Paramagnetic 13C and 15N NMR analyses of the push and pull effects in cytochrome c peroxidase and Coprinus cinereus peroxidase variants: functional roles of highly conserved amino acids around heme.
Paramagnetic (13)C and (15)N nuclear magnetic resonance (NMR) spectroscopy of heme-bound cyanide ((13)C(15)N) was applied to 11 cytochrome c peroxidase (CcP) and Coprinus cinereus peroxidase (CIP) mutants to investigate contributions to the push and pull effects of conserved amino acids around heme. The (13)C and (15)N NMR data for the distal His and Arg mutants indicated that distal His is the key amino acid residue creating the strong pull effect and that distal Arg assists. The mutation of distal Trp of CcP to Phe, the amino acid at this position in CIP, changed the push and pull effects so they resembled those of CIP, whereas the mutation of distal Phe of CIP to Trp changed this mutant to become CcP-like. The (13)C NMR shifts for the proximal Asp mutants clearly showed that the proximal Asp-His hydrogen bonding strengthens the push effect. However, even in the absence of a hydrogen bond, the push effect of proximal His in peroxidase is significantly stronger than in globins. Comparison of these NMR data with the compound I formation rate constants and crystal structures of these mutants showed that (1) the base catalysis of the distal His is more critical for rapid compound I formation than its acid catalysis, (2) the primary function of the distal Arg is to maintain the distal heme pocket in favor of rapid compound I formation via hydrogen bonding, and (3) the push effect is the major contributor to the differential rates of compound I formation in wild-type peroxidases.


trans-Activation of parvovirus P38 promoter by the 76K noncapsid protein.
The autonomously replicating parvoviruses contain a 5-kilobase linear single-stranded DNA genome that produces two noncapsid proteins, N1 and N2, and two overlapping capsid proteins, VP1 and VP2. To characterize the regulation of viral transcription, we began with a study of the promoter for the coat proteins (P38) at map unit 38. Various constructions containing the P38 promoter were fused to the bacterial gene for chloramphenicol acetyltransferase (cat), and the relative efficiency of expression was determined in the presence and absence of parvovirus gene products. Our results show that the P38 promoter is a weak promoter without a trans-activation mediated by the 76,000-molecular-weight (76K) N1 protein. The N1 protein, supplied either by superinfection with virus or cotransfection with the cloned N1 gene, increased greatly the expression of the P38 promoter. In addition, sequences 3' to the promoter, within the region + 127 to + 648 (assuming an mRNA start site at 2008), were required for optimal expression but not for trans-activation. These results suggest that the production of parvovirus capsid proteins is under the indirect control of the P4 promoter and one of its gene products.


Case report: the University of Michigan dioxin exposure study: a follow-up investigation of a case with high serum concentration of 2,3,4,7,8-pentachlorodibenzofuran.
Polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and dioxin-like polychlorinated biphenyls that have toxic equivalency factors (TEFs) were measured in serum of 946 subjects in five Michigan counties. The study was motivated by concerns about human exposure to dioxin-contaminated sediments in the Tittabawassee River (TR). Most of the toxic equivalency in TR sediments is from two furan congeners, 2,3,7,8-tetrachlorodibenzofuran and 2,3,4,7,8-pentachlorodibenzofuran (2,3,4,7,8-pentaCDF). The individual with the highest adjusted (for age, age squared, and body mass index) serum level of 2,3,4,7,8-pentaCDF in the study (42.5 ppt) reported a unique history of raising cattle and vegetables in the floodplain of the TR. Interviews and serum samples were obtained from the index case and 15 other people who ate beef and vegetables raised by the index case. 2,3,4,7,8-pentaCDF in beef lipid was estimated to have been more than three orders of magnitude greater than background (1,780 vs. 1.1 ppt). The mean, median, and 95th percentile for serum 2,3,4,7,8-pentaCDF in the study control population were 6.0, 5.4, and 13.0 ppt, respectively, and were 9.9, 8.4, and 20.5 ppt among beef and vegetable consumers, respectively. Back extrapolation for the index case suggests that his increase in serum concentration of 2,3,4,7,8-pentaCDF above background may have been as high as 146 ppt. Consumption of beef and/or vegetables raised on dioxin-contaminated soil may be an important completed pathway of exposure. RELEVANCE TO PUBLIC HEALTH PRACTICE: Animals and crops should not be raised for human consumption in areas contaminated with dioxins.


A possible role for nerve growth factor in the augmentation of sodium channels in models of chronic pain.
Inflammation induces an upregulation of sodium channels in sensory neurons. This most likely occurs as a result of the retrograde transport of cytochemical mediators released during the inflammatory response. The purpose of this study was to determine the effect of the subcutaneous administration of one such mediator, nerve growth factor (NGF), on the production of sodium channels in neurons of the rat dorsal root ganglion. For this, hindpaw withdrawal from either a thermal or mechanical stimulus was measured in rats at selected intervals for up to 2 weeks following injections of NGF. Sodium channel augmentation was then examined in dorsal root ganglia using site-specific, anti-sodium channel antibodies. Both thermal and mechanical allodynia was observed between 3 and 12 h post-injection. The hyperalgesic response returned to baseline by approximately 24 h post-injection. Sodium channel labeling was found to increase dramatically in the small neurons of the associated dorsal root ganglia beginning at 23 h, reached maximum intensity by 1 week, and persisted for up to 3 months post-injection. Pre-blocking NGF with anti-NGF prevented the NGF-induced decrease in paw withdrawal latencies and significantly reduced the intensity of sodium channel labeling. The results indicate that NGF is an important mediator both in the development of acute hyperalgesia and in the stimulation of sodium channel production in dorsal root ganglia during inflammation.


A phase II trial of sorafenib in first-line metastatic urothelial cancer: a study of the PMH Phase II Consortium.
Sorafenib is an oral multikinase inhibitor that blocks cell proliferation via the ERK pathway and angiogenesis via the VEGF pathway. This phase II trial was conducted to determine the efficacy and tolerability of sorafenib for the treatment of patients with metastatic urothelial cancer (UC) who had not had prior chemotherapy for advanced disease. Seventeen chemo-naïve UC patients with adequate performance status and organ function were treated with sorafenib 400 mg twice daily on a continuous basis until progression or unacceptable toxicity. The primary endpoint was objective tumor response rate as measured by RECIST criteria. Secondary endpoints included rate of prolonged stable disease (>3 months), time to progression, median and 1 yr survival and safety and tolerability. There were no objective responses. Only one patient had stable disease by RECIST criteria and remained on treatment more than 3 months. Three patients had stable disease by RECIST criteria but were on treatment less than 3 months due to progressive disease (PD) or adverse events (AE). Eight patients had PD by RECIST criteria as their best overall response. Two patients had symptomatic PD prior to cycle 2 evaluation, and three patients were inevaluable (1 death, 1 AE, 1 withdrew consent).The time to progression was 1.9 months (range 0.7-8.7 months) and median survival was 5.9 months. The most common grade 3+ toxicities were abdominal pain, back pain, hand-foot reaction and bladder infection. Sorafenib does not show sufficient activity as a single agent in first-line metastatic urothelial cancer to warrant further investigation.


Phase 2, multicenter, open-label study of tigatuzumab (CS-1008), a humanized monoclonal antibody targeting death receptor 5, in combination with gemcitabine in chemotherapy-naive patients with unresectable or metastatic pancreatic cancer.
Tigatuzumab is the humanized version of the agonistic murine monoclonal antibody TRA-8 that binds to the death receptor 5 and induces apoptosis of human cancer cell lines via the caspase cascade. The combination of tigatuzumab and gemcitabine inhibits tumor growth in murine pancreatic xenografts. This phase 2 trial evaluated the efficacy of tigatuzumab combined with gemcitabine in 62 chemotherapy-naive patients with histologically or cytologically confirmed unresectable or metastatic pancreatic cancer. Patients received intravenous tigatuzumab (8 mg/kg loading dose followed by 3 mg/kg weekly) and gemcitabine (1000 mg/m(2) once weekly for 3 weeks followed by 1 week of rest) until progressive disease (PD) or unacceptable toxicity occurred. The primary end point was progression-free survival (PFS) at 16 weeks. Secondary end points included objective response rate (ORR) (complete responses plus partial responses), duration of response, and overall survival (OS). Safety of the combination was also evaluated. Mean duration of treatment was 18.48 weeks for tigatuzumab and 17.73 weeks for gemcitabine. The PFS rate at 16 weeks was 52.5% (95% confidence interval [CI], 39.3-64.1%). The ORR was 13.1%; 28 (45.9%) patients had stable disease and 14 (23%) patients had PD. Median PFS was 3.9 months (95% CI, 2.2-5.4 months). Median OS was 8.2 months (95% CI, 5.1-9.6 months). The most common adverse events related to tigatuzumab were nausea (35.5%), fatigue (32.3%), and peripheral edema (19.4%). Tigatuzumab combined with gemcitabine was well tolerated and may be clinically active for the treatment of chemotherapy-naive patients with unresectable or metastatic pancreatic cancer.


Pharmacological rational of dry ripe fruit of Aegle marmelos L. as an anti-nociceptive agent in different painful conditions.
The aim of study is to investigate central and peripheral analgesic effects of methanolic extract of dry ripe fruit of Aegle marmelos Linn. Corea (Am. Cr) by two methods, tail flick test and acetic acid induced writhing test at 100, 250 and 500mg/kg doses in animal models. Analgesic activity against tail flick test revealed that Am. Cr induced significant increase in latency period in dose dependent manner i.e. 65.38% at 100mg/kg, 395.37% at 250mg/kg (p<0.01) and 459.25% at 500mg/kg (p<0.01) body weight at 1hr after drug delivery while at 2hr effect decreased i.e. 61.53% at 100mg/kg, 161.11% (p<0.01) at 250mg/kg and 165.74% (p<0.01) at 500mg/kg but interestingly again there is an elongation in latency period at 3hr i.e. 106.15% at 100mg/kg dose, 251.85% (p<0.01) at 250mg/kg and 293.51% (p<0.05) at 500mg/kg respectively. The standard drug Diclofenac sodium at the dose of 5mg/kg continuously increased the latency period but less significantly as compared to the test substance i.e. 79.43%, 113.08% and 222.42% (p<0.05) respectively. Acetic acid induced writhing test produced highest significant activity at the dose of 100mg/kg i.e. 89.83% (p<0.01) as compared to Diclofenic sodium (standard drug) at a dose of 5mg/kg body weight i.e 63.63% (p<0.01). It is concluded that dry ripe fruit of A. marmelos possesses significant dual analgesic activities i.e. central and peripheral.


