Treatment of 5 dogs with immune-mediated thrombocytopenia using Romiplostim.
Immune thrombocytopenia (ITP) in dogs is analogous to that in humans. Romiplostim, a novel thrombopoietin receptor (TPO-R) agonist, is currently used for the treatment of refractory ITP in humans, but not in dogs. Here, we describe the response to romiplostim in five dogs with refractory ITP. Five dogs with severe and refractory ITP (three primary and two secondary) received romiplostim subcutaneously. Four dogs were administered 3-5 μg/kg and one dog received 10-13 μg/kg body weight once weekly. Romiplostim was well-tolerated and administration was associated with an increase in platelet counts in all five dogs. Four of the five dogs entered remission and relapses were not observed over a follow-up period of 3-10 months. Romiplostim is effective in the treatment of ITP in dogs at least as well as in humans. This finding may help to develop and use new therapeutics for ITP in dogs and humans.


Influence of different control strategies on muscle activation patterns in trunk muscles.
Adequate training of the trunk muscles is essential to prevent low back pain. Although sit-ups are simple to perform, the perceived high effort is the reason why training the abdominal muscles is seldom continued over a longer period of time. It is well known that the abdominal muscles are inferior to the back muscles in terms of force, but this cannot explain the extreme difference in perceived effort between trunk flexion and extension tasks. Therefore, this study was aimed at the identification of control strategy influences on the muscular stress level. Thirty-nine subjects were investigated. The performed tasks were restricted to the sagittal plane and were implemented with simulated and realized tilt angles. Subjects were investigated in an upright position with their lower bodies fixed and their upper bodies free. Posture-controlled tasks involved graded forward and backward tilting, while force-controlled tasks involved the application of force based on a virtual tilt angle. The Surface EMG (SEMG) was taken from five trunk muscles on both sides. Control strategies seemed to have no systematic influence on the SEMG amplitudes of the back muscles. In contrast, the abdominal muscles exhibited significantly higher stress levels under posture-controlled conditions without relevantly increasing antagonistic co-activation of back muscles. The abdominal muscles' relative differences ranged from an average of 20% for the external oblique abdominal muscle to approximately 40% for the rectus abdominal muscle. The perceived high effort expended during sit-ups can now be explained by the posture-controlled contractions that are required.


Intracranial Recordings of Occipital Cortex Responses to Illusory Visual Events.
Ambiguous visual stimuli elicit different perceptual interpretations over time, creating the illusion that a constant stimulus is changing. We investigate whether such spontaneous changes in visual perception involve occipital brain regions specialized for processing visual information, despite the absence of concomitant changes in stimulation. Spontaneous perceptual changes observed while viewing a binocular rivalry stimulus or an ambiguous structure-from-motion stimulus were compared with stimulus-induced perceptual changes that occurred in response to an actual stimulus change. Intracranial recordings from human occipital cortex revealed that spontaneous and stimulus-induced perceptual changes were both associated with an early transient increase in high-frequency power that was more spatially confined than a later transient decrease in low-frequency power. We suggest that the observed high-frequency and low-frequency modulations relate to initiation and maintenance of a percept, respectively. Our results are compatible with the idea that spontaneous changes in perception originate from competitive interactions within visual neural networks. Ambiguous visual stimuli elicit different perceptual interpretations over time, creating the illusion that a constant stimulus is changing. The literature on the neural correlates of conscious visual perception remains inconclusive regarding the extent to which such spontaneous changes in perception involve sensory brain regions. In an attempt to bridge the gap between existing animal and human studies, we recorded from intracranial electrodes placed on the human occipital lobe. We compared two different kinds of ambiguous stimuli, binocular rivalry and the phenomenon of ambiguous structure-from-motion, enabling generalization of our findings across different stimuli. Our results indicate that spontaneous and stimulus-induced changes in perception (i.e., "illusory" and "real" changes in the stimulus, respectively) may involve sensory regions to a similar extent.


Targeted metabolomic analysis of 33 amino acids and biogenic amines in human urine by ion-pairing HPLC-MS/MS: Biomarkers for tacrolimus nephrotoxicity after renal transplantation.
Calcineurin inhibitor nephrotoxicity, especially for the widely used tacrolimus, has become a major concern in post-transplant immunosuppression. Multiparametric amino acid metabolomics is useful for biomarker identification of tacrolimus nephrotoxicity, for which specific quantitative methods are highlighted as a premise. This article presents a targeted metabolomic assay to quantify 33 amino acids and biogenic amines in human urine by high-performance liquid chromatography coupled with tandem mass spectrometry. Chromatographic separation was carried out on an Agilent Zorbax SB-C18 column (3.0 × 150 mm, 5 μm) with addition of an ion-pairing agent in the mobile phase, and MS/MS detection was achieved in both the positive and negative multiple reaction monitoring modes. Good correlation coefficients (r2 > 0.98) were obtained for most analytes. Intra- and inter-day precision, stability, carryover and incurred sample reanalysis met with the acceptance criteria of the guidance of the US Food and Drug Administration. Analysis on urine from healthy volunteers and renal transplantation patients with tacrolimus nephrotoxicity confirmed symmetric dimethylarginine and serine as biomarkers for kidney injury, with AUC values of 0.95 and 0.81 in receiver operating characteristic analysis, respectively. Additionally, symmetric dimethylarginine exhibited a tight correlation with serum creatinine, and was therefore indicative of renal function. The targeted metabolomic assay was time and cost prohibitive for amino acid analysis in human urine, facilitating the biomarker identification of tacrolimus nephrotoxicity.


The cardioprotective effects of levosimendan: preclinical and clinical evidence.
Levosimendan, a drug used in the treatment of acute and decompensated heart failure, has positive inotropic and antistunning effects mediated by calcium sensitization of contractile proteins, and vasodilatory and antiischemic effects mediated via the opening of ATP-sensitive potassium channels in vascular smooth-muscle cells. Recently, it also has been shown to act on mitochondrial ATP-sensitive potassium (mitoKATP) channels, an action thought to protect the heart against ischemia-reperfusion damage. This finding has suggested a possible application for levosimendan in clinical situations in which preconditioning would be beneficial (eg, in pre- and perioperative settings in cardiac surgery). The demonstration that levosimendan can prevent or limit myocyte apoptosis via the activation of mitoKATP channels provides a potential mechanism whereby this agent might protect cardiac myocytes during episodes of acute heart failure. This finding may explain why short-term treatment with levosimendan may improve longer-term survival. The present article reviews the literature on the cardioprotective actions of levosimendan, with particular emphasis on its recently recognized effects on mitoKATP channels and the putative preconditioning effects of that action. A therapeutic approach to acute heart failure that includes a cardioprotective strategy could have a clinically meaningful benefit on disease progression beyond alleviation of symptoms.


Colonic mucin discharge by a cholinergic agonist, prostaglandins, and peptide YY in the isolated vascularly perfused rat colon.
The model of the isolated, vascularly perfused rat colon was assessed in the present study to investigate the nervous, hormonal, and local/paracrine pathways involved in colonic mucin secretion. A colonic loop was perfused via the superior mesenteric artery with a Krebs-Henseleit buffer containing 25% washed bovine erythrocytes at a rate of 2.5 ml/min. After a 10-min control period, each compound to be tested was infused intra-arterially for 30 min. Tissue samples from the proximal and midsegments of the perfused rat colon were then fixed and stained for mucus cell count. Intra-arterial administration of bethanechol evoked a concentration-dependent decrease in the number of stained mucus cells per crypt section over the range 2 x 10(-6) to 2 x 10(-4) M: 16.6 +/- 1.4 stained mucus cells per crypt in the midportion of the perfused rat colon (n = 5) with bethanechol 2 x 10(-4) M versus 28.8 +/- 1.5 for controls (n = 6). After infusion of 1.25 and 2.5 microM 16,16-dimethyl prostaglandin E2 (dmPGE2), the number of stained mucus cells per crypt section was significantly reduced: 21.6 +/- 0.6 (n = 6) and 20.6 +/- 1.4 (n = 7), respectively. An increase in the number of cavitated mucus cells was also observed (22.1 +/- 6.7 and 38.5 +/- 4.1% of cavitated mucus cells in the midsegment of the perfused rat colon with 1.25 and 2.5 microM dmPGE2, respectively, vs. 12.3 +/- 4.1% for controls). In contrast, prostaglandin F2alpha did not significantly affect mucus discharge from colonic cells. Peptide YY (10(-10), 10(-9) and 10(-8) M) induced a dose-dependent increase in the percentage of cavitated mucus cells (16.7 +/- 2.8, 23.1 +/- 4.2, and 31.2 +/- 3.4% of cavitated mucus cells in the midsegment, respectively). The proximal and midsegments of the perfused rat colon were equally sensitive to each secretagogue. In the isolated, vascularly perfused rat colon, mucus cells strongly respond to the well-known mucin secretagogues, bethanechol and dmPGE2. This approach has already led to the identification of a novel stimulant of mucin secretion: peptide YY. Our ex vivo model, in which goblet cells are submitted to well-defined luminal and blood-borne stimuli is, therefore, reliable to investigate the nervous, hormonal, and local/paracrine pathways involved in the colonic mucin secretion.


Acquired hepatocerebral degeneration in a patient with hepatitis B and hepatitis delta virus coinfection.
Acquired hepatocerebral degeneration is a neurological syndrome with typical clinical (extrapyramidal and neuropsychiatric) symptoms and brain magnetic resonance imaging findings (high T1 signal in the globus pallidus). It occurs mainly in patients with advanced liver disease, such as in patients co-infected with hepatitis B virus (HBV) and hepatitis delta virus (HDV). However, there are no reports relating HBV/HDV coinfection and acquired hepatocerebral degeneration. This report presents the case of a 49-year-old woman with characteristics of acquired hepatocerebral degeneration and liver cirrhosis due to HBV/HDV coinfection, and presents the main theories of the physiopathology of this condition.


Copper-catalyzed oxidative amination and allylic amination of alkenes.
Enamines and enamides are useful synthetic intermediates and common components of bioactive compounds. A new protocol for their direct synthesis by a net alkene C-H amination and allylic amination by using catalytic Cu(II) in the presence of MnO2 is reported. Reactions between N-aryl sulfonamides and vinyl arenes furnish enamides, allylic amines, indoles, benzothiazine dioxides, and dibenzazepines directly and efficiently. Control experiments further showed that MnO2 alone can promote the reaction in the absence of a copper salt, albeit with lower efficiency. Mechanistic probes support the involvement of nitrogen-radical intermediates. This method is ideal for the synthesis of enamides from 1,1-disubstituted vinyl arenes, which are uncommon substrates in existing oxidative amination protocols.


