Analysis of cannabinoids by liquid chromatography-mass spectrometry in milk, liver and hemp seed to ensure food safety.
A method for determining cannabinoids, Δ9-tetrahidrocannabinol (THC), 11-nor-9-carboxy-Δ9-THC (THC-COOH) and 11-hidroxy-Δ9-THC (THC-OH) in milk, liver and hemp seeds based on liquid chromatography tandem mass spectrometry has been optimized and validated. Analytes were extracted with methanol and the extracts cleaned-up by solid-phase extraction using Oasis HLB (60mg). The developed method was validated according to the Commission Decision 2002/657/EC. The decision limit (CCα) and detection capability (CCβ) ranged from 3.10-10.5ngg-1 and 3.52-11.5ngg-1, the recoveries were 76-118% and matrix effect ranged from -17.8% to 19.9% in the three matrices studied. The method was applied to food samples obtaining positive results for THC in hemp seeds (average 0.82μgg-1) and three brands of junior formula milk at concentrations from 4.76 to 56.11ngg-1. The developed method was suitable achieving identification and quantification of cannabinoids in food matrices.


[Daily interruption of sedation: always a quality indicator?].
The Spanish Society of Critical Care Medicine (SEMICYUC) has recently published an updated version of Quality Indicators in Critical Care. Daily sedative interruption is included among them. As this practice is controversial, research studies are revised and guidelines for its implementation are proposed.


Humoral signals mediate the circadian expression of rat period homologue (rPer2) mRNA in peripheral tissues.
Northern blot analysis revealed the circadian expression of rat period homologue (rPer2) mRNA in peripheral mononuclear leukocytes that have no neuronal connections. The mRNA expression in the suprachiasmatic nucleus (SCN), peripheral mononuclear leukocytes, and other peripheral tissues exhibited prominent, synchronous circadian oscillation with peaks at early night. As we have previously shown the peripheral circadian expression of rPer2 mRNA is abolished in SCN-lesioned rats [Sakamoto, K., Nagase, T. Fukui, H., Horikawa, K., Okada, T., Tanaka, H., Sato, K., Miyake, Y., Ohara, O., Kako, K. and Ishida, N., J. Biol. Chem., 273 (1998) 27039-27042], our findings suggest that some humoral signals, driven by the SCN, mediate the circadian expression of mammalian per homologues in peripheral tissues.


Preliminary note on the differentiation of angioblasts and the method by which they produce blood-vessels, blood-plasma and red blood-cells as seen in the living chick. 1917.
Reprinted from The Anatomical Record, Vol. 13, pp. 199-204, 1917.


Bilateral visual loss: more than meets the eye.
A 72-year-old woman presented with acute onset bilateral visual loss. She had no other symptoms or signs, but had a complex past medical history including blood transfusions and immunosuppression. T2-weighted magnetic resonance imaging demonstrated bilateral occipital lobe vasogenic edema, consistent with posterior reversible encephalopathy syndrome (PRES). Her vision improved with conservative management.


Deaths and injuries attributed to infant crib bumper pads.
To document deaths attributed to bumper pads and injuries from their use that are potentially preventable. The US Consumer Product Safety Commission maintains files on cases voluntarily reported to them of deaths and injury related to commercial products. These cases represent an unknown fraction of total occurrences. We searched this database for deaths related to crib bumpers for the years 1985 to 2005. We also searched other Consumer Product Safety Commission databases for crib-related injuries that potentially might have been prevented by bumpers. Additionally, we examined 22 retail crib bumpers and described features that could be hazardous. Twenty-seven accidental deaths reported by medical examiners or coroners were attributed to bumper pads. The mechanism of death included suffocation and strangulation by bumper ties. Twenty-five nonfatal injuries were identified, and most consisted of minor contusions. All retail bumpers had hazardous properties. These findings suggest that crib and bassinet bumpers are dangerous. Their use prevents only minor injuries. Because bumpers can cause death, we conclude that they should not be used.


Time and again: effects of repetition and retention interval on 2 year olds' event recall.
How and what very young children remember is a central question for understanding the course of memory development. In this research, we examined the effects of two factors on 2-year-old children's ability to recall novel events: repetition of the experience and time since experience. Twenty 24-month-old and twenty 28-month-old children participated in unusual laboratory play events. Half of the children returned after a 2-week delay and again after a 3-month delay (repeated experience condition); the remaining children returned only after 3 months (single experience condition). Memory was assessed by asking children to reenact the events. Recall was generally accurate, and there were no significant effects of age. All children recalled more information about the activities associated with the event than about the objects. Surprisingly, children in the repeated experience condition recalled as much about the events at the 3-month retention interval as at the 2-week retention interval. Further, children in this condition recalled more information at the 3-month retention interval than children in the single experience condition, suggesting that reexperiencing an event may guard against long-term forgetting.


Factors influencing left atrial volume in a population with preserved ejection fraction: Left ventricular diastolic dysfunction or clinical factors?
Increased left atrial volume (LAV) predicts a higher incidence of cardiovascular events and is widely recognized as a major surrogate marker of left ventricular (LV) diastolic dysfunction (DD). Although the pathophysiology of LA enlargement is probably multifactorial, few studies have examined comprehensively the clinical factors that lead to LA enlargement in the absence of valvular disease or LV systolic dysfunction. Therefore, we investigated associations between LAV and several clinical and echocardiographic parameters including DD. We enrolled 557 subjects without significant valve disease or LV systolic dysfunction from the health check-up clinic retrospectively. We performed univariable and multivariable linear regression using lnLAV index as the dependent variable and the following independent variables: gender, age, smoking status, drinking habit, hypertension, diabetes, body mass index (BMI), LV ejection fraction, DD, LV mass index, hemoglobin, serum creatinine, serum total cholesterol, serum uric acid, serum sodium, and serum iron. In multivariable analysis, LAV index was independently associated with BMI, lower hemoglobin, and moderate and severe DD compared with normal diastolic function (p<0.001), but not with mild DD (p=0.70). LA enlargement was independently associated with moderate and severe DD, but not with mild DD. Furthermore, obesity and lower hemoglobin were associated with LAV independently of DD.


Magnetic resonance compatibility of intraocular lenses measured at 7 Tesla.
To determine whether intraocular lenses (IOLs) are compatible with magnetic resonance imaging (MRI) at a magnetic field strength of 7 Tesla, the highest field strength at which clinical MRI scans are performed. A set of 23 intraocular lenses was selected based on the presence of dyes and metals and different geometric shapes. MR compatibility was evaluated in a high-field 7-Tesla MRI scanner according to the American Standard Test Method (ASTM). The magnetically induced displacement was measured via the angular deflection method. The degree of magnetic susceptibility artifact formation was evaluated by positioning the IOLs in a phantom gel for scanning, using a three-dimensional gradient echo (GRE) sequence. All images were visually inspected to determine the spatial extent of any signal voids. Fiber-optic temperature probes were deployed to measure radio-frequency (RF) heating using a GRE sequence with powers 10 times higher than clinical settings. No significant displacement was detected with any of the tested IOLs. A significant magnetic susceptibility artifact was caused by the small platinum component of the Worst Platinum Clip IOL. None of the other 22 IOLs caused measurable susceptibility artifacts. Measurements on RF-induced heating showed no significant temperature rise (<0.25°C) of the tested IOLs. MRI did not induce movement or RF heating of any of the IOLs. We conclude that all the tested intraocular lenses are considered safe for MRI up to and including 7 Tesla. One IOL, the Worst Platinum Clip IOL, caused a significant imaging artifact.


Derivation of a clinical prediction rule for chronic post-surgical pain after total knee arthroplasty considering biopsychosocial factors: A prospective cohort study.
Biopsychosocial factors are involved in the occurrence of chronic post-surgical pain (CPSP) after total knee arthroplasty (TKA). The purpose of this study was to develop a clinical prediction rule (CPR) that considers biopsychosocial factors to predict which patients are more likely to develop CPSP after TKA. CPSP after TKA was dichotomized into CPSP and non-CPSP groups using the Likert scale and Minimal clinically important difference, and binomial logistic regression analysis was performed. Cut-off values were then calculated using the extracted factors and dichotomized variables. The cut-off values and dichotomized variables were then used to derive a CPR that discriminates between groups with and without CPSP. Seventy-one TKA patients were included in the study. Binomial logistic regression analysis revealed that Central Sensitization Inventory (CSI) and Pittsburgh Sleep Quality Index (PSQI) were associated with CPSP. The cut-off values for CSI and PSQI were 26 and 7, respectively. The CPSP scale was created using the cut-off values of CSI and PSQI, with a score of 0 for being below the cut-off values of both CSI and PSQI, 1 for being above the cut-off values of either CSI or PSQI, and 2 for being above the cut-off values of both CSI and PSQI. Furthermore, the area under the curve (AUC) for CPR created by the presence of CPSP and using the CPSP scale was significant (AUC = 0.766; P = 0.001). The combination of the two tests, CSI and PSQI, suggested the possibility of predicting CPSP after TKA.


Pre- and postjunctional effects of neuropeptide Y on the rabbit isolated ear artery.
1. In the isolated perfused and superfused rabbit ear artery, neuropeptide Y (NPY, 0.3-100 nmol/l) had no direct vasoconstrictor action, but produced a concentration-dependent and reversible enhancement of vasoconstrictor responses to both sympathetic nerve stimulation and exogenous noradrenaline. 2. In arteries in which the noradrenergic transmitter stores had been radiolabelled with [3H]-noradrenaline, 100 nmol/l NPY inhibited the stimulation-induced (1 Hz for 30 s) release of radioactivity, but the lower concentrations tested (10 and 30 nmol/l) had no effect. NPY (10, 30 and 100 nmol/l) had no effect on the resting release of radioactivity. 3. Thus, NPY in low concentrations enhances vasoconstrictor responses in the rabbit ear artery by a postjunctional action; prejunctionally, NPY inhibits stimulation-induced transmitter release when it is present in high concentrations.