Quantitative schlieren techniques applied to high current arc investigations.
Applications of quantitative schlieren techniques to pulsed high current electric arcs are described. A new formalism for the Abel inversion is derived that does not start from the small angle approximation. The numerical treatment of experimental data is given special attention. A pulsed argon laser was constructed that gives enough background intensity to outshine a 2000-A arc. Several high speed schlieren techniques were tried to obtain quantitative information about the density and temperature in the neighborhood of the arc.


Pathological rupture of the spleen associated with hepatitis A.
Spontaneous rupture of the spleen refers to rupture of a normal spleen in the absence of trauma. Atraumatic rupture of a diseased spleen is termed pathological rupture and has been described in a variety of pathological conditions. This report describes a case of pathological splenic rupture associated with hepatitis A confirmed by positive IgM antibodies. To the best of our knowledge, this case represents the sole verified example of this previously unrecognized life-threatening complication of hepatitis.


Phosphatidylinositol-4,5-bisphosphate phosphodiesterase and phosphomonoesterase activities of rat brain. Some properties and possible control mechanisms.
The phosphatidylinositol-4,5-bisphosphate [PtdIns(4,5)P2] [and to a lesser extent, the phosphatidylinositol-4-phosphate (PtdIns4P)] phosphodiesterase and monoesterase activities of a rat brain supernatant have been studied by using 32P-labelled substrates prepared from human red blood cells. PtdIns(4,5)P2 monoesterase is maximally stimulated by Mg2+, though some activity is detectable in Ca2+/EDTA (Mg2+-free) buffers. The phosphodiesterase, however, is Ca2+-dependent, and in Ca2+/EDTA buffers with the pure lipid as substrate, shows maximal activity at 100 nM-Ca2+. If PtdIns(4,5)P2 is presented as a component of a lipid mixture of similar composition to that of the inner half of the lipid bilayer of a rat liver plasma membrane, the phosphodiesterase shows considerable activity at 1 microM-Ca2+, and is maximal at 100 microM-Ca2+. However, if it is assayed against the same substrate in Ca2+/EGTA buffers with 3mM-Mg2+ and 80 mM-KCl present (as an approximate parallel with the ionic environment in vivo), it shows no detectable activity below 100 microM-Ca2+, and is maximal at 1 mM-Ca2+. The monoesterase can hydrolyse PtdIns(4,5)P2 in such a lipid mixture at all Ca2+ concentrations with 1 or 3 mM-Mg2+ present. PtdIns(4,5)P2 phosphodiesterase can be induced to attack its substrate under ionic conditions similar to those in vivo (0.1-1 microM-Ca2+; 1 mM-Mg2+; 80 mM-KCl) by the conversion of its substrate into a non-bilayer configuration. If given such a substrate [by mixing PtdIns(4,5)P2 with an excess of phosphatidylethanolamine (PtdEtn)] it shows a shallow Ca2+-dependency curve from 0.1 to 100 microM and then a steep rise to 1 mM-Ca2+. Together these observations lead us to the suggestion that a perturbation in a membrane in vivo equivalent to a non-bilayer configuration would be sufficient to induce phosphodiesterase-catalysed PtdIns(4,5)P2 breakdown. When given substrates mixed with excess PtdEtn at pH 7.25 (or 5.5), 1 microM-Ca2+, 1 mM-Mg2+ and 80 mM-KCl, the rat brain supernatant phosphodiesterase activity hydrolysed PtdIns(4,5)P 50-100-fold faster than it hydrolysed phosphatidylinositol (PtdIns). If the supernatant was presented with such a non-bilayer mixture containing a ten-fold excess of PtdIns over PtdIns(4,5)P2, the latter phospholipid was still hydrolysed by phosphodiesterasic cleavage at nearly ten times the rate of the former. Receptor-stimulated phosphodiesterase cleavage of polyphosphoinositides is an early event in cell activation by many agonists. The properties of PtdIns(4,5)P2 phosphodiesterase in vitro suggest that a change in the presentation of its substrate would be a sensitive and sufficient control on the enzyme's activity in vivo.


Propranolol for the management of behavioural and psychological symptoms of dementia.
Propranolol is a β-adrenergic antagonist used in the management of hypertension, cardiac arrhythmia, and angina pectoris. There is some evidence that propranolol may benefit individuals with behavioural and psychological symptoms of dementia (BPSD). A total of three case series, one randomized controlled trial and one case report were identified (from a literature search of three major databases: PubMed, Ovid, and Cochrane collaboration) that assessed the use of propranolol for the management of BPSD. From these studies, it appears that propranolol improves BPSD, including agitation and aggression. Propranolol is also well tolerated with no significant bradycardia or hypotension noted in these studies. Current data on the use of propranolol for the management of BPSD are limited in comparison to other pharmacological agents (atypical antipsychotics, antidepressants, acetylcholinesterase inhibitors, memantine, and cannabinoids) and treatment modalities (repetitive transcranial magnetic stimulation and electroconvulsive therapy). The efficacy and safety of these treatments among individuals with BPSD has been evaluated in multiple controlled studies. In clinical practice, the routine use of propranolol among people with BPSD cannot be recommended at this time given the limited data. However, propranolol can be trialled among individuals with BPSD when symptoms have not responded adequately to other medications. Propranolol may also be used prior to embarking on trials of repetitive transcranial magnetic stimulation and electroconvulsive therapy among people with BPSD given the greater acceptance of this medication in the general population.


News exposure predicts anti-Muslim prejudice.
News coverage of Islamic extremism is reigniting debates about the media's role in promoting prejudice toward Muslims. Psychological theories of media-induced prejudice date to the 1950's, and find support from controlled experiments. However, national-scale studies of media effects on Muslim prejudice are lacking. Orthogonal research investigating media-induced prejudice toward immigrants has failed to establish any link. Moreover, it has been found that people interpret the news in ways that confirm pre-existing attitudes, suggesting that media induced Muslim prejudice in liberal democracies is unlikely. Here, we test the association between news exposure and anti-Muslim prejudice in a diverse national sample from one of the world's most tolerant societies, where media effects are least likely to hold (N = 16,584, New Zealand). In support of media-induced Islamophobia, results show that greater news exposure is associated with both increased anger and reduced warmth toward Muslims. Additionally, the relationship between media exposure and anti-Muslim prejudice does not reliably vary with political ideology, supporting claims that it is widespread representations of Muslims in the news, rather than partisan media biases, that drives anti-Muslim prejudice.


The mucus bundles responsible for airway cleaning are retained in cystic fibrosis and by cholinergic stimulation.
The beneficial effect of anticholinergic therapy for chronic lung diseases such as chronic obstructive pulmonary disease (COPD) is well documented, although cholinergic stimulation paradoxically inhibits liquid absorption, increases ciliary beat frequency and increases airway surface liquid transport.Using pig tracheobronchial explants, we quantified basal mucus transport before as well as after incubation with the clinically used antimuscarinic compound ipratropium bromide (Atrovent) and stimulation with acetylcholine.As expected, surface liquid transport was increased by acetylcholine and carbachol. In contrast, the mucus bundles secreted from the submucosal glands normally transported on the cilia were stopped from moving by acetylcholine, an effect inhibited by ipratropium bromide. Interestingly, in pigs lacking a functional cystic fibrosis (CF) transmembrane conductance regulator (CFTR) channel, the mucus bundles were almost immobile. As in wild-type pigs, CF surface liquid transport increased after carbachol stimulation. The stagnant CF mucus bundles were trapped on the tracheal surface attached to the surface goblet cells. Pseudomonas aeruginosa bacteria were moved by the mucus bundles in wild-type but not CF pigs.Acetylcholine thus uncouples airway surface liquid transport from transport of the surface mucus bundles as the bundles are dynamically inhibited by acetylcholine and the CFTR channel, explaining initiation of CF and COPD, and opening novel therapeutic windows.


[Evolutionary chromosomes and sex dimorphism].
In order to understand the phenomenon of sex, it is not enough to know about its reproductive role and role in recombination. In addition, knowledge of its evolutionary role is required. Sexual differentiation allows us to test evolutionary innovations in the male genome before they are transmitted to the female genome. This is possible in the framework of dichronous evolution, when the evolution of the male sex goes faster than the evolution of the female sex. This implies that there clearly should be male genes, which are already present in the male genome, but are not yet in the female genome; and vice versa, there should exist female genes, which have already been lost by the male sex, but still remain in females. Dichronicity results from different reaction norm of the sexes. This interpretation implies (1) the nomadic behavior of genes in the genome (2) the existence of evolutionary sexual dimorphism, which begins in the form of modificational changes in the female sex followed by selection-derived changes in the male sex; (3) the discovery of evolutionary roles of chromosomes and the proposition of a new concept, according to which the Y-chromosome represents a "gateway" for environment-related information coming to the genome, i.e., place of "birth" and testing of new genes, which comprise the basis of evolutionary sexual dimorphism--its initiator, accelerator, and regulator. The X-chromosome is the "transport" chromosome that transfers new genes from the Y-chromosomes into autosomes and therefore serves as a stabilizer, relaxer, and liquidator of evolutionary sexual dimorphism; it also serves as repository of the outgoing genes that await elimination. This concept provides for a new interpretation of the appearance, localization, and movement of genes along chromosomes and between them, and it sheds new light on the phenomena of chromosomal inactivation, mobile genes, association of Y-chromosome with stress, viruses, etc.


In vitro assessment of anticryptosporidial efficacy and cytotoxicity of adenosine analogues using a SYBR Green real-time PCR method.
The aims of this study were to provide a cost-effective and valuable method for evaluating drug efficacy against Cryptosporidium parvum using a quantitative SYBR Green real-time PCR (qPCR) and to assess the efficacy of adenosine analogues as drug templates. C. parvum HNJ-1 strain growing in human ileocaecal adenocarcinoma cells was employed as an in vitro culture system. To normalize the DNA extraction efficiency, a specific plasmid was added to each sample before DNA purification; the genomic DNA of infected cells was quantified by qPCR using specific primers to confirm drug efficacy and cytotoxicity. To determine the mechanism of action, enzymatic inhibition analyses were conducted using C. parvum S-adenosyl-l-homocysteine hydrolase (CpSAHH) recombinant protein. The dose-dependent growth inhibition of C. parvum was confirmed; 50% effective concentrations of neplanocin A (NPA) and 2-fluoroadenosine (2FA) were 139 μM and 0.842 μM, respectively. Cytotoxicity evaluation showed that the 50% growth inhibition concentration of 2FA was 1.18 μM; NPA did not exhibit any cytotoxicity up to 200 μM. The screening system revealed the specific but marginal efficacy of NPA and showed 2FA to be cytotoxic. Recombinant CpSAHH inhibition analyses showed that NPA competitively inhibited CpSAHH activity (K(i )= 0.395 μM), whereas 2FA did not. This novel qPCR system confirmed not only drug efficacy against C. parvum but also cytotoxicity to host cells. Moreover, since the SYBR Green method is cost effective, it could therefore be used in a wide variety of clinical and research-oriented applications of Cryptosporidium analysis.