Electronic structure and stability of fluorophore-nitroxide radicals from ultrahigh vacuum to air exposure.
Thin film processes of organic radicals remain widely unknown, although these materials may have a significant technological potential. In aiming at their use in applications, we explore the electronic structure of thin films of a nitronyl nitroxide radical attached to a fluorophore core. According to our findings, this molecule maintains its radical function and, consequently, its sensing capabilities in the thin films. The films are characterized by a high structural degree of the molecular arrangement, coupled to strong vacuum and air stability that make this fluorophore-nitroxide radical an extremely promising candidate for application in electronics. Our work also identifies a quantitative correlation between the results obtained by the simultaneous use of X-ray photoemission and electron paramagnetic resonance spectroscopy. This result can be used as a standard diagnostic tool in order to link the (in situ-measured) electronic structure with classical ex situ paramagnetic investigations.


Emergency drugs in general practice.
General practitioners need the knowledge, skills, drugs and equipment for managing medical emergencies. Clinics need treatment rooms and doctor's bags that enable emergencies to be managed onsite and offsite respectively. Rural medical generalists may provide more advanced emergency management in their local hospitals. In managing emergencies, GPs may be working with paramedics, therefore it helps to be familiar with their skills and with the drugs they carry.


Modeling the potential influence of particle deposition on the accumulation of organic contaminants by submerged aquatic vegetation.
Submerged aquatic vegetation can act as both a mitigating factor(e.g., reducing downstream impacts of pesticides following runoff/spray drift) and mobilizing factor (e.g., remobilization of contaminants from sediments) influencing the fate and distribution of organic contaminants in the environment. Consequently, there has been wide scientific and regulatory interest in assessing the role of these plants in different contamination scenarios. Mechanistic models describing the environmental fate of contaminants in submerged aquatic vegetation are useful tools for interpreting laboratory and field measurements in addition to providing valuable information to risk assessors. In this study, we developed a fugacity-based model to investigate the influence of particle deposition to plant surfaces on the fate and distribution of two substances in small ponds. The main motivation for conducting this study was to address the fact that the potential contribution of this process is not typically considered by many types of models describing contaminant dynamics in submerged aquatic vegetation. For the hydrophobic substance included in this evaluation (lambda-cyhalothrin), model performance was greatly improved by including this process. The model was also applied in a generic context to compare the importance of particle deposition versus directwater uptake as a function of chemical properties (log Kow) and concentration of suspended solids in the water column. The generic application demonstrated that contaminant mass transfer is dominated by particle deposition for chemicals with log Kow greater than approximately 5.5--6 across a wide range of suspended solid concentrations and can be important even for low log Kow substances in some circumstances. Further empirical and modeling studies are recommended to explore this process more comprehensively.


Genotyping by"cold single-strand conformation polymorphism" of the UGT1A1 promoter polymorphism in Mexican mestizos.
Since no data have previously been reported concerning both the (TA) n polymorphism at the promoter of the UGT1A1 gene in the Mexican population and the use of single-strand conformation polymorphism (SSCP) for the detection of such polymorphism, genotyping by SSCP in 375 G-6-PD normal (Group A) and 81 G-6-PD-deficient (Group B) mestizos belonging to 14 states was carried out. Allele frequencies for (TA)6 and (TA)7 repeats were 0.654 and 0.334, respectively, in Group A and 0.685 and 0.315 in Group B; in the former group, the (TA)5 allele was also observed with a frequency of 0.012. The frequencies of the genotype (TA)7/(TA)7 were 10.1% (Group A) and 8.6% (Group B). The (TA)7/(TA)8 genotype was also observed in a patient with unconjugated hyperbilirubinemia. Due to the importance of its potential medical implications, the observed high frequency (10%) of the (TA)7/(TA)7 genotype is stressed. Genotyping by SSCP of the (TA) n polymorphism is an adequate methodological option.


Seasonal and nocturnal activities of the rhinoceros borer (Coleoptera: Scarabaeidae) in the north Saharan oases ecosystems.
The rhinoceros borer Oryctes agamemnon Burmeister (Coleoptera: Scarabaeidae) is a date palm insect pest that causes damage to trunk and roots of palm trees in several countries, including Tunisia, the United Arab Emirates (UAE), Oman, and Saudi Arabia. The aim of this study was to monitor the seasonal and nocturnal activities of this beetle. Experiments were performed on a date palm of Rjim Maatoug during a 6-yr period (2004-2007, 2009-2010). Field survey using light traps shows that O. agamemnon is a univoltine, with a single population peak. Adults appear in the field around late May-early June and the population continued to build until maximum numbers are reached between the end of July and the beginning of August in the same year. No adults were found after first 10 d of November. This peak was characterized by female dominance in number. The monitoring of nocturnal activity showed that it starts its activities roughly 40 min after the sundown and continues until approximately 1 h before sunrise. The highest number of trapped beetles was remarked in the two first hours of flight activity, with a dominance of female in the first hour and a dominance of male in the second hour. We remarked that the sex ratio (female:male) of the cumulated number of trapped adults in the different years and nights of survey was in favor of females.


Demographic and psychological correlates of satisfaction with healthcare access in New Zealand.
To explore the distribution of New Zealanders' satisfaction with their healthcare access and identify various demographic and psychological factors associated with this satisfaction. 15,822 participants responded to the 2014/15 New Zealand Attitudes and Values Study (NZAVS) survey. This survey included questions on participants' demographic and psychological characteristics, and an item asking for their level of satisfaction with "your access to healthcare when you need it". The majority of participants were highly satisfied with their access to healthcare when needed (68.4%), while 25.3% were moderately satisfied and 6.1% expressed low satisfaction. Younger individuals, those of Māori, Pacific, or Asian ethnicity, and those with higher deprivation and lower income, exhibited lower satisfaction. Conversely, those with a partner, living in an urban area and non-parents showed greater satisfaction. These effects remained significant even after controlling for various psychological factors. In terms of personality, those high on Extraversion, Agreeableness, Conscientiousness and Honesty-Humility, but low on Neuroticism and Openness also expressed higher satisfaction. Lastly, higher self-rated health was associated with greater satisfaction. The majority of New Zealanders are highly satisfied with their access to healthcare when needed, but a considerable number of people express some degree of dissatisfaction. This is broadly consistent with the New Zealand Health Survey (NZHS), and substantiates the finding that approximately 28-32% of New Zealanders experience low healthcare access. Our findings make a novel contribution to the literature by showing that the Big-Six personality traits are reliably associated with New Zealanders' satisfaction with their healthcare access.


Differential actions of prostaglandins in separate cell populations from fetal rat bone.
Prostaglandins (PGs) may stimulate or inhibit bone cell replication and protein synthesis. These disparities may be concentration or time dependent, or occur in discrete cell types or by different second signals. Cell populations that express progressive degrees of osteoblast-like activity can be obtained by serial collagenase digestion of fetal rat parietal bone. The first (population 1) appears less differentiated, whereas the later (populations 3-5) exhibit biochemical features characteristic of osteoblasts. Within 24 h of treatment, three separate PGs increased DNA synthesis in population 1 with relative potencies of PGE1 < PGE2 < PGF2 alpha. By contrast, PGE1 and PGE2 (both strong cAMP inducers) inhibited basal DNA synthesis in population 3-5. These differences were paralleled by analogous changes in collagen and noncollagen synthesis in each population. The mitogenic effect in population 1 persisted for 72 h, and at later times was sensitive to indomethacin. These changes were unlikely to be cAMP dependent, as PGF2 alpha did not induce cAMP production, and the cAMP inducer forskolin was inhibitory. Moreover, phorbol ester treatment enhanced DNA synthesis to a greater extent in population 1 than in populations 3-5, and cotreatment with H-8 (at Km, approximately 10 microM) and staurosporine (at Km, approximately 0.01 microM) decreased the mitogenic effect of PGs in population 1, consistent with a reduction in protein kinase-C activation. These studies suggest that PGs activate less differentiated bone cells by a protein kinase-dependent event, whereas cAMP (induced by PGE1 and PGE2) decreases DNA and protein synthesis in more differentiated bone cells and tempers the increase in cellular activation found in population 1. Consequently, agents or events that increase the synthesis of specific PGs could differentially regulate, in positive and negative ways, biochemical activities in discrete bone cell populations.


Pharmacological profile of 4-(2',4'-difluorobiphenyl-4-yl)-2-methylbutyric acid (deoxoflobufen).
4-(2',4'- Difluorobiphenyl-4-yl)-2-methylbutyric acid (deoxoflobufen, VUFB 19053, CAS 847475-35-8) has been developed as a new omega-biphenyl-alkanoic acid and studied in comparison with the racemic form of 4-(2',4'-difluorobiphenyl-4-yl)-2-methyl-4-oxobutanoic acid (flobufen, CAS 112344-52-2). The compounds were tested in a series of models including acute inflammation induced by carrageenan, adjuvant arthritis, in vitro inhibition of the leuktotriene B4 (LTB4) production, reaction of the graft versus the host (GVHR), production of specific antibodies against ovalbumin, peritoneal exudate formation induced by thioglycollate and phagocytosis of thioglycollate-stimulated mouse peritoneal macrophages. Deoxoflobufen exhibited strong anti-inflammatory, antiarthritic and immunomodulatory effects in most of the performed tests. Anti-inflammatory and antiarthritic effects are fully comparable with those of flobufen, however, the compound is less toxic and has apparently stronger immunomodulating effects.


Site-specific accumulation of recently activated CD4+ Foxp3+ regulatory T cells following adoptive transfer.
CD4(+) Foxp3(+) regulatory T (Treg) cells are required for the maintenance of self-tolerance, as demonstrated by profound autoimmunity in mice and humans with inactivating Foxp3 mutations. Recent studies demonstrate that Treg cells are anatomically compartmentalized within secondary lymphoid organs based on their TCR repertoire and specific organ-protective function; however, whether this reflects differential homing or in situ selection is not known. Here, using Foxp3-GFP reporter mice, we have examined the ability of polyclonal Treg cells from cervical LNs to return to their site-of-origin following adoptive transfer to nonlymphopenic congenic recipients. We find that bulk cervical LN Treg cells do not home directly to cervical LNs but rather accumulate site specifically over time following transfer. Site-specific enrichment is both more rapid and more pronounced among a population of recently activated (CD69(+) ) Treg cells. These data suggest that compartmentalization of Treg cells within secondary lymphoid organs may be governed by antigen recognition and implicate CD69 as a potential marker of recently activated Treg cells recognizing locally expressed antigens.