Tunable Domain Switching Features of Incommensurate Antiferroelectric Ceramics Realizing Excellent Energy Storage Properties.
An incommensurate modulated antiferroelectric phase is a key part of ideal candidate materials for the next generation of dielectric ceramics with excellent energy storage properties. However, there is less research carried out when considering its relatively low polarization response. Here, the incommensurate phase is modulated by stabilizing the antiferroelectric phase and the energy storage performance of the incommensurate phase under ultrahigh electric field is studied. The tape-casting method is applied to construct dense and thin ceramics. La3+ doping induces a room-temperature incommensurate antiferroelectric orthorhombic matrix. With little Cd2+ , the extremely superior energy storage performances arose as follows: when 0.03, the recoverable energy storage density reaches ≈19.3 J cm-3 , accompanying an ultrahigh energy storage efficiency of ≈91% (870 kV cm-1 ); also, a giant discharge energy density of ≈15.4 J cm-3 emerges during actual operation. In situ observations demonstrate that these superior energy storage properties originate from the phase transition from the incommensurate antiferroelectric orthorhombic phase to the induced rhombohedral relaxor ferroelectric one. The adjustable incommensurate period affects the depolarization response. The revealed phase-transition mechanism enriches the existing antiferroelectric-ferroelectric transition. Attention to the incommensurate phase can provide a reference for the selection of the next generation of high-performance antiferroelectric materials.


Shifting the imbalance: Intentional reuse of Dutch sewage effluent in sub-surface irrigation.
Worldwide, agricultural irrigation currently accounts for 69% of freshwater withdrawal. Countries with a temperate climate, such as the Netherlands, experience periodic freshwater shortages in agriculture. The pressure on available freshwater will increase due to climate change and a growing demand for freshwater by e.g. industrial activities. Possible alternative water resources are considered in order to meet the current and future water demand. In this study we explore where, and how much, sewage treatment plant (STP) effluent can directly be reused in agricultural sub-surface irrigation (SSI) during an average and a dry season scenario, for all active (335) Dutch STPs. SSI systems may have a higher water demand as part of the STP effluent is transported with groundwater flow, although aboveground irrigation has a loss of water due to interception. Furthermore, such aboveground irrigation systems provide direct contact of crops with irrigation water. SSI systems provide a soil barrier which may function as a filter and buffer zone. In the Dutch situation, direct intentional reuse of STP effluent can fulfill up to 25% of croplands SSI water demand present within a five-kilometer transport buffer from the STPs during an average season and 17% during a dry season. Hereto, respectively, 78% and 84% of the total available Dutch STP effluent would be used. Thus, the intentional direct STP effluent reuse in agricultural SSI has the potential to satisfy a significant amount of the agricultural water demand at a national scale, presuming responsible reuse: safe applications for humans and environment and no limiting effects on water availability for other actors.


Quantification of shoulder muscle intramuscular fatty infiltration on T1-weighted MRI: a viable alternative to the Goutallier classification system.
Quantification of rotator cuff intramuscular fatty infiltration is important for clinical decision-making in patients with rotator cuff tear. The semi-quantitative Goutallier classification system is the most commonly used method, but has limited reliability. Therefore, we sought to test a freely available fuzzy C-means segmentation software program for reliability of the quantification of shoulder intramuscular fatty infiltration on T1-weighted MR images and for correlation with fat fraction by six-point Dixon MRI. We performed a prospective cross-sectional study to measure visible intramuscular fat area percentage on oblique sagittal T1 MR images by fuzzy C-means segmentation and fat fraction maps by six-point Dixon MRI for 42 shoulder muscles. Intra- and inter-observer reliability were determined. Correlative analysis for fuzzy C-means and six-point Dixon intramuscular fatty infiltration measures was also performed. We found that inter-observer reliability for the quantification of visible intramuscular fat area percentage by fuzzy C-means segmentation and fat fraction by six-point Dixon MRI was 0.947 and 0.951 respectively. The intra-observer reliability for the quantification of visible intramuscular fat area percentage by fuzzy C-means segmentation and fat fraction by six-point Dixon MRI was 0.871 and 0.979 respectively. We found a strong correlation between fuzzy C-means segmentation and six-point Dixon techniques; r = 0.850, p < 0.001 by individual muscle; and r = 0.977, p < 0.002 by study subject. Quantification of intramuscular fatty infiltration by fuzzy C-means segmentation on T1-weighted sequences demonstrates excellent reliability and strong correlation with fat fraction by six-point Dixon MRI. Quantitative fuzzy C-means segmentation is a viable alternative to the semi-quantitative Goutallier classification system.


Topical high molecular weight hyaluronan reduces radicular pain post laminectomy in a rat model.
A controversy exists about the mechanism of causation of the post-laminectomy pain syndrome. Some believe that epidural scarring, and attendant spinal nerve and nerve root scarring and tethering to the disc or pedicle at the site of surgery contributes to post-laminectomy pain in such patients. However, clinical outcome studies on this question are inconclusive and the assertion remains controversial. Definitive studies to help resolve the question are needed. Previously our laboratory has reported on a preclinical post-laminectomy model that mimics the postoperative proliferative fibrotic response grossly, as well as by biochemical assessment of the collagen content within the spinal canal. The post-laminectomy fibrotic response was attenuated in that study by application of a topical antifibrotic (high molecular weight hyaluronan gel) or by insertion of an absorbable roofing barrier (0.2-mm-thick Macropore sheet material) over the laminectomy defect before wound closure. The question remains of relevance of the attenuation of the fibrotic response to post-laminectomy chronic pain syndromes. The purpose of this study is to evaluate the effect of therapeutic attenuation of proliferative scar within the spinal canal post laminectomy on the pain-related behavioral response in a preclinical rat model. An established L5-L6 rat laminectomy model with a unilateral L5-6 disc injury was employed to assess postoperative proliferative fibrosis of the L5 spinal nerves using quantitative biochemical hydroxyproline assessment of the collagen content in four experimental groups. These observations were correlated with gross descriptions of spinal nerve scarring or tethering. Associated manifestations of a sensory pain-related response in the L5 spinal nerve receptor area of the hind paws was studied using standard tactile allodynia assessment with the von Frey hair technique. The tactile allodynia findings were supplemented by weekly descriptors of behavioral pain manifestations. Bilateral laminectomies at L5 and L6 and a unilateral right disc injury (L5-6) were performed on 35 male adult Sprague-Dawley rats, weighing 400+ grams (approved by the VA Institutional Animal Care Use Committee). The study consisted of four groups: 1) normal nonoperative control; 2) a sham-operated group; 3) an untreated laminectomy-disc injury group; and 4) a laminectomy-disc injury treatment group in which 0.1 cc topical high molecular weight hyaluronan (HMW HA) gel was layered over the dura and into the laminectomy canal before closure. Before animals were entered into the study, they were checked for the presence of abnormal response to the tactile testing procedure of the L5 sensory receptor area. Animals exhibiting anomalous responses were excluded from the study. Behavioral testing for tactile allodynia was performed at weekly intervals post laminectomy beginning at 3 weeks. Pain-related behavior was characterized at weekly intervals. A behavioral test cage with a wire mesh floor allowed for tactile allodynia testing. Graduated von Frey hairs whose stiffness increased logarithmically from 0.41 to 15 g were used for tactile allodynia tests. The animals were killed 8 weeks postoperatively for analysis. The dissected spinal nerve and nerve root specimens were studied biochemically for hydroxyproline content to estimate total collagen in and around the L5 neural structures. Statistical analyses were performed using analysis of variance and a Fisher comparison t test. The major observations on the untreated preclinical post-laminectomy rat model previously described by this laboratory were confirmed. All untreated animals developed a tail contracture concave toward the right (disc injury side) consistent with asymmetrical lumbar muscle spasm. Only one animal in the HA gel treatment group had a tail contracture. It was of mild degree and occurred in an animal that demonstrated slightly increased right L5 tactile sensitivity. Gross inspection of the dissected specimens demonstrated spinal nerve scarring and tethering to the disc and pedicle greater on the right than the left in untreated animals, findings that were markedly reduced in the treatment group. Collagen content of the L5 spinal nerve and nerve roots with attached scar were significantly lower in the HA gel treatment group than in the untreated laminectomy group (p=.0014). Pain behavioral testing of the L5 receptor area of the right hind paw in the untreated laminectomy group showed markedly increased sensitivity to tactile allodynia testing compared with the corresponding limb of the control group (p=.0001), to the corresponding limb of the sham group (p=.0001), and compared with the HMW HA gel treatment group (p=.0010). Comparisons of the pain behavioral data between the sham and the post-laminectomy HA gel treatment group and the control animals lacked statistical significance. This study supports the concept of a relationship between perineural fibrosis and radicular neuropathy in the model described, and emphasizes the role of disc injury and spinal nerve retraction in the post-laminectomy fibrotic process. Furthermore, it shows promise for preliminary assessment of interventions with other anti-inflammatory agents, for characterization of the neurochemical profile of the post-laminectomy pain state, and for exploration of newer pharmaceutical agents potentially useful in the prevention or management of the post-laminectomy syndrome. Post-laminectomy scar is but one of many potential causes of the post-laminectomy pain syndrome. Furthermore, a cautionary note must be emphasized as in all studies using preclinical models, conclusions drawn from the studies cannot be extended directly to patients without confirmatory clinical follow-up studies.


Liberating crystallographers.
In this issue of Structure, DePristo et al. describe a new program, RAPPER, that should automate some of the most time-consuming tasks associated with rebuilding, refining, and completing protein crystal structures at moderate resolutions.


Comparison of light-emitting diode wavelength on activity and migration of rabbit ACL cells.
The purpose of this study was to evaluate the biological response and gene expression of New Zealand White Rabbit anterior cruciate ligament (ACL) fibroblasts for different wave lengths of light-emitting diode (LED) irradiation. In other words, this study was undertaken to evaluate the effects of different wavelengths of LED irradiation on cell growth, expression of extracellular matrix and growth factors, migration, and expression of actin and integrin. Proliferation assay showed that red (630 nm, 9.5 J/cm(2)) and green LED (530 nm, 9.8 J/cm(2)) irradiated cells were more increased than control group but there was no difference between the control group and the blue LED (460 nm, 27 J/cm(2)) irradiated group. Moreover, the expression of insulin-like growth factor, transforming growth factor-beta (TGF-β1), and collagen I were significantly increased in the red and green LED-irradiated group, but the expression of collagen was decreased in the blue LED-irradiated group. The results of staining showed that collagen and TGF-β1 were weaker in the control group and blue LED-irradiated cells, but stronger in the red and green LED-irradiated cells. Also, in the red and green LED-irradiated group, the expression of actin and integrin was not changed compared to the control group, but the expression of actin and integrin was decreased in the blue irradiated group. This study revealed that irradiation with a wavelength of 460 nm (blue LED) is cytotoxic to ACL cells, but irradiation with nontoxic fluencies of 530 (green LED) and 630 nm (red LED) wavelengths induced cell growth in cultured ACL cells.