Pharmacological enrollment of aldehyde dehydrogenase modulators to assist treating ischemia reperfusion-induced intestinal injury: is there a gap to be bridged?
This commentary highlights the research presented by Zhu et al. [1]. In this issue of the Clinical Science, the authors evaluated the protective effect of Alda-1 (a novel class of small molecule aldehyde dehydrogenase (ALDH2) activators) in the intestinal ischemia reperfusion (IR) injury. Remarkably, enhancing the ADLH2 activity by the use of Alda-1 can ameliorate several deleterious effects related to aldehydes, and may provide a better protection against an injury preestablished by IR. Together, an innovative metabolic strategy for treating patients with IR injury could be the use of ALDH modulators in a near future.


[Experiences with the intracutaneous Mendel-Mantoux tuberculin test in routine screening in a public health office].
The intracutaneous tuberculin test according to Mendel-Mantoux is internationally recognised as a standard testing method in all countries of the globe. In routine everyday working of the Public Health Office of the German provincial capital Stuttgart the Mendel-Mantoux tuberculin test was examined in respect of practicability, acceptance and economy and was compared against the multipuncture-stamp test. 2,710 persons for whom the tuberculin test was prescribed according to the German infectious diseases law were tested for tuberculosis by the Stuttgart Public Health Office between August 1995 and February 1996, using the Mendel-Mantoux test with Behring 5 TE GT. Of these, 154 persons did not return for checking the result. Checking was effected on 2,556 persons. The tested persons were 28.4 years old on the average, the majority being female (59.5% against 40.5% males) and of German nationality (60.7% against 39.3% other nationals). A positive reaction was seen in 37% (induration > 6 mm), in 23.3% Germans against 57.5% other nationals. In 65% of the positive results the indurations had a diameter of more than 10 mm, in 29.2% more than 15 mm. 80 probands, i.e. 2.5% of all tested persons, had an excessive reaction; firmation of blisters with local necrosis. The intracutaneous test according to Mendel-Mantoux can be routinely performed by Public Health offices, is well accepted by the persons concerned, and is also economic if 3 tests are conducted per bottle. The proportion of positive findings is similar to that obtained with the multipuncture-stamp test, whereas the significance of the results is more differentiated. The indication should be in accordance with the objective.


Prevalence, diversity and genetic structure of Escherichia coli isolates from septic tanks.
The present study investigated the diversity and genetic structure of Escherichia coli isolates from 100 septic tanks in the Canberra region, Australia. The physicochemical characteristics of the septic tanks were determined to examine the extent to which environmental factors might influence E. coli prevalence, diversity and population structure. The results of this study indicated that the temperature of the septic tank could explain some of the variation observed in the number of E. coli isolates recovered per septic tank, whereas pH was an important driver of E. coli diversity. Conductivity, pH and household size had a significant impact on E. coli population structure, and household size significantly affected the probability of detecting human-associated E. coli lineages [sequence types (STs) 69, 73, 95 and 131] in septic tanks. Phylogroup A and B1 strains were not randomly distributed among septic tanks, and the strong negative association between them may indicate intraspecific competition. The findings of this study suggest that the combination of environmental factors and intraspecific interactions may influence the distribution and genetic structure of E. coli in the environment.


Sugammadex Use in a Patient with Wolff-Parkinson-White (WPW) Syndrome.
Wolff-Parkinson-White (WPW) syndrome is a disease associated with episodes of supraventricular tachycardia and ventricular pre-excitation or atrial fibrillation. WPW is characterized by an aberrant electrical conduction pathway between atria and ventricles. The major anesthetic problem connected with WPW syndrome is the risk of tachyarrhythmias due to accessory pathway. Therefore, it has been proposed that the aim of anesthetic management should be the avoidance of tachyarrhythmia and sympathetic stimulation. Sugammadex was administered as a neuromuscular reversal agent in this case. To our knowledge, this is the first case report of sugammadex use in a patient with WPW. This report presents a case of general anesthesia management in a patient with WPW syndrome. We think that it is appropriate to use sugammadex to reverse rocuronium for the prevention of sudden hemodynamic changes in patients with WPW who underwent general anesthesia.


Circulating levels of CD34+ cells predict long-term cardiovascular outcomes in patients on maintenance hemodialysis.
CD34+ cells maintain vascular homeostasis and predict cardiovascular outcomes. We previously evaluated the association of CD34+ cells with cardiovascular disease (CVD) events over 23 months, but long-term CVD outcomes in relation to levels of CD34+ cells in patients on maintenance hemodialysis are unclear. Herein, we analyzed the long-term predictive potential levels of CD34+ cells for CVD outcomes and all-cause mortality. Between March 2005 and May 2005, we enrolled 215 patients on maintenance hemodialysis at Nagoya Kyoritsu Hospital and followed them up to 12.8 years. According to the CD34+ cell counts, patients were classified into the lowest, medium, and highest tertiles. Levels of CD34+ cells were analyzed in association with four-point major adverse CV events (MACEs), CVD death, and all-cause mortality. In univariate analysis age, smoking habit, lower geriatric nutrition risk index, lower calcium × phosphate product, and lower intact parathyroid hormone were significantly associated with the lowest tertile. Whereas, in multivariate analysis, age and smoking habit were significantly associated with the lowest tertile. Among 139 (64.7%) patients who died during a mean follow-up period of 8.0 years, 39 (28.1%) patients died from CVD. Patients in the lowest tertile had a significantly lower survival rate than those in the medium and highest tertiles (p ≤ 0.001). Using multivariable analyses, the lowest tertile was significantly associated with four-point MACEs (hazard ratio 1.80, p = 0.023) and CVD death (hazard ratio 2.50, p = 0.011). In conclusion, our long-term observational study revealed that a low level of CD34+ cells in the circulation predicts CVD outcomes among patients on maintenance hemodialysis.


Quantitation of transgenic plant DNA in leachate water: real-time polymerase chain reaction analysis.
Roundup Ready (RR) genetically modified (GM) corn and soybean comprise a large portion of the annual planted acreage of GM crops. Plant growth and subsequent plant decomposition introduce the recombinant DNA (rDNA) into the soil environment, where its fate has not been completely researched. Little is known of the temporal and spatial distribution of plant-derived rDNA in the soil environment and in situ transport of plant DNA by leachate water has not been studied before. The objectives of this study were to determine whether sufficient quantities of plant rDNA were released by roots during growth and early decomposition to be detected in water collected after percolating through a soil profile and to determine the influence of temperature on DNA persistence in the leachate water. Individual plants of RR corn and RR soybean were grown in modified cylinders in a growth room, and the cylinders were flushed with rain water weekly. Immediately after collection, the leachate was subjected to DNA purification followed by rDNA quantification using real-time Polymerase Chain Reaction (PCR) analysis. To test the effects of temperature on plant DNA persistence in leachate water, water samples were spiked with known quantities of RR soybean or RR corn genomic DNA and DNA persistence was examined at 5, 15, and 25 degrees C. Differences in the amounts and temporal distributions of root-derived rDNA were observed between corn and soybean plants. The results suggest that rainfall events may distribute plant DNA throughout the soil and into leachate water. Half-lives of plant DNA in leachate water ranged from 1.2 to 26.7 h, and persistence was greater at colder temperatures (5 and 15 degrees C).


Associations of anthropometric factors with KRAS and BRAF mutation status of primary colorectal cancer in men and women: a cohort study.
Obesity is a well-established risk factor for colorectal cancer (CRC), and accumulating evidence suggests a differential influence of sex and anthropometric factors on the molecular carcinogenesis of the disease. The aim of the present study was to investigate the relationship between height, weight, bodyfat percentage, waist- and hip circumference, waist-hip ratio (WHR), body mass index (BMI) and CRC risk according to KRAS and BRAF mutation status of the tumours, with particular reference to potential sex differences. KRAS and BRAF mutations were analysed by pyrosequencing in tumours from 494 incident CRC cases in the Malmö Diet and Cancer Study. Hazard ratios of CRC risk according to anthropometric factors and mutation status were calculated using multivariate Cox regression models. While all anthropometric measures except height were associated with an increased risk of KRAS-mutated tumours, only BMI was associated with an increased risk of KRAS wild type tumours overall. High weight, hip, waist, WHR and BMI were associated with an increased risk of BRAF wild type tumours, but none of the anthropometric factors were associated with risk of BRAF-mutated CRC, neither in the overall nor in the sex-stratified analysis. In men, several anthropometric measures were associated with both KRAS-mutated and KRAS wild type tumours. In women, only a high WHR was significantly associated with an increased risk of KRAS-mutated CRC. A significant interaction was found between sex and BMI with respect to risk of KRAS-mutated tumours. In men, all anthropometric factors except height were associated with an increased risk of BRAF wild type tumours, whereas in women, only bodyfat percentage was associated with an increased risk of BRAF wild type tumours. The results from this prospective cohort study further support an influence of sex and lifestyle factors on different pathways of colorectal carcinogenesis, defined by KRAS and BRAF mutation status of the tumours.


Heterogeneity in gut microbiota drive polyphenol metabolism that influences α-synuclein misfolding and toxicity.
The intestinal microbiota actively converts dietary flavanols into phenolic acids, some of which are bioavailable in vivo and may promote resilience to select neurological disorders by interfering with key pathologic mechanisms. Since every person harbors a unique set of gut bacteria, we investigated the influence of the gut microbiota's interpersonal heterogeneity on the production and bioavailability of flavonoid metabolites that may interfere with the misfolding of alpha (α)-synuclein, a process that plays a central role in Parkinson's disease and other α-synucleinopathies. We generated two experimental groups of humanized gnotobiotic mice with compositionally diverse gut bacteria and orally treated the mice with a flavanol-rich preparation (FRP). The two gnotobiotic mouse groups exhibited distinct differences in the generation and bioavailability of FRP-derived microbial phenolic acid metabolites that have bioactivity towards interfering with α-synuclein misfolding or inflammation. We also demonstrated that these bioactive phenolic acids are effective in modulating the development and progression of motor dysfunction in a Drosophila model of α-synucleinopathy. Lastly, through in vitro bacterial fermentation studies, we identified select bacteria that are capable of supporting the generation of these bioavailable and bioactive phenolic acids. Outcomes from our studies provide a better understanding of how interpersonal heterogeneity in the gut microbiota differentially modulates the efficacy of dietary flavanols to protect against select pathologic mechanisms. Collectively, our findings provide the basis for future developments of probiotic, prebiotic, or synbiotic approaches for modulating the onset and/or progression of α-synucleinopathies and other neurological disorders involving protein misfolding and/or inflammation.


