Emotion dysregulation and reward responsiveness as predictors of autonomic reactivity to an infant cry task among substance-using pregnant and postpartum women.
Maternal substance use may interfere with optimal parenting, lowering maternal responsiveness during interactions with their children. Previous work has identified maternal autonomic nervous system (ANS) reactivity to parenting-relevant stressors as a promising indicator of real-world parenting behaviors. However, less is known about the extent to which individual differences in emotion dysregulation and reward processing, two mechanisms of substance use, relate to maternal ANS reactivity in substance-using populations. The current study examined associations among emotion dysregulation, reward responsiveness, and ANS reactivity to an infant cry task among 77 low-income and substance-using women who were either pregnant (n = 63) or postpartum (n = 14). Two indicators of ANS functioning were collected during a 9 min computerized infant cry task (Crybaby task): respiratory sinus arrhythmia (RSA) and pre-ejection period. Mothers also completed self-reported measures of emotion dysregulation and reward responsiveness. Analyses revealed that trait emotion regulation was associated with RSA reactivity to the Crybaby task, such that greater emotion dysregulation was associated with greater RSA reduction during the infant cry task than lower emotion dysregulation. Reward responsiveness was not significantly associated with either indicator of ANS reactivity to the task. Findings revealed distinct patterns of associations linking emotion dysregulation with ANS reactivity during a parenting-related computerized task, suggesting that emotion regulation may be a key intervention target for substance-using mothers.


The Finally Rewarding Search for A Cytotoxic Isosteviol Derivative.
Acid hydrolysis of stevioside resulted in a 63% yield of isosteviol (1), which served as a starting material for the preparation of numerous amides. These compounds were tested for cytotoxic activity, employing a panel of human tumor cell lines, and almost all amides were found to be non-cytotoxic. Only the combination of isosteviol, a (homo)-piperazinyl spacer and rhodamine B or rhodamine 101 unit proved to be particularly suitable. These spacered rhodamine conjugates exhibited cytotoxic activity in the sub-micromolar concentration range. In this regard, the homopiperazinyl-spacered derivatives were found to be better than those compounds with piperazinyl spacers, and rhodamine 101 conjugates were more cytotoxic than rhodamine B hybrids.


One Size Does Not Fit All: Choosing Practical Cognitive Screening Tools for Your Practice.
Every year, millions of patients worldwide undergo cognitive testing. Unfortunately, new barriers to the use of free open access cognitive screening tools have arisen over time, making accessibility of tools unstable. This article is in follow-up to an editorial discussing alternative cognitive screening tools for those who cannot afford the costs of the Mini-Mental State Examination and Montreal Cognitive Assessment (see www.dementiascreen.ca). The current article outlines an emerging disruptive "free-to-fee" cycle where free open access cognitive screening tools are integrated into clinical practice and guidelines, where fees are then levied for the use of the tools, resulting in clinicians moving on to other tools. This article provides recommendations on means to break this cycle, including the development of tool kits of valid cognitive screening tools that authors have contracted not to charge for (i.e., have agreed to keep free open access). The PRACTICAL.1 Criteria (PRACTIcing Clinician Accessibility and Logistical Criteria Version 1) are introduced to help clinicians select from validated cognitive screening tools, considering barriers and facilitators, such as whether the cognitive screening tools are easy to score and free of cost. It is suggested that future systematic reviews embed the PRACTICAL.1 criteria, or refined future versions, as part of the standard of review. Methodological issues, the need for open access training to insure proper use of cognitive screening tools, and the need to anticipate growing ethnolinguistic diversity by developing tools that are less sensitive to educational, cultural, and linguistic bias are discussed in this opinion piece. J Am Geriatr Soc 68:2207-2213, 2020.


Inadequate Boston Bowel Preparation Scale scores predict the risk of missed neoplasia on the next colonoscopy.
The risks of missed findings after inadequate bowel preparation are not fully characterized in a diverse cohort. We aimed to evaluate the likelihood of missed polyps after an inadequate preparation as assessed by using the Boston Bowel Preparation Scale (BBPS). In this observational study of prospectively collected data within a large, national, endoscopic consortium, we identified patients aged 50 to 75 years who underwent average-risk screening colonoscopy (C1) followed by a second colonoscopy for any indication within 3 years (C2). We determined the polyp detection rates (PDRs) and advanced PDRs during C2 stratified by C1 BBPS scores. Among segment pairs without polyps at C1 (N = 601), those with inadequate C1 BBPS segment scores had a higher PDR at C2 (10%) compared with those with adequate bowel preparation at C1 (5%; P = .04). Among segment pairs with polyps at C1 (N = 154), segments with inadequate bowel preparation scores at C1 had higher advanced PDRs at C2 (20%) compared with those with adequate bowel preparation scores at C1 (4%; P = .03). In multivariable analysis, the presence of advanced polyps at C1 (adjusted odds ratio [OR] 3.5; 95% confidence intervals [CIs], 1.1-10.8) but not inadequate BBPS scores at C1 (adjusted OR 1.8; 95% CI, 0.6-5.1) was associated with a significantly increased risk of advanced polyps at C2. Inadequate BBPS segment scores generally are associated with higher rates of polyps and advanced polyps at subsequent colonoscopy within a short timeframe. The presence of advanced polyps as well as inadequate BBPS segment scores can inform the risk of missed polyps and help triage which patients warrant a timely repeat colonoscopy.


Over Expression of the Cyanobacterial Pgr5-Homologue Leads to Pseudoreversion in a Gene Coding for a Putative Esterase in Synechocystis 6803.
Pgr5 proteins play a major direct role in cyclic electron flow paths in plants and eukaryotic phytoplankton. The genomes of many cyanobacterial species code for Pgr5-like proteins but their function is still uncertain. Here, we present evidence that supports a link between the Synechocystis sp. PCC6803 Pgr5-like protein and the regulation of intracellular redox balance. The knockout strain, pgr5KO, did not display substantial phenotypic response under our experimental conditions, confirming results obtained in earlier studies. However, the overexpression strain, pgr5OE, accumulated 2.5-fold more chlorophyll than the wild type and displayed increased content of photosystems matching the chlorophyll increase. As a result, electron transfer rates through the photosynthetic apparatus of pgr5OE increased, as did the amount of energy stored as glycogen. While, under photoautotrophic conditions, this metabolic difference had only minor effects, under mixotrophic conditions, pgr5OE cultures collapsed. Interestingly, this specific phenotype of pgr5OE mutants displayed a tendency for reverting, and cultures which previously collapsed in the presence of glucose were now able to survive. DNA sequencing of a pgr5OE strain revealed a second site suppression mutation in slr1916, a putative esterase associated with redox regulation. The phenotype of the slr1916 knockout is very similar to that of the strain reported here and to that of the pmgA regulator knockout. These data demonstrate that, in Synechocystis 6803, there is strong selection against overexpression of the Pgr5-like protein. The pseudoreversion event in a gene involved in redox regulation suggests a connection of the Pgr5-like protein to this network.


Tumor-induced injury of primary afferent sensory nerve fibers in bone cancer pain.
Bone is the most common site of chronic pain in patients with metastatic cancer. What remains unclear are the mechanisms that generate this pain and why bone cancer pain can be so severe and refractory to treatment with opioids. Here we show that following injection and confinement of NCTC 2472 osteolytic tumor cells within the mouse femur, tumor cells sensitize and injure the unmyelinated and myelinated sensory fibers that innervate the marrow and mineralized bone. This tumor-induced injury of sensory nerve fibers is accompanied by an increase in ongoing and movement-evoked pain behaviors, an upregulation of activating transcription factor 3 (ATF3) and galanin by sensory neurons that innervate the tumor-bearing femur, upregulation of glial fibrillary acidic protein (GFAP) and hypertrophy of satellite cells surrounding sensory neuron cell bodies within the ipsilateral dorsal root ganglia (DRG), and macrophage infiltration of the DRG ipsilateral to the tumor-bearing femur. Similar neurochemical changes have been described following peripheral nerve injury and in other non-cancerous neuropathic pain states. Chronic treatment with gabapentin did not influence tumor growth, tumor-induced bone destruction or the tumor-induced neurochemical reorganization that occurs in sensory neurons or the spinal cord, but it did attenuate both ongoing and movement-evoked bone cancer-related pain behaviors. These results suggest that even when the tumor is confined within the bone, a component of bone cancer pain is due to tumor-induced injury to primary afferent nerve fibers that innervate the tumor-bearing bone. Tumor-derived, inflammatory, and neuropathic mechanisms may therefore be simultaneously driving this chronic pain state.


[Workers with Ehlers-Danlos syndrome: indications for health surveillance and suitable job assignment].
In Europe 23.5% of the working-age population suffers from chronic or rare diseases, including Ehlers-Danlos syndrome (EDS), an inherited disorder of the connective tissue that frequently leads to impairment also at work. We sought to evaluate the impairment of different functional areas in EDS subjects, using EDS-Disability Test (EDS-DT) (7 visual analogical scales: pain, stiffness, daily activities of life, in the home, outside the home, at work, in social relationships). We administered the EDS-DT to 50 workers with EDS (classic type n=35, hypermobile n=14, vascular n=3) and 150 healthy workers (non-EDS). EDS subjects showed higher perceived disability (median 29.31 vs 7.22, p< 0.0001) than non-EDS. We observed a trend suggesting that the hypermobile type has a higher level of pain (54 vs 42) and work impairment (62.5 vs 42.5) (p>0.20), whereas the classic type has greater impairment of daily living activities (18.57 vs 11.43) and in the home (34.29 vs 25.71) (p>0.20). Subjects reporting moderate-to-vigorous occupational physical activities (OPA) showed a higher disability level in the work area (p=0.04). People with EDS suffer from chronic pain, impaired quality of life and employment difficulties. The hypermobile type seems more compromised in functional areas such as pain and work, while the classic type is more compromised in daily routine and home activities. The employment section was more impaired for EDS subjects who perform tasks with greater physical effort.