A common neural code for meaning in discourse production and comprehension.
How does the brain code the meanings conveyed by language? Neuroimaging studies have investigated this by linking neural activity patterns during discourse comprehension to semantic models of language content. Here, we applied this approach to the production of discourse for the first time. Participants underwent fMRI while producing and listening to discourse on a range of topics. We used a distributional semantic model to quantify the similarity between different speech passages and identified where similarity in neural activity was predicted by semantic similarity. When people produced discourse, speech on similar topics elicited similar activation patterns in a widely distributed and bilateral brain network. This network was overlapping with, but more extensive than, the regions that showed similarity effects during comprehension. Critically, cross-task neural similarities between comprehension and production were also predicted by similarities in semantic content. This result suggests that discourse semantics engages a common neural code that is shared between comprehension and production. Effects of semantic similarity were bilateral in all three RSA analyses, even while univariate activation contrasts in the same data indicated left-lateralised BOLD responses. This indicates that right-hemisphere regions encode semantic properties even when they are not activated above baseline. We suggest that right-hemisphere regions play a supporting role in processing the meaning of discourse during both comprehension and production.


A Heterozygous Variant of FGF13 Caused X-Linked Developmental and Epileptic Encephalopathy 90 in a Chinese Family.
Developmental and epileptic encephalopathy (DEE) refers to a group of severe epilepsy encephalopathy and development disorders, and its typical clinical features include seizures, drug resistance, and developmental delay or regression. To date, limited studies have reported DEEs driven by FGF13. Here, we reported a girl with developmental and epileptic encephalopathy 90 caused by variant of FGF13. Her electroencephalogram (EEG) showed discontinuous hypsarrhythmia, and a heterozygous nonsynonymous variant in FGF13 [NM_004114.4: c.5C>G, p.(Ala2Gly)] was identified from the proband. The variant was not reported in public databases such as gnomAD and Exome Aggregation Consortium (ExAC), and was predicted to be damaging to proteins and classified as likely pathogenic according to the ACMG guidelines. The seizure was finally controlled by a combination of ACTH + zonisamide (10 mg/kg.d) + levetiracetam (52 mg/kg.d) + clonazepam (0.7 mg/kg.d).


In situ estimates of annual net nitrogen mineralization and nitrification in a subarctic watershed.
Annual estimates of surface soil nitrogen transformations were determined using an in situ method in four different subarctic vegetation types within a watershed in southwestern Alaska. The net nitrogen mineralization estimates were 22.5, 0.5, 4.7, and 2.7 kg-N ha-1 yr-1 for the alder, dry tundra, moist tundra, and white spruce sites, respectively. Only the soil from the alder site showed net nitrification (about 10 kg-N ha-1 yr-1). Annual inogranic nitrogen flux from the overlying organic layer to the mineral soil was almost seven times greater than net N production in the surface mineral soil in the alder site, indicating that the alder forest floor is potentially a substantial source for plant-available N. Rates of mobilization of N from the surface organic layers of the other sites were similar to net N production rates in surface mineral soils. In situ rates of N transformations showed a similar trend among sites as did laboratory estimates conducted in a previous study, suggesting a strong substrate control of N transformations in these soils.


Responses of Bunias orientalis to Short-term Fungal Infection and Insect Herbivory are Independent of Nutrient Supply.
Plants have to allocate their resources in both growth and defense under different environmental challenges. Several plant species have become invasive particularly in disturbed fertile habitats, which may influence their resource allocation. We studied the effects of nitrate fertilization (low versus high) on various plant responses towards a pathogenic fungus, Alternaria brassicae, and a herbivorous insect species, Mamestra brassicae, in a population of Bunias orientalis, which is invasive in parts of central Europe. Aboveground biomass and leaf trichome density were enhanced in plants under high fertilization. In contrast, the short-term fungal infection and herbivory had no effect on aboveground biomass. Leaf water, nitrogen content and glucosinolate concentrations were neither affected by fertilization nor in response to antagonist attack. The total soluble sugar content, especially fructose, as well as leaf peroxidase activity increased significantly in leaves upon fungal infection, but independent of fertilization. Larval biomass gain and herbivore survival were likewise unaffected by fertilization. Our findings highlight that under conditions of high fertilization, B. orientalis plants allocate more resources into growth and morphological defenses than chemical defenses. In contrast, induced responses to short-term antagonist attack seem independent of nitrate availability in this population.


Gold-catalyzed cycloisomerization of 1,5-allenynes via dual activation of an ene reaction.
Tris(triphenylphosphinegold) oxonium tetrafluoroborate, [(Ph3PAu)3O]BF4, catalyzes the rearrangement of 1,5-allenynes to produce cross-conjugated trienes. Experimental and computational evidence shows that the ene reaction proceeds through a unique nucleophilic addition of an allene double bond to a cationic phosphinegold(I)-complexed phosphinegold(I) acetylide, followed by a 1,5-hydrogen shift.


The Amoebal Model for Macropinocytosis.
Macropinocytosis is a relatively unexplored form of large-scale endocytosis driven by the actin cytoskeleton. Dictyostelium amoebae form macropinosomes from cups extended from the plasma membrane, then digest their contents and absorb the nutrients in the endo-lysosomal system. They use macropinocytosis for feeding, maintaining a high rate of fluid uptake that makes assay and experimentation easy. Mutants collected over the years identify cytoskeletal and signalling proteins required for macropinocytosis. Cups are organized around plasma membrane domains of intense PIP3, Ras and Rac signalling, proper formation of which also depends on the RasGAPs NF1 and RGBARG, PTEN, the PIP3-regulated protein kinases Akt and SGK and their activators PDK1 and TORC2, Rho proteins, plus other components yet to be identified. This PIP3 domain directs dendritic actin polymerization to the extending lip of macropinocytic cups by recruiting a ring of the SCAR/WAVE complex around itself and thus activating the Arp2/3 complex. The dynamics of PIP3 domains are proposed to shape macropinocytic cups from start to finish. The role of the Ras-PI3-kinase module in organizing feeding structures in unicellular organisms most likely predates its adoption into growth factor signalling, suggesting an evolutionary origin for growth factor signalling.


Cuproptosis-related gene CDKN2A as a molecular target for IPF diagnosis and therapeutics.
Idiopathic pulmonary fibrosis (IPF) is a progressive chronic interstitial lung disease with limited therapeutic options. Cuproptosis is a recently proposed novel form of programmed cell death, which has been strongly implicated in the development of various human diseases. However, the prognostic and therapeutic value of cuproptosis-related genes (CRGs) in IPF remains to be elucidated. In the present study, weighted gene co-expression network analysis (WGCNA) was employed to identify the key CRGs associated with the development of IPF. The subsequent GSEA, immune cell correlation analysis, and single-cell RNA-Seq analysis were conducted to explore the potential role of the identified CRGs in IPF. In addition, ROC curves and survival analysis were used to assess the prognostic value of the key CRGs in IPF. Moreover, we explored the molecular mechanisms of participation of identified key CRGs in the development of pulmonary fibrogenesis through in vivo and in vitro experiments. The expression level of cyclin-dependent kinase inhibitor 2A (CDKN2A) is upregulated in the lung tissues of IPF patients and associated with disease severity. Notably, CDKN2A was constitutively expressed by fibrosis-promoting M2 macrophages. Decreased CDKN2A expression sensitizes M2 macrophages to elesclomol-induced cuproptosis in vitro. Inhibition of CDKN2A decreases the number of viable macrophages and attenuates bleomycin-induced pulmonary fibrosis in mice. These findings indicate that CDKN2A mediates the resistance of fibrosis-promoting M2 macrophages to cuproptosis and promotes pulmonary fibrosis in mice. Our work provides fresh insights into CRGs in IPF with potential value for research in the pathogenesis, diagnosis, and a new therapy strategy for IPF.


The combined Collis-Nissen operation for esophageal reflux strictures.
This report evaluates the efficacy of the combined Collis-Nissen operation in achieving long-term reflux control in patients with reflux strictures. A Collis-Nissen procedure with dilation of a reflux stricture was performed in 64 adults. The strictures were mild (easily dilated) in 37, moderate (requiring some force to dilate) in 17, and severe (requiring very forceful dilation) in 10. Two strictures were perforated intraoperatively. There was 1 postoperative death, and 4 patients have been lost to follow-up. The remaining 59 patients have been followed from 2 to 120 months (average, 43 months) after operation. Subjectively, reflux is absent in 48 (81%), mild in 4 (7%), moderate in 5 (9%), and severe in 2 (3%). Objectively, intraesophageal pH studies show good or excellent reflux control in 94% at 1 year and 66% at 2 to 5 years. Dysphagia has been eliminated in 71%, is mild in 10%, moderate (requiring occasional dilation) in 12%, and severe (requiring regular dilations) in 7%. The combined Collis-Nissen operation provides good long-term reflux control and relief of dysphagia in most patients with reflux strictures. However, patients with reflux strictures after previous repairs are likely to have unsatisfactory results and may best be managed with resectional therapy. Resection may also ultimately prove to be a better option for patients with more severe strictures.


CNS delivery of anti-CD52 antibodies modestly reduces disease severity in an animal model for multiple sclerosis.
Alemtuzumab is a humanized monoclonal antibody that depletes CD52-bearing B and T lymphocytes. Clinical trials defined that systemic administration of alemtuzumab reduces disease severity in the relapsing-remitting phase of multiple sclerosis (MS). However, its efficacy in progressive MS patients is limited, which may reflect the inability of alemtuzumab to cross the reconstituted BBB in these patients. Objective: to study whether central nervous system (CNS) delivery of anti-CD52 antibodies reduces disease severity and the neuroinflammatory burden in the experimental autoimmune encephalomyelitis (EAE) model. Anti-CD52 antibodies were administered intrathecally during the acute and chronic phases of EAE. Flow cytometry and immunohistochemistry were utilized to define immunological and pathological parameters. We show that subcutaneously administrated anti-CD52 antibodies completely abolish EAE disease severity. CNS delivery of anti-CD52 antibodies during both the acute and chronic phases of EAE moderately reduces disease severity and the neuroinflammatory burden. Our findings further suggest that CNS delivery of anti-CD52 antibodies impacts both the peripheral and CNS immune cell compartments in the EAE model but not in healthy mice. Collectively, our findings highlight the therapeutic potential of CNS delivery of alemtuzumab for the treatment of progressive as well as early MS.


