2) represent the first examples of 2-(5-oxotetrahydrofuran-2-yl) chromones with 7-carboxylic group.


Inter-segmental coordination in progressions for the longswing on high bar.
This study focused on identifying the most effective skill progression for developing the longswing on high bar in men's artistic gymnastics. Building on previous work by Irwin and Kerwin, in which a method to rank progressions based on their angular kinematics was developed, this study aimed to use the method to quantify similarities in inter-segmental coordination between selected progressions and the longswing on high bar. Video images of four members of the UK men's national gymnastics squad performing three series of five longswings and eight progressions were recorded at 50Hz. Two-dimensional direct linear transformation techniques were used to determine the real-world coordinates from the digitized data. Inter-segmental coordination of the hip and shoulder joints during the functional phases of the longswing was assessed using continuous relative phase. Similarity between the longswing and each progression was represented by a "specificity score", which was also used to rank the progressions. Each progression's specificity score was calculated by combining a "difference score" (root mean squared difference between the continuous relative phase profiles of the longswing and the progression) and a "variability score" (standard deviation of the continuous relative phase profiles for each progression). The progressions that were most similar to the longswing included the looped bar longswing and layaway swing down (ranked 1st and 2nd), with specificity scores of 9% and 10% respectively. In contrast, the least similar progressions were the looped bar "no action" longswing (51%) and pendulum swing (63%) (ranked 7th and 8th). Establishing effective skill learning pathways is recognized as a key component of the coaching process and ranking progressions based on their specificity score provided a mechanism to identify progressions with similar inter-segmental coordination profiles to the key skill on the high bar, the longswing.


IL-23 Gene and Protein Expression in Childhood Asthma.
Asthma is the chronic inflammation of airways characterized by eosinophilic infiltration, mucus overproduction, airway hyper-responsiveness and airway remodeling. These changes are induced mostly by cytokines which are produced by T helper (Th) 2 cells. Recently, the role of interleukin-23 (IL-23) in the pathogenesis of adult allergic asthma has been studied. To explore IL-23 serum levels and its expression in persistent asthma compared with healthy children younger than five years old. Blood samples of 40 children with mild and severe persistent asthma were compared to 34 healthy children regarding IL-23 serum levels and gene expression using enzyme-linked immunosorbentassay (ELISA) and real time quantitative polymerase chain reaction (PCR). The IL-23 gene expression level was significantly different in the 25 children with mild persistent asthma and the 15 children with severe persistent asthma compared to the control group (p=0.001). There was no significant difference in IL-23 gene expression level between the two groups of patients with mild and severe persistent asthma. A significant difference was seen in IL-23 serum levels between the 25 children with persistent asthma and control group (p=0.002). For pre-school children with history and physical exam in favor of asthma which cannot be tested by spirometry, IL-23 serum levels may be an auxiliary biomarker for the diagnosis of asthma.


A morphometric study on the early stages of dendrite changes in the axotomized motoneuron of the spinal cord in newborn rats.
To study the early effect of axotomy on spinal motoneurons and dendritic trees in the newborn rat. The left sciatic nerve of 50 neonatal Sprague--Dawley rats aged 5 days old was transected and the right area kept as a control. The operated animals were killed 2, 4, 8, 12 and 24 hours after axotomy. The L4--L6 segments of the spinal cord were sampled, and stained either with Golgi-Cox or Cresyl fast violet methods. The number of motoneurons, the largest soma diameter and the different parameters of the dendritic trees such as: number, length and thickness of the primary, secondary and tertiary branches in the axotomized sides were estimated and compared statistically with that of the intact sides. The results indicated that in the axotomized sides, the number of motoneurons and the largest soma diameter were decreased, but these were significant only in 12- and 24-hour groups. The number of branches of the dendritic trees including primary, secondary and tertiary branches was not significantly decreased in the groups. The thickness of the dendrites showed a reduction; however, this was significant only for the tertiary branches in the 24-hour groups. The length of the primary, secondary and tertiary branches of the dendrites--especially the latter--were also decreased significantly in most of the groups. Axotomy at the early stages in newborn rats resulted in noticeable morphometrical changes in motoneurons and their dendrites.


[Acute encephalitis complicating acute polyradiculoneuritis].
We report the case of a 43-year-old man who developed an acute encephalopathy after IVIg therapy for AIDP. Imaging studies showed predominantly posterior leukoencephalopathy. The signs and symptoms of the encephalopathy completely resolved by steroids. Two patients with acute encephalopathy, following IVIg therapy, were reported previously in the literature. However, our observation differed from them by the presence of a marked pleiocytosis of cephalospinal fluid before beginning of IgIV. Two hypothesis may be made: a post-infectious meningo-encephalo-AIDP or a complication of IgIV. This side effect, even rare, has not to be underestimated.


Priming Adipose Stem Cells with Tumor Necrosis Factor-Alpha Preconditioning Potentiates Their Exosome Efficacy for Bone Regeneration.
Mesenchymal stem cells (MSCs) have been widely used for tissue repair and regeneration. However, the inherent drawbacks, including limited cell survival after cell transplantation, have hindered direct MSC transplantation for tissue repair and regeneration. The aim of this study was to investigate if exosomes isolated from MSCs can promote the proliferation and differentiation of human primary osteoblastic cells (HOBs) and be potentially used for bone tissue regeneration. We showed that adipose tissue-derived MSC (ASC)-derived exosomes (ASC-EXO) were able to promote the proliferation and osteogenic differentiation in HOBs; and the trophic effects of ASC-EXO on HOBs were further harnessed when ASCs were preconditioned with tumor necrosis factor-alpha (TNF-α) for 3 days, which mimics the acute inflammatory phase upon bone injury. In addition, we showed that Wnt-3a content was elevated in ASC-EXO when ASCs were preconditioned by TNF-α, and inhibiting Wnt signaling decreased the osteogenic gene expression levels in HOBs which were cultured in TNF-α preconditioned ASCs conditioned medium. In conclusion, it was demonstrated that ASC-EXO, especially primed by TNF-α preconditioning on ASCs, offer a promising approach to replace direct stem cell transplantation for bone repair and regeneration.