Localization of the transmembrane 4 superfamily (TM4SF) member PETA-3 (CD151) in normal human tissues: comparison with CD9, CD63, and alpha5beta1 integrin.
It has recently been shown that several members of the tetraspan superfamily, including CD9 and CD63, associate with each other and with beta1 integrins. In this study, we examined the distribution of a recently identified tetraspan, PETA-3 (CD151), and of CD9, CD63, alpha5beta1, and the integrin beta1 chain in normal human tissues by the indirect immunoperoxidase and alkaline phosphatase-anti-alkaline phosphatase techniques. PETA-3 showed a broad distribution and was expressed by endothelium, epithelium, Schwann cells, and dendritic cells and by skeletal, smooth, and cardiac muscle. Expression in skin was mostly restricted to the basal cells of the epidermis and was downregulated on differentiation. In the small intestine, PETA-3 was expressed by crypt and villous enterocytes with a mostly basolateral distribution, but was not detectable on the brush border. CD9 was expressed on the plasma membrane of enterocytes in crypts and at the bases of the villi whereas CD63 demonstrated a unique granular appearance concentrated in the apical cytoplasm below the brush border. The findings of this study show co-localization of PETA-3 with CD9, CD63, alpha5beta1, and beta1 in particular tissues, demonstrating that tetraspan/integrin complexes may occur. However, the lack of co-localization of these antigens in other tissues also implies distinct roles for these molecules.


Defining the Structures at Risk and an Anatomical Safe Zone for Percutaneous Antegrade Subtalar Joint Fixation With a Single Screw: A Cadaveric Study.
Percutaneous antegrade (anterior to posterior) fixation for subtalar joint (STJ) arthrodesis offers various intraoperative and biomechanical advantages. Currently, the entry point for percutaneous antegrade STJ screw fixation is not clearly described and variable. To our knowledge, there are no publications that evaluate anatomic structures at risk or define an anatomically safe entry point for this fixation. The aim of this investigation was to define an anatomically safe and reproducible entry point for percutaneous antegrade STJ arthrodesis fixation, while also describing anatomic structures at risk when undertaking this method of fixation. We hypothesized that percutaneous single screw antegrade STJ fixation would encroach upon named anatomic structures in more than one cadaveric specimen. Ten cadaver limbs were used in this investigation. A percutaneous guidewire was inserted 5 mm lateral to the tibialis anterior tendon. The midpoint of the talar neck served as the sagittal plane starting point, as seen on the lateral fluoroscopic view. A cannulated 6.5-mm headed screw was inserted antegrade through the STJ into the calcaneus. Each specimen was dissected to assess the distance from the screw to nearby anatomic structures and distance from the tibialis anterior tendon to named structures. Our hypothesis was found to be incorrect, as 0/10 screws invaded neurovascular or tendinous structures. The dorsalis pedis artery and deep peroneal nerve were on average 12.1 ± 2.79 mm and 12.2 ± 2.82 mm lateral to the screw, respectively. These findings are clinically relevant and ultimately allow us to define an anatomic safe starting point for percutaneous antegrade STJ single screw fixation.


Global and local precedence: selective attention in form and motion perception.
This study explores the perception of stimuli at two levels: local parts and the wholes that comprise these parts. Previous research has produced contradictory results. Some studies (e.g., Pomerantz & Sager, 1975) show local precedence, in which the local parts are more difficult to ignore in selective attention tasks. Other studies (e.g., Navon, 1977) have shown the opposite effect, global precedence. The present five experiments trace the causes of this discrepancy by exploring the effects of the relative discriminabilities of the local and global levels of the stimuli and the differences between two different measures of selective attention, namely, Stroop-type interference (attributable to incongruity on the irrelevant dimension) and Garner-type interference (attributable to variability on the irrelevant dimension). The experiments also examine whether the precedence effects previously examined in form perception generalize to motion perception. The results show that (a) some cases of global precedence are due solely to the greater perceptual discriminability of the global level and thus demonstrate only that more discriminable stimuli are harder to ignore; (b) instances of both local and global precedence can be demonstrated for certain types of stimuli, even when the discriminabilities of their local and global levels have been equated; and (c) the Stroop and Garner measures of selective attention are not equivalent but instead measure different types of interference. In addition, a distinction is made between two fundamentally different types of part-whole relationships that exist in visual configurations, one based only on the positions of the parts (Type P) and one based also on the nature of the parts (Type N). Previous research has focused on Type P, which appears to be irrelevant to the broader questions of Gestalt and top-down effects in perception. It is concluded that bona fide cases of both local and global precedence have been amply documented but that no general theory can account for why or when these effects will appear until we better understand both the nature of part-whole relationships and the perceptual processes that are tapped by different measures of selective attention.


A quantitative weight of evidence methodology for the assessment of reproductive and developmental toxicity and its application for classification and labeling of chemicals.
Hazard assessment of chemicals usually applies narrative assessments with a number of weaknesses. Therefore, application of weight of evidence (WoE) approaches are often mandated but guidance to perform a WoE assessment is lacking. This manuscript describes a quantitative WoE (QWoE) assessment for reproductive toxicity data and its application for classification and labeling (C&L). Because C&L criteria are based on animal studies, the scope is restricted to animal toxicity data. The QWoE methodology utilizes numerical scoring sheets to assess reliability of a publication and the toxicological relevance of reported effects. Scores are given for fourteen quality aspects, best practice receives the highest score. The relevance/effects scores (0 to four) are adjusted to the key elements of the toxic response for the endpoint and include weighting factors for effects on different levels of the biological organization. The relevance/effects scores are then assessed against the criteria dose-response, magnitude and persistence of effects, consistency of observations with the hypothesis, and relation of effects to human disease. The quality/reliability scores and the relevance/effect scores are then multiplied to give a numerical strength of evidence for adverse effects. This total score is then used to assign the chemical to the different classes employed in classification.


Characterization of high-Q optical microcavities using confocal microscopy.
Confocal microscopy was initially developed to image complex circuits and material defects. Previous imaging studies yielded only qualitative data about the location and number of defects. In the present study, this noninvasive method is used to obtain quantitative information about the Q factor of an optical resonant cavity. Because the intensity of the fluorescent signal measures the number of defects in the resonant cavity, this signal is a measure of the number of surface scattering defects, one of the dominant loss mechanisms in optical microcavities. The Q of the cavities was also determined using conventional linewidth measurements. Based upon a quantitative comparative analysis of these two techniques, it is shown that the Q can be determined without a linewidth measurement, allowing for a noninvasive characterization technique.


Screening of Regulated and Emerging Mycotoxins in Bulk Milk Samples by High-Resolution Mass Spectrometry.
In this work, a retrospective screening based on ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) based on Orbitrap-Q-Exactive Focus™ was used to check the occurrence of regulated and emerging mycotoxins in bulk milk samples. Milk samples were collected from dairy farms in which corn silage was the main ingredient of the feeding system. The 45 bulk milk samples were previously analyzed for a detailed untargeted metabolomic profiling and classified into five clusters according to the corn silage contamination profile, namely: (1) low levels of Aspergillus- and Penicillium-mycotoxins; (2) low levels of fumonisins and other Fusarium-mycotoxins; (3) high levels of Aspergillus-mycotoxins; (4) high levels of non-regulated Fusarium-mycotoxins; (5) high levels of fumonisins and their metabolites. Multivariate statistics based on both unsupervised and supervised analyses were used to evaluate the significant fold-change variations of the main groups of mycotoxins detected when comparing milk samples from clusters 3, 4, and 5 (high contamination levels of the corn silages) with cluster 1 and 2 (low contamination levels of the corn silages). Overall, 14 compounds showed a significant prediction ability, with antibiotic Y (VIP score = 2.579), bikaverin (VIP score = 1.975) and fumonisin B2 (VIP score = 1.846) being the best markers. The k-means clustering combined with supervised statistics showed two discriminant groups of milk samples, thus revealing a hierarchically higher impact of the whole feeding system (rather than the only corn silages) together with other factors of variability on the final mycotoxin contamination profile. Among the discriminant metabolites we found some Fusarium mycotoxins, together with the tetrapeptide tentoxin (an Alternaria toxin), the α-zearalenol (a catabolite of zearalenone), mycophenolic acid and apicidin. These preliminary findings provide new insights into the potential role of UHPLC-HRMS to evaluate the contamination profile and the safety of raw milk to produce hard cheese.


Double-outlet left ventricle. Echocardiographic diagnosis.
This is a case report of a double-outlet left ventricle associated with tricuspid atresia and hypoplasia of the right ventricle, diagnosed during echocardiography with color-flow imaging, in a three-month-old child who presented with fatigue and cyanosis. The child underwent palliative pulmonary arterial banding without an invasive procedure, and showed sustained improvement during follow-up.


Solid-state 13C NMR study of tyrosine protonation in dark-adapted bacteriorhodopsin.
Solid-state 13C MAS NMR spectra were obtained for dark-adapted bacteriorhodopsin (bR) labeled with [4'-13C]Tyr. Difference spectra (labeled minus natural abundance) taken at pH values between 2 and 12, and temperatures between 20 and -90 degrees C, exhibit a single signal centered at 156 ppm, indicating that the 11 tyrosines are protonated over a wide pH range. However, at pH 13, a second line appears in the spectrum with an isotropic shift of 165 ppm. Comparisons with solution and solid-state spectra of model compounds suggest that this second line is due to the formation of tyrosinate. Integrated intensities indicate that about half of the tyrosines are deprotonated at pH 13. This result demonstrates that deprotonated tyrosines in a membrane protein are detectable with solid-state NMR and that neither the bR568 nor the bR555 form of bR present in the dark-adapted state contains a tyrosinate at pH values between 2 and 12. Deprotonation of a single tyrosine in bR568 would account for 3.6% of the total tyrosine signal, which would be detectable with the current signal-to-noise ratio. We observe a slight heterogeneity and subtle line-width changes in the tyrosine signal between pH 7 and pH 12, which we interpret to be due to protein environmental effects (such as changes in hydrogen bonding) rather than complete deprotonation of tyrosine residue(s).