Exploring the Spectral and Temporal Characteristics of Human Beatbox Sounds: A Preliminary Study.
Human beatbox is a developing hip-hop branch of music wherein impersonations of the percussion drum are done using manipulations of the oro-laryngopharyngeal structures. This study presents a preliminary attempt of exploration and documentation of the spectral and temporal measures of beatbox sounds produced by a single beatbox performer. An analytical observational study design was adopted wherein; audio recordings were taken from a professional beatboxer. The participant produced five different types of beatbox sequences consisting of classic kick, inward Ph snare with /ʃ/, throat bass, and uvular oscillation sounds. The recorded beatbox sounds were segmented into preburst and postburst events and were analyzed acoustically. The scrutiny of the results revealed that the beatbox productions shared characteristic features of linguistic sounds , namely, the following manners of productions, stops, fricatives, and affricates, and further, oro- and laryngopharyngeal regions of the vocal tract as the places of articulation. It is interesting to note that, the art of betboxing involves wide variety of articulatory configurations and also, the use of resonatory sub-system in the vocal tract.. This knowledge could expand the professional realm of Speech-Language Pathology necessitating the professionals to equip themselves with vocal demands of this relatively budding vocal art form called the beatboxing.


FTO promotes tumour proliferation in bladder cancer via the FTO/miR-576/CDK6 axis in an m6A-dependent manner.
The aberrant expression of fat mass and obesity-associated protein (FTO) has been confirmed to be associated with a variety of cancers and participates in the regulation of multiple biological behaviours. FTO plays an oncogenic role in bladder cancer, but few studies have focused on how FTO promotes bladder cancer progression by regulating miRNA synthesis. Here, we confirmed that FTO expression was significantly increased in bladder cancer and was associated with a poor prognosis. FTO overexpression promoted bladder cancer cell proliferation, whereas FTO knockdown inhibited bladder cancer cell proliferation. We also demonstrated that FTO promoted bladder cancer cell proliferation via the FTO/miR-576/CDK6 pathways. Taken together, our work revealed that FTO plays a critical role in bladder cancer and could be a potential diagnostic or prognostic biomarker for this disease.


The rate of reply and nature of responses to suicide-related posts on Twitter.
The social media platform Twitter has been used by individuals to communicate suicidal thoughts and intentions. Currently, the nature and rate of reply to this type of Twitter content is unknown. This brief report aimed to understand how Twitter users respond to suicide-related content as compared to non-suicide related content. Using a dataset of suicide and non-suicide related posts, replies, retweets and likes were analysed and compared. The content of the first replies to suicide-related posts were also reviewed. When compared to non-suicide related posts, those that were suicide-related received a significantly greater number of replies, with fewer retweets and likes. The rate of reply to the suicide-related posts was also significantly faster than that of the non-suicide related posts, with the average reply occurring within 1 h. Thematic analysis revealed that 62% of the first replies to suicidal posts were of a potentially helpful nature (e.g. discouraging suicide, caring, or clarifying), while 23% were dismissive or encouraging of the suicide. These findings indicate that Twitter users respond differently to suicidal content. Further research is needed to determine the effects of the replies on suicidal intentions or ideations, and whether this platform can be used to intervene, increase help-seeking, or provide anti-stigma campaigns.


Oral carriage of yeasts in two villages in Papua New Guinea.
Oral swabs were taken from 194 subjects in two villages in Papua New Guinea. Yeasts were isolated from 103 (53.1%) individuals of which 41 (21.1%) were Candida albicans. A wide variety of other yeasts were also identified.


Production of modified crosslinked cell-free hemoglobin for human use: the role of quantitative determination of endotoxin contamination.
In vivo toxicity remains a major barrier to the successful use of cell-free hemoglobin (Hb) as an oxygen carrier in humans. Bacterial endotoxin (lipopolysaccharide, LPS) is known to contribute to the in vivo toxicity of Hb preparations, and the prevention of LPS contamination is a critical aspect of the effort to create an efficacious Hb blood substitute. Limulus amebocyte lysate assays for endotoxin were performed on multiple Hb samples from 26 independent production runs for the preparation of human crosslinked cell-free hemoglobin (alpha alpha Hb). High levels of LPS contamination (1- > 100 ng/mL) of alpha alpha Hb solutions were detected in multiple samples during many of the initial production runs. It was observed that LPS contamination of alpha alpha Hb solutions could occur at any step during the production sequence. Substantial enhancement by alpha alpha Hb of the biologic effects of LPS was demonstrated by two independent assays for endotoxin (the Limulus amebocyte lysate test and a mononuclear cell procoagulant assay), whereas LPS biologic activity was only slightly increased by human serum albumin and substantially diminished by IgG. These results suggest that the prevention of LPS-related toxicities in vivo may be more important to the clinical use of Hb solutions than to the use of other intravenous protein products. Therefore, it was encouraging to note that, with the careful monitoring for LPS in the production facility and in multiple samples during cell-free Hb production, sources of LPS contamination were recognized and the appropriate sites were made endotoxin-free.(ABSTRACT TRUNCATED AT 250 WORDS)


The persistence of antibodies induced by meningococcal polysaccharides of groups A and C in human volunteers.
Ninety-nine college students (58 males and 41 females) aged 17 to 23 years were each injected subcutaneously with 50 micrograms of meningococcal polysaccharides A and C. Titrations of antisera obtained at various time intervals during the two subsequent years were made by passive haemagglutination microtitration using human O Rh-negative red blood cells from a single source. The percentage of responders (those who developed a fourfold increase in titre) to polysaccharide A was 97.9% and that to polysaccharide C was 94.8%. Pre-immunization titres of 16 to polysaccharide A and 8 to polysaccharide C were considered to be threshold values above which the response might be impaired. There was a significant (P less than 0.01) difference in the geometric mean HA titre between vaccinees and control groups after vaccination at each time interval studied. The majorities of both the vaccinees and the controls had a higher peak titre to polysaccharide A than to polysaccharide C.


ADC Histogram Analysis of Cervical Cancer Aids Detecting Lymphatic Metastases-a Preliminary Study.
Apparent diffusion coefficient (ADC) histogram analysis has been used to some extent in cervical cancer (CC) to distinguish between low-grade and high-grade tumors. Although this differentiation is undoubtedly helpful, it would be even more crucial in the presurgical setting to determine whether a tumor already gained the potential to metastasize via the lymphatic system. So far, no studies investigated the potential of 3T ADC histogram analysis in CC to differentiate between nodal-positive and nodal-negative entities. Therefore, the principal aim of our study was to investigate the potential of 3T ADC histogram analysis to differentiate between CC with and without lymph node metastasis. The second aim was to elucidate possible differences in ADC histogram parameters between CC with limited vs. advanced tumor stages and well-differentiated vs. undifferentiated lesions. Finally, correlations of p53 expression and Ki-67 index with ADC parameters were analyzed. Eighteen female patients (mean age 55.4 years, range 32-79 years) with histopathologically confirmed cervical squamous cell carcinoma of the uterine cervix were prospectively enrolled. Tumor stages, tumor grading, status of metastatic dissemination, Ki67-index, and p53 expression were assessed in these patients. Diffusion weighted imaging (DWI) was obtained in a 3T scanner using the following b values: b0 and b1000 s/mm2. Group comparisons using Mann-Whitney U test revealed the following findings: nodal-positive CC had statistically significant lower ADC parameters (ADCmin, ADCmean, median ADC, Mode, p10, p25, p75, and p90) in comparison to nodal-negative CC (all p < 0.05). ADCentropy was significantly elevated (p = 0.046) in tumors with advanced T stages (T3/4) compared to tumors with limited T stage (T2). ADCmin values were different in a statistically significant manner comparing G1/G2 and G3 tumors (40.45 ± 18.63 vs. 65.0 ± 23.63 × 10-5 mm2 s-1, p = 0.035). Furthermore, Spearman Rho calculation identified an inverse correlation between ADCentropy and p53 expression (r = -0.472, p = 0.048). The main finding of our study is the discriminability of nodal-positive from nodal-negative CC using ADC histogram analysis in 3T DWI. This information is crucial for the gynecological surgeon to identify the optimal treatment strategy for patients suffering from CC. Furthermore, ADCentropy was identified as a potential imaging biomarker for tumor heterogeneity and might be able to indicate further molecular changes like loss of p53 expression, which is associated with EMT and consequentially indicates a poor prognosis in CC. Finally, our study confirmed the findings of previous works, which indicated that histogram analysis of ADC maps can distinguish between low-grade and high-grade CC. In conclusion, it can be stated that ADC histogram analysis provides additional, prognostically important information on tumor biology in CC.


Poison exon annotations improve the yield of clinically relevant variants in genomic diagnostic testing.
Neurodevelopmental disorders (NDDs) often result from rare genetic variation, but genomic testing yield for NDDs remains below 50%, suggesting that clinically relevant variants may be missed by standard analyses. Here, we analyze "poison exons" (PEs), which are evolutionarily conserved alternative exons often absent from standard gene annotations. Variants that alter PE inclusion can lead to loss of function and may be highly penetrant contributors to disease. We curated published RNA sequencing data from developing mouse cortex to define 1937 conserved PE regions potentially relevant to NDDs, and we analyzed variants found by genome sequencing in multiple NDD cohorts. Across 2999 probands, we found 6 novel clinically relevant variants in PE regions. Five of these variants are in genes that are part of the sodium voltage-gated channel alpha subunit family (SCN1A, SCN2A, and SCN8A), which is associated with epilepsies. One variant is in SNRPB, associated with cerebrocostomandibular syndrome. These variants have moderate to high computational impact assessments, are absent from population variant databases, and in genes with gene-phenotype associations consistent with each probands reported features. With a very minimal increase in variant analysis burden (average of 0.77 variants per proband), annotation of PEs can improve diagnostic yield for NDDs and likely other congenital conditions.


Activatable Two-Photon Near-Infrared Fluorescent Probe Tailored toward Peroxynitrite In Vivo Imaging in Tumors.
A malignant tumor remains one of the leading causes of deaths across the world. Thus, diagnosis of tumor development with noninvasive visualizing methods is significant for tumor therapy. Herein, an activatable two-photon NIR fluorescent probe DHQ-Rd-PN for in vivo imaging of peroxynitrite in a tumor was elaborately designed. The probe demonstrated an increased NIR emission in response to peroxynitrite in vitro, which ensured that the probe detects ONOO- in cell and in vivo. Cellular imaging results disclosed that the probe was competent to detect adscititious ONOO- level change in HeLa cells, as well as endogenous ONOO- concentration in lipopolysaccharides (LPS) and IFN-γ-stimulated RAW 264.7 cells. Additionally, zebrafish in vivo imaging revealed that the probe accumulated in the pancreas and was lightened up by the addition of ONOO-. Remarkably, the probe can be harnessed to image an ONOO- production profile in xenograft 4T1 tumor mice by both one-photon and two-photon in vivo fluorescence imaging. Benefiting with the two-photon excitable properties and NIR emissive properties, the probe can be used for noninvasive in vivo imaging of ONOO- in the onset and development of tumors for the first time. This work provided a noninvasive and efficient detection method for ONOO- in a tumor, which would find more applications in tumor diagnosis and therapies.