Opportunism and the resilience of barnacles (Cirripedia: Thoracica) to environmental change.
Cirripede-like organisms have their origins in the Palaeozoic, but until the Cainozoic, were represented primarily by pedunculated forms, such as the Scalpelliformes. Acorn barnacles (Balanomorpha) are first recorded after the Cretaceous-Tertiary extinction event. During the late Palaeogene, rapid radiation of cirripedes resulted in sufficient diversification for them to occupy most marine environments. That they survived both the Palaeocene-Eocene Thermal Maximum and the Pleistocene glaciation is testament to their ability to rapidly adapt to opportunities. The wide habitat distribution of balanomorphs in particular is unparalleled; they are known from the upper littoral (Chthamalus) to depths of 3600 m (Tetrachaelasma) and within this attached to rock, wood and miscellaneous flotsam, plus in symbiosis or commensalism with larger marine organisms. Darwin's (1854) view of the Tertiary as the age of barnacles is reflected in this diversity, distribution and biomass. All cirripedes are, nonetheless, at risk, from rapid habitat change, competition, pollution and, especially in light of their sessile habit, from predation. This paper assesses the viability of a number of cirripedes and concludes that the Lepadiformes, Scalpelliformes and Balanomorpha are the most resilient, and will most quickly adapt to occupy new niches when opportunities arise.


Surfactant protein C mutations in sporadic forms of idiopathic interstitial pneumonias.
Interstitial pneumonias have recently been associated with mutations in the gene encoding surfactant protein C (SFTPC). In particular, SFTPC mutations have been reported in a number of familial forms of pulmonary fibrosis and in infants with interstitial lung diseases. The present study searched for SFTPC mutations in adult patients with sporadic idiopathic interstitial pneumonia. In total, 35 adult patients with sporadic idiopathic interstitial pneumonia and 50 healthy subjects were investigated for SFTPC mutations by direct DNA sequencing. Of the patients with sporadic idiopathic interstitial pneumonia, 25 suffered from idiopathic pulmonary fibrosis and 10 patients from nonspecific interstitial pneumonia. Only two frequent nonsynonymous variants, T138N and S186N, were detected. Allele frequencies of both variations as well as of other identified noncoding alterations did not differ significantly between the diverse patient groups and control subjects. In conclusion, mutations in the gene encoding surfactant protein C are not common in sporadic cases of idiopathic pulmonary fibrosis and nonspecific interstitial pneumonia, suggesting that the mutated gene does not play an important role in the pathogenesis of these forms of idiopathic interstitial pneumonia.


Pu-erh tea hot-water extract activates Akt and induces insulin-independent glucose transport in rat skeletal muscle.
Skeletal muscle is a major organ that is important for whole-body glucose metabolism. We found that when isolated rat epitrochlearis muscle was incubated with a Pu-erh tea hot-water extract (PTE) for 30 min, the rate of 3-O-methyl-D-glucose (3MG) transport increased in the absence of insulin. This activation was associated with an increase in Ser(473) phosphorylation of Akt, a signaling intermediary leading to insulin-dependent glucose transport, but not Tyr(458) phosphorylation of phosphoinositide 3-kinase p85, an upstream molecule of Akt. PTE-stimulated 3MG transport was also not accompanied by Thr(172) phosphorylation of the catalytic α-subunit of 5'-AMP-activated protein kinase (AMPK). Gallic acid, a water-soluble ingredient in Pu-erh tea, stimulated Akt phosphorylation, but not AMPK phosphorylation. These results suggest that Pu-erh tea potentially promotes skeletal muscle glucose transport at least in part by activating Akt.


Arthroscopy of the elbow.
Arthroscopy of the elbow is a technically demanding surgical procedure and attention to detail is essential. The surgeon needs a thorough knowledge of the extraarticular portal anatomy to avoid damaging nearby neurovascular structures. Several technical points need to be considered at all times, especially maintaining the elbow at 90 degrees of flexion to relax the neurovascular structures and maximally distended to move them farther away from the entering arthroscopic instruments.


Dynamic changes in methylome and transcriptome patterns in response to methyltransferase inhibitor 5-azacytidine treatment in citrus.
DNA methylation is known to play an important role in various developmental processes in plants. However, there is a general lack of understanding about the possible functions of DNA methylation in fruit trees. Using callus as a model, methylome, transcriptome and metabolite changes were assessed after treatment with the DNA methyltransferase inhibitor 5-azacytidine (5azaC). Genome-wide methylome analysis revealed the demethylation of a diverse of genes, including many genes encoding transcription factors (TFs), genes involved in biological processes, and the up-regulation of a wide range of transposable elements (TEs). Combined with the RNA-seq data, we observed no obvious genome-wide correlation between the changes in methylation status and expression levels. Furthermore, 5azaC treatment induced carotenoid degradation along with strong activation of carotenoid cleavage dioxygenases 1 (CpCCD1). Functional complementation analysis in bacterial system showed that CpCCD1 exhibited strong catalytic activities toward zeaxanthin, β-carotene and lycopene. In summary, 5azaC treatments induced carotenoid degradation by CpCCD1 activation and led to a genome-wide demethylation effect.


Effect of topographic cone location on outcomes of corneal collagen cross-linking for keratoconus and corneal ectasia.
To assess the effect of preoperative topographic cone location on 1-year outcomes of corneal collagen cross-linking (CXL). In this prospective, randomized, controlled clinical trial, 99 eyes (66 keratoconus, 33 ectasia) from 76 patients underwent CXL. Cone location was defined by the coordinates of preoperative maximum keratometry (maximum K) using the anterior sagittal curvature topography map (Pentacam, Oculus Optikgeräte GmbH). Patients were divided into three groups: those with a maximum K located within the central 3-mm (central cone group), 3- to 5-mm (paracentral cone group), and outside the 5-mm (peripheral cone group) optical zones. Topography and visual acuity data were obtained preoperatively and at 1 year. In the combined cohort, maximum K and uncorrected and corrected distance visual acuity significantly improved by -1.60±3.40 diopters (D) (P<.001), -0.08±0.25 logMAR (P=.001), and -0.10±0.18 log-MAR (P<.001), respectively. Comparing cone groups, maximum K decreased by 2.60±4.50 D (P<.001) in the central cone group, 1.10±2.50 D (P=.02) in the paracentral cone group, and 0.40±1.20 D (P=.08) in the peripheral cone group. Differences among groups were statistically significant (P<.001). Uncorrected distance visual acuity improved by -0.07±0.3 logMAR (P=.1) (central cone group), -0.1±0.17 logMAR (P=.004) (paracentral cone group), and -0.1±0.25 logMAR (P=.04) (peripheral cone group). Corrected distance visual acuity improved by -0.14±0.21 logMAR (P<.001) (central cone group), -0.08±0.17 logMAR (P=.01) (paracentral cone group), and -0.08±0.12 logMAR (P=.002) (peripheral cone group). For both UDVA and CDVA outcomes, these differences among groups were not statistically significant. After CXL, more topographic flattening occurs in eyes with centrally located cones and the least flattening effect occurs when the cone is located peripherally. This cone-location effect is found in eyes with both keratoconus and ectasia.


Description of a new HLA-E (E*01031) allele and its frequency in the Spanish population.
An HLA-E polymorphism study by oligotyping and DNA sequencing was carried out in the Spanish population. As a result, a new HLA-E allele (E*01031) initially assigned by polymerase chain reaction oligotyping as E*0104 was found. This allele presents a synonymous change at codon 77 (AAT-->AAC; Asn) when compared with the E*01032 allele. This position is located in the alpha-helix (alpha 1-domain) and is involved in the peptide binding region of the hypothetical HLA-E molecule. Among 60 Spanish individuals, HLA-E*0101 presents the highest phenotype frequency, followed in decreasing order by E*01032, E*01031 (new allele), and E*0102. Also, new partial intron 1 and complete intron 2 sequences from E*0101, E*01031, and E*01032 are described; the sequences are identical among the three forms. However, the intron 2 sequence of the E*0102 allele bears a two-base deletion not found in apes.


Zoledronic acid suppresses callus remodeling but enhances callus strength in an osteoporotic rat model of fracture healing.
In this study, we demonstrated that the use of zoledronic acid does not impair fracture healing, but results in superior callus size and resistance at the fracture site, which could be the consequence of a lower rate of bone turnover due to its anti-catabolic effect. To investigate the effect of inhibition of bone remodeling by the bisphosphonate, zoledronic acid, on callus properties in an osteoporotic rat model of fracture healing. Ovariectomized (OVX) rats were randomly divided into four treatment groups (n=24 per group): saline control (CNT); and three systemic zoledronic acid-injected groups (0.1mg/kg), administered 1 day (ZOLD1), 1 week (ZOLW1), and 2 weeks (ZOLW2) after fracture. Rats were killed at either 6 or 12 weeks postoperatively. Postmortem analyses included radiography, microcomputed tomography, histology, histomorphometry, biomechanical tests, and nanoindentation tests. Treatment with zoledronic acid led to a significant increase in trabecular bone volume within the callus, as well as in callus resistance, compared to those in the saline control rats; delayed administration (ZOLW2) reduced intrinsic material properties, including ultimate stress and elastic modulus, and microarchitecture parameters, including bone volume/total volume (BV/TV), trabecular thickness (Tb.Th), and connectivity density (Conn.D), compared with ZOLD1 at 12 weeks after surgery. OVX had a negative effect on the progression of endochondral ossification at 6 weeks. Zoledronic acid administration at an early stage following fracture may bind to early callus, and thus not affect subsequent callus formation and endochondral ossification, while delayed administration (ZOLW2) mildly suppresses bony callus remodeling. The superior results obtained with zoledronic acid (ZOLD1, ZOLW1, and ZOLW2) compared to CNT in terms of callus size and resistance could be the consequence of a lower rate of bone turnover at the fracture site due to the anti-catabolic effect of zoledronic acid. Mild suppression of callus remodeling by delayed administration did not impair the initial phase of the fracture healing process.


Alternative methods for cleaning membranes in water and wastewater treatment.
Membrane fouling is caused by foulant deposition or adsorption through physical or chemical interactions on the membrane surface, causing the reduction of flux through the membrane. The main drawbacks of chemical agents used for cleaning are cost, damage caused on the membrane, and waste stream making the process unattractive. Alternative, methods such as ultrasound, enzymatic process, and osmotic backwashing were explored for membrane cleaning. Among all mentioned methods, micronanobubbles have been reported as a promising and emergent method for membrane surface cleaning; unfortunately, the information is limited, but preliminary studies have shown it as an efficient, cheap, and environmentally friendly technique. Other methods like electrically and vibratory-enhanced membrane cleaning also could be interesting but currently are unexplored and information is limited. PRACTITIONER POINTS: Chemical cleaning is an efficient option; however, from an environmental point of view, it is not attractive, and high concentrations could cause damage to the membrane. Micronanobubbles are an emergent and suitable technology for membrane and surface cleaning. Membrane modification and functionalization avoid membrane fast fouling, and the cleaning process is easier, but the manufacture cost could be expensive.