Stress-related transcription factor AtfB integrates secondary metabolism with oxidative stress response in aspergilli.
In filamentous fungi, several lines of experimental evidence indicate that secondary metabolism is triggered by oxidative stress; however, the functional and molecular mechanisms that mediate this association are unclear. The basic leucine zipper (bZIP) transcription factor AtfB, a member of the bZIP/CREB family, helps regulate conidial tolerance to oxidative stress. In this work, we investigated the role of AtfB in the connection between oxidative stress response and secondary metabolism in the filamentous fungus Aspergillus parasiticus. This well characterized model organism synthesizes the secondary metabolite and carcinogen aflatoxin. Chromatin immunoprecipitation with specific anti-AtfB demonstrated AtfB binding at promoters of seven genes in the aflatoxin gene cluster that carry CREs. Promoters lacking CREs did not show AtfB binding. The binding of AtfB to the promoters occurred under aflatoxin-inducing but not under aflatoxin-noninducing conditions and correlated with activation of transcription of the aflatoxin genes. Deletion of veA, a global regulator of secondary metabolism and development, nearly eliminated this binding. Electrophoretic mobility shift analysis demonstrated that AtfB binds to the nor-1 (an early aflatoxin gene) promoter at a composite regulatory element that consists of highly similar, adjacent CRE1 and AP-1-like binding sites. The five nucleotides immediately upstream from CRE1, AGCC(G/C), are highly conserved in five aflatoxin promoters that demonstrate AtfB binding. We propose that AtfB is a key player in the regulatory circuit that integrates secondary metabolism and cellular response to oxidative stress.


Immune gene expression in the mosquito vector Culex quinquefasciatus during an avian malaria infection.
Plasmodium relictum is the most widespread avian malaria parasite in the world. It is listed as one of the 100 most dangerous invasive species, having been responsible for the extinction of several endemic bird species, and the near-demise of several others. Here we present the first transcriptomic study focused on the effect of P. relictum on the immune system of its vector (the mosquito Culex quinquefasciatus) at different times post-infection. We show that over 50% of immune genes identified as being part of the Toll pathway and 30%-40% of the immune genes identified within the Imd pathway are overexpressed during the critical period spanning the parasite's oocyst and sporozoite formation (8-12 days), revealing the crucial role played by both these pathways in this natural mosquito-Plasmodium combination. Comparison of infected mosquitoes with their uninfected counterparts also revealed some unexpected immune RNA expression patterns earlier and later in the infection: significant differences in expression of several immune effectors were observed as early as 30 min after ingestion of the infected blood meal. In addition, in the later stages of the infection (towards the end of the mosquito lifespan), we observed an unexpected increase in immune investment in uninfected, but not in infected, mosquitoes. In conclusion, our work extends the comparative transcriptomic analyses of malaria-infected mosquitoes beyond human and rodent parasites and provides insights into the degree of conservation of immune pathways and into the selective pressures exerted by Plasmodium parasites on their vectors.


Expression of KNUCKLES in the Stem Cell Domain Is Required for Its Function in the Control of Floral Meristem Activity in Arabidopsis.
In the model plant Arabidopsis thaliana, the zinc-finger transcription factor KNUCKLES (KNU) plays an important role in the termination of floral meristem activity, a process that is crucial for preventing the overgrowth of flowers. The KNU gene is activated in floral meristems by the floral organ identity factor AGAMOUS (AG), and it has been shown that both AG and KNU act in floral meristem control by directly repressing the stem cell regulator WUSCHEL (WUS), which leads to a loss of stem cell activity. When we re-examined the expression pattern of KNU in floral meristems, we found that KNU is expressed throughout the center of floral meristems, which includes, but is considerably broader than the WUS expression domain. We therefore hypothesized that KNU may have additional functions in the control of floral meristem activity. To test this, we employed a gene perturbation approach and knocked down KNU activity at different times and in different domains of the floral meristem. In these experiments we found that early expression in the stem cell domain, which is characterized by the expression of the key meristem regulatory gene CLAVATA3 (CLV3), is crucial for the establishment of KNU expression. The results of additional genetic and molecular analyses suggest that KNU represses floral meristem activity to a large extent by acting on CLV3. Thus, KNU might need to suppress the expression of several meristem regulators to terminate floral meristem activity efficiently.


Stochastic simulations of cargo transport by processive molecular motors.
We use stochastic computer simulations to study the transport of a spherical cargo particle along a microtubule-like track on a planar substrate by several kinesin-like processive motors. Our newly developed adhesive motor dynamics algorithm combines the numerical integration of a Langevin equation for the motion of a sphere with kinetic rules for the molecular motors. The Langevin part includes diffusive motion, the action of the pulling motors, and hydrodynamic interactions between sphere and wall. The kinetic rules for the motors include binding to and unbinding from the filament as well as active motor steps. We find that the simulated mean transport length increases exponentially with the number of bound motors, in good agreement with earlier results. The number of motors in binding range to the motor track fluctuates in time with a Poissonian distribution, both for springs and cables being used as models for the linker mechanics. Cooperativity in the sense of equal load sharing only occurs for high values for viscosity and attachment time.


Reactive Extrusion and Magnesium (II) N-Heterocyclic Carbene Catalyst in Continuous PLA Production.
Reactive extrusion and magnesium (II) N-heterocyclic carbene catalyst are successfully employed in continuous polylactide synthesis. The possibility of using six-membered N-heterocyclic carbene adducts to act as efficient catalysts towards the sustainable synthesis of poly(l-lactide) through ring-opening polymerization of l-lactide (LA) is first investigated in bulk batch reactions. Under optimized solvent-free conditions, polylactide (PLA) of moderate to high molecular weights and excellent optical activities are successfully achieved. These promising results are further applied in the continuous production of PLA in an extruder.


X-ray characterization, electronic band structure, and thermoelectric properties of the cluster compound Ag2Tl2Mo9Se11.
We report on a detailed investigation of the crystal and electronic band structures and of the transport and thermodynamic properties of the Mo-based cluster compound Ag2Tl2Mo9Se11. This novel structure type crystallizes in the trigonal space group R3̅c and is built of a three-dimensional network of interconnected Mo9Se11 units. Single-crystal X-ray diffraction indicates that the Ag and Tl atoms are distributed in the voids of the cluster framework, both of which show unusually large anisotropic thermal ellipsoids indicative of strong local disorder. First-principles calculations show a weakly dispersive band structure around the Fermi level as well as a semiconducting ground state. The former feature naturally explains the presence of both hole-like and electron-like signals observed in Hall effect. Of particular interest is the very low thermal conductivity that remains quasi-constant between 150 and 800 K at a value of approximately 0.6 W·m(-1)·K(-1). The lattice thermal conductivity is close to its minimum possible value, that is, in a regime where the phonon mean free path nears the mean interatomic distance. Such extremely low values likely originate from the disorder induced by the Ag and Tl atoms giving rise to strong anharmonicity of the lattice vibrations. The strongly limited ability of this compound to transport heat is the key feature that leads to a dimensionless thermoelectric figure of merit ZT of 0.6 at 800 K.


Cutting edge: dendritic cell-restricted antigen presentation initiates the follicular helper T cell program but cannot complete ultimate effector differentiation.
Follicular helper T (T(FH)) cells are critical for germinal center (GC) formation. The processes that drive their generation and effector potential remain unclear. In this study, we define requirements for MHC class II APCs in murine T(FH) cell formation by either transiently ablating or restricting Ag presentation to dendritic cells (DCs). We find that cognate interactions with DCs are necessary and sufficient to prime CD4(+) T cells toward a CXCR5(+)ICOS(+)Bcl6(+) T(FH) cell intermediate. However, in the absence of additional APCs, these T(FH) cells fail to produce IL-21. Furthermore, in vitro priming of naive T cells by B cells engenders optimal production of IL-21, which induces a GC B cell transcriptional profile. These results support a multistep model for effector T(FH) cell priming and GC initiation, in which DCs are necessary and sufficient to induce a T(FH) cell intermediate that requires additional interactions with distinct APCs for full effector function.


Recombination rates across porcine autosomes inferred from high-density linkage maps.
Studies of the variation in recombination rate across the genome provide a better understanding of evolutionary genomics and are also an important step towards mapping and dissecting complex traits in domestic animals. With the recent completion of the porcine genome sequence and the availability of a high-density porcine single nucleotide polymorphism (SNP) array, it is now possible to construct a high-density porcine linkage map and estimate recombination rate across the genome. A total of 416 animals were genotyped with the Porcine SNP60BeadChip, and high-density chromosome linkage maps were constructed using CRI-MAP, assuming the physical order of the Sscrofa10 assembly. The total linkage map length was 2018.79 cM, using 658 meioses and 14,503 SNPs. The estimated average recombination rate across the porcine autosomes was 0.86 cM/Mb. However, a large variation in recombination rate was observed among chromosomes. The estimated average recombination rates (cM/Mb) per chromosome ranged from 0.48 in SSC1 to 1.48 in SSC10, displaying a significant negative correlation with the chromosome sizes. In addition, the analysis of the variation in the recombination rates taking 1-Mb sliding windows has allowed us to demonstrate the variation in recombination rates within chromosomes. In general, a larger recombination rate was observed in the extremes than in the centre of the chromosome. Finally, the ratio between female and male recombination rates was also inferred, obtaining a value of 1.38, with the heterogametic sex having the least recombination.


A novel pathway involving progesterone receptor, 12/15-lipoxygenase-derived eicosanoids, and peroxisome proliferator-activated receptor gamma regulates implantation in mice.
The 12/15-lipoxygenases (12/15-LOX) catalyze the stereo-specific oxygenation of arachidonic and linoleic acids into a complex series of signaling molecules, including the hydroxyeicosatetraenoic acids (HETEs) and hydroxyoctadecadienoic acids (HODEs). Our previous studies, using high density oligonucleotide microarrays, suggested a novel link between progesterone receptor (PR) signaling and 12/15-LOX-mediated fatty acid metabolism in preimplantation mouse uterus. In this paper, using PR knockout mice, we established that the transcripts encoding leukocyte-12/15-LOX (L-12/15-LOX) and epidermal-12/15-LOX (E-12/15-LOX) are indeed downstream targets of regulation by PR in the uterine surface epithelium. Maximal induction of both L- and E-12/15-LOX on the day of implantation resulted in a marked increase in the uterine levels of the eicosanoids, 12-HETE, 15-HETE, and 13-HODE. Mice with null mutation in L-12/15-LOX had significantly reduced uterine levels of arachidonic acid metabolites and exhibited a partial impairment in implantation. Complete blockade of uterine 12/15-LOX activity by a specific inhibitor led to greater than 80% reduction in a number of implantation sites relative to untreated controls. Cell-based assays indicated that 12-HETE, 15-HETE, and 13-HODE function as activating ligands of peroxisome proliferator-activated receptor gamma (PPARgamma), suggesting that this nuclear receptor could be a downstream target of 12/15-LOX-derived metabolites in the preimplantation uterus. Consistent with this hypothesis, administration of rosiglitazone, a potent PPARgamma-selective agonist, efficiently reversed inhibition of implantation by the 12/15-LOX-specific inhibitor. Rosiglitazone also induced a number of potential target genes of 12/15-LOX-derived metabolites in the pregnant uterus, indicating their regulation by PPARgamma. Collectively, our results uncovered a novel signaling pathway in which progesterone-induced synthesis of the 12/15-LOX-derived lipid mediators activates PPARgamma and its downstream gene networks, which in turn function as critical regulators of implantation in the mouse.