A.D.A.M. test (Antibiofilm Dressing's Activity Measurement) - Simple method for evaluating anti-biofilm activity of drug-saturated dressings against wound pathogens.
In the present article, we propose a simple Antibiofilm Dressing's Activity Measurement (A.D.A.M.) test that allows to check in vitro a dressing's suitability against biofilm-related wound infections. To perform the test, three agar discs are covered with biofilm formed by the tested pathogen after which they are assembled one over another in the form of an agar plug and placed in the well of a 24-well plate. The top disc is covered with the analyzed dressing and the entire set is incubated for 24h. During this time, the investigated antimicrobial substance is released from the dressing and penetrates to subsequent biofilm-covered agar discs. Biofilm reduction is measured using 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) spectrometric assay and the results are compared to untreated control samples (agar plug covered with biofilm and without the dressing/or with a passive dressing placed on the top disc). Furthermore, in order to standardize the differences in penetrability of the drugs released from active dressings the results can be expressed as a dimensionless value referred to as the Penetrability Index. In summary, A.D.A.M. test is simple, cheap, can be performed practically in every clinical laboratory and takes no more time than routine microbiological diagnostics. Apart from measuring the released drug's activity, the A.D.A.M. test allows to assess drug penetrability (across three agar discs), reflecting real wound conditions, where microbes are frequently hidden under the necrotic tissue or cloth. In conclusion, the A.D.A.M. test produces a high volume of data that, when analyzed, can provide a researcher with a valuable hint concerning the applicability of active dressings against specific biofilm pathogens in a particular setting.


Functional and antioxidant properties of cookies incorporated with foxtail millet and ginger powder.
The functional, sensory, nutritive, phytochemical and antioxidant properties of flours derived from wheat, foxtail millet and ginger along with the cookies formulated from these ingredients was studied. The foxtail millet (20, 30 and 40%) and ginger powder (5, 10 and 15%) blended wheat flours were investigated and compared to control (100% wheat flour). The increase in substitution levels of both the alternatives improved the functional properties of the composite flour and cookies formulated. Sensory analysis revealed that cookies prepared from 30% foxtail millet and 10% ginger powder had comparable scores. These were further subjected to nutritive and phytochemical analyses. The results revealed that cookies formulated using both the alternatives had reasonable proximate composition comparable to control. The substituted cookies had higher total soluble phenolic content and antioxidant capacities in terms of DPPH and ABTS scavenging potential and a strongly positive correlation was observed between the total soluble phenolic content and antioxidant capacities in composite flour and the cookies formulated from the flours. Overall the addition of foxtail millet flour (30%) and ginger powder (10%) improved the nutritional and phenolic linked antioxidant potential which can lead to shelf life extension and enhanced health benefits.


Analysis of cervical spine alignment in currently asymptomatic individuals: prevalence of kyphotic posture and its relationship with other spinopelvic parameters.
Many studies tend to characterize cervical kyphosis as a significant clinical condition that needs to be treated. Moreover, opinions vary on whether cervical kyphosis should be considered a pathologic status or a natural occurrence in asymptomatic people. This study aimed to determine the frequency of kyphotic posture of the cervical spine in currently asymptomatic individuals and to ascertain its relation with other spinopelvic parameters. A cross-sectional radiographic study was carried out. This study targeted 1,026 currently asymptomatic adult volunteers who agreed to participate in this study from January 2010 to March 2016. Only 958 were eligible for the study. Radiographic images, including the C-spine dynamic view and whole-spine lateral view, were measured. The sagittal parameters of the cervical spine and other parts of the spine and pelvis, such as the C2-C7 angle, C0-C2 range of motion (ROM), C2-C7 ROM, and C0-C7 ROM, thoracic kyphosis, lumbar lordosis, sacral slope, pelvic tilt, and pelvic incidence, were measured. Based on the C-spine neutral lateral X-ray, a C2-C7 Cobb angle greater than 0 degree was defined as lordosis and an angle less than 0 degree was defined as kyphosis. Patients who showed kyphosis were further classified into the reducible or non-reducible group, depending on the ability of recovering neck motions (lordosis) in extension. The cervical and other global spine parameters between the two groups were analyzed, and the relation between the cervical alignment and other parts of the spine and pelvis were also examined. This study was not supported by any funding and had no conflicts of interest. Nearly one-fourth of the asymptomatic participants (26.3%) have kyphotic cervical posture, and almost one-sixth of the kyphotic individuals (16.7%) have non-reducible kyphosis. The prevalence increases with advanced age; non-reducible cases are mostly kyphotic, kyphosis stems from the C2-C7 region, and kyphosis is not correlated with any of the radiological parameters of the other parts of the spine except lumbar lordosis. Cervical kyphosis can be observed in normal healthy adults.


In vivo interaction of bilirubin with the cells of the immune system in mice: an ultrastructural electronmicroscopic study.
Morphological evidence revealed the presence of round-shaped particles after application of bilirubin. The possibility of polymerization of bilirubin-albumin complexes is discussed.


A two-dimensional grating surface made of quaternary Huygens elements excited by a real source.
In this article, a new method to create an anomalous reflection in the desired direction is proposed. Two-dimensional grating surfaces consisting of four particles with the properties of a Huygens source are employed in each period. Then, this method is extended to the problem in which the grating surface is illuminated by a real source such as a horn. In this case, the designed grating surface has different periods in both directions to collimate the reflected wave and give an in-phase wavefront. Using our method, a high-efficiency reflectarray (RA) based on quaternary Huygens grating is designed. This RA is distinguished from common RAs due to its beam squint capability. This array offers more aperture efficiency and thus more gain in comparison to the leaky waves that inherently have low aperture efficiency. Therefore, our designed RA can compete with leaky wave antennas in many applications. The mentioned RA is designed to have the main beam in the direction of [Formula: see text], at the frequency of 12 GHz. The simulation results show the realized gain and SLL of this antenna are 24.8 dB and [Formula: see text] dB, respectively. Also, by changing the frequency in the range of 12-15 GHz, the direction of the main beam varies from [Formula: see text] to [Formula: see text].


Distance-dependent inhibition facilitates focality of gamma oscillations in the dentate gyrus.
Gamma oscillations (30-150 Hz) in neuronal networks are associated with the processing and recall of information. We measured local field potentials in the dentate gyrus of freely moving mice and found that gamma activity occurs in bursts, which are highly heterogeneous in their spatial extensions, ranging from focal to global coherent events. Synaptic communication among perisomatic-inhibitory interneurons (PIIs) is thought to play an important role in the generation of hippocampal gamma patterns. However, how neuronal circuits can generate synchronous oscillations at different spatial scales is unknown. We analyzed paired recordings in dentate gyrus slices and show that synaptic signaling at interneuron-interneuron synapses is distance dependent. Synaptic strength declines whereas the duration of inhibitory signals increases with axonal distance among interconnected PIIs. Using neuronal network modeling, we show that distance-dependent inhibition generates multiple highly synchronous focal gamma bursts allowing the network to process complex inputs in parallel in flexibly organized neuronal centers.Perisomatic-inhibitory interneurons (PIIs) contribute to the generation of gamma oscillations in the hippocampus. Here the authors demonstrate distance-dependent inhibition between PIIs in freely moving mice, and use computational analysis to show that distance-dependent inhibition supports the emergence of focal gamma bursts.


A comparison between two methods of chronological dating of human endometrial biopsies during the luteal phase, and their correlation with histologic dating.
This prospective study was performed on 61 infertile women to examine the correlation between histologic dating using the same criteria by two independent observers and chronological dating by two different methods: (1) determination of luteinizing hormone (LH) peak by daily LH assay, (2) calculation based on the onset of the next menstrual period (NMP). The correlation between histologic dating and chronological dating was found to be significantly better if the LH peak was used to determine the chronological date than if the NMP was used (r = 0.70 and 0.37, respectively).


The role of individual difference in determining the nature of comprehension defects in aphasia.
This study was concerned with the relationship between locus of lesion and subtypes of language comprehension defect in aphasia. Previous studies had indicated that performance in sound recognition identified two subtypes of aural comprehension deficit and that performance in pantomime recognition identified two subtypes of reading comprehension defect. Defects in sound recognition and pantomime recognition were found in association with a variety of lesion loci. However, comprehension defective aphasics with normal sound recognition or pantomime recognition had lesions closely similar in location and size to those of aphasics who performed poorly on these tests. Findings suggest that whether an aphasic with a language comprehension defect is impaired in sound recognition or pantomime recognition depends, at least in part, on individually variable predisposing factors. It would appear possible that this phenomenon reflects a separate dimension of hemispheric cerebral dominance.


Subunit M2 of mammalian ribonucleotide reductase. Characterization of a homogeneous protein isolated from M2-overproducing mouse cells.
The M2 subunit of mammalian ribonucleotide reductase was purified to homogeneity from hydroxyurea-resistant, M2-overproducing mouse cells. The purification procedure involved affinity chromatography on an anti-tubulin antibody-Sepharose column and high performance gel permeation chromatography. The pure protein is a dimer of Mr = 88,000, containing stoichiometric amounts of a non-heme iron center and a tyrosyl free radical. The radical is destroyed by hydroxyurea but can readily be regenerated on incubation of the radical-free protein alone with iron-dithiothreitol in the presence of air. The ability to spontaneously regenerate the tyrosyl radical distinguishes protein M2 from the corresponding subunit of Escherichia coli ribonucleotide reductase, protein B2, but apart from that the two proteins are very similar.


The impact of fetal echocardiography on the prevalence of liveborn congenital heart disease.
Fetal echocardiography allows for early detection of congenital heart disease, and pregnancy termination may be an option in cases of complex defects. In the current study, the most important factors contributing to the diagnosis and termination of affected pregnancies are reviewed and their combined effect on the future prevalence of liveborn congenital heart disease is evaluated. The relative reduction of the prevalence of the most severe forms of congenital heart disease is estimated as the product of the probability that (1) a fetal cardiac screening is performed (p (evaluation)), (2) an affected pregnancy is detected (P (detection)), (3) pregnancy termination is decided following antenatal diagnosis (P (decision)). In areas where termination of pregnancy is a realistic and supported option, a universal sonographic screening of all pregnancies (P (evaluation) = 1), with an average reported sensitivity of 35% and a termination rate of 43% following antenatal diagnosis, would result in a 15% overall reduction of the prevalence of the most severe forms of congenital heart disease. However, wide variability exists regarding the defect-specific estimates (2-50% prevalence relative reduction) due to considerable differences in the reported diagnostic sensitivity and termination rates associated with each heart defect. If an earlier diagnosis could be achieved, which is reported to be associated with an average 1.4-fold increased probability of termination, the overall reduction of the prevalence of congenital heart disease could approach 21%. As the skills of obstetric and pediatric cardiology sonographers improve, fetal echocardiography is expected to have a substantial impact on the future epidemiology of liveborn congenital heart disease.


