Anthropogenic Infection of Cats during the 2020 COVID-19 Pandemic.
COVID-19 is a severe acute respiratory syndrome (SARS) caused by a new coronavirus (CoV), SARS-CoV-2, which is closely related to SARS-CoV that jumped the animal-human species barrier and caused a disease outbreak in 2003. SARS-CoV-2 is a betacoronavirus that was first described in 2019, unrelated to the commonly occurring feline coronavirus (FCoV) that is an alphacoronavirus associated with feline infectious peritonitis (FIP). SARS-CoV-2 is highly contagious and has spread globally within a few months, resulting in the current pandemic. Felids have been shown to be susceptible to SARS-CoV-2 infection. Particularly in the Western world, many people live in very close contact with their pet cats, and natural infections of cats in COVID-19-positive households have been described in several countries. In this review, the European Advisory Board on Cat Diseases (ABCD), a scientifically independent board of experts in feline medicine from 11 European Countries, discusses the current status of SARS-CoV infections in cats. The review examines the host range of SARS-CoV-2 and human-to-animal transmissions, including infections in domestic and non-domestic felids, as well as mink-to-human/-cat transmission. It summarises current data on SARS-CoV-2 prevalence in domestic cats and the results of experimental infections of cats and provides expert opinions on the clinical relevance and prevention of SARS-CoV-2 infection in cats.


Hepatitis B and C viruses in infants and young children.
Advances during the past 20 years have led to a better understanding of the prevention, diagnosis, and treatment of acute and chronic hepatitis B (HBV) and hepatitis C (HCV) infections in the pediatric population. Universal vaccination and prenatal testing for HBV have decreased the incidence rate of acute HBV infections from more than 3/100,000 to 0.34/100,000 in all children. Diagnosis of chronic HBV is confirmed with positive serologic testing on two occasions at least 6 months apart. Current approved therapies with interferon alpha and lamivudine for children with chronic HBV infection have shown some efficacy, but results have been variable. In contrast, the lack of an effective HCV vaccine and the risk of mother-to-child transmission may increase the number of children with vertically acquired HCV that ultimately go on to develop liver fibrosis or cirrhosis. Diagnosis of HCV in the neonate should be postponed until after the child reaches 1 year of age because infants may have transient viremia. Treatment for HCV infected children has not been studied extensively. Peginterferon alpha-2a and Ribavirin are not currently approved for pediatric use; however, recent studies in children have shown potential benefit. More effective and less toxic therapies for young patients with HBV and HCV are needed, as are methods to interrupt perinatal transmission of HBV and HCV.


C/EBPβ-LAP*/LAP Expression Is Mediated by RSK/eIF4B-Dependent Signalling and Boosted by Increased Protein Stability in Models of Monocytic Differentiation.
The transcription factor C/EBPβ plays a key role in monocytic differentiation and inflammation. Its small isoform LIP is associated with proliferation at early premonocytic developmental stages and regulated via mTOR-dependent signalling. During later stages of (pre)monocytic differentiation there is a considerable increase in the large C/EBPβ isoforms LAP*/LAP which inhibit proliferation thus supporting terminal differentiation. Here, we showed in different models of monocytic differentiation that this dramatic increase in the LAP*/LAP protein and LAP/LIP ratio was accompanied by an only modest/retarded mRNA increase suggesting an important role for (post)translational mechanisms. We found that LAP*/LAP formation was induced via MEK/RSK-dependent cascades, whereas mTOR/S6K1 were not involved. Remarkably, LAP*/LAP expression was dependent on phosphorylated eIF4B, an acceleratory protein of RNA helicase eIF4A. PKR inhibition reduced the expression of eIF4B and C/EBPβ in an eIF2α-independent manner. Furthermore, under our conditions a marked stabilisation of LAP*/LAP protein occurred, accompanied by reduced chymotrypsin-like proteasome/calpain activities and increased calpastatin levels. Our study elucidates new signalling pathways inducing LAP*/LAP expression and indicates new alternative PKR functions in monocytes. The switch from mTOR- to RSK-mediated signalling to orchestrate eIF4B-dependent LAP*/LAP translation, accompanied by increased protein stability but only small mRNA changes, may be a prototypical example for the regulation of protein expression during selected processes of differentiation/proliferation.