Massively Parallel Implementation of Steered Molecular Dynamics in Tinker-HP: Comparisons of Polarizable and Non-Polarizable Simulations of Realistic Systems.
Steered molecular dynamic (SMD) is a powerful technique able to accelerate rare event sampling in Molecular Dynamics (MD) simulations by applying an external force to a set of chosen atoms. Despite generating nonequilibrium simulations, SMD remains capable of reconstructing equilibrium properties such as the Potential of Mean Force (PMF). Of course, one would like to use all types of force fields (FF) ranging from classical ones to more advanced polarizable models using point induced dipoles and distributed multipoles such as AMOEBA. To enable such studies, the SMD methodology has been implemented in the framework of the massively parallel Tinker-HP software allowing for both long polarizable and non-polarizable MD simulations of large proteins. To validate this new implementation, we first compared the Tinker-HP SMD results to the literature. Tests have been performed on three different benchmark systems: the M-A deca-alanine (112 atoms), the ubiquitin (9737 atoms), and the CD2CD58 complex (97594 atoms). Non-polarizable (AMBER99, AMBER99SB, CHARMM22/CMAP, and OPLS-AA/L) and polarizable (AMOEBAPRO13 and AMOEBABIO18) force fields have been used. For each one of them, PMFs have been reconstructed and compared in terms of free energy barrier and hydrogen bonding fluctuations behavior over time. Using a SMD velocity of 0.01 Å/ps applied to a set of 20 trajectories, we show that polarizable and non-polarizable force fields do not always agree. As it could be anticipated, strong discrepancies are noticed between polarizable and non-polarizable models when considered in vacuum, whereas results are more comparable when a water environment is added. However, for the largest system, i.e., the CD2CD58 complex, strong differences related to the modeling of a salt bridge are noticed exhibiting some potential issues with classical FFs. Overall, such simulations highlight the importance of the inclusion of polarization effects as PMF free energy barriers computed with AMOEBA always decrease compared to non-polarizable force fields.


Neuroendocrine-immune system interactions in amphibians: implications for understanding global amphibian declines.
Amphibians are ancient creatures valued by biologists and naturalists around the world. They share with all other vertebrates a complex neuroendocrine system that enables them to flourish in a variety of aquatic and semiaquatic environments. Studies from a number of laboratories have demonstrated that the immune system of amphibian species is nearly as complex as that of mammals. Yet for reasons that are not well understood, amphibian species are facing greater survival challenges than in the recent past. This article will review our current understanding of the neuroendocrine immune system interactions in amphibians and address the question of whether environmental stressors may contribute to immunosuppression and amphibian declines.


Molecular hydrogen alleviates asphyxia-induced neuronal cyclooxygenase-2 expression in newborn pigs.
Cyclooxygenase-2 (COX-2) has an established role in the pathogenesis of hypoxic-ischemic encephalopathy (HIE). In this study we sought to determine whether COX-2 was induced by asphyxia in newborn pigs, and whether neuronal COX-2 levels were affected by H2 treatment. Piglets were subjected to either 8 min of asphyxia or a more severe 20 min of asphyxia followed by H2 treatment (inhaling room air containing 2.1% H2 for 4 h). COX-2 immunohistochemistry was performed on brain samples from surviving piglets 24 h after asphyxia. The percentages of COX-2-immunopositive neurons were determined in cortical and subcortical areas. Only in piglets with more severe HIE, we observed significant, region-specific increases in neuronal COX-2 expression within the parietal and occipital cortices and in the CA3 hippocampal subfield. H2 treatment essentially prevented the increases in COX-2-immunopositive neurons. In the parietal cortex, the attenuation of COX-2 induction was associated with reduced 8'-hydroxy-2'-deoxyguanozine immunoreactivity and retained microglial ramifcation index, which are markers of oxidative stress and neuroinfiammation, respectively. This study demonstrates for the first time that asphyxia elevates neuronal COX-2 expression in a piglet HIE model. Neuronal COX-2 induction may play region-specific roles in brain lesion progression during HIE development, and inhibition of this response may contribute to the antioxidant/anti-infiammatory neuroprotective effects of H2 treatment.


Systematic review: human gut dysbiosis induced by non-antibiotic prescription medications.
Global prescription drug use has been increasing continuously for decades. The gut microbiome, a key contributor to health status, can be altered by prescription drug use, as antibiotics have been repeatedly described to have both short-term and long-standing effects on the intestinal microbiome. To summarise current findings on non-antibiotic prescription-induced gut microbiome changes, focusing on the most frequently prescribed therapeutic drug categories. We conducted a systematic review by first searching in online databases for indexed articles and abstracts in accordance with PRISMA guidelines. Studies assessing the intestinal microbiome alterations associated with proton pump inhibitors (PPIs), metformin, nonsteroidal anti-inflammatory drugs (NSAIDs), opioids, statins and antipsychotics were included. We only included studies using culture-independent molecular techniques. Proton pump inhibitors and antipsychotic medications are associated with a decrease in α diversity in the gut microbiome, whereas opioids were associated with an increase in α diversity. Metformin and NSAIDs were not associated with significant changes in α diversity. β diversity was found to be significantly altered with all drugs, except for NSAIDs. PPI use was linked to a decrease in Clotridiales and increase in Actinomycetales, Micrococcaceae and Streptococcaceae, which are changes previously implicated in dysbiosis and increased susceptibility to Clostridium difficile infection. Consistent results showed that PPIs, metformin, NSAIDs, opioids and antipsychotics were either associated with increases in members of class Gammaproteobacteria (including Enterobacter, Escherichia, Klebsiella and Citrobacter), or members of family Enterococcaceae, which are often pathogens isolated from bloodstream infections in critically ill patients. We also found that antipsychotic treatment, usually associated with an increase in body mass index, was marked by a decreased ratio of Bacteroidetes:Firmicutes in the gut microbiome, resembling trends seen in obese patients. Non-antibiotic prescription drugs have a notable impact on the overall architecture of the intestinal microbiome. Further explorations should seek to define biomarkers of dysbiosis induced by specific drugs, and potentially tailor live biotherapeutics to counter this drug-induced dysbiosis. Many other frequently prescribed drugs should also be investigated to better understand the link between these drugs, the microbiome and health status.


Early development of the human area postrema and subfornical organ.
The first appearance and early development of two circumventricular organs, the area postrema (AP) and the subfornical organ (SFO), were investigated in human embryos and fetuses from the 4th to the 40th gestational weeks (GW). The AP appears very early in development, during the GW 10; its high vascularization can be seen from GW14, and differentiated neurons are observed from GW 16. The SFO is characterized by a late onset of development. It can first be distinguished at GW 17, but it does not attain cytological differentiation until the last weeks of gestation. It is suggested that the AP has important functions during fetal life, which are related to normal fetal weight and growth; in contrast the SFO, which is connected with drinking behavior and salt/water balance, seems to play a less essential role in early fetal life.


Increase of skeletal muscle relaxation speed by direct injection of parvalbumin cDNA.
Parvalbumin (PV) is a high affinity Ca(2+)-binding protein found at high concentration in fast-contracting/relaxing skeletal muscle fibers of vertebrates. It has been proposed that PV acts in the process of muscle relaxation by facilitating Ca2+ transport from the myofibrils to the sarcoplasmic reticulum. However, on the basis of metal-binding kinetics of PV in vitro, this hypothesis has been challenged. To investigate the function of PV in skeletal muscle fibers, direct gene transfer was applied in normal and regenerating rat soleus muscles which do not synthesize detectable amounts of PV. Two weeks after in vivo transfection with PV cDNA, considerable levels of PV mRNA and protein were detected in normal muscle, and even higher amounts were detected in regenerating muscle. Twitch half-relaxation time was significantly shortened in a dose-dependent way in transfected muscles, while contraction time remained unaltered. The observed shortening of half-relaxation time is due to PV and its ability to bind Ca2+, because a mutant protein lacking Ca(2+)-binding capacity did not promote any change in physiology. These results directly demonstrate the physiological function of PV as a relaxing factor in mammalian skeletal muscle.


Interferon-alpha/beta can impede development of carcinogen-induced squamous-cell tumors in the esophagus of C57B1 mice.
The effect of treatment with murine interferon-alpha/beta preparations on diethylnitrosamine-induced squamous-cell tumors in the esophagus of C57B1 mice was investigated. Diethylnitrosamine was administered in the drinking water for 18 weeks. Interferon was given intraperitoneally during the same 18 weeks or from the end of the period of carcinogen exposure until termination of the experiment. In mice given interferon and diethylnitrosamine synchronously, a significantly lower tumor index (number of tumors/cm of esophageal mucosa) was observed as compared to all control groups. Treatment with interferon after the administration of the carcinogen was terminated had no effect on the appearance of tumors. These data suggest that interferon therapy can exert certain effects on carcinogenesis.


Determination of chemical compositions and antibacterial effects of selected essential oils against human pathogenic strains.
Increasing the rates of drug resistant bacteria, having adverse effects and also high costs of antibiotics lead to essential oils (EOs) with antibacterial properties have gained importance. The present study was predicted to evaluate antibacterial activity of cinnamon, lavender, tea tree, lemon, coconut, oregano, mint, laurel and eucalyptus EOs alone and in combination. Chemical components of effective EOs were examined through gas chromatography/mass spectrometry (GC/MS). Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays were used to identify antibacterial effects of EOs against bacterial strains. The Fractional Inhibitory Concentration index (FICI) of the binary combinations of EOs was determined by checkerboard method. Carvacrol, linalool, linalyl acetate, 1,8-cineole, cinnamaldehyde, terpinen-4-ol and p-cymene were found main components of EOs. Oregano, cinnamon and tea tree EOs exhibited the strongest antibacterial activity with the MIC range between 0.03125-1.00% (v/v). Tea tree/lavender and cinnamon/lavender mixtures showed a synergistic effect against Streptococcus pyogenes and Streptococcus agalactiae. Oregano with tea tree and laurel exhibited a synergistic effect against Staphylococcus aureus. Oregano showed a synergistic effect when combined with cinnamon, lavender and tea tree against S.agalactiae. Our findings indicated that EOs either alone or in combination against pathogens should be preferred as potential antibacterial agents.