[The Investigation of Acanthamoeba and Other Free Living Amoeba in Swab Samples Obtained from Conjunctiva and Eye Lid].
In the study, it is aimed to determine the prevalence of Acanthamoeba and other free-living amoeba (FLA) species in the swab samples obtained from conjunctiva and lower eye lid. For this purpose, swab samples from the 500 patients'eye lid and conjunctiva were obtained who admitted to Cumhuriyet University, Research and Application Hospital, Department of Ophthalmology with variety of reasons. Swab samples were carried out using sterile cotton swab in steril tubes. The swab samples were inoculated onto non-nutrient agar (NNA). Live Escherichia coli was used as food source for the growth of the FLA. The NNA plates were incubated at 300C and examined daily using ligth microscope for two weeks. For morphotyping of the trophozoites and cysts of the FLA were used taxonomic keys. Two of the 500 swab samples (0.4%) were positive for FLA. One of them (0.2%) were identified as Acanthamoeba spp. and other was identified as Hartmannella spp. However, these patients did not reveal any complaints yet. FLA both themselves and bacteria carrying in their body as reservoirs are potential pathogen. The rapid spread of Acanthamoeba keratitis in recent years reveal that these microorganisms are in contact with the eyes.


Emergence of community-associated methicillin-resistant Staphylococcus aureus USA 300 clone as a cause of health care-associated infections among patients with prosthetic joint infections.
Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has emerged as an important cause of staphylococcal infections, but there have been little data on whether CA-MRSA causes health care-associated infections. A case-control study was performed to identify risk factors for prosthetic joint infections (PJI). Antibiograms of isolates associated with PJI were reviewed. Molecular typing of available MRSA isolates was done using pulsed field gel electrophoresis (PFGE). Nares cultures of health care workers who provided care to those orthopedic patients were obtained. Over a 13-month period (January 2003-January 2004), 9.5% of patients with prosthetic hip (THA) or knee (TKA) joint surgery developed PJI (7 TKA and 2 THA). The mean time to development of PJI was 20 days. Five infections were caused by CA-MRSA and 3 by methicillin-susceptible S aureus; one was culture negative. All CA-MRSA isolates had identical antibiograms (resistant to beta-lactams and erythromycin; susceptible to clindamycin, trimethoprim-sulfamethoxazole, rifampin, gentamicin, levofloxacin, and vancomycin). Molecular typing of 2 available CA-MRSA isolates revealed that these were the USA300 clone; these isolates were PVL+ and carried SCCmec IV. CA-MRSA was not recovered from nares cultures from 31 health care workers. In multivariate analysis, TKA (OR, 8.1; 95% CI: 1.3-48.1) and surgery time >180 minutes (OR, 7.4; 95% CI: 1.4-39.6) were associated with PJI. We have demonstrated that the CA-MRSA USA300 clone is no longer just a cause of community-acquired infections but has also emerged as a cause of health care-associated infections, causing PJI at our institution.


The process of evaluation of a new technology: genetic services and the introduction of DNA probes.
This paper gives a summary of the main results and recommendations, contained in reports for the Department of Health, of an evaluation of the introduction of DNA probes in the service activity of clinical genetics. The reports analysed cost and benefits of this technology, the potential demand for its use, the characteristics of the families referred to the service in terms of social and geographical distribution, the relative impact of DNA probes on the work-load of services in comparison with other activities, and the communication skills required by geneticists in relation to patients and other colleagues. However, the major element of this paper is the discussion of issues related to the process of evaluation. There has recently been discussion about ways to improve dissemination of results of health services research. We discuss here the major barriers that public health physicians and health service researchers need to address when playing the role of independent evaluators of services or new technologies. During the course of the evaluation of DNA testing, major changes occurred in the NHS. The evaluator will have to consider not only the costs and effectiveness of the new technology, but also how the findings are interpreted by the providers within the constraints of the new organizational structure.


Liver fibrosis during clinical ascertainment of glycogen storage disease type III: a need for improved and systematic monitoring.
In glycogen storage disease type III (GSD III), liver aminotransferases tend to normalize with age giving an impression that hepatic manifestations improve with age. However, despite dietary treatment, long-term liver complications emerge. We present a GSD III liver natural history study in children to better understand changes in hepatic parameters with age. We reviewed clinical, biochemical, histological, and radiological data in pediatric patients with GSD III, and performed a literature review of GSD III hepatic findings. Twenty-six patients (median age 12.5 years, range 2-22) with GSD IIIa (n = 23) and IIIb (n = 3) were enrolled in the study. Six of seven pediatric patients showed severe fibrosis on liver biopsy (median [range] age: 1.25 [0.75-7] years). Markers of liver injury (aminotransferases), dysfunction (cholesterol, triglycerides), and glycogen storage (glucose tetrasaccharide, Glc4) were elevated at an early age, and decreased significantly thereafter (p < 0.001). Creatine phosphokinase was also elevated with no significant correlation with age (p = 0.4). Liver fibrosis can occur at an early age, and may explain the decrease in aminotransferases and Glc4 with age. Our data outlines the need for systematic follow-up and specific biochemical and radiological tools to monitor the silent course of the liver disease process.


A precise measurement of the [Formula: see text] meson oscillation frequency.
The oscillation frequency, [Formula: see text], of [Formula: see text] mesons is measured using semileptonic decays with a [Formula: see text] or [Formula: see text] meson in the final state. The data sample corresponds to 3.0[Formula: see text] of pp collisions, collected by the LHCb experiment at centre-of-mass energies [Formula: see text] = 7 and 8[Formula: see text]. A combination of the two decay modes gives [Formula: see text], where the first uncertainty is statistical and the second is systematic. This is the most precise single measurement of this parameter. It is consistent with the current world average and has similar precision.


A Novel Genetic Screen Identifies Modifiers of Age-Dependent Amyloid β Toxicity in the Drosophila Brain.
The accumulation of amyloid β peptide (Aβ) in the brain of Alzheimer's disease (AD) patients begins many years before clinical onset. Such process has been proposed to be pathogenic through the toxicity of Aβ soluble oligomers leading to synaptic dysfunction, phospho-tau aggregation and neuronal loss. Yet, a massive accumulation of Aβ can be found in approximately 30% of aged individuals with preserved cognitive function. Therefore, within the frame of the "amyloid hypothesis", compensatory mechanisms and/or additional neurotoxic or protective factors need to be considered and investigated. Here we describe a modifier genetic screen in Drosophila designed to identify genes that modulate toxicity of Aβ42 in the CNS. The expression of Aβ42 led to its accumulation in the brain and a moderate impairment of negative geotaxis at 18 days post-eclosion (d.p.e) as compared with genetic or parental controls. These flies were mated with a collection of lines carrying chromosomal deletions and negative geotaxis was assessed at 5 and 18 d.p.e. Our screen is the first to take into account all of the following features, relevant to sporadic AD: (1) pan-neuronal expression of wild-type Aβ42; (2) a quantifiable complex behavior; (3) Aβ neurotoxicity associated with progressive accumulation of the peptide; and (4) improvement or worsening of climbing ability only evident in aged animals. One hundred and ninety-nine deficiency (Df) lines accounting for ~6300 genes were analyzed. Six lines, including the deletion of 52 Drosophila genes with human orthologs, significantly modified Aβ42 neurotoxicity in 18-day-old flies. So far, we have validated CG11796 and identified CG17249 as a strong candidate (whose human orthologs are HPD and PRCC, respectively) by using RNAi or mutant hemizygous lines. PRCC encodes proline-rich protein PRCC (ppPRCC) of unknown function associated with papillary renal cell carcinoma. HPD encodes 4-hydroxyphenylpyruvate dioxygenase (HPPD), a key enzyme in tyrosine degradation whose Df causes autosomal recessive Tyrosinemia type 3, characterized by mental retardation. Interestingly, lines with a partial Df of HPD ortholog showed increased intraneuronal accumulation of Aβ42 that coincided with geotaxis impairment. These previously undetected modifiers of Aβ42 neurotoxicity in Drosophila warrant further study to validate their possible role and significance in the pathogenesis of sporadic AD.


Ca transients from Ca channel activity in rat cardiac myocytes reveal dynamics of dyad cleft and troponin C Ca binding.
The properties of the dyad cleft can in principle significantly impact excitation-contraction coupling, but these properties are not easily amenable to experimental investigation. We simultaneously measured the time course of the rise in integrated Ca current (ICa) and the rise in concentration of fura 2 with Ca bound ([Ca-fura 2]) with high time resolution in rat myocytes for conditions under which Ca entry is only via L-type Ca channels and sarcoplasmic reticulum (SR) Ca release is blocked, and compared these measurements with predictions from a finite-element model of cellular Ca diffusion. We found that 1) the time course of the rise of [Ca-fura 2] follows the time course of integrated ICa plus a brief delay (1.36 +/- 0.43 ms, n = 6 cells); 2) from the model, high-affinity Ca binding sites in the dyad cleft at the level previously envisioned would result in a much greater delay (>/=3 ms) and are therefore unlikely to be present at that level; 3) including ATP in the model promoted Ca efflux from the dyad cleft by a factor of 1.57 when low-affinity cleft Ca binding sites were present; 4) the data could only be fit to the model if myofibrillar troponin C (TnC) Ca binding were low affinity (4.56 microM), like that of soluble troponin C, instead of the high-affinity value usually used (0.38 microM). In a "good model," the rate constants for Ca binding and dissociation were 0.375 times the values for soluble TnC; and 5) consequently, intracellular Ca buffering at the rise of the Ca transient is inferred to be low.


Relative strength of the mitogenic and interleukin-2-production-inducing activities of staphylococcal exotoxins presumed to be causative exotoxins of toxic shock syndrome: toxic shock syndrome toxin-1 and enterotoxins A, B and C to murine and human T cells.
Several observations suggest that staphylococcal enterotoxins A, B and C (SEA, SEB and SEC, respectively), in addition to toxic shock syndrome toxin-1 (TSST-1), are causative exotoxins of toxic shock syndrome (TSS). Based on the view that polyclonal T cell activation with the causative exotoxins, resulting in over-production of lymphokines, is involved in the development of the pathological changes observed in TSS, we investigated the activities of these four exotoxins to induce proliferation and interleukin 2 production in murine and human lymphocytes by using in vitro culture systems. The results showed that all these exotoxins are strong polyclonal inducers of proliferation and interleukin 2 production in human T cells, whereas TSST-1 and SEA are strong and SEB and SEC are weak polyclonal inducers in murine T cells. These results suggest that SEA, SEB and SEC, in addition to TSST-1, are possibly involved as causative exotoxins in the development of the pathological changes observed in TSS.