Time motion analysis of football (soccer) referees during official matches in relation to the type of fluid consumed.
The aim of the present study was to determine the effect of volume and composition of fluid replacement on the physical performance of male football referees. Ten referees were evaluated during three official matches. In one match the participants were asked to consume mineral water ad libitum, and in the others they consumed a pre-determined volume of mineral water or a carbohydrate electrolyte solution (6.4% carbohydrate and 22 mM Na+) equivalent to 1% of their baseline body mass (half before the match and half during the interval). Total water loss, sweat rate and match physiological performance were measured. When rehydrated ad libitum (pre-match and at half time) participants lost 1.97 ± 0.18% of their pre-match body mass (2.14 ± 0.19 L). This parameter was significantly reduced when they consumed a pre-determined volume of fluid. Sweat rate was significantly reduced when the referees ingested a pre-determined volume of a carbohydrate electrolyte solution, 0.72 ± 0.12 vs 1.16 ± 0.11 L/h ad libitum. The high percentage (74.1%) of movements at low speed (walking, jogging) observed when they ingested fluid ad libitum was significantly reduced to 71% with mineral water and to 69.9% with carbohydrate solution. An increase in percent movement expended in backward running was observed when they consumed a pre-determined volume of carbohydrate solution, 7.7 ± 0.5 vs 5.5 ± 0.5% ad libitum. The improved hydration status achieved with the carbohydrate electrolyte solution reduced the length of time spent in activities at low-speed movements and increased the time spent in activities demanding high-energy expenditure.


Decomposition reaction rate of BCl3-C3H6(propene)-H2 in the gas phase.
The decomposition reaction rate in the BCl(3)-C(3)H(6)-H(2) gas phase reaction system in preparing boron carbides was investigated based on the most favorable reaction pathways proposed by Jiang et al. [Theor. Chem. Accs. 2010, 127, 519] and Yang et al. [J. Theor. Comput. Chem. 2012, 11, 53]. The rate constants of all the elementary reactions were evaluated with the variational transition state theory. The vibrational frequencies for the stationary points as well as the selected points along the minimum energy paths (MEPs) were calculated with density functional theory at the B3PW91/6-311G(d,p) level and the energies were refined with the accurate model chemistry method G3(MP2). For the elementary reaction associated with a transition state, the MEP was obtained with the intrinsic reaction coordinates, while for the elementary reaction without transition state, the relaxed potential energy surface scan was employed to obtain the MEP. The rate constants were calculated for temperatures within 200-2000 K and fitted into three-parameter Arrhenius expressions. The reaction rates were investigated by using the COMSOL software to solve numerically the coupled differential rate equations. The results show that the reactions are, consistent with the experiments, appropriate at 1100-1500 K with the reaction time of 30 s for 1100 K, 1.5 s for 1200 K, 0.12 s for 1300 K, 0.011 s for 1400 K, or 0.001 s for 1500 K, for propene being almost completely consumed. The completely dissociated species, boron carbides C(3)B, C(2)B, and CB, have very low concentrations, and C(3)B is the main product at higher temperatures, while C(2)B is the main product at lower temperatures. For the reaction time 1 s, all these concentrations approach into a nearly constant. The maximum value (in mol/m(3)) is for the highest temperature 1500 K with the orders of -13, -17, and -23 for C(3)B, C(2)B, and CB, respectively. It was also found that the logarithm of the overall reaction rate and reciprocal temperature have an excellent linear relationship within 700-2000 K with a correlation coefficient of 0.99996. This corresponds to an apparent activation energy 337.0 kJ/mol, which is comparable with the energy barrier 362.6 kJ/mol of the rate control reaction at 0 K but is higher than either of the experiments 208.7 kJ/mol or the Gibbs free energy barrier 226.2 kJ/mol at 1200 K.


Health belief systems and the psychobiology of war.
Belief systems overlie powerful biological and psychological forces that are root causes of war. Much as in medicine where an appreciation of health belief systems is necessary in the control of illness and disease, so the paths to the control of war may lie in an understanding of belief systems and ways to circumvent them. Such understanding gives strong theoretical support to many time-honored but underutilized international initiative and educational ventures. The effort of the medical community to educate the public about biomedical aspects of nuclear war should gain more balance and sophistication with an appreciation of belief systems in the psychobiology of war.


Hepatoprotective effects of Poly-[hemoglobin-superoxide dismutase-catalase-carbonic anhydrase] on alcohol-damaged primary rat hepatocyte culture in vitro.
We have prepared a novel nanobiotherapeutic, Poly-[hemoglobin-superoxide dismutase-catalase-carbonic anhydrase], which not only transports both oxygen and carbon dioxide but also a therapeutic antioxidant. Our previous study in a severe sustained 90 min hemorrhagic shock rat model shows that it has a hepatoprotective effect. We investigate its hepatoprotective effect further in this present report using an alcohol-damaged primary hepatocyte culture model. Results show that it significantly reduced ethanol-induced AST release, lipid peroxidation, and ROS production in rat primary hepatocytes culture. It also significantly enhanced the viability of ethanol-treated hepatocytes. Thus, the result shows that Poly-[hemoglobin-superoxide dismutase-catalase-carbonic anhydrase] also has some hepatoprotective effects against alcohol-induced injury in in vitro rat primary hepatocytes cell culture. This collaborate our previous observation of its hepatoprotective effect in a severe sustained 90-min hemorrhagic shock rat model.


An uncommon mimic of an acute asthma exacerbation.
Foreign body aspiration in adults has a variety of clinical presentations and often goes unrecognized. We describe the case of a patient who experienced crack cocaine aspiration and presented with symptoms of an acute asthma exacerbation requiring mechanical ventilation until the eventual diagnosis and bronchoscopic removal of the foreign body.


The role of interventional radiology in the treatment of the diabetic foot.
Although the pathologic features of arterial disease in diabetes do not differ from those of simple atheromatous lesions, the plurisegmental involvement, the predominant subpopliteal location of lesions and the association with impaired cardiac and renal function in these patients imply peculiar problems in the selection of an adequate therapy. At present, in interventional radiology a number of procedures are available for intravascular therapy (angioplasty, stent, locoregional fibrinolisis, mechanical atherectomy), particularly suitable for diabetics who are affected by multisystem impairment. These procedures are not in opposition to the conventional ones (surgical and medical) but rather integrate them, allowing to extend the indications and improve the results in the treatment of the vasculogenic diabetic foot. Based on the personal experience the potential of intravascular procedures in this disease, is analyzed.


The relationship between immunological responsiveness controlled by T-helper 2 lymphocytes and infections with parasitic helminths.
It should have been difficult until relatively recently for immunologists to ascribe a sound biological reason for the continued possession of the allergic phenotype in human populations. Nevertheless, for the past 20 years or so textbooks of immunology have routinely exhibited fanciful and perhaps exaggerated diagrams as to how IgE and eosinophils killed all helminth parasites. These diagrams were largely based on perhaps selective in vitro observations, and it is only now that immunoparasitologists, working on human populations under arduous conditions in the field, are able to provide data to corroborate these findings, and perhaps ascribe a useful purpose for a generally pathological immune response termed Type I hypersensitivity. The present paper reviews much of this recent literature, and asks a number of pertinent questions relating to the relationship between what we now know to be T-helper 2 lymphocyte-driven immunological responsiveness and infections with parasitic helminths.