Perioperative Chemotherapy and Chemoradiotherapy for Patients With Resectable and Borderline Resectable Pancreatic Adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDA) is the third most common cause of cancer death in the United States. Most patients who undergo resection develop recurrence. Standard treatment confers a median overall survival (OS) of 24 months. Exposure to alternate regimens may prevent chemoresistance. This study evaluated multiagent perioperative therapy for potentially resectable PDA patients to improve OS. A single center, phase 2, trial of patients with resectable or borderline resectable PDA. Patients received neoadjuvant therapy with induction chemotherapy (gemcitabine, docetaxel, capecitabine) for 3 cycles, chemoradiation (intensity-modulated radiation therapy with capecitabine and oxaliplatin) followed by surgery, and 2 months of adjuvant gemcitabine and oxaliplatin and 2 months of gemcitabine. The primary endpoint was OS. The secondary endpoint was recurrence-free survival (RFS). Thirty-two eligible patients were enrolled. Twenty-two patients underwent surgical resection. After a median follow-up of 56.8 months, mOS was 31.6 months (95% confidence interval [CI], 14.2-58.1) for all patients, 58.1 months (95% CI, 31.6 to NR) for those who completed surgery. The mRFS was 31.3 months (95% CI, 12.5 to NR). Perioperative therapy with GTX, chemoradiotherapy, and adjuvant GemOx/Gem resulted in promising survival of 58 months for patients who underwent resection and may represent another treatment option for PDA.


Pesticides in groundwater at a national scale (France): Impact of regulations, molecular properties, uses, hydrogeology and climatic conditions.
Contaminants in groundwater are a major issue worldwide. Temporal trends of such occurrences in French groundwaters were evaluated for several active substances of pesticides belonging to different chemical classes, to identify key factors explaining groundwater contamination. Our study relied on exploitation of a French national database (ADES, created in the mid-1990s and remarkable for the available data, including over 88 million analyses). Temporal changes in the frequency of exceeding a reference value of 0.1 μg/L for several substances were determined at yearly and monthly scales. Such trends were examined by distinguishing different periods according to changes in regulations (new approval, withdrawal, or dose reduction), and were combined with data on effective rainfall as a proxy for groundwater recharge, on aquifer lithology, and on sales of active substances as a proxy for actual applications. A review of monthly data shows that a rapid transfer of pesticides with contrasting physico-chemical properties can occur after application in many aquifers, regardless of their lithology. For substances such as metolachlor, showing a sharp increase in sales, a clear relationship exists between quantities sold and frequency of exceeding the reference value. For other active substances, such as isoproturon or chlortoluron, frequencies of exceedance are governed by both sales and effective rainfall. Finally, the occurrence of active substances in groundwater several years after their withdrawal from the market is explained by at least three major mechanisms: the transfer time from soil into groundwater, processes of remobilization from soil and/or unsaturated zone, and no or low degradation in the saturated zone. While these processes are well documented for atrazine and different types of aquifers, they can be virtually unknown for other active substances.


Partially capitated managed care versus FFS for special needs children.
Little research has examined whether Medicaid managed care plans (MCPs) that incorporate case management are effective in coordinating services for children with special health care needs (CSHCN). This study evaluates the effects of enrollment of special needs children into a partially capitated MCP (with ongoing case management) versus the fee-for-service (FFS) option on use of therapeutic services, specifically speech, occupational, and physical therapy by site of service (school versus health care sector). Results show that special needs children enrolled in the partially capitated MCP are significantly more likely to obtain occupational and physical therapy at school relative to their FFS counterparts. Moreover, children enrolled in FFS are significantly less likely to be either regular or frequent users of each type of therapy relative to children enrolled in managed care. We attribute much of these disparities in use of therapeutic services at school to the availability of case management and coordination that is an integral component of the partially capitated MCP.


[Radioimmunodetection of prostatic cancer: in vivo 131I-gamma-seminoprotein for diagnosis of prostatic cancer by nuclear imaging].
69 patients with prostatic cancer were subjected to radioimmunodetection (RAID) with 131I-labeled antibody against gamma-seminoprotein (gamma-Sm), the images of malignant tumor sites was obtained by single photon emission computed tomography (SPECT) with dual radionuclide and computer processing. Of the 69 patients 66 were confirmed by RAID. The ratio of tumor tissue to normal tissue (T/N) 6.9 and the best time for incaging was 96 hours after injection of anti-gamma-Sm. The diameter of the minimum tumor in RAID was 0.5 cm. metastatic prostatic cancer in the pelvis or bone location as well as the origin tumors were also detected in 13 cases. Anti-gamma-SmRAID can differentiate benign from malignant prostatic neoplasms. In 37 cases of benign prostate hyperplasia, only two were positive. The positive detective rate of B-ultrasound and CT was 70.8% and 73.1% respectively.


The influence of food texture and liquid consistency modification on swallowing physiology and function: a systematic review.
Texture modification has become one of the most common forms of intervention for dysphagia, and is widely considered important for promoting safe and efficient swallowing. However, to date, there is no single convention with respect to the terminology used to describe levels of liquid thickening or food texture modification for clinical use. As a first step toward building a common taxonomy, a systematic review was undertaken to identify empirical evidence describing the impact of liquid consistency and food texture on swallowing behavior. A multi-engine search yielded 10,147 non-duplicate articles, which were screened for relevance. A team of ten international researchers collaborated to conduct full-text reviews for 488 of these articles, which met the study inclusion criteria. Of these, 36 articles were found to contain specific information comparing oral processing or swallowing behaviors for at least two liquid consistencies or food textures. Qualitative synthesis revealed two key trends with respect to the impact of thickening liquids on swallowing: thicker liquids reduce the risk of penetration-aspiration, but also increase the risk of post-swallow residue in the pharynx. The literature was insufficient to support the delineation of specific viscosity boundaries or other quantifiable material properties related to these clinical outcomes. With respect to food texture, the literature pointed to properties of hardness, cohesiveness, and slipperiness as being relevant both for physiological behaviors and bolus flow patterns. The literature suggests a need to classify food and fluid behavior in the context of the physiological processes involved in oral transport and flow initiation.


Investigation of dynamic properties of erbium fiber laser for ultrasonic sensing.
Dynamic properties of an erbium fiber laser (EFL) is researched and demonstrated for ultrasonic sensing in this research. The EFL has ring cavity incorporated with a phase-shifted fiber Bragg grating. A numerical model is used to analyze its dynamic responses to quasi-static change, continuous wave and burst wave. The ultrasonic behavior of the EFL resembles the forced single degree of freedom vibration with damping. Corresponding experimental results fit the simulation results well, showing some interesting ultrasonic properties of this EFL. After certain data process method, this EFL can be used in practical ultrasonic nondestructive testing.


Diagnosis of twin zygosity by mailed questionnaire.
A deterministic questionnaire method for zygosity determination is developed for use in epidemiological studies of adult twins. It is based on the answers of both members of a twin pair to two questions on similarity and confusion in childhood. The algorithm of the method is used to determine the zygosity status of a twin pair at two different levels of certainty. The validity of the method is tested by making blood marker determinations of 11 polymorphic marker systems fro a random sample of 104 twin pairs. The agreement between questionnaire and blood marker diagnosis was 100%, but the stricter level of certainty left 8.7% in the nonclassified group. The genetical representativeness of the sample is tested by the allele distribution of the markers as compared to the Finnish population data as well as by the distribution of the number of intra-pair differences in blood markers.


Insights into the Functions of eIF4E-Biding Motif of VPg in Potato Virus A Infection.
The interaction between the viral protein genome-linked (VPg) and eukaryotic initiation factor 4E (eIF4E) or eIF(iso)4E of the host plays a crucial role in potyvirus infection. The VPg of potato virus A (PVA) contains the Tyr-X-X-X-X-Leu-phi (YXXXLΦ) binding motif for eIF(iso)4E. In order to investigate its role in PVA infection, we substituted the conserved tyrosine and leucine residues of the motif with alanine residues in the infectious cDNA of PVA (PVAVPgmut). PVAVPgmut RNA replicated in infiltrated leaves, but RNA accumulation remained low. Systemic infection occurred only if a reversion to wild type PVA occurred. VPg was able to stabilize PVA RNA and enhance the expression of Renilla luciferase (3'RLUC) from the 3' end of the PVA genome. VPgmut could not support either PVA RNA stabilization or enhanced 3'RLUC expression. The RNA silencing suppressor helper-component proteinase (HCPro) is responsible for the formation of PVA-induced RNA granules (PGs) during infection. While VPgmut increased the number of PG-like foci, the percentage of PVA RNA co-localization with PGs was reduced from 86% to 20%. A testable hypothesis for future studies based on these results is that the binding of eIF(iso)4E to PVA VPg via the YXXXLΦ motif is required for PVA RNA stabilization, as well as the transfer to the RNA silencing suppression pathway and, further, to polysomes for viral protein synthesis.


Layer-by-layer transfer of multiple, large area sheets of graphene grown in multilayer stacks on a single SiC wafer.
Here we report a technique for transferring graphene layers, one by one, from a multilayer deposit formed by epitaxial growth on the Si-terminated face of a 6H-SiC substrate. The procedure uses a bilayer film of palladium/polyimide deposited onto the graphene coated SiC, which is then mechanically peeled away and placed on a target substrate. Orthogonal etching of the palladium and polyimide leaves isolated sheets of graphene with sizes of square centimeters. Repeating these steps transfers additional sheets from the same SiC substrate. Raman spectroscopy, scanning tunneling spectroscopy, low-energy electron diffraction and X-ray photoelectron spectroscopy, together with scanning tunneling, atomic force, optical, and scanning electron microscopy reveal key properties of the materials. The sheet resistances determined from measurements of four point probe devices were found to be ∼2 kΩ/square, close to expectation. Graphene crossbar structures fabricated in stacked configurations demonstrate the versatility of the procedures.