[Contrast-enhanced ultrasonography as the most perspective diagnostic method for unstable atherosclerotic plaque of carotid artery].
Problem of internal carotid artery disease diagnosis appears to be crucial today. Complications of this pathology are strokes and transient ischemic attacks. There is no technology for their prediction or at least stratifying risks. Some recent researches are devoted to a new diagnostic method. This new technology is called Contrast Enhanced Ultrasonography (CEUS) and followed by outstanding results in studying the morphological peculiarities of internal carotid artery plaques and predicting the probability of complications. CEUS is a new way for atherosclerotic process analysis because it is able to detect intraplaque neovascularization and vascular wall inflammation.


[Pharmacological treatment options for prevention of alcohol relapse].
Many studies address the neurobiological mechanisms of alcohol craving. The study results obtained so far show that there may be at least three subtypes of craving, which may be linked to changes in different neurotransmitter systems. Actually in Germany acamprosate and disulfiram are approved for clinical use for the prevention of alcohol relapse. Also off-label therapy with naltrexone is possible. However studies show that these substances are not effective in all patients. In the light of hypothetical changes in different neurotransmitter systems, subtype specific therapy may be necessary for successful prevention of alcohol relapse. Therefore new therapy options are needed in order to treat alcohol dependent persons. Substances that seem to have efficacy in preclinical or preliminary clinical studies are baclofen, topiramate, odansetron, rimonabant and memantine.


[New prospects in the therapeutic use of somatostatin and its derivatives].
The discovery and subsequent clinical application of somatostatine, a polypeptide neurohormone of 14 amino acids, and of its analogs, opens a novel chapter of neuroendocrinology that is still in full evolution and to a large extent unknown. The isolation of an octapeptide, a selective somatostatine analog, permits to prolong its action, in fact it has a halflife of about 140 min in old subjects and about 100 min. in the young. Thanks to its excellent tolerability, the synthetic hormone can be usefully applied in the treatment of acromegaly, gigantism and hypersomatotropinemic conditions in general, and even in other clinical branches, such as treatment of esophageal hemorrhage due to the rupture of varices in liver cirrhosis or to erosion of gastric blood vessels in patients suffering from peptic ulcer disease.


Phospholipids chiral at phosphorus. Absolute configuration of chiral thiophospholipids and stereospecificity of phospholipase D.
Separate diastereomers of 1,2-dipalmitoyl-sn-glycero-3- thiophosphoethanolamine ( DPPsE ) were prepared in 97% diastereomeric purity and characterized by 31P, 13C, and 1H nuclear magnetic resonance (NMR). The isomers hydrolyzed by phospholipases A2 and C specifically were designated as isomer B (31P NMR delta 59.13 in CDCl3 + Et3N ) and isomer A (59.29 ppm), respectively, analogous to the isomers B and A of 1,2-dipalmitoyl-sn-glycero-3- thiophosphocholine ( DPPsC ) [ Bruzik , K., Jiang , R.-T., & Tsai, M.-D. (1983) Biochemistry 22, 2478-2486]. Phospholipase D from cabbage was shown to be specific to isomer A of DPPsC in transphosphatidylation . The product DPPsE was shown to be isomer A. The absolute configuration of chiral DPPsE at phosphorus was elucidated by bromine-mediated desulfurization in H2 18O to give chiral 1,2-dipalmitoyl-sn-glycero-3-[18O]phosphoethanolamine ( [18O]DPPE) followed by 31 P NMR analysis [ Bruzik , K., & Tsai, M.-D. (1984) J. Am. Chem. Soc. 106, 747-754]. The absolute configuration of chiral DPPsC was elucidated by desulfurization in H2 18O mediated by bromine or cyanogen bromide to give chiral 1,2-dipalmitoyl-sn-glycero-3-[18O]phosphocholine ( [18O]DPPC), which was then converted to [18O]DPPE by phospholipase D with retention of configuration [ Bruzik , K., & Tsai, M.-D. (1984) Biochemistry (preceding paper in this issue)]. The results indicate that isomer A of both DPPsE and DPPsC is SP whereas isomer B is RP.


Detection of growth factors and proto-oncogene mRNA in the growing tip of red deer (Cervus elaphus) antler using reverse-transcriptase polymerase chain reaction (RT-PCR).
Deer antler is a unique mammalian organ that has an annual cycle of regeneration. The antler grows very rapidly from the tip at up to 1 cm/day in red deer for a 90- to 120-day period. It is hypothesised that locally produced growth factors are required to control and stimulate this growth. The tip of the growing antler from animals whose antlers had been growing for 30, 60, or 90 days was dissected into four zones: epidermis/dermis, reserve mesenchyme, precartilaginous, and cartilaginous. Total RNA was extracted, and the presence of various growth factors and proto-oncogenes was detected using RT-PCR, IGF-I, IGF-II, TGF beta 1, TGF beta 2, c-fos, c-myc, and beta-actin were all present as single bands of the expected molecular weight in the four zones of the antler at each stage of growth. There were higher levels of IGF-I, TGF beta 2, and c-myc relative to beta-actin in the epidermis/dermis layer than in the other three zones. There were no differences in the expression of any of the genes between the three stages of growth. The presence of TGF beta 3 cannot be confirmed since multiple bands were seen in all antler tissues. A single band of the expected size for TGF alpha was seen only in the epidermal/dermal layer of the antler, with multiple bands of different molecular weight being detected in the other zones of the antler. This work has demonstrated the presence of multiple growth factors in the growing deer antler and supports the hypothesis that paracrine/autocrine stimulation is important for regulating antler growth.


Direct Formation of Electronic Excited NO2 Contributes to the High Yield of HONO during Photosensitized Renoxification.
Photosensitized renoxification of HNO3 is found to produce HONO in an unexpectedly high yield, which has been considered an important source for atmospheric HONO. Conventionally, the production of HONO is ascribed to the secondary photolysis of the primarily formed NO2. In this study, by using humic acid (HA) as a model environmental photosensitizer, we provide evidence of the direct formation of NO2 in its electronic excited state (NO2*) as a key intermediate during the photosensitizing renoxification of HNO3. Moreover, the high HONO yield originates from the heterogeneous reaction of the primarily formed NO2* with the co-adsorbed water molecules on HA. Such a mechanism is supported by the increase of the product selectivity of HONO with relative humidity. Further luminescence measurements demonstrate clearly the occurrence of an electronic excited state (NO2*) from photolysis of adsorbed HNO3 on HA. This work deepens our understanding of the formation of atmospheric HONO and gives insight into the transformation of RNS.


Biodegradation Kinetics of 1,4-Dioxane in Chlorinated Solvent Mixtures.
This study investigated the impacts of individual chlorinated solvents and their mixtures on aerobic 1,4-dioxane biodegradation by Pseudonocardia dioxanivorans CB1190. The established association of these co-occurring compounds suggests important considerations for their respective biodegradation processes. Our kinetics and mechanistic studies demonstrated that individual solvents inhibited biodegradation of 1,4-dioxane in the following order: 1,1-dichloroethene (1,1-DCE) > cis-1,2-diochloroethene (cDCE) > trichloroethene (TCE) > 1,1,1-trichloroethane (TCA). The presence of 5 mg L(-1) 1,1-DCE completely inhibited 1,4-dioxane biodegradation. Subsequently, we determined that 1,1-DCE was the strongest inhibitor of 1,4-dioxane biodegradation by bacterial pure cultures exposed to chlorinated solvent mixtures as well as in environmental samples collected from a site contaminated with chlorinated solvents and 1,4-dioxane. Inhibition of 1,4-dioxane biodegradation rates by chlorinated solvents was attributed to delayed ATP production and down-regulation of both 1,4-dioxane monooxygenase (dxmB) and aldehyde dehydrogenase (aldH) genes. Moreover, increasing concentrations of 1,1-DCE and cis-1,2-DCE to 50 mg L(-1) respectively increased 5.0-fold and 3.5-fold the expression of the uspA gene encoding a universal stress protein. In situ natural attenuation or enhanced biodegradation of 1,4-dioxane is being considered for contaminated groundwater and industrial wastewater, so these results will have implications for selecting 1,4-dioxane bioremediation strategies at sites where chlorinated solvents are present as co-contaminants.


[A severe episode in a patient with recurrent disseminated acute encephalitis due to vaccination against hepatitis B. For or against vaccination?].
Several reports of new cases of CNS demyelination or reactivation of MS after hepatitis B vaccination have raised the possibility of a causal relationship. Conversely, some authors have concluded that the risk of developing a demyelinating CNS event after a hepatitis B vaccination is unknown. We have observed a 40 year old man, with diabetes mellitus (DM) type 1 and a previous history of recurrent acute demyelinating encephalomyelitis (rADEM). The patient had experienced three episodes of neurological dysfunction and he fulfilled the criteria for definite clinical MS but MRI showed demyelinating lesions in the pons and cerebellum without MRI criteria of MS. CSF analysis showed oligoclonal banding. The patient had been clinically stable during the last 6 years. Yearly MRI during this period had not shown any new disease activity. He was admitted in our MS Clinic due to dizziness, nausea, vomiting and diplopia, 6 weeks after the first of the two injections for hepatitis B vaccine after participating in the national programme of vaccination in DM type 1 patients. Clinical examination showed intranuclear ophtalmoplegia, visual loss in the left eye and worsening of the previous cerebellar and pyramidal signs. MRI showed an increase in the old lesions with high intensity signals on T2 weighted sequences with post gadolinium enhancement on T1 weighted sequences located in the brainstem and mesencephalon. The patient s diabetes mellitus deteriorated with ketoacidosis that needed increased doses of insulin. His condition worsened and he developed partial motor seizures. He improved 15 days later but he still had involvement of the cerebellar and pyramidal systems and occasional dizziness. As pointed out by some authors and in view of this observation, it would seem reasonable, as a precautionary measure, to avoid hepatitis B vaccination in patients with a personal or family history of symptoms suggestive of a demyelinating disease of the CNS.


Tonal and orthographic analysis in a Cantonese-speaking individual with nonfluent/agrammatic variant primary progressive aphasia.
Clinical understanding of primary progressive aphasia (PPA) has been established based on English-speaking population. The lack of linguistic diversity in research hinders the diagnosis of PPA in non-English speaking patients. This case report describes the tonal and orthographic deficits of a multilingual native Cantonese-speaking woman with nonfluent/agrammatic variant PPA (nfvPPA) and progressive supranuclear palsy. Our findings suggest that Cantonese-speaking nfvPPA patients exhibit tone production impairments, tone perception deficits at the lexical selection processing, and linguistic dysgraphia errors unique to logographic script writer. These findings suggest that linguistic tailored approaches offer novel and effective tools in identifying non-English speaking PPA individuals.