Integrin signaling and mechanotransduction in regulation of somatic stem cells.
Somatic stem cells are characterized by their capacity for self-renewal and differentiation, making them integral for normal tissue homeostasis. Different stem cell functions are strongly affected by the specialized microenvironment surrounding the cells. Consisting of soluble signaling factors, extracellular matrix (ECM) ligands and other cells, but also biomechanical cues such as the viscoelasticity and topography of the ECM, these factors are collectively known as the niche. Cell-ECM interactions are mediated largely by integrins, a class of heterodimeric cell adhesion molecules. Integrins bind their ligands in the extracellular space and associate with the cytoskeleton inside the cell, forming a direct mechanical link between the cells and their surroundings. Indeed, recent findings have highlighted the importance of integrins in translating biophysical cues into changes in cell signaling and function, a multistep process known as mechanotransduction. The mechanical properties of the stem cell niche are important, yet the underlying molecular details of integrin-mediated mechanotransduction in stem cells, especially the roles of the different integrin heterodimers, remain elusive. Here, we introduce the reader to the concept of integrin-mediated mechanotransduction, summarize current knowledge on the role of integrin signaling and mechanotransduction in regulation of somatic stem cell functions, and discuss open questions in the field.


Smart bandwidth allocation for next generation networks adopting software-defined network approach.
This data article contains information on a new intelligent bandwidth allocation model for future network (Smart Allocation). The included data describe the topology of the network testbed and the obtained results. Obtained data show the effectiveness of the proposed model in comparison with the MAM and RDM bandwidth allocation models. In relation to the performances evaluation, a variety of flows are used such as: voice over IP (VoIP), video, HTTP, and Internet Control Message Protocol (ICMP). The evaluation criteria are: VoIP latency and jitter, Peak Signal to Noise Ratio (PSNR) video, retransmission video, goodput, HTTP response page, and the Round-Trip Time (RTT) ICMP delay. The presented data are extracted based on simulation.


Predictive resistance factors in lung cancer patients treated with Nivolumab. Retrospective study.
Immunotherapy is the current treatment in non-small cell lung cancer (NSCLC). 20% of patients treated with immunotherapy have a prolonged response. What about the remaining 80%? How can we explain that some patients get no benefit from immunotherapy? We retrospectively analyzed predictive factors of primary or secondary resistance to immunotherapy in NSCLC patients from 2 French hospitals between 2015 and 2018. Moreover, we evaluated whether PD1 inhibitor had an impact on the antitumor effects of salvage chemotherapy administered after immunotherapy. We chose to focus on taxanes. Ninety-six patients were included in this cohort, 65(68%) patients were considered as having primary resistance and 31(32%) secondary resistance. Resistant populations did not differ. At immunotherapy initiation, median survival was 4.6 months for primary resistant patients (95%CI-4.6-6.8) and 15.6 months (95%CI-9.8-NA) for secondary resistant patients. The disease control rates with taxane were 15% in pre immunotherapy conditions vs 50% in post immunotherapy. Response rates improved regardless of the status of resistance. This study enriches data about immunotherapy in real-life in NSCLC. Prognostic resistance factors still seem complicated to identify. The high rate of taxane responders in post immunotherapy in this retrospective cohort support the use of taxane in therapeutic escape.


Action of foot activator on growth and differentiation of cells in hydra.
Foot activator is a small peptide found in hydra and specifically activates foot formation. I present a method for the further purification of foot activator by high-pressure liquid chromatography. The morphogenetically active fractions were assayed for their effect at the cellular level. Foot activator acts as a mitogen by pushing epithelial and interstitial cells, which are arrested in G2, into mitosis. In the presence of foot activator, epithelial stem cells are stimulated to differentiate into foot mucus cells, and interstitial nerve precursor cells differentiate into mature nerve cells. The interaction of foot activator with head activator in the development of hydra is discussed.


Vitamin D3-mediated resistance to a multiple sclerosis model disease depends on myeloid cell 1,25-dihydroxyvitamin D3 synthesis and correlates with increased CD4+ T cell CTLA-4 expression.
Microglial cell activation is the earliest biomarker of the inflammatory processes that cause central nervous system (CNS) lesions in multiple sclerosis. We hypothesized that 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) production by activated microglia and macrophages in the CNS inhibits these inflammatory processes. To test this hypothesis, we targeted the Cyp27b1 gene specifically in myeloid cells, then analyzed the influence of disrupted myeloid cell 1,25-(OH)2D3 synthesis on vitamin D3-mediated resistance to experimental autoimmune encephalomyelitis (EAE). Myeloid cell 1,25-(OH)2D3 synthesis was essential for vitamin D3-mediated EAE resistance. Increased CTLA-4 expression in the CNS-infiltrating CD4+ Tconv and Treg cells and decreased splenic B cell CD86 expression correlated with resistance. These new data provide solid support for the view that vitamin D3 reduces MS risk in part through a mechanism involving myeloid cell 1,25-(OH)2D3 production and CTLA-4 upregulation in CNS-infiltrating CD4+ T cells. We suggest that CTLA-4 serves as a vitamin D3-regulated immunological checkpoint in multiple sclerosis prevention.


Optic nerve gliomas. I. Clinical diagnosis and pathology.
At the National Institute of Neurosurgery a team of ophthalmologists and neurosurgeons has for 20 years been collaborating in the research and treatment of optic nerve gliomas. Among 34 patients, 30 were under the age of 20 years, and 20 under the age of 10 years. Of the 20 patients younger than 10 years, 16 were girls. The clinical symptoms were classified into three groups: I. Unilateral visual impairment with unilateral axial proptosis (16 patients), strabismus (6 cases), and typical X-ray findings of the skull. II. Uni- or bilateral visual impairment without proptosis (12 patients), depressed vision in the homolateral eye (9 patients), hemianopia (2 out of 4 examined patients) and hemiparesis (6 cases). III. Uni- or bilateral visual impairment without proptosis but with hormonal or metabolic disturbances (6 patients). Carotid angiography and PEG were performed in groups II and III and revealed a picture that is characteristic of suprasellar tumours. 99mTc pertechnetate scanning was done in six cases and showed isotope accumulation in the tumour in five. Pathologically, considering also the clinical aspects, two types of localization merit consideration: 1. The relationship between the tumour and the optic nerve (tumours growing within and extending beyond the optic pathways, respectively) and 2. The site of the tumour within the optic nerve (intraorbital, intraorbital and intracranial, intracranial only, chiasmal, and diffuse). Our data of histology and tissue culture prove the benign nature of the tumour even in the recurrences.


Effect of mast cell granules on the gene expression of nitric oxide synthase and tumour necrosis factor-alpha in macrophages.
Previous studies have shown that mast cell granules (MCG) inhibit numerous macrophage functions including tumour cytotoxicity, superoxide and nitric oxide (NO) production, and FCgamma2a receptor-mediated phagocytosis. In this study, the effect of MCG on macrophage TNF alpha and nitric oxide synthase (iNOS) mRNA expression, and the production and fate of TNF alpha were examined. Upon activation with LPS+IFN gamma, macrophages expressed both TNF alpha and iNOS mRNA and produced both TNF alpha and NO. Co-incubation of LPS+IFN gamma-activated macrophages with MCG resulted in dose-dependent inhibition of iNOS mRNA expression. TNF alpha production in the activated macrophages was decreased by MCG, which was associated with a reduction in TNF alpha mRNA expression. MCG were also capable of degrading both macrophage-generated and recombinant TNF alpha. The direct effect of MCG on TNF alpha was partially reversed by a mixture of protease inhibitors. These results demonstrate that MCG decrease the production of NO and TNF alpha by inhibiting macrophage iNOS and TNF alpha gene expression. Furthermore, MCG post-transcriptionally alter TNF alpha levels via proteolytic degradation.


Microwave-accelerated bioassay technique for rapid and quantitative detection of biological and environmental samples.
Quantitative detection of molecules of interest from biological and environmental samples in a rapid manner, particularly with a relevant concentration range, is imperative to the timely assessment of human diseases and environmental issues. In this work, we employed the microwave-accelerated bioassay (MAB) technique, which is based on the combined use of circular bioassay platforms and microwave heating, for rapid and quantitative detection of Glial Fibrillary Acidic Protein (GFAP) and Shiga like toxin (STX 1). The proof-of-principle use of the MAB technique with the circular bioassay platforms for the rapid detection of GFAP in buffer based on colorimetric and fluorescence readouts was demonstrated with a 900W kitchen microwave. We also employed the MAB technique with a new microwave system (called the iCrystal system) for the detection of GFAP from mice with brain injuries and STX 1 from a city water stream. Control bioassays included the commercially available gold standard bioassay kits run at room temperature. Our results show that the lower limit of detection (LLOD) of the colorimetric and fluorescence based bioassays for GFAP was decreased by ~1000 times using the MAB technique and our circular bioassay platforms as compared to the commercially available bioassay kits. The overall bioassay time for GFAP and STX 1 was reduced from 4h using commercially available bioassay kits to 10min using the MAB technique.


Phthalates (PAEs) and reproductive toxicity: Hypothalamic-pituitary-gonadal (HPG) axis aspects.
Phthalates (PAEs) are widely used for their excellent ability to improve plastic products. As an essential endocrine axis that regulates the reproductive system, whether dysfunction of the hypothalamic-pituitary-gonadal (HPG) axis is involved in reproductive toxicity mediated by environmental endocrine disruptors PAEs has become a hot topic of widespread concern. This study systematically reviewed the adverse effects of multiple PAEs on the HPG axis in different models and objectively discussed the possible underlying mechanisms. The abnormal release of gonadotropin-releasing hormone and gonadotropin, dysfunction of sex hormone receptors and steroid hormone synthesis, and general damage, including cell proliferation, oxidative stress, apoptosis, and autophagy have been confirmed to be involved in this process. Although it is widely established that PAEs induce HPG axis dysfunction, the specific mechanisms involved remain unclear. From a systematic review of relevant publications, it appears that the abnormal expression of peroxisome proliferator-activated, aryl hydrocarbon, and insulin receptors mediated by PAEs is key upstream event that induces these adverse outcomes; however, this inference needs to be further verified. Overall, this study aimed to provide reliable potential biomarkers for future environmental risk assessment and epidemiological investigation of PAEs.