Variants in several genomic regions associated with asperger disorder.
Asperger disorder (ASP) is one of the autism spectrum disorders (ASD) and is differentiated from autism largely on the absence of clinically significant cognitive and language delays. Analysis of a homogenous subset of families with ASP may help to address the corresponding effect of genetic heterogeneity on identifying ASD genetic risk factors. To examine the hypothesis that common variation is important in ASD, we performed a genome-wide association study (GWAS) in 124 ASP families in a discovery data set and 110 ASP families in a validation data set. We prioritized the top 100 association results from both cohorts by employing a ranking strategy. Novel regions on 5q21.1 (P = 9.7 × 10(-7) ) and 15q22.1-q22.2 (P = 7.3 × 10(-6) ) were our most significant findings in the combined data set. Three chromosomal regions showing association, 3p14.2 (P = 3.6 × 10(-6) ), 3q25-26 (P = 6.0 × 10(-5) ) and 3p23 (P = 3.3 × 10(-4) ) overlapped linkage regions reported in Finnish ASP families, and eight association regions overlapped ASD linkage areas. Our findings suggest that ASP shares both ASD-related genetic risk factors, as well as has genetic risk factors unique to the ASP phenotype.


Mechanisms of host wasting induced by administration of cytokines in rats.
This study determined the effects of chronic administration of the two principal proximate cytokines, interleukin-1 (IL-1) and tumor necrosis factor (TNF), on host wasting in rats. The effects were compared with those observed in a matched pair-fed group to distinguish the contribution from anorexia. Both TNF and IL-1 produced weight loss, net nitrogen loss, and skeletal muscle catabolism and increased liver weight. Such effects were independent from and additive to those resulting from semistarvation. However, under equivalent nutritional conditions, TNF infusion led to a greater effect on muscle protein catabolism and liver weight and caused liver protein anabolism, whereas only the group receiving IL-1 had altered glucose metabolism in the postabsorptive state. Tachyphylaxis was seen in the response of food intake over time after administration of IL-1. These actions define the two principal mechanisms for the development of protein calorie malnutrition that occur because of cytokine action, anorexia producing semistarvation and an increased net protein catabolic rate reducing anabolic efficiency.


Performance Changes of Elite Paralympic Judo Athletes During a Paralympic Games Cycle: A Case Study with the Brazilian National Team.
The aim of this study was to describe the variations in power performance of elite Paralympic judo athletes across three consecutive training cycles of preparation for the ParaPan American Games, the World Championship and the Paralympic Games. Eleven Paralympic judokas from the Brazilian National team participated in this study. They were repeatedly assessed using squat and countermovement jumps, mean propulsive power (MPP) in the jump-squat (JS), the bench press and prone bench pull at several moments of the preparation. Training supervision based on the optimum power zone (range of loads where power production is maximized) was provided in the final cycle, prior to the Paralympic Games. Magnitude-based inference was used to compare the repeated measurements of power performance. Lower and upper limb muscle power gradually increased throughout the cycles; however, the best results in all exercises were observed prior to the Paralympic Games, during which the team won four silver medals. As an illustration, prior to participation in the Paralympic Games the MPP in the JS was likely to very likely higher than prior to the World Championship (effect size [ES] = 0.77) and ParaPan American Games (ES = 0.53), and in January and March 2016 (ES = 0.98 and 0.92, respectively; months preceding the Paralympic Games). Power performance assessments can provide information about the evolution of Paralympic judokas, and training at the optimum power zone seems to constitute an effective method to improve lower and upper limb power in these athletes.


Antiangiogenic cancer therapy: monitoring with molecular US and a clinically translatable contrast agent (BR55).
To develop and test human kinase insert domain receptor (KDR)-targeted microbubbles (MBs) (MB(KDR)) for imaging KDR at the molecular level and for monitoring antiangiogenic therapy in a human colon cancer xenograft tumor model in mice. Animal studies were approved by the Institutional Administrative Panel on Laboratory Animal Care. A heterodimeric peptide that binds to human KDR with low nanomolar affinity (K(D) = 0.5 nmol/L) was coupled onto the surface of perfluorobutane-containing lipid-shelled MBs (MB(KDR)). Binding specificity of MB(KDR) to human KDR and cross-reactivity with murine vascular endothelial growth factor (VEGF) receptor 2 (VEGFR2) were tested in cell culture under flow shear stress conditions (at 100 sec(-1)). In vivo binding specificity of MB(KDR) to VEGFR2 was tested in human LS174T colon cancer xenografts in mice with a 40-MHz ultrasonographic (US) transducer. Targeted contrast material-enhanced US imaging signal by using MB(KDR) was longitudinally measured during 6 days in tumors with (n = 6) and without (n = 6) antiangiogenic treatment (anti-VEGF antibody). Ex vivo VEGFR2 staining and microvessel density analysis were performed. Significant differences were evaluated (t, Mann-Whitney, or Wilcoxon test). Cell culture experiments showed four times greater binding specificity of MB(KDR) to human KDR and cross-reactivity to murine VEGFR2 (P < or = .01). In vivo imaging signal was more than three times higher (P = .01) with MB(KDR) compared with control MBs and decreased significantly (approximately fourfold lower, P = .03) following in vivo receptor blocking with anti-VEGFR2 antibody. One day after initiation of antiangiogenic therapy, imaging signal was significantly decreased (approximately 46% lower, P = .02) in treated versus untreated tumors; it remained significantly lower (range, 46%-84% decreased; P = .038) during the following 5 days. Microvessel density was significantly reduced (P = .04) in treated (mean, 7.3 microvessels per square millimeter +/- 4.7 [standard deviation]) versus untreated tumors (mean, 22.0 microvessels per square millimeter +/- 9.4); VEGFR2 expression was significantly decreased (>50% lower, P = .03) in treated tumors. Human MB(KDR) allow in vivo imaging and longitudinal monitoring of VEGFR2 expression in human colon cancer xenografts.


How does pulmonary rehabilitation survive in a managed care market?
As managed care enrollment increases, pulmonary rehabilitation coordinators need to become well versed in the terminology and grasp the way managed care organizations operate. Maintaining access to pulmonary rehabilitation services for chronic lung disease patients is imperative. Learning how to obtain treatment referrals and authorizations has become challenging. Using offered resources can aid program coordinators in their struggle to survive in an ever-changing health care delivery system. Preventative and restorative rehabilitation intervention saves utilization of expensive health care resources.


NaF iontophoresis, an alternative to cavity lining--an in vivo study.
The study carried out on 30 teeth in 28 patients of 12-16 years to assess NaF iontophoresis as an alternative to cavity lining revealed that out of 25 cavities which were sensitive to air, water jet and exploration (14 mild, 9 moderate and 2 severe), in 20 cavities the sensitivity decreased to nil, in 9 to mild and in one to moderate following iontophoresis. It was found that in all the cases, the first application of iontophoresis decreased the sensitivity by 50-60 percent and the second by 70-90 percent. Immediately after silver filling there was no sensitivity in 25 teeth, whereas 4 were mildly and one moderately sensitive. After a six month follow up in 24 patients who reported, all the teeth were comfortable.


Relaxation dynamics in fluids of platelike colloidal particles.
The relaxation dynamics of a model fluid of platelike colloidal particles is investigated by means of a phenomenological dynamic density functional theory. The model fluid approximates the particles within the Zwanzig model of restricted orientations. The driving force for time dependence is expressed completely by gradients of the local chemical potential, which in turn is derived from a density functional-hydrodynamic interactions are not taken into account. These approximations are expected to lead to qualitatively reliable results for low densities like those within the isotropic-nematic two-phase region. The formalism is applied to model an initially spatially homogeneous stable or metastable isotropic fluid which is perturbed by switching a two-dimensional array of Gaussian laser beams. Switching on the laser beams leads to an accumulation of colloidal particles in the beam centers. If the initial chemical potential and the laser power are large enough, a preferred orientation of particles occurs, breaking the symmetry of the laser potential. After switching off the laser beams again, the system can follow different relaxation paths: It either relaxes back to the homogeneous isotropic state or it forms an approximately elliptical high-density core which is elongated perpendicular to the dominating orientation in order to minimize the surface free energy. For large supersaturations of the initial isotropic fluid, the high-density cores of neighboring laser beams of the two-dimensional array merge into complex superstructures.


Characterization of wear debris in total elbow arthroplasty.
The purpose of this study was to evaluate wear debris in periprosthetic tissues at the time of revision total elbow arthroplasty. Polyethylene, metallic, and bone cement debris were characterized, and the tissue response was quantified. Capsular and medullary tissue samples were collected during revision surgery. Polyethylene debris was characterized by scanning electron microscopy after tissue digestion. The concentrations of metal and cement debris were quantified by inductively coupled plasma mass spectrometry. Tissue response was graded with a semiquantitative histologic method. Polyethylene particle size varied from the submicron range to over 100 μm. The mean diameter ranged from 0.6 μm to about 1 μm. Particles in the synovial tissues were larger and less abundant than those in tissues from the medullary canal. Cement, titanium alloy, and low levels of cobalt-chrome debris were also present, with cement predominating over metal debris. Histiocyte response was associated with small polyethylene particles (0.5-2 μm), and giant cells were associated with large polyethylene particles (>2 μm). Histiocyte scores positively correlated with the polyethylene particle number and the presence of metal. We have shown that periprosthetic tissues of total elbow patients who have undergone revision for loosening and osteolysis contain polyethylene, cement, and metal debris. Although the polyethylene particles were of a size and shape that have been previously shown to result in activation of phagocytic cells, osteolysis after total elbow arthroplasty is a multimodal process. Because of the presence of multiple wear particle sources, a cause-and-effect relationship between polyethylene debris and osteolysis cannot be established with certainty.