Clinical values of circulating tumor cells count in localized renal cell carcinoma.
Renal cancer is one of the most common malignant tumors of the urinary system, with distant metastasis occurring 30% of patients. Therefore, early detection and monitoring of tumor progression are of great significance in the diagnosis and treatment of renal cancer. However, current biomarkers used to diagnose, monitor recurrence and assess prognosis of renal cancer are still uncertain. Circulating tumor cells (CTCs) are tumor cells detached from the primary tumor or metastasis, invaded and existing in the peripheral blood, and are one of the most promising liquid biopsy targets because they can provide complete cell biological information. Microfluidic chip has advantages of miniaturization, high integration, and fast analysis, which has advantages in CTC separation and enrichment. In this study, 1 mL peripheral blood of each 30 patients with early localized renal cancer was collected before and 1 day after surgery. CTC enrichment was performed by microfluidic chip and CTCs were identified by immunofluorescence staining. All patients were followed up for a median of 17 months. The number of CTCs before surgery was higher than that after surgery (P<0.001), and the number was positively correlated with tumor-node-metastasis (TNM) stage and International Society of Urological Pathology (ISUP) grade. Patients in group CTC ≤2 had a longer progression-free survival (PFS) than those in group CTC ≥3 (P<0.05). Surgical treatment can remarkably reduce the number of CTCs in patients, and CTC counts can also play a role in monitoring tumor load and predicting prognosis in renal cancer.


Fiber-based platform for synchronous imaging of endogenous and exogenous fluorescence of biological tissue.
Endogenous and exogenous fluorescence emission from biological samples encodes complementary information. Here we report, to the best of our knowledge, the first results from an optical imaging platform with interleaved excitation and detection of exogenous and endogenous fluorescence from tissue samples using a single flexible multimode fiber that delivers the excitation beam and collects the emitted light. A custom-built reflective optical chopper wheel with synchronized rotation temporally multiplexes an autofluorescence lifetime imaging apparatus with an intensity-based fluorescence module tailored to imaging green fluorescent protein. We demonstrate the functionality of such platform imaging dyes of varying fluorescence signatures and resolving cellularized areas on bio-engineered tissue constructs.


Influence of the Flushing Response in the Relationship between Alcohol Consumption and Cardiovascular Disease Risk.
The purpose of this study was to examine the relationship between cardiovascular disease risk and alcohol consumption according to facial flushing after drinking among Korean men. The subjects were 1,817 Korean men (non-drinker group, 283 men; drinking-related facial flushing group, 662 men; non-flushing group, 872 men) >30 years who had undergone comprehensive health examinations at the health promotion center of a Chungnam National University Hospital between 2007 and 2009. Alcohol consumption and alcohol-related facial flushing were assessed through a questionnaire. Cardiovascular disease risk was investigated based on the 2008 Framingham Heart Study. With the non-drinker group as reference, logistic regression was used to analyze the relationship between weekly alcohol intake and cardiovascular disease risk within 10 years for the flushing and non-flushing groups, with adjustment for confounding factors such as body mass index, diastolic blood pressure, low density lipoprotein cholesterol, triglycerides, and exercise patterns. Individuals in the non-flushing group with alcohol consumption of ≤4 standard drinks (1 standard drink = 14 g of alcohol) per week had significantly lower moderate or high cardiovascular disease risk than individuals in the nondrinker group (adjusted odds ratio, 0.51; 95% confidence interval, 0.37 to 0.71). However, no significant relationship between the drinking amount and cardiovascular disease risk was observed in the flushing group. Cardiovascular disease risk is likely lowered by alcohol consumption among non-flushers, and the relationship between the drinking amount and cardiovascular disease risk may differ according to facial flushing after drinking, representing an individual's vulnerability.


Targeted therapy for malignant melanoma.
Treatment of advanced melanoma has undergone a paradigm shift over the last 10-15 years. The frustrating results of studies on medical treatment ten years ago have been replaced by studies constantly improving survival in patients with advanced melanoma. Immune checkpoint inhibitors belong to one group of treatments and targeted therapy to another. Fifty percent of melanomas are BRAF mutation positive. Normally, the mitogen activated protein kinase or MAP kinase (Ras-BRAF-MEK-Erk chain) pathways translate external signals to intracellular growth and proliferation. In BRAF mutated melanoma cells, the mutated BRAF kinase is excessively active leading to autonomous proliferation and cancerous growth. This kinase can be blocked by BRAF-inhibitors. If given to BRAF negative melanoma patients, it may lead to disease progression because Ras is not inhibited in these cells. Development of Squamous cell carcinomas as a serious adverse event to BRAF inhibition may be caused by similar mechanisms in non BRAF mutated keratinocytes. A spontaneous and paradoxical loss of effect is seen with BRAF inhibitors due to various ways melanoma cells bypass BRAF. This is somewhat counteracted by the addition of a MEK1/2 inhibitor. Overall progression-free survival has increased from a median of two months for chemotherapy, via 7-8 months for BRAF inhibitor to 10-14 months for newer BRAF and MEK inhibitor combination therapy.