The 26S proteasome in Schistosoma mansoni: bioinformatics analysis, developmental expression, and RNA interference (RNAi) studies.
The 26S proteasome is a proteolytic complex responsible for the degradation of the vast majority of eukaryotic proteins. Regulated proteolysis by the proteasome is thought to influence cell cycle progression, transcriptional control, and other critical cellular processes. Here, we used a bioinformatics approach to identify the proteasomal constituents of the parasitic trematode Schistosoma mansoni. A detailed search of the S. mansoni genome database identified a total of 31 putative proteasomal subunits, including 17 subunits of the regulatory (19S) complex and 14 predicted catalytic (20S) subunits. A quantitative real-time RT-PCR analysis of subunit expression levels revealed that the S. mansoni proteasome components are differentially expressed among cercaria, schistosomula, and adult worms. In particular, the data suggest that the proteasome may be downregulated during the early stages of schistosomula development and is subsequently upregulated as the parasite matures to the adult stage. To test for biological relevance, we developed a transfection-based RNA interference method to knockdown the expression of the proteasome subunit, SmRPN11/POH1. Transfection of in vitro transformed S. mansoni schistosomula with specific short-interfering RNAs (siRNAs) diminished SmRPN11/POH1 expression nearly 80%, as determined by quantitative RT-PCR analysis, and also decreased parasite viability 78%, whereas no significant effect could be seen after treatment with the same amount of an irrelevant siRNA. These results indicate that the subunit SmRPN11/POH1 is an essential gene in schistosomes and further suggest an important role for the proteasome in parasite development and survival.


The role of Sertoli cells in the differentiation and exclusion of rat testicular germ cells in primary culture.
We have previously established a primary co-culture of spermatogenic and somatic cells of the rat testis, in which spermatogenic cells differentiate to some extent in terms of the occurrence of testis-specific gene expression. In the present study, the interaction between spermatogenic and somatic Sertoli cells was investigated in this culture system. Spermatogenic differentiation did not proceed when these two cell types were placed on opposite sides of a permeable membrane, thus avoiding direct contact. Further, a significant proportion of spermatogenic cells died by apoptosis during culture, and Sertoli cells engulfed and digested the degenerating spermatogenic cells. These results indicate that Sertoli cells participate both in the differentiation of spermatogenic cells and in the exclusion of degenerating spermatogenic cells, by directly attaching to those cells.


Influence of growth conditions on the production of a bacteriocin by Lactococcus lactis subsp. lactis ST34BR, a strain isolated from barley beer.
Bacteriocin ST34BR, a small polypeptide of 2.9 kDa produced by Lactococcus lactis subsp. lactis ST34BR, inhibits the growth of Enterococcus faecalis, Escherichia coli, Lactobacillus plantarum, Lactobacillus casei, Pseudomonas aeruginosa and Staphylococcus aureus. MRS broth, adjusted to pH 6.0 yielded 6,400 AU/ml, compared to 400 AU/ml recorded in BHI broth, M17 broth, 10% (w/v) soy milk, and 8% and 10% (w/v) molasses. At pH of 4.5 only 800 AU/ml was produced. Based on comparative studies in MRS broth, without organic nitrogen, supplemented with different combinations of tryptone, meat extract and yeast extract, tryptone was identified as the stimulating nitrogen compound. Growth in the presence of 20 g/l glucose, maltose, mannose or sucrose yielded bacteriocin levels of 6,400 AU/ml, whereas the same concentration of lactose and fructose yielded 3,200 AU/ml and 1,600 AU/ml, respectively. No difference in bacteriocin ST34BR activity was recorded in MRS broth supplemented with 2 g/l K2HPO4 and 2 g/l, 5 g/l, 10 g/l or 50 g/l KH2PO4. However, 20 g/l KH2PO4 increased bacteriocin ST34BR production to 12,800 AU/ml. Glycerol at 1g/l to 10 g/l in MRS broth reduced bacteriocin activity to 3,200 AU/ml, whilst 20 g/l and 50 g/l yielded only 1,600 AU/ml. The presence of cyanocobalamin, L-ascorbic acid, thiamine and DL-6,8-thioctic acid in MRS broth at 1.0 ppm, respectively, did not result in increased activity levels.


Microablative fractional radiofrequency for sexual dysfunction and vaginal Trophism: A randomized clinical trial.
To evaluate Microablative Fractional Radiofrequency (MAFRF) as a possible option in treating vaginal atrophy. This was a randomized, controlled clinical trial with postmenopausal women diagnosed with vaginal atrophy. The treatment consisted of three sessions of MAFRF, compared to vaginal estrogen administration and an untreated control group. Assessments occurred at baseline and 90 days. The primary endpoints were sexual function, evaluated by the Female Sexual Function Index (FSFI), and vaginal health, assessed by the Vaginal Health Index (VHI). Secondary outcomes included vaginal microbiota composition (Nugent score) and epithelial cell maturation (Maturation Value ‒ MV). One hundred and twenty women (40 in each group) were included. Concerning the FSFI, both groups, MAFRF (median 4.8 [3.6‒6.0]) and vaginal estrogen (mean 4.7 ± 1.1), experienced improved sexual desire when compared to the control group (median 3.6 [2.4‒4.8]). Regarding the total score of VHI, the authors observed an improvement in the mean of the MAFRF (23.7 ± 2.0) and vaginal estrogen groups (23.5 ± 1.9) when compared to the control (14.8 ± 2.9). The Nugent score was reduced in the MAFRF and estrogen groups (p < 0.01) compared to the control group. Lastly, the MV was modified after treatment with MAFRF (p < 0.01) and vaginal estrogen (p < 0.001). No differences existed between the MAFRF and vaginal estrogen groups in the studied variables. No adverse effects were reported following the MAFRF protocol. Radiofrequency was comparable in efficacy to estrogen administration for treating vulvovaginal atrophy. It deserves consideration as a viable option in managing this condition.


Porous ZnO Ultrathin Nanosheets with High Specific Surface Areas and Abundant Oxygen Vacancies for Acetylacetone Gas Sensing.
In this study, porous ZnO ultrathin nanosheets with abundant surface oxygen vacancies were prepared by a hydrothermal technique followed by an annealing method using graphene oxide (GO) as a template. The high specific surface area of GO with ultrathin thickness provided an important template for the ZnO ultrathin nanosheets. The as-prepared porous ZnO ultrathin nanosheets exhibited superior acetylacetone sensing performance. The sensor response of the porous ZnO ultrathin nanosheets was 191.1 for 100 ppm acetylacetone, which was approximately 4 times higher than that of ZnO clusters (prepared without GO template) at 340 °C. The porous ZnO ultrathin nanosheets also exhibited excellent selectivity and operational stability. The excellent gas sensing performance of the porous ZnO ultrathin nanosheets was due to their high specific surface area (130.5 m2/g) and abundant surface oxygen vacancy.


[Study on identification and traceability of tea material cultivar by combined analysis of multi-partial least squares models based on near infrared spectroscopy].
The present study attempted to achieve the identification and traceability of tea material cultivar through combination of multi-partial least squares models and Euclidean distance, etc. The results indicate that with the samples manufactured with tea fresh leaves of cultivar Longjing 43, Qunti, Yingshuang and Wuniuzao as the analysis objects, 4 models were established in this study so as to identify the tea with material cultivar being tea fresh leaves of cultivar Longjing 43, Qunti, Yingshuang and Wuniuzao separately by PLS. Their accuracy rate of identification of samples in the calibration set were 89.8%, 90.9%, 96.1% and 99.5%, respectively, while those in test set were 87.1%, 84.2%, 96.1% and 97.5%, respectively. After the "first identification" through the combined analysis of the four models for identification of tea material cultivar and the "second identification" adopting the Euclidean distance, the accuracy rate of material cultivar recognition for the tea samples was 90.3% (calibration set) and 83.5% (test set), respectively. This study provided a reference method for the identification of tea manufactured with a specific material cultivar and the material cultivar traceability of the manufactured tea.


A chemosterilized antigen-extracted autodigested alloimplant for bone banks.
Limited chemical extraction of hydrophobic glycopeptides and subtotal autodigestion of the donor's cells and plasma membranes in undemineralized cortical bone in vitro reduces the putative quantity of haptenic substances absorbed by the recipient. Iodoacetic acid and sodium azide or other sulfhydryl group enzyme inhibitors added to the buffer solutions during in vitro autodigestion and estraction of intracellular alloantigens protects the bone matrix morphogenetic property against enzymatic degradation. The delayed hypersensitivity reaction induced by aseptically collected freeze-dried bone and the destruction of the bone morphogenetic property caused by radiation-sterilization is avoidable by sequential chemodigestion and chemosterilization of bone that preserves the maximum morphogenetic potential while transferring a minimum quantity of alloantigen.


Wearable/disposable sweat-based glucose monitoring device with multistage transdermal drug delivery module.
Electrochemical analysis of sweat using soft bioelectronics on human skin provides a new route for noninvasive glucose monitoring without painful blood collection. However, sweat-based glucose sensing still faces many challenges, such as difficulty in sweat collection, activity variation of glucose oxidase due to lactic acid secretion and ambient temperature changes, and delamination of the enzyme when exposed to mechanical friction and skin deformation. Precise point-of-care therapy in response to the measured glucose levels is still very challenging. We present a wearable/disposable sweat-based glucose monitoring device integrated with a feedback transdermal drug delivery module. Careful multilayer patch design and miniaturization of sensors increase the efficiency of the sweat collection and sensing process. Multimodal glucose sensing, as well as its real-time correction based on pH, temperature, and humidity measurements, maximizes the accuracy of the sensing. The minimal layout design of the same sensors also enables a strip-type disposable device. Drugs for the feedback transdermal therapy are loaded on two different temperature-responsive phase change nanoparticles. These nanoparticles are embedded in hyaluronic acid hydrogel microneedles, which are additionally coated with phase change materials. This enables multistage, spatially patterned, and precisely controlled drug release in response to the patient's glucose level. The system provides a novel closed-loop solution for the noninvasive sweat-based management of diabetes mellitus.


Root canal morphology of mandibular premolars.
Four hundred mandibular first premolars and 400 mandibular second premolars were decalcified, injected with dye, and made transparent to determine the number of root canals, their type, the ramifications of the main root canal, the location of apical foramina and transverse anastomoses, and the frequency of apical deltas. The mandibular first premolar had one canal at the apex in 74.0% of the teeth, two canals at the apex in 25.5%, and three canals at the apex in 0.5% of the teeth. The mandibular second premolar had one canal at the apex in 97.5% and two canals at the apex in 2.5% of the teeth.