Carboplatin and vinorelbine in elderly patients with non-small cell lung cancer.
Many lung cancers are diagnosed in patients over 70 years of age, but there are little published data on chemotherapy in elderly patients. We therefore activated a phase II study in order to assess the tolerance and activity of carboplatin (80 mg/m(2)) and vinorelbine (25 mg/m(2)) administered weekly in patients aged greater than or equal to 70 with advanced non-small cell lung cancer. Twenty-five patients (22 males, 3 females; performance status ECOG, 0-2; median age, 75 years, range 70-85) were included in the study and are assessable for response and side effects. A total of 162 cycles of therapy have been delivered (median/patient, 6 cycles). Seven partial remissions (28%; 95% confidence interval 5-36%), 8 disease stabilizations, and 10 progressions have been observed. Median time to disease progression was 4 months, and median survival was 5 months (range, 2-25+). Grade III/IV toxicity consisted mainly of leukopenia and neutropenia observed respectively in 5 and 7 patients. In conclusion, the schedule demonstrated activity and good tolerability in this subset of patients. Elderly patients with good performance status and adequate organ function can be safely treated with systemic chemotherapy.


Micro- to Nanoscale Bio-Hybrid Hydrogels Engineered by Ionizing Radiation.
Bio-hybrid hydrogels consist of a water-swollen hydrophilic polymer network encapsulating or conjugating single biomolecules, or larger and more complex biological constructs like whole cells. By modulating at least one dimension of the hydrogel system at the micro- or nanoscale, the activity of the biological component can be extremely upgraded with clear advantages for the development of therapeutic or diagnostic micro- and nano-devices. Gamma or e-beam irradiation of polymers allow a good control of the chemistry at the micro-/nanoscale with minimal recourse to toxic reactants and solvents. Another potential advantage is to obtain simultaneous sterilization when the absorbed doses are within the sterilization dose range. This short review will highlight opportunities and challenges of the radiation technologies to produce bio-hybrid nanogels as delivery devices of therapeutic biomolecules to the target cells, tissues, and organs, and to create hydrogel patterns at the nano-length and micro-length scales on surfaces.


Using machine learning improves predictions of herd-level bovine tuberculosis breakdowns in Great Britain.
In the United Kingdom, despite decades of control efforts, bovine tuberculosis (bTB) has not been controlled and currently costs ~ £100 m annually. Critical in the failure of control efforts has been the lack of a sufficiently sensitive diagnostic test. Here we use machine learning (ML) to predict herd-level bTB breakdowns in Great Britain (GB) with the aim of improving herd-level diagnostic sensitivity. The results of routinely-collected herd-level tests were correlated with risk factor data. Four ML methods were independently trained with data from 2012-2014 including ~ 4700 positive herd-level test results annually. The best model's performance was compared to the observed sensitivity and specificity of the herd-level test calculated on the 2015 data resulting in an increased herd-level sensitivity from 61.3 to 67.6% (95% confidence interval (CI): 66.4-68.8%) and herd-level specificity from 90.5 to 92.3% (95% CI: 91.6-93.1%). This approach can improve predictive capability for herd-level bTB and support disease control.


Replica-Exchange Enveloping Distribution Sampling: Calculation of Relative Free Energies in GROMOS.
Molecular dynamics (MD) simulations have become an important tool to investigate biological systems. Free-energy calculations based on MD are playing an increasingly important role for computer-aided drug design and material discovery in recent years. Free-energy differences between pairs of end-states can be estimated using well-established methods such as thermodynamic integration (TI) or Bennett's acceptance ratio (BAR). An attractive alternative is the recently developed replica-exchange enveloping distribution sampling (RE-EDS) method, which enables estimating relative free-energy differences between multiple molecules from a single simulation. Here, we provide an introduction to the principles underlying RE-EDS and give an overview of the RE-EDS pipeline. In addition, we provide a description of the two complementary tools RestraintMaker and amber2gromos. We briefly discuss the findings of three recent applications of RE-EDS to calculate relative binding or hydration free energies. In all three studies, good agreement was found between the results obtained using RE-EDS and experimental values as well as values obtained using other free-energy methods.


Mitotic bookmarking of genes: a novel dimension to epigenetic control.
Regulatory machinery is focally organized in the interphase nucleus. The information contained in these focal nuclear microenvironments must be inherited during cell division to sustain physiologically responsive gene expression in progeny cells. Recent results suggest that focal mitotic retention of phenotypic transcription factors at promoters together with histone modifications and DNA methylation--a mechanism collectively known as gene bookmarking--is a novel parameter of inherited epigenetic control that sustains cellular identity after mitosis. The epigenetic signatures imposed by bookmarking poise genes for activation or suppression following mitosis. We discuss the implications of phenotypic transcription factor retention on mitotic chromosomes in biological control and disease.


Hepatitis B vaccine in developing countries: problems and prospects.
Hepatitis B vaccines are highly immunogenic. To determine the efficacy of low doses and of the intradermal route of vaccination, 197 Nigerian children were given 3 monthly doses of Hevac B. Of these, 96 had 2 micrograms subcutaneously and 101 had 2 micrograms intradermally. One month after completing the schedule, 82.3% and 74.3% of the respective groups had become anti-HBs positive without adverse side-effects. In the second part of the study, 50 chronic HBsAg carriers were vaccinated. Compared to placebo-treated carriers, no effect was demonstrated with regard to HBsAg clearance or anti-HBs production. Immune complexes were not attributable to Hevac B. No untoward effects were noted. Booster vaccination of 50 initial non-responders resulted in the development of significant levels of anti-HBs in 20 (40%) of the recipients. None of the 'non-responders' developed clinical or virological evidence of HBV infection. It is concluded that low-dose vaccination is effective and that the intradermal route may be useful in developing countries.


Synergistic inhibition by verapamil and quinine of P-glycoprotein-mediated multidrug resistance in a human myeloma cell line model.
In an effort to develop a clinically useful approach to overcoming P-glycoprotein-mediated multidrug resistance (MDR1), we evaluated combined chemosensitization with verapamil and quinine in a multidrug-resistant (MDR) human myeloma cell line model. In clonogenic assay, verapamil was used at concentrations from 0.1 to 1.0 micrograms/mL, bracketing the plasma levels achieved by oral administration and high-dose intravenous (IV) infusion, respectively. The dose of quinine was held constant at 1.0 micrograms/mL, a plasma concentration readily achieved by oral administration. At each dose level of verapamil tested, the combination with quinine proved more effective than either drug individually in reversing resistance to doxorubicin and vinblastine and synergistic chemosensitizing interaction was observed. Verapamil at 0.1 microgram/mL combined with quinine was capable of restoring sensitivity to doxorubicin fully and reduced resistance to vinblastine as effectively as verapamil alone at 1.0 micrograms/mL. Furthermore, the combination of 1.0 mumol verapamil with 10 mumols quinine increased accumulation and retention of anthracycline in the resistant cells to a greater extent than did either drug individually (P less than .001) and inhibited drug efflux as effectively as verapamil alone at 10 mumols. Our findings suggest that combined chemosensitization with verapamil and quinine may prove useful for overcoming MDR1 in patients with drug-refractory B-cell neoplasms such as multiple myeloma or non-Hodgkin's lymphomas.


Effect of infectious dose and season on development of hemorrhagic pneumonia in mink caused by Pseudomonas aeruginosa.
Hemorrhagic pneumonia is an acute and fatal disease of farmed mink caused by Pseudomonas aeruginosa. The pathogenesis of this disease has not yet been resolved. Mink are the only animals known to be susceptible to acute, contagious, and fatal lung infections caused by P. aeruginosa. The purpose of this study was to investigate the correlation between dose-response and season of infection and to clarify whether Danish mink are carriers of P. aeruginosa on their nasal mucosa during the season for hemorrhagic pneumonia. To elucidate the pathogenesis of the disease, an infectious dose-response trial was carried out on adult mink and mink kits, both in the season for hemorrhagic pneumonia (November) as well as out of season (July). It proved difficult to infect mink via the intra-nasal route. Only 4 out of 60 infected mink developed clinical disease and were euthanized, all of them in November, illustrating that predisposing factors in the mink itself and not infectious dose might be crucial for disease development. We were able to culture P. aeruginosa from the nasal cavity of the clinically healthy experimental mink 8 d after inoculation. This indicated that the mink can carry P. aeruginosa on their nasal mucosa without developing the disease. It was not possible, however, to culture P. aeruginosa from the nasal cavity of clinically healthy mink obtained from farms in November, which indicates that the organism is not a normal part of the nasal mucosal flora of mink.


Allosteric gating of a large conductance Ca-activated K+ channel.
Large-conductance Ca-activated potassium channels (BK channels) are uniquely sensitive to both membrane potential and intracellular Ca2+. Recent work has demonstrated that in the gating of these channels there are voltage-sensitive steps that are separate from Ca2+ binding steps. Based on this result and the macroscopic steady state and kinetic properties of the cloned BK channel mslo, we have recently proposed a general kinetic scheme to describe the interaction between voltage and Ca2+ in the gating of the mslo channel (Cui, J., D.H. Cox, and R.W. Aldrich. 1997. J. Gen. Physiol. In press.). This scheme supposes that the channel exists in two main conformations, closed and open. The conformational change between closed and open is voltage dependent. Ca2+ binds to both the closed and open conformations, but on average binds more tightly to the open conformation and thereby promotes channel opening. Here we describe the basic properties of models of this form and test their ability to mimic mslo macroscopic steady state and kinetic behavior. The simplest form of this scheme corresponds to a voltage-dependent version of the Monod-Wyman-Changeux (MWC) model of allosteric proteins. The success of voltage-dependent MWC models in describing many aspects of mslo gating suggests that these channels may share a common molecular mechanism with other allosteric proteins whose behaviors have been modeled using the MWC formalism. We also demonstrate how this scheme can arise as a simplification of a more complex scheme that is based on the premise that the channel is a homotetramer with a single Ca2+ binding site and a single voltage sensor in each subunit. Aspects of the mslo data not well fitted by the simplified scheme will likely be better accounted for by this more general scheme. The kinetic schemes discussed in this paper may be useful in interpreting the effects of BK channel modifications or mutations.