Cytogenetic analysis in the examination of solid tumors in children.
Although pediatric solid tumors are cytogenetically less well characterized than childhood leukemias, an understanding of the role of chromosomal changes in the development of these neoplasms is emerging. The major clinical importance of chromosome analysis today is diagnostic. Especially in small cell round cell tumors of childhood, the unique karyotypic patterns that characterize some of the differential diagnostic entities make it possible to determine with a high degree of certainty which type of cancer the child has. Molecular studies have revealed that almost all retinoblastomas show homozygous loss of function of the RB1 gene in 13q14. At the cytogenetic level, however, aberrations of 13q are seen in less than 25% of retinoblastomas; instead, the presumably progression-related i(6p) and aberrations leading to gain of 1q predominate, each being present in one-third of the tumors. Twenty percent of cytogenetically aberrant Wilms' tumors show structural rearrangements, often deletions, of 11p13 and 11p15, where the WT1 and WT2 genes map. Other frequent changes are trisomy 12 and duplication of 1q. The most common (80%) cytogenetic abnormality in neuroblastoma is loss of distal 1p, a chromosome segment thought to harbor at least two tumor-suppressor genes of importance in tumorigenesis. Double minute chromosomes or homogeneously staining regions are present in one-third of all neuroblastomas and are associated with MYCN amplification. Loss of 1p material or MYCN amplification predicts a poor outcome. The most common (30%) chromosomal aberration in primitive neuroectodermal tumors of the central nervous system is i(17q). The formation of this isochromosome may help inactivate a tumor-suppressor gene located distal to the TP53 locus on 17p. No specific chromosome abnormality has been detected in gliomas, but monosomy 22 and rearrangements leading to loss of 1p and gain of 1q are recurrent. Few hepatoblastomas with chromosomal changes have been reported, but several potential primary aberrations have been described, including +2, +20, and duplication 8q. In Ewing's sarcoma, t(11;22)(q24;q12) is the primary aberration, with trisomy 8 and gain of 1q being frequent secondary changes. Fibrosarcomas in children often carry only numeric aberrations, especially trisomy for chromosomes 11, 20, 17, and 8. Most osteosarcomas are cytogenetically complex, and no specific abnormality has been detected; the single most common change is loss of chromosome 13, which is observed in half the tumors. In contrast, the low-malignancy parosteal osteosarcomas often display supernumerary ring chromosomes as the sole karyotypic deviation. The cytogenetic profiles of rhabdomyosarcomas differ among the various morphologic subtypes.(ABSTRACT TRUNCATED AT 400 WORDS)


Comparison of injury epidemiology between the Wenchuan and Lushan earthquakes in Sichuan, China.
We aimed to compare injury characteristics and the timing of admissions and surgeries in the Wenchuan earthquake in 2008 and the Lushan earthquake in 2013. We retrospectively compared the admission and operating times and injury profiles of patients admitted to our medical center during both earthquakes. We also explored the relationship between seismic intensity and injury type. The time from earthquake onset to the peak in patient admissions and surgeries differed between the 2 earthquakes. In the Wenchuan earthquake, injuries due to being struck by objects or being buried were more frequent than other types of injuries, and more patients suffered injuries of the extremities than thoracic injuries or brain trauma. In the Lushan earthquake, falls were the most common injury, and more patients suffered thoracic trauma or brain injuries. The types of injury seemed to vary with seismic intensity, whereas the anatomical location of the injury did not. Greater seismic intensity of an earthquake is associated with longer delay between the event and the peak in patient admissions and surgeries, higher frequencies of injuries due to being struck or buried, and lower frequencies of injuries due to falls and injuries to the chest and brain. These insights may prove useful for planning rescue interventions in trauma centers near the epicenter.


Non-ideal mixing behaviour of hydrogen bonding in mixtures of protic ionic liquids.
Ionic liquids (ILs) attract interest in science and technology as a result of their unique properties. Binary and ternary mixtures of ILs significantly increase the number of possible cation/anion combinations, resulting in targeted physical and chemical properties. In this work, we study the mixing behaviour of two protic ILs: triethyl ammonium methylsulfonate [Et3 NH][CH3 SO3 ] and triethylammonium triflate [Et3 NH][CF3 SO3 ]. We find a characteristic deviation from ideal mixing by means of low-frequency infrared spectroscopy. By using molecular dynamics simulations, we explain this behaviour as being the result of different strengths of anion/cation hydrogen bonding. This non-ideality of non-random H-bond mixing is also reflected in macroscopic properties such as the viscosity. Mixing suitable ILs may, thus, result in new ILs with targeted physical properties.


Mechanism of an inhibitory effect of nipradilol on rat vascular smooth muscle cell growth.
The aim of this study was to clarify the mechanism of an inhibitory effect of nipradilol on cultured rat vascular smooth muscle cell (VSMC) growth. After being starved, cultured VSMCs were stimulated by 5% fetal bovine serum with various concentrations of nipradilol. Nipradilol dose-dependently decreased the values of [(3)H]-thymidine incorporation, cell numbers and total cellular protein content, and the levels of phosphorylated extracellular signal-regulated protein kinase 1/2 and p38. It also suppressed the level of proliferative cell nuclear antigen in a dose-dependent manner. In contrast, nipradilol did not change the level of the phosphorylated value of c-jun NH(2)-terminal protein kinase or cytoplasmic histone-associated DNA fragments in VSMCs. These results indicate that nipradilol suppresses cell growth without apoptosis in rat VSMCs, suggesting that it could be effective for preventing the progression of restenosis after angioplasty.


Elevated Wall Shear Stress in Aortic Type B Dissection May Relate to Retrograde Aortic Type A Dissection: A Computational Fluid Dynamics Pilot Study.
Retrograde aortic type A dissection (RTAD) is a known complication in patients with aortic type B dissection. The purpose of this computational fluid dynamics (CFD) study was to identify haemodynamic risk factors for the occurrence of RTAD. Computed tomographic angiography (CTA) images of 10 patients with type B dissections, who subsequently developed a RTAD, were retrospectively analysed together with patients constituting a control group (n = 10) where no further vascular events after the initial type B dissection occurred. CFD simulations were conducted based on 3D surface models of the aortic lumen derived from CTA datasets. For both groups, pressures, velocity magnitudes and wall shear stress (WSS) were compared at the site of the future RTAD entry tear and the surrounding aortic wall. WSS at the site of the future entry tear was significantly elevated compared with the surrounding wall (15.10 Pa vs. 5.15 Pa, p < .001) and was significantly higher in the RTAD group than in the control group (6.05 Pa, p < .002). Pressures and velocity magnitudes were not significantly elevated at the entry tear (3825.8 Pa, 0.63 m/s) compared with the aortic arch (3549.8 Pa, 0.50 m/s) or control group (3501.7 Pa, 0.62 m/s). Increased WSS accompanies the occurrence of RTAD. The results merit the design for a prospective study to confirm whether WSS is a risk factor for the occurrence of RTAD.