Polycyclic aromatic hydrocarbons associated with total suspended particles and surface soils in Kunming, China: distribution, possible sources, and cancer risks.
The concentrations, distribution, possible sources, and cancer risks of polycyclic aromatic hydrocarbons (PAHs) in total suspended particles (TSPs) and surface soils collected from the same sampling spots were compared in Kunming, China. The total PAH concentrations were 9.35-75.01 ng/m(3) and 101.64-693.30 ng/g dry weight (d.w.), respectively, in TSPs and surface soils. Fluoranthene (FLA), pyrene (PYR), chrysene (CHR), and phenanthrene (PHE) were the abundant compounds in TSP samples, and phenanthrene (PHE), fluorene (FLO), fluoranthene (FLA), benzo[b]fluoranthene (BbF), and benzo[g,h,i]perylene (BghiP) were the abundant compounds in surface soil samples. The spatial distribution of PAHs in TSPs is closely related to the surrounding environment, which varied significantly as a result of variations in source emission and changes in meteorology. However, the spatial distribution of PAHs in surface soils is supposed to correlate with a city's urbanization history, and high levels of PAHs were always observed in industry district, or central or old district of city. Based on the diagnostic ratios and principal component analysis (PCA), vehicle emissions (especially diesel-powered vehicles) and coal and wood combustion were the main sources of PAHs in TSPs, and the combustion of wood and coal, and spills of unburnt petroleum were the main sources of PAHs in the surface soils. The benzo[a]pyrene equivalent concentration (BaPeq) for the TSPs and surface soil samples were 0.16-2.57 ng/m(3) and 11.44-116.03 ng/g d.w., respectively. The incremental lifetime cancer risk (ILCR) exposed to particulate PAHs ranged from 10(-4) to 10(-3) indicating high potential of carcinogenic risk, and the ILCR exposed to soil PAHs was from 10(-7) to 10(-6) indicating virtual safety. These presented results showed that particle-bound PAHs had higher potential carcinogenic ability for human than soil PAHs. And, the values of cancer risk for children were always higher than for adults, which demonstrated that children were sensitive to carcinogenic effects of PAHs.


Designing and structure evaluation of multi-epitope vaccine against ETEC and EHEC, an in silico approach.
Diarrheal diseases represent a major health problem in developing countries. Several viruses and bacterial agents, such as Enterotoxigenic Escherichia coli (ETEC) and Enterohemorrhagic Escherichia coli (EHEC) are responsible for human enteric infections. In humans, EHEC infections result in bloody or non-bloody diarrhea, which may be complicated by haemorrhagic colitis and haemolytic uraemic syndrome (HUS). Infection by ETEC is accompanied by a non inflammatory watery diarrhea. E. coli follows a common strategy of infection: colonization on a mucosal site, evasion of host defenses, multiplication, and host damage. Intimin, Stx, Lt and Cfa proteins are the virulence factors expressed by these strains. Antibiotic treatment is generally not recommended for most cases of diarrhea, since antibiotic usage may lead to antibiotic resistance in ETEC and may also change the intestinal flora. We hypothesized that the chimeric forms of these effectors as vaccine candidates would reduce the colonization of bacteria. This study is based on an in silico analysis of chimeric protein structure and its stability and solubility. The secondary and tertiary structures of selected domains were also predicted. Moreover, T and B cell epitopes were mapped. Protein structure Prediction showed that each domain of antigen was separated completely also stable for recombinant expression. We believe that this chimeric vaccine candidate is effective for prevention of bacteria caused diarrheal diseases.


A qualitative analysis of the effectiveness of telehealthcare devices (ii) barriers to uptake of telehealthcare devices.
Monitoring health and care needs through the use of telehealthcare devices has been proposed to help alleviate funding concerns in a climate of limited budgets. As well as improving cost effectiveness, such an approach could be used to help individuals live at home for longer. In practice however, these devices often go unused. A qualitative study was carried out to determine the barriers to uptake of these devices from both the perspective of the end user and from key players in the healthcare supply chain. A qualitative approach was used involving focus groups and interviews. Two UK-based focus groups were held with users and potential users, to assess their views on the wide array of devices available. 27 individuals were involved in the groups, all over the age of 60. Additionally 27 telephone interviews were conducted with key supply chain players to ascertain their views on the barriers to uptake of these devices. A semi-structured interview guide was used. All data were audio-recorded, transcribed verbatim and analysed using a thematic approach. Users were generally unaware of the wide array of devices available and when shown a selection, were often unclear as to their purpose. The interviews revealed extensive barriers to uptake due to lack of awareness, unfamiliar terminology, complex supply routes and costs, resistance from professionals to device usage and lack of expertise. Public and professional awareness campaigns are required with appropriate funding mechanisms for users to gain access to devices. The numerous barriers identified require systematically addressing, so that device usage is better promoted, enabling individuals to live at home successfully for longer.


MAP kinase modules: the excursion model and the steps that count.
MAP kinase modules propagate diverse extracellular signals to downstream effectors. The two dual phosphorylation reactions catalyzed by the modules are thought to control the switch behavior of the pathway. Here we review recent approaches to understand these pathways through signal-to-response studies in cells and in vitro. These data are reconciled with physical models as well as predictions made on mathematical and theoretical grounds. Biochemical analysis has shown recently that the dual phosphorylation reactions catalyzed by MAP kinase modules are sequential at both levels of the cascade. The observed order of phosphorylation events suggests an excursion from the Ser/Thr kinase activity of the MAP3K into Tyr kinase activity of the central dual specificity MAP2K. How the order of events might be encoded in the structures and interactions is discussed. The ordered mechanism confirms predictions that reactions should be sequential to generate the steep signal-to-response curves and delayed responses observed in cells.


Transitioning to adult health care facilities for young adults with a chronic condition.
Youth with chronic conditions and special health care needs are surviving into adulthood. In certain circumstances, these youth may choose to remain with their pediatricians for their health care needs. However, most youth will require complex coordinated care in order for them to live full and productive lives to the best that they are able. Issues such as independence and dependence, educational and vocational choices, and insurance are important in transitioning to an adult-focused system of health care. This article addresses strategies that will assist young adults with the process of transitioning from pediatric care centers to adult health care centers. Despite the importance of providing a transition program, few transition programs exist for young people with chronic conditions (Sawyer, Blair, & Bowes, 1997). This article will also discuss barriers to the transition process.


Superovulation and intrauterine insemination in the treatment of male factor infertility.
Recent studies report that superovulation combined with intrauterine insemination (IUI) is more successful than superovulation alone, IUI alone or superovulation with intracervical insemination in couples with male subfertility. Our study evaluated two superovulation protocols in the management of male factor infertility using IUI: (A) clomiphene citrate and human chorionic gonadotropin (HCG) and (B) human menopausal gonadotropin and HCG. Fifteen couples with severe oligoasthenozoospermia (OAS) were treated with protocol A in 54 cycles, and no pregnancies were achieved. Eight of the 15 couples with severe OAS subsequently received protocol B for 24 cycles and elicited no pregnancies. Thirty-seven couples with moderate OAS received protocol A for 169 cycles, and 2 pregnancies ensued (5.4% per couple and 1.12% per cycle). Twelve of the 35 nonpregnant couples with moderate OAS then received protocol B for 31 cycles, and 4 pregnancies were recorded (33.3% per couple and 12.9% per cycle).


Ordering in asymmetric block copolymer films by a compressible fluid.
We examine the morphological structures of asymmetric poly(ethylene oxide)-b-poly(1,1'-dihydroperflurooctyl methacrylate) (PEO-b-PFOMA) thin films upon annealing in a compressible fluid, supercritical CO2 (Sc-CO2). The strong affinity between PFOMA and CO2 is found to induce phase segregation when annealing PEO-b-PFOMA films at the same temperature as compared with vacuum. In vacuum, PEO-b-PFOMA films remain disordered from 80 to 180 degrees C, whereas, in Sc-CO2 at 13.9 MPa, an upper order-disorder transition (UODT) between 116 and 145 degrees C is found. In Sc-CO2, the observed ordered structure is layers of PEO spheres embedded in the matrix of PFOMA, followed by a brush layer, in which PEO wets the substrate. The swelling isotherms of PFOMA and PEO in CO2 are correlated with the Sanchez-Lacombe equation of state (SLEOS) to estimate the interaction parameters, XPFOMA-CO2 and XPEO-CO2. The phase segregation (order) induced by CO2 relative to vacuum at a given temperature is explained in terms of two factors: (1) copolymer volume fraction upon dilution with CO2, phi, and (2) the relative interaction parameter, DeltaX= XPEO-CO2 - XPFOMA-CO2. The latter factor favors order and is dominant at low temperatures over the phi factor, which always favors disorder. At high temperatures (above the T(ODT)), the preferential swelling of PFOMA by CO2 is less pronounced ( DeltaX decreases), and the copolymer is disordered.


Predicting Colloidal Stability of High-Concentration Monoclonal Antibody Formulations in Common Pharmaceutical Buffers Using Improved Polyethylene Glycol Induced Protein Precipitation Assay.
Colloidal stability is an important consideration when developing high concentration mAb formulations. PEG-induced protein precipitation is a commonly used assay to assess the colloidal stability of protein solutions. However, the practical usefulness and the current theoretical model for this assay have yet to be verified over a large formulation space across multiple mAbs and mAb-based modalities. In the present study, we used PEG-induced protein precipitation assays to evaluate colloidal stability of 3 mAbs in 24 common formulation buffers at 20 and 5 °C. These prediction assays were conducted at low protein concentration (1 mg/mL). We also directly characterized high concentration (100 mg/mL) formulations for cold-induced phase separation, turbidity, and concentratibility by ultrafiltration. This systematic study allowed analysis of the correlation between the results of low concentration assays and the high concentration attributes. The key findings of this study include the following: (1) verification of the usefulness of three different parameters (Cmid, μB, and Tcloud) from PEG-induced protein precipitation assays for ranking colloidal stability of high concentration mAb formulations; (2) a new method to implement PEG-induced protein precipitation assay suitable for high throughput screening with low sample consumption; (3) improvement in the theoretical model for calculating robust thermodynamic parameters of colloidal stability (μB and εB) that are independent of specific experimental settings; (4) systematic evaluation of the effects of pH and buffer salts on colloidal stability of mAbs in common formulation buffers. These findings provide improved theoretical and practical tools for assessing the colloidal stability of mAbs and mAb-based modalities during formulation development.