Effects of intravenous glucose infusion and nutritional balance on serum concentrations of nonesterified fatty acids, glucose, insulin, and progesterone in nonlactating dairy cows.
The objective of this study was to evaluate serum concentrations of nonesterified fatty acids, glucose, insulin, and progesterone in nonlactating dairy cows according to nutritional balance and glucose infusion. Ten nonlactating, ovariectomized Gir x Holstein cows were stratified by body weight (BW) and body condition score (BCS) on d -28 of the study, and randomly assigned to 1) negative nutrient balance (NB) or 2) positive nutrient balance (PB). From d -28 to d 0, cows were allocated according to nutritional treatment (5 cows/treatment) into 2 low-quality pastures with reduced forage availability. However, PB cows individually received, on average, 3 kg/cow per day (as-fed) of a concentrate during the study. All cows had an intravaginal progesterone releasing device inserted on d -14, which remained in cows until the end of the study. Cow BW and BCS were assessed again on d 0. On d 0, cows within nutritional treatment were randomly assigned to receive, in a crossover design containing 2 periods of 24h each, 1) intravenous glucose infusion (GLU; 0.5 g of glucose/kg of BW, as a 5% glucose solution administered, on average, at 32 mL/min over a 3-h period), or 2) intravenous saline infusion (SAL; 0.9% solution infused on average at 32 mL/min over a 3-h period). Prior to the beginning of each period, all cows were fasted for 12h. Blood samples were collected, relative to the beginning of the infusion, at -12 and -11.5h (beginning of fasting), and at -0.5, 0, 0.5, 1, 2, 3, 4, 5, and 6h. Following the last blood collection of period 1, cows received (PB) or not (NB) concentrate and were returned to their respective pastures. Changes in BCS and BW were greater in NB cows compared with PB cows (-0.60 and -0.25+/-0.090 for BCS, respectively; -22.4 and 1.2+/-6.58 kg for BW, respectively). Cows receiving GLUC had greater glucose concentrations from 0.5 to 3h relative to infusion compared with SAL cows. Insulin concentrations were greater in PB cows assigned to GLUC compared with SAL cohorts at 0.5 and 3h following infusion, whereas NB cows assigned to GLUC had greater insulin concentrations compared with SAL cohorts at 0.5, 1, 2, and 3h. Progesterone concentrations were greater in PB cows assigned to GLUC at 2, 3, and 4h following infusion compared with SAL cohorts. In conclusion, the effects of glucose infusion on serum concentrations of insulin and progesterone in nonlactating dairy cows were dependent on cow nutritional status.


Cafestol increases serum cholesterol levels in apolipoprotein E*3-Leiden transgenic mice by suppression of bile acid synthesis.
Cafestol, a diterpene present in unfiltered coffee, potently increases serum cholesterol levels in humans. So far, no suitable animal model has been found to study the biochemical background of this effect. We determined the effect of cafestol on serum cholesterol and triglycerides in different mouse strains and subsequently studied its mechanism of action in apolipoprotein (apo) E*3-Leiden transgenic mice. ApoE*3-Leiden, heterozygous low density lipoprotein-receptor (LDLR+/-) knockout, or wild-type (WT) C57BL/6 mice were fed a high- (0.05% wt/wt) or a low- (0.01% wt/wt) cafestol diet or a placebo diet for 8 weeks. Standardized to energy intake, these amounts are equal to 40, 8, or 0 cups of unfiltered coffee per 10 MJ per day in humans. In apoE*3-Leiden mice, serum cholesterol was statistically significantly increased by 33% on the low- and by 61% on the high-cafestol diet. In LDLR+/- and WT mice, the increases were 20% and 24%, respectively, on the low-cafestol diet and 55% and 46%, respectively, on the high-cafestol diet. These increases were mainly due to a rise in very low density lipoprotein (VLDL) and intermediate density lipoprotein cholesterol in all 3 mouse strains. To investigate the mechanism of this effect, apoE*3-Leiden mice were fed a high-cafestol or a placebo diet for 3 weeks. Cafestol suppressed enzyme activity and mRNA levels of cholesterol 7alpha-hydroxylase by 57% and 58%, respectively. mRNA levels of enzymes involved in the alternate pathway of bile acid synthesis, ie, sterol 27-hydroxylase and oxysterol 7alpha-hydroxylase, were reduced by 32% and 48%, respectively. The total fecal bile acid output was decreased by 41%. Cafestol did not affect hepatic free and esterified cholesterol, but it decreased LDLR mRNA levels by 37%. The VLDL apoB and triglyceride production rates, as measured after Triton injection, were 2-fold decreased by cafestol, indicating that the number of particles secreted had declined and that there was no change in the amount of triglycerides present in the VLDL particle during cafestol treatment. However, the VLDL particles contained a 4-times higher amount of cholesteryl esters, resulting in a net 2-fold increased secretion of cholesteryl esters. The decrease in triglyceride production was the result of a reduction in hepatic triglyceride content by 52%. In conclusion, cafestol increases serum cholesterol levels in apoE*3-Leiden mice by suppression of the major regulatory enzymes in the bile acid synthesis pathways, leading to decreased LDLR mRNA levels and increased secretion of hepatic cholesterol esters. We suggest that suppression of bile acid synthesis may provide an explanation for the cholesterol-raising effect of cafestol in humans.


The role of the Strait of Gibraltar in shaping the genetic structure of the Mediterranean Grenadier, Coryphaenoides mediterraneus, between the Atlantic and Mediterranean Sea.
Population genetic studies of species inhabiting the deepest parts of the oceans are still scarce and only until recently we started to understand how oceanographic processes and topography affect dispersal and gene flow patterns. The aim of this study was to investigate the spatial population genetic structure of the bathyal bony fish Coryphaenoides mediterraneus, with a focus on the Atlantic-Mediterranean transition. We used nine nuclear microsatellites and the mitochondrial cytochrome c oxidase I gene from 6 different sampling areas. No population genetic structure was found within Mediterranean with both marker types (mean ΦST = 0.0960, FST = -0.0003, for both P > 0.05). However, within the Atlantic a contrasting pattern of genetic structure was found for the mtDNA and nuclear markers (mean ΦST = 0.2479, P < 0.001; FST = -0.0001, P > 0.05). When comparing samples from Atlantic and Mediterranean they exhibited high and significant levels of genetic divergence (mean ΦST = 0.7171, FST = 0.0245, for both P < 0.001) regardless the genetic marker used. Furthermore, no shared haplotypes were found between Atlantic and Mediterranean populations. These results suggest very limited genetic exchange between Atlantic and Mediterranean populations of C. mediterraneus, likely due to the shallow bathymetry of the Strait of Gibraltar acting as a barrier to gene flow. This physical barrier not only prevents the direct interactions between the deep-living adults, but also must prevent interchange of pelagic early life stages between the two basins. According to Bayesian simulations it is likely that Atlantic and Mediterranean populations of C. mediterraneus were separated during the late Pleistocene, which is congruent with results for other deep-sea fish from the same region.


1-C-6 epitope in cartilage proteoglycan G2 domain is masked by keratan sulphate.
The presence of the protein epitope recognized by monoclonal antibody 1-C-6 was investigated on the globular G1 and G2 domains of pig cartilage proteoglycan core protein. After reduction of disulphide bonds and removal of keratan sulphate chains with keratanase, both G1 and G2 domains were shown to contain the epitope. However, without keratanase digestion the epitope on the G2 domain was poorly detected. The results suggest that a keratan sulphate chain substituted close to the epitope sequence in the G2 domain prevents antibody access to the epitope and thus masks its detection. This shows the 1-C-6 epitope to be a conserved protein sequence in the G2 domain of proteoglycans from different species, but its detection may be masked by glycosylation.


A qualitative quantum rate model for hydrogen transfer in soybean lipoxygenase.
The hydrogen transfer reaction catalysed by soybean lipoxygenase (SLO) has been the focus of intense study following observations of a high kinetic isotope effect (KIE). Today high KIEs are generally thought to indicate departure from classical rate theory and are seen as a strong signature of tunnelling of the transferring particle, hydrogen or one of its isotopes, through the reaction energy barrier. In this paper, we build a qualitative quantum rate model with few free parameters that describes the dynamics of the transferring particle when it is exposed to energetic potentials exerted by the donor and the acceptor. The enzyme's impact on the dynamics is modelled by an additional energetic term, an oscillatory contribution known as "gating." By varying two key parameters, the gating frequency and the mean donor-acceptor separation, the model is able to reproduce well the KIE data for SLO wild-type and a variety of SLO mutants over the experimentally accessible temperature range. While SLO-specific constants have been considered here, it is possible to adapt these for other enzymes.


[The effect of supplying rats with vitamin K and administration of pelentane on Na,K- and spectrin-dependent ATPase in erythrocyte ghosts].
It was found that the activity of spectrin-dependent ATPase of erythrocyte ghosts isolated from rats with alimentary deficiency of vitamin K was significantly increased as compared with control animals, whereas in rats kept on a vicasol-rich diet this parameter was unchanged. In vitamin K-deficient rats the amount of proteins loosely bound to erythrocyte membranes was significantly reduced. At the same time, the activity of the integral enzyme (Na, K-ATPase) did not depend on the vitamin K provision despite the fact that in vitamin K-deficient animals kept on a vicasol-rich diet the enzyme affinity for ouabain was strongly decreased as compared with control. It was suggested that this effect might be due to the changes in the lipid and protein environment of the membrane-bound enzyme. Administration of the antivitamin K, pelentane, did not induce any conspicuous changes in the enzyme activities. It was concluded that antivitamin K does not induce any modification of the properties of erythrocyte-linked enzymes observed under conditions of vitamin K deficiency.


Citizens' expectations and likely use of a NHS Walk-in Centre: results of a survey and qualitative methods of research.
To find out which groups of people would use a National Health Service walk-in centre that would offer mainly health care advice, staffed by nurses. To understand the circumstances in which people would use a walk-in centre and to ascertain to what extent it would meet patients' expressed health-care needs. A postal survey of 2400 people plus 27 semi-structured interviews and one focus group. The study was conducted in Wakefield, Yorkshire UK, and included both white and ethnic minority groups. Most people reported that they would use a walk-in centre. It would be more attractive to young as compared with older people, ethnic minority as compared with white people, people who are dissatisfied with access to NHS services and people with urgent health-care problems. People want a wide range of services, including diagnosis, treatment, prevention, and general information. People also want access to both doctors and nurses, to male as well as female practitioners, to counsellors and interpreters. The type of service planned for this walk-in centre will meet some of the expressed needs. However, patients' expectations of the walk-in centre exceed planned provision in a number of key respects. Walk-in centres without GPs and with limited services will disappoint the public. It is important that walk-in centres are evaluated and attention paid to 'local voices' before additional money is allocated for such centres elsewhere.