Co-morbidity and acute decompensations of COPD requiring non-invasive positive-pressure ventilation.
To assess the prevalence and the impact of chronic and/or acute non-respiratory co-morbidity on short and longer-term outcome of non-invasive positive pressure ventilation (NIPPV) in acute decompensations of chronic obstructive pulmonary disease (COPD) with acute hypercapnic respiratory failure (AHRF). An observational study in a three-bed respiratory monitoring unit in a respiratory ward of a non-university hospital. We grouped 120 consecutive COPD patients requiring NIPPV for AHRF (pH 7.28+/-0.05, PaO2/FIO2 ratio 192+/-63, PaCO2 78.3+/-12.3 mmHg) according to whether NIPPV succeeded (n=98) or failed (n=22) in avoiding the need for endotracheal intubation and whether alive (n=77) or dead (n=42) at 6 months. The prevalence of chronic and acute co-morbidity was, respectively, 20% and 41.7%; most of the cases were cardiovascular. In-hospital NIPPV failure was greater in patients with than in those without chronic (33.3% vs. 14.6%) or acute co-morbidity (32% vs. 8.6%). Six-month mortality was worse in patients with than in those without chronic (54.2% vs. 30.5%) or more than one acute co-morbidity (66.7% vs. 30.8%). Multiple regression analysis predicted in-hospital NIPPV failure by acute co-morbidity and forced expiratory volume in 1 s, while death at 6 months was predicted by having more than one acute co-morbidity, non-cardiovascular chronic co-morbidity and Activities of Daily Living score. Chronic and acute co-morbidities are common in COPD patients with AHRF needing NIPPV and their presence influences short and longer-term outcome.


An Insight into Glyco-Microheterogeneity of Plasma von Willebrand Factor by Mass Spectrometry.
Human plasma von Willebrand Factor (VWF) plays essential roles in primary hemostasis in cooperation with other coagulations factors. There is ample indication that glycosylation affects many biological phases during the protein life cycle. However, comprehensive characterization of all probable N-glycosites simultaneous with O-glycosites is still not fully revealed. Thus, the intention of this exploration was to estimate the occupancy of all canonical N-glycosites besides simultaneous characterization of N- and O-glycoforms. An RP-LC-MS/MS system functionalized with CID and HCD tandem mass was utilized to analyze VWF. N-Glycosite occupancy varied along the protein backbone chain. Out of 257 HCD spectra, 181 characterized glycoforms were specified as either N- or O-glycosites. Sequential cleavage of glycosidic bonds along with Human Database mass matching have confirmed the glycoform structures. A total of 173 glycoforms represented most commonly biantennary and infrequently tri- and tetra-antennary N-glycans beside high mannose, hybrid, ABH antigen-terminated, and sulfated N-glycans. Many glycoforms were common across all N-sites. Noteworthy, previously unreported N-glycosites within domain D'(TIL'-E') showed glycosylation. Moreover, sialylated core 1 and core 2 O-glycans were detected on 2298T. Given subtle characterization of site-specific glycoforms, we can attain a profound understanding of the biological roles of VWF as well as facilitate the production of VWF-based therapeutics.


Residual feed intake and its effect on Salmonella enteritidis infection in growing layer hens.
Previous phenotypic selection on residual feed intake (RFI) identified 20 efficient R- chickens and 20 nonefficient R+ chickens. Residual feed intake was defined as the difference between observed feed intake (FI) and expected FI based on metabolic BW and BW gain, and was used as a measure for feed efficiency. Body weight and BW gain were similar for both groups. Feed intake and RFI were significantly higher in R+ birds. It is hypothesized that nonefficient R+ birds are more flexible to divert resources from production processes toward maintenance processes, thus being better capable of handling a bacterial challenge. Chickens of both groups were randomly allocated to immunization with heat-inactivated Salmonella enteritidis bacteria, or inoculation with live Salmonella bacteria. Transportation to the isolation units caused a decrease in FI in R+ birds. This may reflect a particular way of coping with stress in R+ birds. More R+ birds stopped bacterial shedding considering a nonshedding interval of 10 or 11 d (P = 0.041). Nonantigen-specific antibody responses against keyhole limpet hemocyanin (KLH) were higher in R- birds. We conclude that R+ birds are able to keep their metabolism at a higher level, as indicated by higher heart and liver weights, and that Salmonella infection leads to reduced heart, liver, and gizzard weights. Oviduct weight and number of small yellow follicles were reduced in infected birds. Antigen-specific antibody responses were not different between the groups, indicating high priority for this parameter as a life trait. Possible differences in stress susceptibility between efficient and nonefficient chickens need further examination.


Variant APOL1 protein in plasma associates with larger particles in humans and mouse models of kidney injury.
Genetic variants in apolipoprotein L1 (APOL1), a protein that protects humans from infection with African trypanosomes, explain a substantial proportion of the excess risk of chronic kidney disease affecting individuals with sub-Saharan ancestry. The mechanisms by which risk variants damage kidney cells remain incompletely understood. In preclinical models, APOL1 expressed in podocytes can lead to significant kidney injury. In humans, studies in kidney transplant suggest that the effects of APOL1 variants are predominantly driven by donor genotype. Less attention has been paid to a possible role for circulating APOL1 in kidney injury. Using liquid chromatography-tandem mass spectrometry, the concentrations of APOL1 were measured in plasma and urine from participants in the Seattle Kidney Study. Asymmetric flow field-flow fractionation was used to evaluate the size of APOL1-containing lipoprotein particles in plasma. Transgenic mice that express wild-type or risk variant APOL1 from an albumin promoter were treated to cause kidney injury and evaluated for renal disease and pathology. In human participants, urine concentrations of APOL1 were correlated with plasma concentrations and reduced kidney function. Risk variant APOL1 was enriched in larger particles. In mice, circulating risk variant APOL1-G1 promoted kidney damage and reduced podocyte density without renal expression of APOL1. These results suggest that plasma APOL1 is dynamic and contributes to the progression of kidney disease in humans, which may have implications for treatment of APOL1-associated kidney disease and for kidney transplantation.


Evaluation of Efficacy of Intraligamentary Injection Technique for Extraction of Mandibular Teeth-A Prospective Study.
Fear of dental pain is one of the most common reasons for delaying dental treatment. Local Anaesthesia (LA) is the most commonly employed technique of achieving pain control in dentistry. Pterygomandibular Nerve Block (PNB), for achieving mandibular anaesthesia has been the traditional technique used and is associated with a few set of complications which include pain, nerve injury, trismus, and rarely facial nerve palsy, and sustained soft tissue anaesthesia. These complications have resulted in a rapid need for research on alternative local anaesthetic techniques. This study was undertaken with the objective to determine pain, duration, profoundness and complications associated with administration of Intraligamentary Injection Technique (ILT). This study was conducted on 194 patients (male=122, female=72) who reported for dental extractions in mandibular posteriors. The ILT was administered with ligajet intraligamentary jet injector using cartridge containing lignocaine hydrochloride 2% with adrenaline 1:80000 and a 30 gauge needle at buccal (mesiobuccal), lingual, mesial and distal aspect of the mandibular molars. The data was analyzed by using statistical computer software SPSS 11.0 (Statistical package for social sciences 11.O version of SPSS Inc.). Median was derived for Pain on Injection (PI) and Pain during Procedure (PP). Mean and standard deviation was derived for Duration of Anaesthesia (DA). Various advantages were seen such as, localized soft tissue anaesthesia, decreased PI (SD=0.83), and minimal PP (SD=0.94). The DA (SD=4.62) and mean value of 24.06 minutes. This study is one of its kinds where intraligamentary injection has been used for extraction of mandibular molars. It was also successfully used in patients with exaggerated gag reflex and patients suffering from trismus due to oral submucous fibrosis. The intraligamentary injection technique can thus be used effectively to anaesthetize mandibular molars, as a primary technique for extraction of mandibular posterior teeth.