The augmented Roothaan-Hall method for optimizing Hartree-Fock and Kohn-Sham density matrices.
We present a novel method for the optimization of Hartree-Fock and Kohn-Sham energies that does not suffer from the flaws of the conventionally used two-step Roothaan-Hall (RH) with direct inversion in iterative subspace (DIIS) acceleration scheme, improving the reliability of the optimization while reducing its cost. The key to its success is the replacement of the two separate steps of each RH/DIIS iteration by a single concerted step that fully exploits the Hessian information available from the previous iterations. It is a trust-region based method and therefore by design converges to an energy minimum. Numerical examples are given to illustrate that the algorithm is robust and cost efficient, converging smoothly to a minimum also in cases when the RH/DIIS algorithm fails to converge or when it converges to a saddle point rather than to a minimum. The algorithm is based on matrix multiplications and becomes linearly scaling for sufficiently large systems.


Analysis of a wooden specimen's mechanical properties through acoustic measurements in the very near field.
This paper presents the method for prediction of some basic mechanical parameters of a wood specimen by using simple instrumentation. The standard acoustic method for assessment of modulus of elasticity using the impulse excitation is extended by using a recording in the very near field. In this way, visualization of vibration patterns of the analyzed sample was accomplished for all modes below 5 kHz. This enables detection of signals with a good signal-to-noise ratio over a wide frequency range which is then used for loss factor assessment.


MRI-Based Assessment of Lower-Extremity Muscle Volumes in Patients Before and After ACL Reconstruction.
Study of muscle volumes in patients after anterior cruciate ligament (ACL) injury and reconstruction (ACL-R) is largely limited to cross-sectional assessment of the thigh musculature, which may inadequately describe posttraumatic and postsurgical muscle function. No studies have prospectively examined the influence of ACL injury and reconstruction on lower-extremity muscle volumes. Assess magnetic resonance imaging-derived lower-extremity muscle volumes, and quantify quadriceps strength and activation in patients following ACL injury and reconstruction. Prospective case series. Research laboratory and magnetic resonance imaging facility. Patients (or Other Participants): Four patients (2 men and 2 women; age = 27.4 (7.4) y, height = 169.2 (8.1) cm, and mass = 74.3 (18.5) kg) scheduled for ACL-R. Thirty-five muscle volumes were obtained from a bilateral lower-extremity magnetic resonance imaging before and after ACL-R. Muscle volumes expressed relative to (1) a normative database presurgery and postsurgery, (2) limb symmetry presurgery and postsurgery, and (3) percentage change presurgery to postsurgery. Quadriceps function was quantified by normalized knee extension maximal voluntary isometric contraction torque and central activation ratio. Involved vastus lateralis and tibialis anterior were consistently smaller than healthy individuals (z < -1 SD) presurgery and postsurgery in all patients. Involved rectus femoris and vastus lateralis were more than 15% smaller than the contralateral limb presurgery, whereas the involved rectus femoris, gracilis, vastus medialis, vastus intermedius, and vastus lateralis muscle volumes exceeded 20% asymmetry postoperatively. Involved gracilis and semitendinosus atrophied more than 30% from presurgery to postsurgery. Involved maximal voluntary isometric contraction torque and central activation ratio increased by 12.7% and 12.5%, respectively, yet strength remained 33.2% asymmetric postsurgery. Adaptations in lower-extremity muscle volumes are present following ACL injury and reconstruction. Anterior thigh and shank muscles were smaller than healthy individuals, and large asymmetries in quadriceps volumes were observed presurgery and postsurgery. Selective atrophy of the semitendinosus and gracilis occurred following surgery. Volumetric deficits of the quadriceps musculature may exist despite improvements in muscle strength and activation.


A Convenient Method for Extraction and Analysis with High-Pressure Liquid Chromatography of Catecholamine Neurotransmitters and Their Metabolites.
The extraction and analysis of catecholamine neurotransmitters in biological fluids is of great importance in assessing nervous system function and related diseases, but their precise measurement is still a challenge. Many protocols have been described for neurotransmitter measurement by a variety of instruments, including high-pressure liquid chromatography (HPLC). However, there are shortcomings, such as complicated operation or hard-to-detect multiple targets, which cannot be avoided, and presently, the dominant analysis technique is still HPLC coupled with sensitive electrochemical or fluorimetric detection, due to its high sensitivity and good selectivity. Here, a detailed protocol is described for the pretreatment and detection of catecholamines with high pressure liquid chromatography with electrochemical detection (HPLC-ECD) in real urine samples of infants, using electrospun composite nanofibers composed of polymeric crown ether with polystyrene as adsorbent, also known as the packed-fiber solid phase extraction (PFSPE) method. We show how urine samples can be easily precleaned by a nanofiber-packed solid phase column, and how the analytes in the sample can be rapidly enriched, desorbed, and detected on an ECD system. PFSPE greatly simplifies the pretreatment procedures for biological samples, allowing for decreased time, expense, and reduction of the loss of targets. Overall, this work illustrates a simple and convenient protocol for solid-phase extraction coupled to an HPLC-ECD system for simultaneous determination of three monoamine neurotransmitters (norepinephrine (NE), epinephrine (E), dopamine (DA)) and two of their metabolites (3-methoxy-4-hydroxyphenylglycol (MHPG) and 3,4-dihydroxy-phenylacetic acid (DOPAC)) in infants' urine. The established protocol was applied to assess the differences of urinary catecholamines and their metabolites between high-risk infants with perinatal brain damage and healthy controls. Comparative analysis revealed a significant difference in urinary MHPG between the two groups, indicating that the catecholamine metabolites may be an important candidate marker for early diagnosis of cases at risk for brain damage in infants.


Effectiveness of melatonin on aflatoxicosis in chicks.
The efficacy of melatonin co-administration on aflatoxicosis in chicks was investigated. Ross PM3 breed chicks were divided into groups of 10 and given conventional feed. One of the groups was kept as a control (C), and the others were given 150ppb aflatoxin (AF1), 300ppb aflatoxin (AF2), 150ppb aflatoxin plus 10mg/kg/bwt melatonin (AF1+M), 300ppb aflatoxin plus 10mg/kg/bwt melatonin (AF2+M), 10mg/kg/bwt melatonin (M), and 1% ethanol (E). After 21 day-treatment period, the chicks were sacrificed, liver and kidney tissues were collected, processed for immuno-histochemical staining, in situ TUNEL method, and biochemical analyses. Vacuolar degeneration, necrosis, bile duct hyperplasia in liver, and mild tubular degeneration in kidney were detected in AF groups. Pathological changes were markedly reduced in AF+M groups, and a microscopic view similar to group C was observed. Increased immunoreactivity against inducible nitric oxide synthase (iNOS) and nitrotyrosine was detected in AF groups compared to weak immunoreactivity in group C. Immunoreactivity in AF+M groups was markedly reduced compared to AF groups and was similar to group C in liver and kidney. Many apoptotic cells were detected in the livers of AF groups, whereas there were no apoptotic cells in AF+M groups. While reduced glutathione (GSH) levels in liver and kidney of AF groups were greatly reduced, malondialdehyde (MDA) levels increased. With melatonin co-administration, the levels of GSH and MDA approached to the values of group C. These results indicated that nitrosative tissue degeneration caused by aflatoxin could be greatly reduced by melatonin supplementation in chicks.


[From the early history of roentgen documentation].
By chance, eight X-ray plates dating from 1899 were found in 1986. Each of these glass plates shows the patient's name, the date of the examination were performed in Vienna by Prof. Franz Exner, who was a friend of W.C. Roentgen. The glass plates are 25 x 13 cm in area and 3 mm thick. One side is coated with silver bromide. Probably these are some of the earliest X-rays records in the history of radiology. Their quality is comparable with the quality that can be achieved today, even though the materials and the imaging method used were somewhat unsophisticated.


CD44 SNPrs187115: A Novel Biomarker Signature that Predicts Survival in Resectable Pancreatic Ductal Adenocarcinoma.
Although pancreatic ductal adenocarcinoma (PDAC) is an aggressive tumor, like other common cancers, it displays a wide range of biology. However, at present, there are no reliable tests to predict patients' cancer-specific outcomes and guide personalized treatment decisions. In this study, we aim to identify such biomarkers in resectable PDAC by studying SNPs in the CD44 gene, which drives the progression of pancreatic cancer. A total of 348 PDAC patients from three independent cohorts [Switzerland, Germany, The Cancer Genome Atlas (TCGA)] who underwent pancreatic resection are included in the study. Information on the haplotype structure of the CD44 gene is obtained using 1000 Genomes Project data, and the genotypes of the respective tagging SNPs are determined. Cox proportional hazards models are utilized to analyze the impact of SNP genotype on patients' survival. We identify an SNP in the CD44 gene (SNPrs187115) that independently associates with allelic differences in prognosis in all study cohorts. Specifically, in 121 Swiss patients, we observe an up to 2.38-fold (P = 0.020) difference in tumor-related death between the genotypes of SNPrs187115 We validate those results in both the German (HR = 2.32, P = 0.044, 101 patients) and the TCGA cohort (HR = 2.36, P = 0.044, 126 patients). CD44 SNPrs187115 can serve as a novel biomarker readily available at the time of PDAC diagnosis that identifies patients at risk for faster tumor progression and guide personalized treatment decisions. It has the potential to significantly expand the pool of patients that would benefit from tumor resection. Clin Cancer Res; 22(24); 6069-77. ©2016 AACR.


Clonal spread of Acinetobacter baumannii in a general intensive care unit.
The epidemiological characterization of multiply resistant Acinetobacter baumannii isolates from a six-bed Intensive Care Unit (ICU) is described. Investigations for A. baumannii were performed in three subsequent surveillance studies. In the first study, surveillance cultures were taken from patients, health care personnel and the environment; in the second study surveillance cultures were taken at 0, 4, and 7 days from all patients admitted consecutively to the ward; and in the third study surveillance cultures were taken from patients, health care personnel and the environment. During the first study all four hospitalized patients were found to harbour A. baumannii. Hand cultures did not grow any A. baumannii when staff entered the ward from home, but 7 positive health care workers were identified out of 25 samples taken during work, and two cultures of environmental specimens grew A. baumannii. During the second study, 4 of 86 (4.6%) patients resulted colonized with A. baumannii. In the third epidemiological study, no A. baumannii was cultured from either patients, health care personnel or the environment. All isolates recovered from various patients or sources produced conserved macrorestriction Pulsed-Field Gel Electrophoresis (PFGE) patterns and showed the same antibiotic resistance; therefore, they can be considered indistinguishable. The same antibiotic resistance and macrorestriction patterns were observed in previously isolated A. baumannii strains in the ward during May 1997, suggesting the persistence of a single A. baumannii in the ICU. The present study confirms that molecular typing is an essential tool in the epidemiology and control of nosocomial infections, showing here the persistence of a single A. baumannii clone in the ICU. The origin of this strain remains unknown but, when basic infection control measures were reinforced, emphasizing the importance of hand antisepsis and judicious use of gloves, control of A. baumannii spread in the ward was achieved.