Effect of brachytherapy technique and patient characteristics on cervical cancer implant dosimetry.
Our purpose was to evaluate the relationship between brachytherapy technique and patient characteristics on dose to organs-at-risk (OARs) in patients undergoing high dose rate (HDR) brachytherapy for cervical cancer. From 1998 to 2008, 31 patients with cervical cancer with full dosimetric data were identified who received definitive external-beam radiation and HDR brachytherapy with tandem and ovoid applicators. Doses were recorded at point A, the International Commission on Radiation Units and Measurements (ICRU)-38 rectal point, the ICRU-38 bladder point, the vaginal surface, and the pelvic sidewall. Generalized estimating equations were used to determine the significance of changes in OAR to point A dose ratios with differences in brachytherapy technique or patient characteristics. Patients underwent a median of 5 brachytherapy procedures (range, 3 to 5), with a total of 179 procedures for 31 patients. For all brachytherapy treatments, the average ratios between the doses for the rectal, bladder, vaginal surface, and pelvic sidewall reference points to those at point A were 0.49, 0.59, 1.15, and 0.17, respectively. In general, decreased OAR dose was associated with a lower stage, younger age, increased ovoid size, increased tandem length, and earlier implant number. Increased tandem curvature significantly increased bladder dose and decreased rectal dose. Intravenous anesthesia usage was not correlated with improved dosimetry. This study allowed identification of patient and procedure characteristics influencing OAR dosing. Although the advent of 3-dimensional (3D) image-guided brachytherapy will bring new advances in treatment optimization, the actual technique involved at the time of the brachytherapy implant procedure will remain important.


Multiple concentric ring sign in the ultrasonographic diagnosis of intussusception.
Gray scale ultrasound examination of two patients with abdominal space-occupying lesions demonstrated characteristic ring shadows on longitudinal scans. In one case a multiple concentric ring shadow resulted from an ileocolic intussusception, and in the other a "bull's eye" image was due to an infiltrating carcinoma of the pyloric antrum. A study of these patients confirms the suspicion [1] that there is a characteristic ultrasonic appearance of intussusception in bowel loops.


The genus Neoseiulus Hughes (Mesostigmata: Phytoseiidae) in the Espinhao Range, a great reservoir of biodiversity in Brazil.
Several surveys were conducted between 2010 and 2012 in the Brazilian range Serra do Espinhao, a highly biodiverse region extending more than 1000 km in the states of Bahia and Minas Gerais. We report in this paper eight species of the genus Neoseiulus Hughes collected on natural, non-cultivated, vegetation. Two of these species are described, illustrated and proposed as new taxa, N. cipoensis sp. nov. and N. diamantinus sp. nov. New morphological information as well as host-plants and geographical distribution are provided for six additional species, N. benjamini (Schicha), N. californicus (McGregor), N. goiano Demite, Cavalcante Lofego, N. idaeus Denmark Muma, N. melinis Lofego Moraes, and N. tunus (De Leon).


Gas-pressurized torrefaction of biomass wastes: Roles of pressure and secondary reactions.
A gas-pressurized (GP) torrefaction method, proposed in our resent work, can significantly promote the upgrading and oxygen removal of biomass wastes, compared to the traditional torrefaction (AP). However, the mechanism of the GP torrefaction process is not clear. In present work, semi-closed (SC) torrefaction, GP torrefaction, and AP torrefaction were conducted to reveal the roles of pressure and secondary reactions during GP torrefaction quantitatively. The results showed that the pressure significantly promoted the upgrading of biomass during GP torrefaction at 200 °C. The contribution of pressure on the oxygen removal of GP torrefaction at 200 °C was 63.87%. At relatively high temperature of around 250 °C, the promotions were caused by the synergistic effect of pressure and secondary reactions. The contribution of secondary reactions on the oxygen removal was 53.99%. Thus, the process of the GP torrefaction of biomass wastes was preliminarily understood.


Recent development of small-molecular inhibitors against Clostridioides difficile infection.
Clostridioides difficile infection is one of the leading causes of antibiotic-associated infectious diarrhea, and is associated with increased incidence and severity worldwide. While antibiotics have traditionally been used for prophylaxis and treatment of C. difficile infection, elevated antibiotic resistance has promoted the development and spread of C. difficile infection. Since the current standard-of-care antibiotics are ineffective for treating infections, there is an urgent need for novel antibacterial drugs or strategies to target C. difficile infection. C. difficile virulence and vital physiological functions are considered to be ideal targets. Thus, several promising lead compounds have been identified through screening both synthetic and natural product libraries. The goal of this review is to provide an update of the current scientific knowledge of C. difficile infection, focusing on small molecule inhibitors, which can effectively inhibit C. difficile by suppressing virulence or destroying vital physiological structures.


Mechanism of the synergic effects of calcium chloride and electroporation on the in vitro enhanced skin permeation of drugs.
We have already reported the substantial synergic effects of CaCl(2) and electroporation (EP) on in vitro skin permeation of calcein and FITC dextrans. In the present paper, we investigated the mechanisms for these effects by considering changes in lamellar structure and barrier recovery time of the biggest skin barrier, the stratum corneum, by this combined treatment. The change in skin lamellar structure was evaluated by lipid mobility in the stratum corneum using ATR-FTIR, calcein release from stratum corneum-lipid liposomes (SCLL), in vitro skin permeation of calcein and transepidermal water loss (TEWL). The ATR-FTIR measurement, in vitro skin permeation and changes in TEWL were also used for examining the barrier recovery time. The C-H stretching band of skin lipids produced with EP was blue-shifted when compared to that without EP. Asymmetric C-H stretching was highest with EP in CaCl(2) solution. Little release of calcein was observed from SCLL without EP, whereas higher releases were observed after EP with or without NaCl or CaCl(2). Particularly high calcein release (>20%) was observed over 60 min with EP in CaCl(2) solution. The in vitro permeation study of calcein was conducted through excised hairless rat skin that was pretreated with EP before skin excision. Permeation rate was highest in skin excised immediately after in vivo EP, and this rate decreased with time after EP treatment. TEWL recovered to control levels within 2 h after EP in distilled water or NaCl solution, whereas high TEWL was maintained after EP in CaCl(2) solution. These results suggest that at least lamellar destruction of stratum corneum must be related to the enhanced skin permeation of drugs by the combination of CaCl(2) and EPF. On the other hand, a prolonged enhancing effect on the skin permeation of calcein by this combination may be due to a high lamellar destruction and/or delayed barrier repair of stratum corneum.


Some properties of mammalian cardiac cytochrome c1.
Investigations into the nature of the axial heme ligands, the strength of the heme crevice, the reactivity with cyanide, and the ascorbate reducibility of cytochrome c1 were performed to explore structure-function relationships of cytochrome c1. The existence of an absorbance band at 690 nm, which was quenched by raising the pH with a pK of 9.2 corresponding to a low spin-low transition, suggested that a methionine residue probably functioned as one of the axial heme iron ligands in this cytochrome. Spectral titrations of cytochrome c1 in the low pH range showed a markedly elevated pK for the low spin-high spin transition relative to cytochrome c. Denaturation studies with urea, the absence of any reaction with cyanide, and the evidence from other lines would appear to indicate that the heme group of cytochrome c1 was reduced by ascorbate at approximately 5% of the rate of reduction of cytochrome c but this rate dramatically increased with increasing pH concomitant with the disappearance of the 690 nm absorbance band. Circular dichroic spectra substantiated that elevated pH produced conformational changes localized to the heme crevice and probably also the regions containing aromatic residues. The enhanced rate of ascorbate reduction was perhaps a consequence of the increased accessibility of the heme iron to ascorbate. Major unfolding of the protein in 8 M urea, however, completely abolished the ascorbate reducibility of cytochrome c1. The buried nature of the heme group of cytochrome c1 would probably preclude transfer of an electron from cytochrome c1 to cytochrome c through a direct Fe-Fe or a heme-heme interaction. This poses an important question concerning the mechanism of this electron transfer between these two cytochromes not only in mitochondria but also in solution.


Healing and revascularization of apical periodontium and dental pulps in apicoectomized and nonapicoectomized tooth replants in monkeys. A study of fully developed teeth.
This report primarily concerns an experimental study of the effect of apicoectomy on the healing and revascularization of the periodontal membrane and dental pulps in tooth replants. Five rhesus monkeys (Macaca mulatta) were subjected to surgical procedures in which twenty incisor teeth were extracted and immediately replanted. Ten of these incisors were subjected to apicoectomy. Healing and revascularization of periapical tissues and dental pulps were evaluated by histologic and India-ink-perfusion techniques. Apicoectomized replants were characterized by (1) loss of dental substance (tooth apex) and creation of large periapical spaces, (2) exposure of dental pulps and production of periapical inflammatory processes with destruction of alveolar bone, and (3) blocking of normal healing because of persistence of chronic periapical inflammatory processes. In contrast, nonapicoectomized replants depicted short-lived primary inflammatory responses and healing processes characterized by variable histologic responses.


Assumed white blood cell count of 8,000 cells/μL overestimates malaria parasite density in the Brazilian Amazon.
Quantification of parasite density is an important component in the diagnosis of malaria infection. The accuracy of this estimation varies according to the method used. The aim of this study was to assess the agreement between the parasite density values obtained with the assumed value of 8,000 cells/μL and the automated WBC count. Moreover, the same comparative analysis was carried out for other assumed values of WBCs. The study was carried out in Brazil with 403 malaria patients who were infected in different endemic areas of the Brazilian Amazon. The use of a fixed WBC count of 8,000 cells/μL to quantify parasite density in malaria patients led to overestimated parasitemia and resulted in low reliability when compared to the automated WBC count. Assumed values ranging between 5,000 and 6,000 cells/μL, and 5,500 cells/μL in particular, showed higher reliability and more similar values of parasite density when compared between the 2 methods. The findings show that assumed WBC count of 5,500 cells/μL could lead to a more accurate estimation of parasite density for malaria patients in this endemic region.


[Antigenic relationship among Candida guilliermondii strains (author's transl)].
In this short communication the results of cross aglutination and absorption tests among three sandard Candida guilliermondii strains (C. guilliermondii CBS--566, C. guilliermondii var. carpophila CBS--5256, C. guilliermondii var. japonica CBS--6021) are given. It showed that these microorganisms have the same antigenic structure.