A standardized approach to assessing physician expectations and perceptions of continuing medical education.
Rapidly expanding science and mandates for maintaining credentials place increasing demands on continuing medical education (CME) activities to provide information that is current and relevant to patient care. Quality may be seen as the perceived level of service measured against consumer expectations. Standard tools have not been developed to determine how well CME activities meet consumer expectations. A widely used approach for evaluating perceptions of service quality in other fields, SERVQUAL, was adapted for CME by eliciting perspectives from physician consumers of CME and CME providers through nominal group techniques. These perspectives were used to develop a CMEQUAL evaluation survey instrument. Feasibility testing was conducted. Data were analyzed and items were tested for internal consistency. CME participants were individuals willing to complete items related to expectations before participation and perceptions after participation in a CME activity. Of the CME activity participants who provided CMEQUAL rating data for the study, 43% rated their overall perceptions of the CME activity below their overall expectations. CME activities most clearly met participant expectations in providing fair and balanced evidence-based content. Areas of lower priority for participants included opportunities for self-assessment, solving cases, and interactive learning. Two areas highly valued by participants but not adequately addressed by CME activities were (1) translating trial data to patient seen in practice and (2) addressing barriers to optimal patient management. Developing standards for evaluating physician perceptions of the quality of CME activities may assist CME providers in improving the effectiveness of CME activities in meeting physician learning needs.


Phactr3/scapinin, a member of protein phosphatase 1 and actin regulator (phactr) family, interacts with the plasma membrane via basic and hydrophobic residues in the N-terminus.
Proteins that belong to the protein phosphatase 1 and actin regulator (phactr) family are involved in cell motility and morphogenesis. However, the mechanisms that regulate the actin cytoskeleton are poorly understood. We have previously shown that phactr3, also known as scapinin, localizes to the plasma membrane, including lamellipodia and membrane ruffles. In the present study, experiments using deletion and point mutants showed that the basic and hydrophobic residues in the N-terminus play crucial roles in the localization to the plasma membrane. A BH analysis (http://helixweb.nih.gov/bhsearch) is a program developed to identify membrane-binding domains that comprise basic and hydrophobic residues in membrane proteins. We applied this program to phactr3. The results of the BH plot analysis agreed with the experimentally determined region that is responsible for the localization of phactr3 to the plasma membrane. In vitro experiments showed that the N-terminal itself binds to liposomes and acidic phospholipids. In addition, we showed that the interaction with the plasma membrane via the N-terminal membrane-binding sequence is required for phactr3-induced morphological changes in Cos7 cells. The membrane-binding sequence in the N-terminus is highly conserved in all members of the phactr family. Our findings may provide a molecular basis for understanding the mechanisms that allow phactr proteins to regulate cell morphogenesis.


Synergistic effect of aqueous removal of fluoroquinolones by a combined use of peroxymonosulfate and ferrate(VI).
This paper examines for the first time the combined use of ferrate(VI) (FeVIO42-, Fe(VI)) with peroxymonosulfate (PMS) to oxidize four fluoroquinolones (FQs) (flumequine (FLU), enrofloxacin (ENR), marbofloxacin (MAR), and ofloxacin (OFL)) at pH 7.0 and 25.0 °C. The results demonstrate that a combination of PMS and Fe(VI) synergistically enhance removal of FQs compared to the removal seen with either PMS or Fe(VI) alone. The oxidized products (OPs) of the reaction of FLU with the PMS-Fe(VI) system, identified by liquid chromatography high-resolution mass spectrometry (LC-HRMS) techniques, showed transformation products resulted from hydroxylation, ring cleavage, and de-fluorination processes. Thus, the combination of PMS and Fe(VI) can be developed as an innovative oxidative technology to remove contaminants from water.


Glycosidic linkages of swede cell walls and their residues recovered from the terminal ileum of the pig.
The glycosidic linkage pattern of unfractionated swede cell walls was determined by methylation analysis. The main linkages in descending order of concentration were: (1----4)-linked glucose, (1----4)-linked xylose, terminal galactose, (1----4)-linked mannose, terminal arabinose and (1----2)-linked rhamnose. The major branch points were to the O-2 or O-3 atoms of xylose, O-4 of rhamnose and O-6 of glucose. The subsequent degradation of swede cell walls by passage through the small intestine of the pig was studied by quantitative methylation analysis of material recovered from animals cannulated 150 mm before the ileo-caecal junction. In addition to a substantial loss of uronic acid and phenolic material, the proportions of the different glycosidic linkages recovered from the digesta varied greatly from those of the parent material. Pectic polysaccharides had apparent digestibilities in the range of 0.60-0.75 while those of hemicellulosic components varied between 0.15 and 0.57. Glucose units linked 1----4 (with an apparent digestibility of 0.24) accounted for the greatest weight loss of neutral sugars from the cell wall, followed by terminal galactose, terminal arabinose and 2-linked arabinose.


A case of the Carney triad.
To report a case of mediastinal paraganglioma in the setting of the Carney triad (gastric leiomyosarcoma, extra-adrenal paraganglioma, and pulmonary chondroma). A case of the Carney triad is presented, and the related literature is discussed. A 47-year-old woman with a history of gastric leiomyosarcoma and retroperitoneal paraganglioma sought medical attention because of progressive shortness of breath. She was found to have a mediastinal mass in the aortopulmonary window. The patient had normal blood pressure and normal urinary catecholamines, but metaiodobenzylguanidine scintigraphy showed an intense uptake in the region of the mass, consistent with paraganglioma. She subsequently underwent resection of the mediastinal mass, and the surgical pathology report indicated paraganglioma. Patients with the Carney triad can have multicentric paragangliomas. Metaiodobenzylguanidine scanning may be helpful in the diagnostic workup.