N-3 Polyunsatured Fatty Acids in Menopausal Transition: A Systematic Review of Depressive and Cognitive Disorders with Accompanying Vasomotor Symptoms.
Depression is one of the most important health problems worldwide. Women are 2.5 times more likely to experience major depression than men. Evidence suggests that some women might experience an increased risk for developing depression during "windows of vulnerability", i.e., when exposed to intense hormone fluctuations, such as the menopause transition. Indeed, this period is associated with different symptoms, including vasomotor, depressive, and cognitive symptoms, which have all been shown to worsen as women approach menopause. Even though hormonal therapy represents the most effective treatment, side effects have been reported by several studies. Therefore, an increased number of women might prefer the use of alternative medicine for treating menopausal symptoms. N-3 long-chain polyunsaturated fatty acids (n-3 LCPUFAs) are included among these alternative treatments. We here provide a review of studies investigating the effects of n-3 LCPUFAs on hot flashes and depressive and cognitive disorders in menopausal women. The reported results are scattered and heterogeneous. In conclusion, a beneficial role of n-3 LCPUFAs in hot flashes, and depressive and cognitive symptoms related to menopausal transition is still far from conclusive.


Characterization of human interleukin 2 receptor (Tac antigen) in normal and leukemic T cells: co-expression of normal and aberrant receptors on Hut-102 cells.
Human interleukin 2 receptors ( IL2R ) from various cell sources such as mitogen-activated normal human T cells, adult T cell leukemia (ATL)-derived cell line cells (MT-1, ATL-6, Hut-102), and a natural killer-like cell line YT cells, were studied both by one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), with a monoclonal anti- IL2R antibody (anti-Tac). In synthetic labeling with [35S]methionine, anti-Tac specifically precipitated two glycoproteins, one with m.w. of 60,000 to 65,000 and pI 4.2 to 4.7, and the other with m.w. about 40,000 and pI 6.2 to 6.5. A study of surface iodination revealed that the former component was membrane-associated mature IL2R , and a pulse-chase study showed that the latter component was a precursor for IL2R that already possessed tunicamycin-sensitive N-linked sugar side chain(s). This precursor was found to be posttranslationally processed into mature IL2R by further glycosylation within 240 min. Among the cells studied, a human T cell leukemia virus-positive leukemic cell line, Hut-102, was distinctive in that it possessed an additional membrane glycoprotein (m.w. 55,000 to 60,000, pI 4.2 to 5.0) also defined by anti-Tac. This unique glycoprotein of Hut-102 cells appeared to be an aberrant IL2R .


Cardiac and Skeletal Muscle Myosin Exert Procoagulant Effects.
Trauma-induced coagulopathy (TIC) and the tissue injury-provoked procoagulant profile are prevalent in severely injured patients, but their mechanisms remain unclear. Myosin, exposed by or released from tissue injury, may play a role in promoting thrombin generation and attenuating fibrinolysis. The objective of the study is to examine the effects of cardiac and skeletal muscle myosins on coagulation in whole blood using thrombelastography (TEG). Whole blood was collected from healthy adult volunteers (n=8) and native TEGs were performed to evaluate the global coagulation response in the presence of cardiac or skeletal muscle myosin by measuring reaction (R) time (minutes), clot angle (), and maximum amplitude (MA, mm). TEG measurements were compared using paired t tests. Cardiac and skeletal muscle myosins decreased R, from 10.8 min to 8.0 min (P<0.0001) and 6.9 min (P =0.0007), respectively. There were no effects observed on clot propagation (angle) or clot strength (MA) with myosin addition. In the presence of tPA, both cardiac and skeletal muscle myosins shortened R from 11.1 min to 8.62 min (P=0.0245) and 7.75 min (P =0.0027), respectively), with no changes on angle or MA. Cardiac and skeletal muscle myosins exhibit procoagulant effects in TEG assays. These whole blood TEG results support the hypothesis that cardiac and skeletal muscle myosins may be either pro-hemostatic or prothrombotic depending on context.


Unraveling the Role of Flavor Structure and Physicochemical Properties in the Binding Phenomenon with Commercial Food Protein Isolates.
Food protein-flavor binding influences flavor release and perception. The complexity of the binding phenomenon lies in the flavor and protein properties. Thus, molecular interactions between commercial whey- or plant-based protein isolates (PI) such as pea, soy, and lupin, with carbonyl and alcohol flavor compounds were assessed by static headspace (HS) GC-MS. HS results showed that not only the displacement of the carbonyl group from the inner part of the flavor structure toward the edge promoted binding up to 52.76% ± 4.65 but also the flavor's degree of unsaturation. Similarly, thermal treatment led to a slight increase in hexanal-protein binding because of possible protein conformational changes. Protein's residual fat (<1%) seemed insufficient to promote significant flavor binding to PI. Despite the complexity of commercial food protein isolates, the results displayed that binding is predominantly influenced by the flavor structure and physicochemical properties, with the protein source and residual fat playing a secondary role.


Lithium chloride-induced taste aversion in C57BL/6J and DBA/2J mice.
Male C57BL/6J and DBA/2J mice (Mus musculus) were tested in a one-bottle conditioned taste aversion paradigm with 15% sucrose as the conditioned stimulus and lithium chloride (LiCl) as the unconditioned stimulus. The results of a dose-response experiment yielded evidence that higher doses of LiCl than used in past studies were required to produce profound aversions in these mice, with a 6.0 mEq/kg dose appearing as optimal. When the mice were conditioned with the 6.0 mEq/kg dose in one-bottle tests, there was little evidence of a strain difference in the acquisition of a taste aversion. During extinction trials, however, DBA/2J mice exhibited stronger resistance to extinction than did C57BL/6J mice. Additional analyses of fluid consumption suggested that strain differences in drinking behavior were evident in the paradigm and that these must be considered in interpreting the strain differences observed in conditioned taste aversion.


Impact of soil salinity on the microbial structure of halophyte rhizosphere microbiome.
The rhizosphere microbiome plays a significant role in the life of plants in promoting plant survival under adverse conditions. However, limited information is available about microbial diversity in saline environments. In the current study, we compared the composition of the rhizosphere microbiomes of the halophytes Urochloa, Kochia, Salsola, and Atriplex living in moderate and high salinity environments (Khewra salt mines; Pakistan) with that of the non-halophyte Triticum. Soil microbiomes analysis using pyrosequencing of 16S rRNA gene indicated that Actinobacteria were dominant in saline soil samples whereas Proteobacteria predominated in non-saline soil samples. Firmicutes, Acidobacteria, Bacteriodetes and Thaumarchaeota were predominant phyla in saline and non-saline soils, whereas Cyanobacteria, Verrucomicrobia, Gemmatimonadetes and the unclassified WPS-2 were less abundant. Sequences from Euryarchaeota, Ignavibacteriae, and Nanohaloarchaeota were identified only from the rhizosphere of halophytes. Dominant halophilic bacteria and archaea identified in this study included Agrococcus, Armatimonadetes gp4, Halalkalicoccus, Haloferula and Halobacterium. Our analysis showed that increases in soil salinity correlated with significant differences in the alpha and beta diversity of the microbial communities across saline and non-saline soil samples. Having a complete inventory of the soil bacteria from different saline environments in Pakistan will help in the discovery of potential inoculants for crops growing on salt-affected land.


Vagal Blocking for Obesity Control: a Possible Mechanism-Of-Action.
Recently, the US FDA has approved "vagal blocking therapy or vBLoc® therapy" as a new treatment for obesity. The aim of the present study was to study the mechanism-of-action of "VBLOC" in rat models. Rats were implanted with VBLOC, an intra-abdominal electrical device with leads placed around gastric vagal trunks through an abdominal incision and controlled by wireless device. Body weight, food intake, hunger/satiety, and metabolic parameters were monitored by a comprehensive laboratory animal monitoring system. Brain-gut responses were analyzed physiologically. VBLOC reduced body weight and food intake, which was associated with increased satiety but not with decreased hunger. Brain activities in response to VBLOC included increased gene expression of leptin and CCKb receptors, interleukin-1β, tumor necrosis factor, and transforming growth factor β1 in the brainstem; increased CCK, somatostatin, and tyrosine hydroxylase in the hippocampus; increased NPY, AgRP, and Foxa2 in the hypothalamus; and reduced CCKb receptor, melanocortin 4 receptor, and insulin receptor in the hypothalamus. Plasma concentrations of CCK, gastrin, glucagon, GLP-1, and PYY and gastric acid secretion were unchanged in response to VBLOC. Based on the present study, we may suggest that VBLOC induces satiety through vagal signaling, leading to reduced food intake and loss of body weight.


Clonal analysis of tumorigenicity and paratumorigenic phenotypes in rat liver epithelial cells chemically transformed in vitro.
Clonal subpopulations of a chemically induced tumorigenic rat liver epithelial cell line were analyzed for their cellular, biochemical, and in vitro growth properties and their tumorigenicity after injection into day-old newborn isogeneic rats. The phenotypic properties studied included DNA content; growth rate in culture; activities of gamma-glutamyl transpeptidase, NADH diaphorase, pyruvate kinase, glucose-6-phosphate dehydrogenase, and lactate dehydrogenase; ability to grow in calcium-poor medium; and ability to form colonies in soft agar. The results show that none of these phenotypes cosegregates with tumorigenicity and therefore is not reliable as a "marker" phenotype for neoplastic transformation in cultured rat liver epithelial cells. The poor correlations, either qualitatively or quantitatively, between paratumorigenic phenotypes and tumorigenicity suggest that neoplastic transformation in these cells involves a specific transforming gene locus or loci and that in vitro paratumorigenic phenotypes are merely epiphenomena of neoplastic transformation and progression. This study further reveals that the efficiency of the tumorigenicity assay of cultured rat liver epithelial cells in isogeneic newborn rats can be considerably improved by incubating the cells in medium containing only trace amounts of serum prior to transplantation into the host animals.


Breaking bad news about cancer: patients' preferences for communication.
The goal of this study was to assess patients' preferences regarding the way in which physicians deliver news about their cancer diagnosis and management. A sample of 351 patients with a variety of cancers completed a measure assessing their preferences for how they would like to be told news about their cancer. Patients rated characteristics of the context and content of the conversation as well as physician characteristics. Factor analysis indicated that patients' preferences for how they would like to be told news regarding their cancer can be grouped into the following three categories: (1) content (what and how much information is told); (2) facilitation (setting and context variables); and (3) support (emotional support during the interaction). Women (P =.02) and patients with higher education (P =.05) had significantly higher scores on the Content scale, women (P =.02) had higher scores on the Support scale, and younger patients (P =.001) and those with more education (P =.02) had higher scores on the Message Facilitation scale. Medical variables were not associated with patients' ratings of the importance of the three subscales. Patients rated items addressing the message content as most important, though the supportive and facilitative dimensions were also rated highly. Understanding what is important to patients when told news about their cancer provides valuable information that may help refine how this challenging task is best performed.