Protein tyrosine phosphatase nonreceptor type 2 exerts antimetastatic functions in pancreatic ductal adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is a highly invasive tumor with a dismal prognosis. Recent studies have demonstrated PTPN2 (protein tyrosine phosphatase nonreceptor type 2) as a potential target for cancer therapy. However, the functions of PTPN2 in PDAC progression remain poorly understood. In this study, we found PTPN2 expression was downregulated in PDAC tissues, and decreased PTPN2 expression was associated with unfavorable prognosis. Functional studies indicated that PTPN2 knockdown promoted the migration and invasion abilities of PDAC cells in vitro, and the liver metastasis in vivo through epithelial-mesenchymal transition process. Mechanistically, MMP-1 was identified as a downstream target of PTPN2 via RNA-seq data and was responsible for the enhanced metastasis of PDAC cells upon PTPN2 knockdown. Moreover, according to chromatin immunoprecipitation and electrophoretic mobility shift assay, PTPN2 depletion transcriptionally activated MMP-1 via regulating the interaction of p-STAT3 with its distal promoter. This study, for the first time, demonstrated that PTPN2 inhibited PDAC metastasis, and presented a novel PTPN2/p-STAT3/MMP-1 axis in PDAC progression.


Trends of Sex Differences in Clinical Outcomes After Myocardial Infarction in the United States.
Female patients have been shown to experience worse clinical outcomes after acute myocardial infarction (AMI) compared with male patients. However, it is unclear what trend these differences followed over time. Data from patients hospitalized with AMI between 2004 and 2015 in the National Inpatient Sample were retrospectively analyzed, stratified according to sex. Multivariable logistic regression analyses were performed to examine the adjusted odds ratios (aORs) of invasive management and in-hospital outcomes according to sex. The Mantel-Haenszel extension of the χ2 test was performed to examine the trend of management and in-hospital outcomes over the study period. Of 7,026,432 AMI hospitalizations, 39.7% (n = 2,789,494) were women. Overall, women were older (median: 77 vs 70 years), with a higher prevalence of risk factors such as diabetes, hypertension, and depression. Women were less likely to receive coronary angiography (aOR, 0.92; 95% confidence interval [CI], 0.91-0.93) and percutaneous coronary intervention (aOR, 0.82; 95% CI, 0.81-0.83) compared with men. Odds of all-cause mortality were higher in women (aOR, 1.03; 95% CI, 1.02-1.04; P < 0.001) and these rates have not narrowed over time (2004 vs 2015: aOR, 1.07 [95% CI, 1.04-1.09] vs 1.11 [95% CI, 1.07-1.15), with similar observations recorded for major adverse cardiovascular and cerebrovascular events. In this temporal analysis of AMI hospitalizations over 12 years, we showed lower receipt of invasive therapies and higher mortality rates in women, with no change in temporal trends. There needs to be a systematic and consistent effort toward exploring these disparities to identify strategies to mitigate them.


Multiple endosomal recycling pathways in rat adipose cells.
Adipose and skeletal-muscle cells can translocate several membrane proteins from intracellular compartment(s) to the cell surface in an insulin-dependent fashion. Among these proteins is Glut4, a physiologically important glucose transporter which mediates insulin's effect on blood glucose clearance. Under basal conditions, Glut4 is localized in uniform, intracellular membrane vesicles with an average diameter of 50-70 nm and a sedimentation coefficient of 100-120 S. The nature of this compartment and its trafficking pathway to the plasma membrane is still unresolved. We show here that, in addition to Glut4, the aminopeptidase gp160 or insulin-responsive aminopeptidase ('IRAP'), sortilin, and an acutely recycling population of the insulin-like growth factor-II/mannose 6-phosphate receptor, this compartment includes 60% of the intracellular population of the transferrin receptor. We used subcellular fractionation, cell-surface biotinylation, and radioactive-ligand (125I-transferrin) uptake to demonstrate that the transferrin receptor recycles between this compartment and the plasma membrane in response to insulin along with Glut4 and other protein components of these vesicles. The co-localization of Glut4 and several endosomal markers in the terminally differentiated fat-cells during several stages of their cycling pathways suggests that the 'Glut4 pathway' may derive from the hormone-insensitive endosomes of undifferentiated preadipocytes. The insulin receptor is excluded from Glut4-containing vesicles in both insulin-stimulated and unstimulated adipocytes, and thus it is likely to traffic independently from Glut4 through different intracellular compartments. Our data show that, in adipose cells, the ligand-dependent recycling pathway of the insulin receptor is structurally separated from the ligand-independent pathway of the transferrin receptor, and that Glut4 is specifically targetted to the latter.


Role of the promoter in activation of transcription by nitrogen regulator I phosphate in Escherichia coli.
The protein nitrogen regulator I (NRI)-phosphate is known to activate the initiation of transcription of the Escherichia coli glnA gene. This activation is facilitated by the binding of the protein to NRI-specific sites located upstream of the sigma 54-dependent glnA promoter. To determine whether binding of NRI-phosphate to upstream sites is sufficient for activation, we placed several promoters not normally activated by NRI-phosphate downstream of NRI binding sites and measured activation in intact cells and in an in vitro transcription system. We found that the sigma 70-dependent lac promoter was not activated, that the sigma 54-dependent Klebsiella pneumoniae nifH promoter was weakly activated, and that a nifH promoter altered in the RNA polymerase binding site was almost as well activated as the glnA promoter. We conclude that the sensitivity of the susceptible promoter depends on the presence of NRI binding sites, but that the presence of bound NRI-phosphate upstream of a promoter is not sufficient for activation of transcription by RNA polymerase. This activation is determined by the structure of the RNA polymerase binding site. We suggest that sigma 54-but not sigma 70-dependent promoters are susceptible to activation by NRI-phosphate and that the nucleotide sequence of the sigma 54-RNA polymerase binding site is an important determinant of the efficiency of activation.


A complex network regulates expression of eps and other virulence genes of Pseudomonas solanacearum.
We have discovered an unusual and complex regulatory network used by the phytopathogen Pseudomonas solanacearum to control transcription of eps, which encodes for production of its primary virulence factor, the exopolysaccharide EPS I. The major modules of this network were shown to be three separate signal transduction systems: PhcA, a LysR-type transcriptional regulator, an dual two-component regulatory systems, VsrA/VsrD and VsrB/VsrC. Using lacZ fusions and RNA analysis, we found that both PhcA and VsrA/VsrD control transcription of another network component, xpsR, which in turn acts in conjunction with vsrB/vsrC to increase transcription of the eps promoter by > 25-fold. Moreover, gel shift DNA binding assays showed that PhcA specifically binds to the xpsR promoter region. Thus, the unique XpsR protein interconnects the three signal transduction systems, forming a network for convergent control of EPS I in simultaneous response to multiple environmental inputs. In addition, we demonstrate that each individual signaling system of the network also acts independently to divergently regulate other unique sets of virulence factors. The purpose of this complex network may be to allow this phytopathogen to both coordinately or independently regulate diverse virulence factors in order to cope with the dynamic situations and conditions encountered during interactions with plants.


Barcoding Queensland Fruit Flies (Bactrocera tryoni): impediments and improvements.
Identification of adult fruit flies primarily involves microscopic examination of diagnostic morphological characters, while immature stages, such as larvae, can be more problematic. One of the Australia's most serious horticultural pests, the Queensland Fruit Fly (Bactrocera tryoni: Tephritidae), is of particular biosecurity/quarantine concern as the immature life stages occur within food produce and can be difficult to identify using morphological characteristics. DNA barcoding of the mitochondrial Cytochrome Oxidase I (COI) gene could be employed to increase the accuracy of fruit fly species identifications. In our study, we tested the utility of standard DNA barcoding techniques and found them to be problematic for Queensland Fruit Flies, which (i) possess a nuclear copy (a numt pseudogene) of the barcoding region of COI that can be co-amplified; and (ii) as in previous COI phylogenetic analyses closely related B. tryoni complex species appear polyphyletic. We found that the presence of a large deletion in the numt copy of COI allowed an alternative primer to be designed to only amplify the mitochondrial COI locus in tephritid fruit flies. Comparisons of alternative commonly utilized mitochondrial genes, Cytochrome Oxidase II and Cytochrome b, revealed a similar level of variation to COI; however, COI is the most informative for DNA barcoding, given the large number of sequences from other tephritid fruit fly species available for comparison. Adopting DNA barcoding for the identification of problematic fly specimens provides a powerful tool to distinguish serious quarantine fruit fly pests (Tephritidae) from endemic fly species of lesser concern.


Pathophysiological and behavioral deficits in developing mice following rotational acceleration-deceleration traumatic brain injury.
Abusive head trauma (AHT) is the leading cause of death from trauma in infants and young children. An AHT animal model was developed on 12-day-old mice subjected to 90° head extension-flexion sagittal shaking repeated 30, 60, 80 and 100 times. The mortality and time until return of consciousness were dependent on the number of repeats and severity of the injury. Following 60 episodes of repeated head shakings, the pups demonstrated apnea and/or bradycardia immediately after injury. Acute oxygen desaturation was observed by pulse oximetry during respiratory and cardiac suppression. The cerebral blood perfusion was assessed by laser speckle contrast analysis (LASCA) using a PeriCam PSI system. There was a severe reduction in cerebral blood perfusion immediately after the trauma that did not significantly improve within 24 h. The injured mice began to experience reversible sensorimotor function at 9 days postinjury (dpi), which had completely recovered at 28 dpi. However, cognitive deficits and anxiety-like behavior remained. Subdural/subarachnoid hemorrhage, damage to the brain-blood barrier and parenchymal edema were found in all pups subjected to 60 insults. Proinflammatory response and reactive gliosis were upregulated at 3 dpi. Degenerated neurons were found in the cerebral cortex and olfactory tubercles at 30 dpi. This mouse model of repetitive brain injury by rotational head acceleration-deceleration partially mimics the major pathophysiological and behavioral events that occur in children with AHT. The resultant hypoxia/ischemia suggests a potential mechanism underlying the secondary rotational acceleration-deceleration-induced brain injury in developing mice.


Current evidence on the potential therapeutic applications of transcranial magnetic stimulation in multiple sclerosis: A systematic review of the literature.
A growing number of studies have evaluated the effects of transcranial magnetic stimulation (TMS) for the symptomatic treatment of multiple sclerosis (MS). We performed a PubMed search for articles, recent books, and recommendations from the most relevant clinical practice guidelines and scientific societies regarding the use of TMS as symptomatic treatment in MS. Excitatory electromagnetic pulses applied to the affected cerebral hemisphere allow us to optimise functional brain activity, including the transmission of nerve impulses through the demyelinated corticospinal pathway. Various studies into TMS have shown statistically significant improvements in spasticity, fatigue, lower urinary tract dysfunction, manual dexterity, gait, and cognitive deficits related to working memory in patients with MS; however, the exact level of evidence has not been defined as the results have not been replicated in a sufficient number of controlled studies. Further well-designed, randomised, controlled clinical trials involving a greater number of patients are warranted to attain a higher level of evidence in order to recommend the appropriate use of TMS in MS patients across the board. TMS acts as an adjuvant with other symptomatic and immunomodulatory treatments. Additional studies should specifically investigate the effect of conventional repetitive TMS on fatigue in these patients, something that has yet to see the light of day.