In vivo evaluation of the effects of interleukins 2, 4 and 7 on enhancing the immunotherapeutic efficacy of anti-tumor cytotoxic T lymphocytes.
The draining lymph nodes of mice injected with viable syngeneic tumor cells contain lymphoid cells capable of generating anti-tumor cytotoxic T lymphocytes (CTL) during a 4-day in vitro culture period and intravenous infusion of relatively low numbers of these CTL can effectively eliminate a challenge of fibrosarcoma cells at distal skin sites that would otherwise result in the death of the host. Using this model system the effects of addition of interleukin (IL) 2, IL4 or IL7 to the culture medium on the therapeutic efficacy of the anti-tumor CTL generated was investigated. In this regard, IL7 was found to be the most potent of the cytokines tested. Addition of IL7 either alone, or in combination with low doses of IL2, resulted in the generation of CTL with significantly (6-8-fold) enhanced therapeutic efficacy in vivo. Anti-tumor effector cells generated in the presence of IL7 were also found to be at least 4-fold more effective at eliminating established tumors than CTL generated in medium alone. Of the other cytokines tested, addition of IL2 resulted in elevated CTL activity in vitro, but only a modest (approximately 2-3-fold) enhancement of the therapeutic efficacy in vivo. Addition of IL4, either alone or in combination with IL2, also resulted in the generation of effector cells with enhanced tumoricidal activity in vitro and yet the therapeutic efficacy of these cells was decreased compared to that of CTL generated in medium alone. These observations indicate that (a) inclusion of IL7 in the medium in which tumor-reactive T cells are cultured can markedly enhance the immunotherapeutic efficacy of the resulting effector cell populations; and (b) in vitro assays of tumoricidal activity cannot be used as a reliable predictor of therapeutic efficacy in vivo.


Positron emission tomography to study the effect of eye closure and optic nerve damage on human cerebral glucose metabolism.
We used 18F-2-fluoro-2-deoxyglucose and positron emission tomography to evaluate the effect of visual deprivation on brain glucose metabolism. In experiment 1, we compared local cerebral metabolic rates for glucose in seven normal volunteers studied with eyes closed to 11 age- and sex-matched normal volunteers studied with eyes open. Whole brain metabolism was similar in the two groups, and region/whole brain analysis of metabolic data showed that metabolism in the calcarine posterior cortex was decreased by 14% (P less than .05) with eye closure. Glucose metabolism in other regions was not different between the two groups. In experiment 2, we compared glucose metabolism in six patients with severe bilateral optic neuropathies to 12 age- and sex-matched normal controls. Whole brain glucose metabolism was unchanged in the optic neuropathy group compared to controls. However, statistically significant reductions in glucose metabolism in the optic neuropathy group were found in anterior calcarine cortex (17%), posterior calcarine cortex (27%), peristriate cortex (27%), and lateral occipital cortex (15%). The metabolic effects of damage to the pregeniculate visual system went well beyond those of simple eye closure.


Systemic administration of human leukocyte interferon to melanoma patients. II. Cellular events associated with changes in natural killer cytotoxicity.
Treatment of patients with malignant melanoma with daily doses of human leukocyte interferon (IFN) resulted in natural killer (NK) cytotoxicity that increased during the 1st week of treatment and subsequently declined to pretreatment levels despite continued IFN administration; events involved in these changes were examined. The increased cytotoxicity was associated with an increased number of cells able to bind K562 targets and an increased proportion of cells able to mediate cytolysis of K562 targets. The period of increasing NK activity was also associated with an increased ability to generate in vitro cytotoxic effectors against K562 targets by stimulation with allogeneic lymphoblastoid cells. The responding cells in this system were depleted of NK activity by adherence to immune complexes. These results suggested that increased NK activity during week 1 of in vivo treatment was associated with augmentation of the development of NK cells from noncytotoxic precursors in addition to direct effects on NK cells. In contrast, the decline in NK activity appeared to be primarily a direct effect on NK cells, because these cells could no longer be augmented in vitro by additional IFN. The refractory state to IFN augmentation of cytotoxicity was limited to the period of IFN treatment in vivo, and responsiveness to IFN was promptly regained upon cessation of treatment. No evidence for the development of suppressor cells of NK effector function could be obtained.


DNA Fiber Assay for the Analysis of DNA Replication Progression in Human Pluripotent Stem Cells.
Human pluripotent stem cells (PSC) acquire recurrent chromosomal instabilities during prolonged in vitro culture that threaten to preclude their use in cell-based regenerative medicine. The rapid proliferation of pluripotent cells leads to constitutive replication stress, hindering the progression of DNA replication forks and in some cases leading to replication-fork collapse. Failure to overcome replication stress can result in incomplete genome duplication, which, if left to persist into the subsequent mitosis, can result in structural and numerical chromosomal instability. We have recently applied the DNA fiber assay to the study of replication stress in human PSC and found that, in comparison to somatic cells states, these cells display features of DNA replication stress that include slower replication fork speeds, evidence of stalled forks, and replication initiation from dormant replication origins. These findings have expanded on previous work demonstrating that extensive DNA damage in human PSC is replication associated. In this capacity, the DNA fiber assay has enabled the development of an advanced nucleoside-enriched culture medium that increases replication fork progression and decreases DNA damage and mitotic errors in human PSC cultures. The DNA fiber assay allows for the study of replication fork dynamics at single-molecule resolution. The assay relies on cells incorporating nucleotide analogs into nascent DNA during replication, which are then measured to monitor several replication parameters. Here we provide an optimized protocol for the fiber assay intended for use with human PSC, and describe the methods employed to analyze replication fork parameters. © 2020 Wiley Periodicals LLC. Basic Protocol 1: DNA fiber labeling Basic Protocol 2: DNA fiber spreading Basic Protocol 3: Immunostaining Support Protocol 1: Microscopy/data acquisition Support Protocol 2: Data analysis.


Deoxyribonuclease I purification from protease contaminations.
Affinity chromatography as a tool in purification of commercial DNA-ase I preparation from protease contaminations by the use of insoluble--protease inhibitors is described.


Three-dimensional aspects of the structural characteristics and kidney angioarchitecture of adult and aged Wistar rats: a scanning electron microscopy study.
The aim of this study was to describe the structural morphological characteristics and organization of the kidney angioarchitecture in adult and aged Wistar rats using different scanning electron microscopy (SEM) methods. Twenty male Wistar rats (Rattus norvegicus) were divided into two groups: the Adult Group (n = 10), which included animals at 3 months of age, and the Aged Group (n = 10), which included animals at 24 months of age. Kidney samples from both groups were analyzed using three distinct SEM methods: coronal sections, cryofracturing and vascular corrosion casts/angioarchitecture. This study demonstrates the specificity of each technique for three-dimensional morphological analyses of the kidney using SEM. In the coronal sections, it was possible to view the structures on a given plane; the stratigraphy was best observed in the cryofracture method, and the vascular corrosion casts showed the organization and arrangement of the angioarchitecture. Thus, we concluded that the SEM methods together provide the best description of the morphological characteristics of normal and pathological kidneys of adult and aged Wistar rats.