Bethanechol and phenylephrine responses in canine inferior vena cava and abdominal aorta.
Both canine inferior vena cava and abdominal aorta showed a concentration dependent increase of contraction in response to phenylephrine; however, inferior vena cava contracted in response to bethanechol but abdominal aorta did not respond to bethanechol. When the inferior vena cava was preconstricted with 1 x 10(-3)M bethanechol, low concentrations of phenylephrine (1 x 10(-7)M to 1 x 10(-6)M) induced relaxation, but relaxation by phenylephrine of abdominal aorta pretreated with 1 x 10(-3)M bethanechol did not occur.


Extremely preterm births: end-of-life decisions in European countries.
To explore the influence of end-of-life decisions (EoL-D) on survival and mortality data in the light of differences reported among European countries. We collected the published data of several epidemiological studies: Epicure, Epipage, Epibel and the Norwegian study performed in the UK, France, Belgium and Norway, respectively. The data concerning the Epibel and Norwegian studies are considered for a preliminary analysis, while the data relating to the Epicure and Epipage studies are compared based on the total published data. The statistical analysis was performed through the class of generalised linear models, and more specifically, through log-linear models. The data considered were the number of babies who died in neonatal intensive care units after active withdrawal classified according to the country and gestational age. RESULTS The probability of death after active withdrawal was significantly higher at 22 and 24 weeks' gestational age compared with week 25, when considering both countries (OR, 2.35 and 1.29, respectively). For the week 23(0)-->(6), the probability of death after active withdrawal was not significant; however, it is relevant when considering the OR (1.31). When considering the country, there was a higher probability of death after active withdrawal in France than in the UK, or alternatively, with the assumed baseline French parameter, in the UK, there was a lower probability of death after active withdrawal, with a significant OR=0.69. CONCLUSIONS The attitude towards EoL-D could in part explain the differences in survival data of extremely preterm infants and should be taken in mind when comparing international survival rates.


Changes in cerebral glucose utilization in patients with panic disorder treated with cognitive-behavioral therapy.
Several neuroanatomical hypotheses of panic disorder have been proposed focusing on the significant role of the amygdala and PAG-related "panic neurocircuitry." Although cognitive-behavioral therapy is effective in patients with panic disorder, its therapeutic mechanism of action in the brain remains unclear. The present study was performed to investigate regional brain glucose metabolic changes associated with successful completion of cognitive-behavioral therapy in panic disorder patients. The regional glucose utilization in patients with panic disorder was compared before and after cognitive-behavioral therapy using positron emission tomography with (18)F-fluorodeoxyglucose. In 11 of 12 patients who showed improvement after cognitive-behavioral therapy, decreased glucose utilization was detected in the right hippocampus, left anterior cingulate, left cerebellum, and pons, whereas increased glucose utilization was seen in the bilateral medial prefrontal cortices. Significant correlations were found between the percent change relative to the pretreatment value of glucose utilization in the left medial prefrontal cortex and those of anxiety and agoraphobia-related subscale of the Panic Disorder Severity Scale, and between that of the midbrain and that of the number of panic attacks during the 4 weeks before each scan in all 12 patients. The completion of successful cognitive-behavioral therapy involved not only reduction of the baseline hyperactivity in several brain areas but also adaptive metabolic changes of the bilateral medial prefrontal cortices in panic disorder patients.


Regulation of IgE-receptor expression, IgE occupancy and secretory capacity of mast cells.
Mast cells play an important role in initiating and modulating allergic and inflammatory reactions. Their responsiveness is determined by three important factors: the expression of IgE receptors on the cell surface, the IgE occupancy of these receptors, and the intrinsic secretory capacity of the cells. In this review, we will summarise some findings relevant to these three aspects of mast cell function, and discuss possible regulatory mechanisms. It appears that the genetic background as well as environmental factors influence all three of these components of the response. T cells appear to play an important role in regulating the IgE-receptor expression and also, independently, the intrinsic secretory capacity of mast cells via an unidentified route, possibly involving the secretory signal transduction chain directly. IgE itself appears to have an important role in the regulation of IgE-receptor expression, as indicated by the upregulation of receptors in vitro in the presence of IgE, and the absence of IgE-binding capacity of mast cells in IL-4 gene knockout mice, lacking IgE production. The IgE-receptors of mast cells are saturated to a high degree under different normal conditions, without an obvious relation to antigenic stimulation, also in athymic animals. We have suggested that this basal IgE content on mast cells may be the result of an antigen-independent production of IgE directed by the mast cells themselves and serving regulatory purposes, modifying the secretory response and preventing a massive possibly harmful degranulation.


Large intracranial intradural mucocele as a complication of frontal sinus osteoma.
Paranasal sinus osteomas are mostly asymptomatic; however, secondary mucocele can develop if they impede the natural sinus drainage. Such a mucocele can destroy the bone and extend into the adjacent structures. We report on an unusual case of frontal sinus osteoma in a 27-year-old patient, complicated by large secondary mucocele that eroded the bone and extended into the frontal lobe of the brain. Unexpectedly, the patient did not report any visual or other symptoms attributable to central nervous system deficit. The mucocele was completely resected through bifrontobasal osteoplastic craniotomy, whereas osteoma was evacuated in its entirety by both drilling and mobilizing. Open surgical approach remains the main treatment for complicated paranasal sinus osteoma, and radical removal of intracranial mucocele is mandatory to prevent the development of life-threatening infections. Although intradural extension of a secondary mucocele is extremely unusual, head and neck surgery specialists should take this severe complication in consideration.


Intima-media thickness of brachial artery, vascular function, and cardiovascular risk factors.
Cardiovascular diseases are associated with impaired flow-mediated vasodilation (FMD) and increase in carotid intima-media thickness (IMT). Both FMD and IMT are independent predictors for cardiovascular outcomes. When measuring FMD and nitroglycerine-induced vasodilation in the brachial artery, IMT can also be simultaneously assessed in the same brachial artery. The purpose of this study was to determine the relationships between IMT of the brachial artery, vascular function, and cardiovascular risk factors. We measured brachial IMT, FMD, and nitroglycerine-induced vasodilation by ultrasound in 388 subjects who underwent health examination (mean age, 45±22 years; age range, 19-86), including patients with cardiovascular diseases. Univariate regression analysis revealed that brachial IMT significantly correlated with age (r=0.71; P<0.001), body mass index (r=0.27; P<0.001), systolic blood pressure (r=0.40; P<0.001), diastolic blood pressure (r=0.31; P<0.001), heart rate (r=0.15; P=0.002), glucose level (r=0.18; P=0.01), and smoking pack-years (r=0.42; P<0.001), as well as Framingham risk score, a cumulative cardiovascular risk index for heart attack (r=0.49; P<0.001). FMD and nitroglycerine-induced vasodilation were inversely associated with brachial IMT (r=-0.39, P<0.001; r=-0.32, P<0.001, respectively). In addition, there was a significant relationship between brachial IMT and carotid IMT (r=0.58; P<0.001). Multivariate analysis revealed that age, sex, hypertension, and brachial artery diameter were independent predictors of brachial IMT. These findings suggest that brachial IMT may be a marker of the grade of atherosclerosis and may be used as a marker of vascular function, providing additive information for stratifying subjects with cardiovascular risk factors.


Facing future climate change: is the past relevant?
From a socio-economic perspective, the 'sharp end' of climate research is very much about looking forward in time. As far as possible, we need to know what to expect and approximately when to expect it. However, it is argued here that our approach to climate change (including its scientific basis and its policy implications) is firmly linked to our understanding of the past. This is mainly due to the role played by palaeoclimate reconstructions in shaping our expectations of the climate system, in particular via their ability to test the accuracy of our climate models. Importantly, this includes the intuitive models that each of us carries around in our mind, as well as the more complex numerical models hiding inside supercomputers. It is through such models that palaeoclimate insights may affect the scientific and political judgements that we must make in the face of persistent and ultimately irreducible predictive uncertainty. Already we can demonstrate a great deal of confidence in our current understanding of the global climate system based specifically on insights from the geological record. If further advances are to be made effectively, climate models should take advantage of both past and present constraints on their behaviour, and should be given added credence to the extent that they are compatible with an increasingly rich tapestry of past climatic phenomena. Furthermore, palaeoclimate data should be accompanied by clearly defined uncertainties, and organized in arrays that are capable of speaking directly to numerical models, and their limitations in particular.


Analytical Method Development and Chemometric Approach for Evidencing Presence of Plasticizer Residues in Nectar Honey Samples.
Over the years, anthropogenic sources have increasingly affected food quality. One of the most sensitive and nutritional matrices affected by chemical contamination is honey, due to the use of acaricides. Recently, the attention has moved to the presence of phthalates (PAEs) and bisphenol A (BP-A), molecules present in plastic materials used both in the production phase and in the conservation of honey. In this study, an analytical method for the simultaneous determination of PAEs (dimethyl phthalate DMP, diethyl phthalate DEP, diisobutyl phthalate DiBP, dibutyl phthalate DBP, bis(2-ethylhexyl) phthalate DEHP, and di-n-octyl-phthalate DnOP) and BP-A was developed. The extraction technique is the ultrasound-vortex-assisted dispersive liquid-liquid microextraction (UVA-DLLME), using 150 µL of toluene as an extraction solvent, followed by the gas chromatography coupled with ion trap mass spectrometry analysis (GC-IT/MS). The developed method is sensitive, reliable, and reproducible: it shows high correlation coefficients (R > 0.999); limits of detection (LODs) less than 11 ng·g-1; limits of quantification (LOQs) less than 16 ng·g-1; repeatability below 3.6%, except BP-A (11.6%); and accuracy below 4.8%, except BP-A (17.6%). The method was applied to 47 nectar honey samples for evidencing similarities among them. The chemometric approach based on Hierarchical Cluster Analysis and Principal Component Analysis evidenced some similitudes about sample origin as well as marked differences between PAE and BP-A sources.