Interleukin-1 generates transmembrane signals from phospholipids through novel pathways in cultured rat mesangial cells.
Although IL-1 stimulates cellular responses in both lymphoid and nonlymphoid cells, the second messengers by which IL-1 activates cells are unknown. Recombinant IL-1 alpha (rIL-1) is a comitogen for glomerular mesangial cells. Using this model we explored potential transmembrane signals by which IL-1 stimulates cellular responses. Certain mitogens hydrolyze inositol phospholipids by phospholipase C to generate 1,2-diacylglycerol, a cofactor for protein kinase C, and inositol (1,4,5)-trisphosphate, which mobilizes intracellular calcium. rIL-1 induced a peak increase in [3H]1,2-diacylglycerol formation at 1 min. Production of 1,2-diacylglycerol often parallels the generation of phosphatidic acid; however, rIL-1 stimulated [32P]phosphatidate formation only after 60 min. rIL-1 did not change the inositol phosphate or cytosolic free calcium concentrations, demonstrating that rIL-1 does not activate an inositol phospholipid-specific phospholipase C. [3H]Phosphorylethanolamine, but not [3H]phosphorylserine or [3H]phosphorylcholine, was maximally elevated at 1 min in mesangial cells incubated with rIL-1. Radioactivity incorporated into phosphatidylethanolamine but not phosphatidylcholine was also decreased in IL-1-stimulated mesangial cells compared with control at 1 min. These data suggest that rIL-1 activates a phospholipase C predominantly linked to phosphatidylethanolamine. In contrast to other mitogens, rIL-1 did not alter intracellular pH. Both 12-0-tetradecanoyl-phorbol-13-acetate, a homologue of 1,2-diacylglycerol, and phosphatidate but not phosphatidylcholine in the presence of 0.5% fetal bovine serum stimulated mesangial cell proliferation. rIL-1-induced cellular activation may be mediated, at least in part, by phospholipid-derived second messengers generated through novel pathways.


Glutamine synthetase immunostaining correlates with pathologic features of hepatocellular carcinoma and better survival after radiofrequency thermal ablation.
Activation of the wnt pathway identifies a subgroup of hepatocellular carcinomas (HCC) with specific epidemiologic and genetic profiles. Wnt activation is predicted by mutation and/or nuclear translocation of beta-catenin and by glutamine synthetase (GS) immunoreactivity. We investigated whether GS staining associates with specific pathologic features of HCC and with survival after radiofrequency thermal ablation. Monoistitutional retrospective-prospective study in a tertiary hospital setting. Two hundred and seven cirrhotics (mean age, 70 years; 63% males, 82.1% hepatitis C virus positive) with early HCC were consecutively treated with radiofrequency thermal ablation (RFTA). Mean tumor size was 2.7 cm; 20.3% of patients had multiple nodules; and median follow-up was 36 months with 54.6% overall mortality. Tumor samples were mainly obtained by biopsy (92,5%) and examined by H&E and immunostaining for beta-catenin and GS. Main outcome measures were overall and tumor-specific mortality by Kaplan-Meier analysis and Cox proportional hazard models corrected for competing risks. Ninety-one patients (43.9%) had GS-positive HCCs by immunostaining. These tumors had larger size (P = 0.012) and characteristic histology (low grade, pseudoacini, hydropic changes, bile staining, lack of steatosis, and fibrosis). Other clinical or treatment variables were similar between groups. Variables correlating with tumor-specific and overall mortality by univariate analysis were tumor recurrence, advanced disease, posttreatment alpha-fetoprotein levels, and GS staining. Yearly, overall mortality rate was lower in GS-positive patients (12.4 versus 20% yearly; P = 0.006). By multivariate analysis, GS immunostaining correlated with reduced specific (hazard ratio, 0.58; 95% confidence interval, 0.34-0.97) and overall mortality (hazard ratio, 0.62; 95% confidence interval, 0.40-0.96). Standard histology and GS status identify a HCC subset with distinct clinical and pathologic features.


Clinical and radiographic features of sarcoidosis in menopausal women--the impact on therapeutic approach and prognosis.
Sarcoidosis is a multisystemic granulomatous disease of unknown etiology. The aim of the study was to determine are there significant differences in clinical manifestations, radiographic and lung function findings and therapeutic approach in menopausal female sarcoidosis pa-tients compared to premenopausal ones. Seventy seven Caucasian women (average age 43.71 years, range 38-54) with sarcoidosis diagnosed at the University Hospital from January to October 2006, were included in the study. They were divided into two groups according to their menstrual period status. The group I included 42 women with normal menstrual cycle, while the group II included 35 menopausal women (either spontaneous or after hysterectomy). The patients were not under hormonal therapy. We found significantly higher proportion of the first radiographic stage (66.7% vs 34.2%, p < 0.05) and acute form of sarcoidosis (57.2% vs 17.1%; p < 0.01) in the group I in relation to the group II. Extrapulmonary sarcoidosis was more frequent in the group II than in the group I (p < 0.01). Disturbances of lung ventilation were registered in 50.8% of all the patients, and decrease of one or both of diffusion parameters was found in 63.6%, but without significant differences between the groups (p > 0.05). Hypercalciuria was found in 19.1% of the patients in the group I and 42.8% of the patient in the group II (p < 0.05). A difference in the therapy approach was also found to be significant with methotrexate more frequently applied in the group II than in the group I (p < 0.01). Menopausal women with sarcoidosis may represent a group of patients that requires special attention in diagnostic procedure, therapeutic approach and follow-up, to prevent unfavourable course of the disease. Attention should be particularly focused on the detection of extrapulmonary sites involvement in this group of the patients. Further prospective studies are needed to reveal the role of hormones, and especially plasma estrogen level in sarcoidosis appearance.


Langerhans Cell Histiocytosis in a Three-Year-Old Girl in Bosnia and Herzegovina.
To present a rare disease, and to point out that clinical manifestations treated for a long period of time without an adequate response to therapy may be a manifestation of a rare disease. We present the case of a 3-year-old girl who had been drinking a large amount of water for the previous ten days with frequent urination, and who experienced the worsening of symptoms of scalp dermatitis that had been treated for a year without success. Physical examination revealed a maculopapular rash on the scalp, neck and both ear shells, and exophthalmos of the right eye with periorbital edema. Magnetic resonance imaging of the orbits showed extensive lesions of the skull bones. Further diagnostic evaluation revealed similar lesions in other bone structures. Biopsy of the affected region, microscopic and immunohistochemical analysis led to diagnosis of Langerhans cell histiocytosis. Langerhans cell histiocytosis mostly occurs in the first three years of life. The incidence is 4-5 patients per million children under 15 years of life. The clinical presentation is highly variable, and can range from isolated, self-healing skin and bone lesions to life-threatening multisystem diseases. Due to the diverse clinical picture, that is often unrecognized, these patients are often referred to other specialists, resulting in the treatment of individual symptoms rather than the underlying disease.


[Urinary infection in adolescents]
OBJECTIVE: Urinary tract infections are caused by various gram-negative bacteria. Escherichia coli is the most common causative agent in all groups of patients. Staphylococcus saprophyticus is now recognized as a common cause of urinary tract infection in adolescents and young adult women. METHODS: Review of literature and clinical experience with urinary tract infections in adolescents at the Pediatric Nephrology Department of Hospital Santa Casa de São Paulo. RESULTS: Lower urinary tract infection, hematuria and dysuria were the most frequent symptoms observed in infections caused by Staphylococcus saprophyticus. Some asymptomatic cases were observed. The infection may be associated with adverse effects of maternal and fetal health during pregnancy. CONCLUSIONS: This article reviews the diagnosis, etiology, treatment, and complications associated with urinary tract infections in adolescents.


Total synthesis of (+)-schweinfurthins B and E.
The first total synthesis of (+)-schweinfurthin B, a potent and differentially active cytotoxic agent, has been accomplished. Completion of the synthesis required just 16 steps in the longest linear sequence from commercially available vanillin. Key synthetic transformations included a Shi epoxidation and an efficient cascade cyclization initiated by treatment of the resulting epoxide with BF(3).OEt(2). Furthermore, use of a methyl ether as a stable protecting group for benzylic alcohols dramatically increased the efficiency of the overall sequence. The benzylic ether can be removed from this electron-rich aromatic system through oxidation with DDQ that provided the desired aldehyde intermediate in quantitative yield and shortened the synthetic sequence. Introduction of the A-ring diol in the required cis stereochemistry then became viable through a short sequence highlighted by an aldol condensation with benzaldehyde to introduce an olefin as a latent carbonyl group at the C-3 position. This synthesis established for the first time the absolute stereochemistry of the natural product, and provides access to material on a scale that will advance biological studies. The total synthesis of the closely related compound (+)-schweinfurthin E also is reported.


Transcutaneous hydrogen clearance--a new non-invasive technique for assessing blood flow in human skin.
A new technique for the non-invasive assessment of skin blood flow is described using a modified, commercially available transcutaneous oxygen electrode. Initially, experiments were carried out on normal volunteers at skin temperatures of 40 degrees C and 44 degrees C. Transcutaneous H2 (tcpH2) clearances were measured after the subject had breathed a mixture of 2% H2 in air until a saturation value in the skin had been reached. tcpH2 clearance results were compared with laser Doppler flux values measured simultaneously on adjacent sites and at the same temperatures under control and reactive hyperaemic conditions. Good reproducibility for the tcpH2 technique and significant correlation (p < 0.002) between it and laser Doppler flux values at the same temperatures was demonstrated. In further experiments, (I125) 4-iodoantipyrine clearances (IAP) were carried out immediately prior to and at the same sites as tcpH2 clearances in critically ischaemic lower limbs. The correlation between the methods and the differences between tcpH2 values observed in normal and critically ischaemic skin were highly significant (p < 0.002).