Erythema chronicum migrans in three soldiers.
Three cases of erythema chronicum migrans in soldiers who had returned from active duty in Central Europe were seen within a ten-week period in San Franciso. Typically, the lesions cleared in less than five days with penicillin in two cases and erythromycin in another.


The inhibition by hydrocortisone of prostaglandin biosynthesis in rat peritoneal leucocytes is correlated with intracellular macrocortin levels.
Hydrocortisone inhibits prostaglandin generation by rat peritoneal leucocytes by releasing the polypeptide phospholipase inhibitor, macrocortin. The susceptibility of these cells to hydrocortisone is directly correlated with their intracellular macrocortin content. Cells depleted of the peptide by prior incubation with steroid cannot respond to the steroid, until a fresh intracellular store has been synthesized. In vitro, this process requires 4-5 h. Cells remain sensitive to the inhibitory action of the peptide at all times.


[The functional characteristics of the enzymatic antioxidant system in the erythrocytes and neutrophils of the blood of patients with generalized forms of meningococcal infection].
Patients with meningococcal infection, meningitis and with a mixed form of the disease were demonstrated to have unbalance in the redox system of glutathione during the all disease periods till the clinical recovery. Activation of glutathione peroxidases to hydrogen peroxide and tertiary butyl hydroperoxide in erythrocytes was coupled, during the whole disease, with unbalance of the time-course of changes in the interrelated enzymes--superoxide dismutase and glutathione peroxidase to hydrogen peroxide, while in neutrophils, the balance of those enzymes remained unimpaired. Glutathione transferase activity appeared reduced both in erythrocytes and neutrophils. Potential mechanisms by which the redox system of glutathione and superoxide dismutase may be deranged in meningococcal infection are under discussion.


The Fontan epidemic: Population projections from the Australia and New Zealand Fontan Registry.
The number and age demographic of the future Fontan population is unknown. Population projections were calculated probabilistically using microsimulation. Mortality hazard rates for each Fontan recipient were calculated from survivorship of 1353 Fontan recipients in the Australia and New Zealand Fontan Registry, based on Fontan type, age at Fontan, gender and morphology. Projected rates of new Fontan procedures were generated from historical rates of Fontan procedures per population births. At the end of 2014, the living Fontan population of Australia and New Zealand was 1265 people from an Australian and New Zealand regional population of 28 million (4.5 per 100,000 population). Of those, 165 (13%) received an atrio-pulmonary (AP) procedure, 262 (21%) a lateral tunnel (LT) procedure and 838 (66%) an extra-cardiac conduit (ECC) procedure. This population is expected to grow to 1917 (95% CI: 1846: 1986) by 2025 (5.8 per 100,000 population), with 149 (8%) AP procedures, 254 (13%) LT procedures, and 1514 (79%) ECC procedures. By 2045, the living Fontan population is expected to reach 2986 (95% CI: 2877: 3085; 7.2 per 100,000 population). The average age of the Fontan population is expected to increase from 18years in 2014 to 23years (95% CI: 22-23) by 2025, and 31years (95% CI: 30-31) by 2045. The Australian and New Zealand population of patients alive after a Fontan procedure will double over the next 20years increasing the demand for heart-failure services and cardiac transplantation. Greater consideration for the needs of this mostly adult Fontan population will be necessary.


Prevalence of Clostridium Difficile-Associated Diarrhoea in Hospitalised Patients (Results of a Russian Prospective Multicentre Study).
The objective of the study was to evaluate the prevalence of Clostridium difficile-associated diarrhoea (CDAD) among hospitalised patients with antibiotic-associated diarrhoea (AAD) in general and by specific types of medical care and hospital units. A prospective, cross-sectional, non-interventional, multicentre study. The main inclusion criteria were: patient age ≥ 18 years, hospital stay of at least 48 h, current antibiotic therapy or antibiotic therapy within the previous 30 days, loose stools (Bristol stool types 5-7 and stool frequency ≥ 3 within ≤ 24 consecutive hours or exceeding normal for the patient) and signed informed consent form. The stool sample was taken to the local (study site) microbiology laboratory for detection of glutamate dehydrogenase (GDH) and toxins A/B using enzyme immunoassay (EIA) stool test. From April 2016 to April 2017, a total of 1245 patients from 12 large hospitals were enrolled in the study. Data on 81 patients were excluded from the analysis for different reasons. Data on 1164 patients (45.2% males and 54.8% females) with a mean age of 54.9 years (range 18-95 years) were analysed. Length of hospitalisation was 2-188 days (median, 8 days). The EIA stool test showed CDAD-positive results in 21.7% (253/1164) patients. The patients were from surgery units (546/1164), internal medicine units (510/1164) and intensive care units (108/1164). The prevalence of CDAD among patients from surgery, internal medicine and intensive care units was 26.2, 17.8 and 17.6%, respectively. Oncology, gastroenterology, septic surgery, oncohaematology and general medical hospital units accounted for more than 75% of all patients included; the prevalence of CDAD by those hospital units was 11.3, 15.0, 39.2, 17.6, and 27.2%, respectively. The proportion of GDH-positive and toxin A/B-negative patients by the rapid stool test result was 16.8% (196/1164). The prevalence of CDAD varied widely between the hospitals (from 0 to 44.3%). The prevalence of CDAD among hospitalised patients with AAD in this study was 21.7% (95% confidence interval: 14.8 and 28.7%). The percentage of CDAD varied widely between hospitals and by specific types of medical care and hospital units.


Aerobic and facultatively anaerobic cellulolytic bacteria from the gut of the termite Zootermopsis angusticollis.
To demonstrate the occurrence of cellulolytic bacteria in the termite Zootermopsis angusticollis. Applying aerobic cultivation conditions we isolated 119 cellulolytic strains from the gut of Z. angusticollis, which were assigned to 23 groups of aerobic, facultatively anaerobic or microaerophilic cellulolytic bacteria. 16S rDNA restriction fragment pattern and partial 16S rDNA sequence analysis, as well as numerical taxonomy, were used for the assignment of the isolates. The Gram-positive bacteria of the actinomycetes branch could be assigned to the order Actinomycetales including the genera Cellulomonas/Oerskovia, Microbacterium and Kocuria. The Gram-positive bacteria from the order Bacillales belonged to the genera Bacillus, Brevibacillus and Paenibacillus. Isolates related to the genera Afipia, Agrobacterium/Rhizobium, Brucella/Ochrobactrum, Pseudomonas and Sphingomonas/Zymomonas from the alpha-proteobacteria and Spirosoma-like from the "Flexibacteriaceae" represented the Gram-negative bacteria. A cell titre of up to 10(7) cellulolytic bacteria per ml, determined for some isolates, indicated that they may play a role in cellulose digestion in the termite gut in addition to the cellulolytic flagellates and termite's own cellulases. The impact of bacteria on cellulose degradation in the termite gut has always been a matter of debate. In the present survey we investigated the aerobic and facultatively anaerobic cellulolytic bacteria in the termite gut.


Hyaluronic acid spacer in focal prostate reirradiation: A single centre experience.
The optimal management of locally recurrent prostate cancer after curative radiotherapy is still unknown. In this study, we evaluated the preliminary results of reirradiation using stereotactic body radiotherapy for locally recurrent prostate cancer after initial definitive local radiotherapy. Between April 2016 and February 2019, 11 patients with recurrent disease at the previously irradiated prostate were treated. Local recurrence was detected by radiological with or without functional imaging modalities including prostate multiparametric/pelvic MRI or positron-emission tomography-computerised tomography with (68Ga)-labelled prostate-specific membrane antigen performed after rising prostate specific antigen serum level during follow-up. All patients received stereotactic body radiotherapy to the recurrent nodule to a total dose of 30Gy in five fractions. Hyaluronic acid spacer was injected between prostate and rectum in seven patients to decrease the rectal dose. Acute toxicity was evaluated by using Common Terminology Criteria for Adverse Events version 4.0, and late toxicity was evaluated by using Radiation Therapy Oncology Group/European Organisation for Research and Treatment of Cancer late radiation morbidity scoring schema. At the diagnosis, the median age was 64 years, and the mean prostate specific antigen serum concentration was 17.7ng/mL. The median interval time between local recurrence and initial definitive radiotherapy was 63 months. Mean prostate specific antigen concentration nadir value during follow-up was 0.43ng/mL. With a median follow up of 19 months, three patients developed either local or distant relapse. One patient had grade 3 acute rectal toxicity, and one patient had grade 2 late urinary toxicity. We did not observe any acute or late toxicity due to hyaluronic acid spacer injection. Reirradiation after local recurrence following initial definitive radiotherapy together with hyaluronic acid spacer use seems to be effective and safe.


The effect of BMI on long-term outcomes after operatively treated ankle fractures: a study with up to 16 years of follow-up.
Ankle fractures are a common injury and the main cause of post-traumatic ankle arthritis. The prevalence of obesity is increasing worldwide, and this population is known to have poorer short and midterm outcomes after ankle fractures. Our objective is to assess long-term patient-reported outcomes in patients with operatively treated ankle fractures, and the effect of BMI on these results using the new and validated patient-reported outcome questionnaire, the Manchester Oxford foot and ankle questionnaire (MOXFQ). We performed a retrospective review of all ankle fractures treated operatively in a ten-year period from 2002-2012. The MOXFQ and SF-12 were sent to all patients and were obtained, on average, 11.1 years after surgery (range 5.3-16.2 years). Two thousand fifty-five ankle fractures were reviewed, of which 478 (34%) patients completed the questionnaires. The mean age was 48.1 ± 15.5 years, 52% were men and the mean BMI was 26.1 ± 4.5 kg/m2. Of the 478, 47% were of normal weight, 36% were overweight, and 17% were obese. Overall, 2.1% were type A, 69.9% B, and 24.9% type C fractures. There were no significant differences in the type of fracture between the BMI groups. Comparing obese and non-obese patients, there were large differences in MOXFQ pain (33 ± 29 vs. 18.7 ± 22.1, effect size 0.55), and function scores (27.3 ± 29 vs. 12.5 ± 21.1, effect size 0.58). No differences in complications and reoperations rates were observed. The BMI value at surgery correlated more strongly with the MOXFQ pain score than the BMI at follow-up (Spearman's Rho 0.283 vs. 0.185, respectively). These findings reveal that obese patients have significant worse long-term outcomes, namely increased pain, poorer function, and greater impairment in everyday life after an operatively treated ankle fracture. Moreover, pain and function linearly declined with increasing BMI. Our findings appear to indicate that increased BMI at surgery is an important contributor to adverse outcome in the operative management of rotational ankle fractures. III.