ABA-mediated ROS in mitochondria regulate root meristem activity by controlling PLETHORA expression in Arabidopsis.
Although research has determined that reactive oxygen species (ROS) function as signaling molecules in plant development, the molecular mechanism by which ROS regulate plant growth is not well known. An aba overly sensitive mutant, abo8-1, which is defective in a pentatricopeptide repeat (PPR) protein responsible for the splicing of NAD4 intron 3 in mitochondrial complex I, accumulates more ROS in root tips than the wild type, and the ROS accumulation is further enhanced by ABA treatment. The ABO8 mutation reduces root meristem activity, which can be enhanced by ABA treatment and reversibly recovered by addition of certain concentrations of the reducing agent GSH. As indicated by low ProDR5:GUS expression, auxin accumulation/signaling was reduced in abo8-1. We also found that ABA inhibits the expression of PLETHORA1 (PLT1) and PLT2, and that root growth is more sensitive to ABA in the plt1 and plt2 mutants than in the wild type. The expression of PLT1 and PLT2 is significantly reduced in the abo8-1 mutant. Overexpression of PLT2 in an inducible system can largely rescue root apical meristem (RAM)-defective phenotype of abo8-1 with and without ABA treatment. These results suggest that ABA-promoted ROS in the mitochondria of root tips are important retrograde signals that regulate root meristem activity by controlling auxin accumulation/signaling and PLT expression in Arabidopsis.


Multiple and exact soliton solutions of the perturbed Korteweg-de Vries equation of long surface waves in a convective fluid via Painlevé analysis, factorization, and simplest equation methods.
In this research, the surface waves of a horizontal fluid layer open to air under gravity field and vertical temperature gradient effects are studied. The governing equations of this model are reformulated and converted to a nonlinear evolution equation, the perturbed Korteweg-de Vries (pKdV) equation. We investigate the latter equation, which includes dispersion, diffusion, and instability effects, in order to examine the evolution of long surface waves in a convective fluid. Dispersion relation of the pKdV equation and its properties are discussed. The Painlevé analysis is applied not only to check the integrability of the pKdV equation but also to establish the Bäcklund transformation form. In addition, traveling wave solutions and a general form of the multiple-soliton solutions of the pKdV equation are obtained via Bäcklund transformation, the simplest equation method using Bernoulli, Riccati, and Burgers' equations as simplest equations, and the factorization method.


[Orbital hydatid cyst. Apropos of a case].
The authors report a new case of orbital hydatid cyst diagnosed in 3 year old children who had presented an insidious exophtalmia explored by CT Scan. The authors recall the CT Scan features of the orbital hydatid cyst and the contribution of the others imaging methods.


Risk adjustment in studies using administrative data.
Administrative data are increasingly used to evaluate service use and cost of treatments in "real world" settings. However, the degree to which administrative data can be used to risk-adjust for differences between nonrandomized groups receiving different treatments has not been studied. This study used data from a large randomized trial to determine whether administrative data describing previous utilization and cost are as good as clinical data in predicting future resource utilization and cost. Clinical data (symptoms, quality of life, pharmacological side effects) and administrative data (inpatient utilization and total cost) were obtained from a large randomized clinical trial comparing clozapine and haloperidol. The combination of administrative measures prior to randomization was the best predictor in a multiple regression model of service use in the year after randomization (adjusted r2 = 0.27-0.31). Models using administrative data alone predicted 24 to 28 percent of the variance of service use and costs, while clinical data predicted only 6 to 7 percent of the variation in the same dependent variables. In this analysis, administrative data about previous utilization and costs were far more successful than clinical data alone at predicting future utilization and costs and are therefore likely to be better risk adjusters in studies with nonrandomized comparison groups.


[Clinical efficacy and safety of huperzine Alpha in treatment of mild to moderate Alzheimer disease, a placebo-controlled, double-blind, randomized trial].
To evaluate the clinical efficacy and safety of huperzine Alpha in treatment of patients with mild to moderate Alzheimer disease (AD). Two hundred and two patients with the diagnosis of possible or probable AD from 15 centers the nationwide were randomly divided into two groups: huperzine Alpha group (n = 100, given huperzine Alpha 400 micro g/day for 12 weeks) and placebo group (n = 102 ). Different scales were used to evaluate the cognitive function, activity of daily life (ADL), non-cognitive disorders, and overall clinical efficacy. Safety evaluation was conducted every 6 weeks. In comparison with the baseline data, there was an improvement of 4.6 points in cognition assessed by ADAS-Cog (P = 0.000); an improvement of 2.7 points by MMSE (P = 0.000), an improvement of 1.5 points in behavior and mood by ADAS-non-Cog (P = 0.008) with 59.2% of the patients being on the mend clinically; and an improvement of 2.4 points by ADL (P = 0.001) with the capacity of ADL improved by at least 10% among 32.75% of the patients. 70% of the patients in huperzine Alpha group scored 1 approximately 3 points, and 27.8% of them scored 1 approximately 2 points by CIBIC-plus. The proportions of patients with an improvement of >/= 4 points by ADAS-Cog were 56.1% and 12.5% in the huperzine Alpha group and placebo group respectively (P = 0.000). The proportions of patients with an improvement of >/= 4 points by MMSE were 37.8% and 10.1% in the huperzine Alpha group and placebo group respectively (P = 0.000). The proportions of patients with an improvement of 1 approximately 3 points in global rating by CIBIC-plus were 59.2% and 40.6% in the huperzine Alpha group and placebo group respectively (P = 0.01). The proportions of patients with an improvement of >/= 10% points by ADL were 32.7% and 17.2% in the huperzine Alpha group and placebo group respectively (P = 0.01). The proportions of patients with an improvement of > 0 points by ADS-non-C0g were 70.0% and 36.3% in the huperzine Alpha group and placebo group respectively (P = 0.000). Mild and transient adverse events (edema of bilateral ankles and insomnia) were observed in 3% of huperzine Alpha treated patients. A safe and effective medicine, huperzine Alpha remarkably improves the cognition, behavior, ADL,and mood of AD patients.


Pediatric Dermatology.
Skin changes are common in children. Common concerns are birthmarks (e.g., hemangiomas and port wine stains), atopic and contact dermatitis, acne, and alopecia areata. The authors review advances in common and not so common skin changes in pediatric patients.