Challenging the Norm: A Multidisciplinary Perspective on Intravenous to Subcutaneous Bridging Strategies for Biologics.
The transition from intravenous (IV) to subcutaneous (SC) administration of biologics is a critical strategy in drug development aimed at improving patient convenience, compliance, and therapeutic outcomes. Focusing on the increasing role of model-informed drug development (MIDD) in the acceleration of this transition, an in-depth overview of the essential clinical pharmacology and regulatory considerations for successful IV to SC bridging for biologics after the IV formulation has been approved are presented. Considerations encompass multiple aspects beginning with adequate pharmacokinetic (PK) and pharmacodynamic (i.e., exposure-response) evaluations which play a vital role in establishing comparability between the IV and SC routes of administrations. Selected key recommendations and points to consider include: 1) PK characterization of the SC formulation, supported by the increasing preclinical understanding of the SC absorption, and robust PK study design and analyses in human; 2) A thorough characterization of the exposure-response profiles including important metrics of exposure for both efficacy and safety; 3) Comparability studies designed to meet regulatory considerations and support approval of the SC formulation, including non-inferiority studies with PK and/or efficacy and safety as primary endpoints; 4) Comprehensive safety package addressing assessments of immunogenicity and patients' safety profile with the new route of administration. Recommendations for successful bridging strategies are evolving and MIDD approaches have been used successfully to accelerate the transition to SC dosing, ultimately leading to improved patient experiences, adherence, and clinical outcomes.


Incorporating a Prognostic Gene Expression Profile Test into the Management of Cutaneous Squamous Cell Carcinoma: An Expert Consensus Panel Report.
Cutaneous squamous cell carcinoma (cSCC) is a growing health concern with a rapidly increasing incidence. Disease-specific mortality is typically preceded by a metastasis, but current staging systems have significant limitations in predicting this event. The 40-gene expression profile (40-GEP) test is a validated method of further stratifying patients based on the risk of regional or distant metastasis, but limited guidelines exist for incorporating this test into clinical practice. To review the available literature on the use of gene expression profile (GEP) testing to assess prognosis in cSCC and create consensus statements to guide dermatology clinicians on its use. A comprehensive literature search of PubMed, EMBASE, and Scopus was completed for English-language original research articles on the use of GEP testing to assess cSCC prognosis. A panel of 8 dermatologists with significant expertise in diagnosing and managing cSCC gathered to review the articles and create consensus statements. A modified Delphi process was used to approve each statement and a strength of recommendation was assigned using the Strength of Recommendation Taxonomy (SORT) criteria. The literature search produced 157 articles that met the search criteria. A thorough screening of the studies for relevance to the research question resulted in 21 articles that were distributed to the panelists for review prior to the roundtable discussion. The panel unanimously voted to adopt 7 consensus statements and recommendations, 6 of which were given a strength of "A" and 1 of which was given a strength of "C". The 40-GEP test provides accurate and independent prognostic information beyond standard staging systems that only incorporate pathologic data. Incorporation of GEP testing into national guidelines can help further stratify patients based on risk of metastasis and thus may improve morbidity and mortality. J Drugs Dermatol. 2023;22(12): doi:10.36849/JDD.7691e.


IL-23 inhibits osteoclastogenesis indirectly through lymphocytes and is required for the maintenance of bone mass in mice.
IL-23 stimulates the differentiation and function of the Th17 subset of CD4(+) T cells and plays a critical role in chronic inflammation. The IL-23 receptor-encoding gene is also an inflammatory disease susceptibility gene. IL-23 shares a common subunit with IL-12, a T cell-dependent osteoclast formation inhibitor, and we found that IL-23 also dose-dependently inhibited osteoclastogenesis in a CD4(+) T lymphocyte-dependent manner. When sufficiently enriched, gammadelta T cells also mediated IL-23 inhibition. Like IL-12, IL-23 acted synergistically with IL-18 to block osteoclastogenesis but, unlike IL-12, IL-23 action depended on T cell GM-CSF production. IL-23 did not mediate IL-12 action although IL-12 induced its expression. Male mice lacking IL-23 (IL-23p19(-/-)) had approximately 30% lower bone mineral density and tibial trabecular bone mass (bone volume (BV)/total volume (TV)) than wild-type littermates at 12 wk and 40% lower BV/TV at 26 wk of age; male heterozygotes also had lower bone mass. Female IL-23p19(-/-) mice also had reduced BV/TV. IL-23p19(-/-) mice had no detectable osteoclast defect in trabecular bone but IL-23p19(-/-) had thinner growth plate hypertrophic and primary spongiosa zones (and, in females, less cartilage remnants) compared with wild type. This suggests increased osteoclast action at and below the growth plate, leading to reduced amounts of mature trabecular bone. Thus, IL-23 inhibits osteoclast formation indirectly via T cells in vitro. Under nonpathological conditions (unlike inflammatory conditions), IL-23 favors higher bone mass in long bones by limiting resorption of immature bone forming below the growth plate.


Some patients with antiphospholipid syndrome express hitherto undescribed antibodies to cardiolipin-binding proteins.
Contrary to infective anticardiolipin (aCL) antibodies, autoimmune aCL antibodies react with phospholipids (PL) mainly via binding to the plasma glycoprotein cofactor beta2-Glycoprotein I (beta2GPI). While there is a well-documented link between the risk of thrombosis and the presence of beta2GPI-dependent anticardiolipin antibodies, the pathological impact of other antiphospholipid antibodies is less clear. By means of cardiolipin affinity-chromatography, we isolated and identified 3 CL-binding proteins, complement component C4, complement factor H and a kallikrein-sensitive glycoprotein, and tested for the presence of autoantibodies against these proteins in patients with antiphospholipid syndrome (APS), systemic lupus erythematosus (SLE) and other autoimmune diseases. High titers of autoantibodies to C4 as compared to age- and sex-matched healthy controls were present in 3 of 26 patients with APS, and weak titers were found in 2 of 26 patients with SLE and in none of 26 patients with other autoimmune diseases. Autoantibodies to complement factor H were found in 4 APS, 3 SLE and none of the other autoimmune patients. Autoantibodies to kallikrein-sensitive glycoprotein were detected in 6 APS patients, 1 SLE patient, and 1 patient with another autoimmune disease. A close relationship between these antibodies was found, suggesting their origin from a common macromolecular complex. However, no relationship with anti-beta2GPI antibodies was found, with the three patients with higher levels of autoantibodies having a low titer of anti-beta2GPI antibodies. In conclusion, some patients with APS harbor circulating antibodies to other CL-binding proteins which might be useful to further characterize these patients.


[Modern approaches to treatment of thrombosis in pregnant women, carriers of hereditary anomalies--F-V-Leiden and G20210A mutation].
studying of efficiency low-molecular heparin for preventive maintenance and treatment of thromboembolic disorders in pregnant women, carriers of F-V- L and F-II-20210 G-A mutation. Under supervision, there were 27 women with thrombosis during pregnancy from 19 to 39 years old. The lower extremities veins thromboses was diagnosed by duplex sonography. F-V- Leiden, F-II-20210 G-A and MTHFR C 677T gene mutation was defined by PCR-PELP method. To all pregnant women Fraxiparin and vitamins of B group was appointed during pregnancy period; at presence of MTHFR a folic acid was appointed in addition. It is concluded that application of Fraxiparin is an effective method of treatment of thrombosis during pregnancy and women--carriers F-V- Leiden, F-II-20210 G-A and MTHFR C 677T gene mutation. High efficiency of Fraxiparin confirms with change of parameters of various parts of a hemostasis and return development of thrombosis evidence.


Algorithm guided outlining of 105 pancreatic cancer liver metastases in Ultrasound.
Manual segmentation of hepatic metastases in ultrasound images acquired from patients suffering from pancreatic cancer is common practice. Semiautomatic measurements promising assistance in this process are often assessed using a small number of lesions performed by examiners who already know the algorithm. In this work, we present the application of an algorithm for the segmentation of liver metastases due to pancreatic cancer using a set of 105 different images of metastases. The algorithm and the two examiners had never assessed the images before. The examiners first performed a manual segmentation and, after five weeks, a semiautomatic segmentation using the algorithm. They were satisfied in up to 90% of the cases with the semiautomatic segmentation results. Using the algorithm was significantly faster and resulted in a median Dice similarity score of over 80%. Estimation of the inter-operator variability by using the intra class correlation coefficient was good with 0.8. In conclusion, the algorithm facilitates fast and accurate segmentation of liver metastases, comparable to the current gold standard of manual segmentation.


Effect of S-adenosyl-1,12-diamino-3-thio-9-azadodecane, a multisubstrate adduct inhibitor of spermine synthase, on polyamine metabolism in mammalian cells.
The effects of the potent spermine synthase inhibitor S-adenosyl-1,12-diamino-3-thio-9-azadodecane (AdoDatad) on polyamine biosynthesis have been studied in transformed mouse fibroblasts (SV 3T3 cells) and in mouse leukemia cells (L1210). A dose-dependent decrease in intracellular spermine concentration was observed in both cell lines when grown in the presence of the inhibitor. A major difference in the effects seen in these two cell lines was the cytotoxicity observed in L1210 cells exposed to the inhibitor, which contrasted with little or no effects on growth of SV 3T3 cells treated similarly. Oxidative metabolism of the drug in L1210 cells was suggested by the fact that addition of aminoguanidine, an amine oxidase inhibitor, to the cell cultures ablated the cytotoxic effects of the inhibitor. Complete analysis of intracellular polyamines was carried out, together with analysis of S-adenosylmethionine, decarboxylated S-adenosylmethionine, and the inhibitor. These analyses revealed that, although the inhibitor had a dramatic effect on spermine biosynthesis in the cells studied, a compensatory increase in spermidine biosynthesis was observed. This resulted in no change in total polyamine concentrations in cells treated with inhibitors of either spermine synthase or spermidine synthase (Pegg et al., 1982) alone or in combination. In all cases, the concentration of the aminopropyl donor decarboxylated S-adenosylmethionine increased dramatically, thus allowing for the observed maintenance of total polyamine levels even in the presence of either one or both potent inhibitors of the aminopropyltransferases. Oxidative metabolism of the inhibitor complicates the interpretation of experiments carried out in the absence of amine oxidase inhibitors such as aminoguanidine.(ABSTRACT TRUNCATED AT 250 WORDS)


Occupational risks and risk perception among Hispanic adolescents.
Although child labor laws are designed to protect youth from serious injury in the workplace, there are a number of other factors that influence the effectiveness of occupational injury prevention including adolescent risk perception, failure to understand worker rights, and lack of experience. The growing adolescent Hispanic population in the U.S. is of particular concern because cultural and economic barriers have the potential to further influence these young workers' safety. A greater emphasis on research among Hispanic adolescents is crucial to minimizing adolescent worker injury. To propose effective age and culturally appropriate intervention programs, it will be necessary to understand the knowledge, attitude and beliefs regarding occupational risks within the working adolescent Hispanic community.