Children with autism spectrum disorders: a comparison of those who regress vs. those who do not.
While autism spectrum disorders (ASD) constitute a group of similar conditions, considerable heterogeneity in symptoms of these neurodevelopmental disorders have been noted. One of the most important, yet least studied, of these factors is developmental regression. One-hundred and twenty-five children were studied and broken down into the following three groups: ASD children with and without substantial regression and typically developing children. In study one, the three groups were compared on global measures of ASD symptomatology, comorbid psychopathology, challenging behaviour and social skills. In study two, the two ASD groups were compared on each individual item from the dependent measures. Mean age when regression occurred was 27.76 months. The ASD children as a whole differed from the typically developing controls, showing more symptoms of ASD, as would be expected, and poorer social skills, while differences were also noted between the two ASD groups. It was determined that children with ASD who regress present with a distinct behavioural profile when compared to children with ASD who do not regress, which included greater levels of impairment on global measures of ASD symptomatology, comorbid psychopathology, challenging behaviour and social skills.


Structure and function of the yeast listerin (Ltn1) conserved N-terminal domain in binding to stalled 60S ribosomal subunits.
The Ltn1 E3 ligase (listerin in mammals) has emerged as a paradigm for understanding ribosome-associated ubiquitylation. Ltn1 binds to 60S ribosomal subunits to ubiquitylate nascent polypeptides that become stalled during synthesis; among Ltn1's substrates are aberrant products of mRNA lacking stop codons [nonstop translation products (NSPs)]. Here, we report the reconstitution of NSP ubiquitylation in Neurospora crassa cell extracts. Upon translation in vitro, ribosome-stalled NSPs were ubiquitylated in an Ltn1-dependent manner, while still ribosome-associated. Furthermore, we provide biochemical evidence that the conserved N-terminal domain (NTD) plays a significant role in the binding of Ltn1 to 60S ribosomal subunits and that NTD mutations causing defective 60S binding also lead to defective NSP ubiquitylation, without affecting Ltn1's intrinsic E3 ligase activity. Finally, we report the crystal structure of the Ltn1 NTD at 2.4-Å resolution. The structure, combined with additional mutational studies, provides insight to NTD's role in binding stalled 60S subunits. Our findings show that Neurospora extracts can be used as a tool to dissect mechanisms underlying ribosome-associated protein quality control and are consistent with a model in which Ltn1 uses 60S subunits as adapters, at least in part via its NTD, to target stalled NSPs for ubiquitylation.


Mangostanaxanthones III and IV: advanced glycation end-product inhibitors from the pericarp of Garcinia mangostana.
Advanced glycation end-products (AGEs) are associated with a non-enzymatic reaction between the amino group of a protein and the carbonyl group of a sugar during hyperglycemia. The precipitation of AGEs in different tissues leads to many complications, such as endothelial dysfunction, cardiovascular complications, atherosclerosis, retinopathy, neuropathy, and Alzheimer's disease. Garcinia mangostana L. (Clusiaceae) (GM) was selected owing to the ability of its polar and non-polar fractions to inhibit AGE formation. For the first time, the bioguided fractionation of its pericarp MeOH extract (GMT) gave rise to two new xanthones, namely, mangostanaxanthones III (1) and IV (3), in addition to six known compounds, β-mangostin (2), garcinone E (4), rubraxanthone (5), α-mangostin (6), garcinone C (7), and 9-hydroxycalabaxanthone (8), from the non-polar faction. Their structures were verified by various spectroscopic methods, including 1D and 2D NMR studies and high-resolution MS data. All of the isolated xanthones significantly inhibited both sugar (ribose) and dicarbonyl compound (methylglyoxal)-induced protein glycation in a dose-dependent manner. This is explained by the ability of the isolated xanthones to inhibit protein oxidation, as indicated by the decreases in dityrosine and N'-formylkynurenine formation.


L-Histidinol Dehydrogenase as a New Target for Old Diseases.
Bacterial infections constitute an always growing health problem worldwide. The resistance to antibiotics of an increasing number of bacterial pathogens necessitates a permanent search for new molecules with different mechanisms of action. Histidine biosynthesis is an ancient pathway found in bacteria, archaebacteria, fungi and plants but absent in mammals. This feature makes it very interesting for the study of new strategies aimed to develop novel classes of antibacterial agents. In particular, one of the enzymes involved in the histidine biosynthesis, i.e. L-histidinol dehydrogenase (HDH), has been demonstrated to be essential for the survival of bacteria associated to several infections, such as brucellosis and tubercolosis. HDH is a Zn2+ enzyme which catalyzes the last two steps in the biosynthesis of Lhistidine: sequential NAD-dependent oxidations of L-histidinol to L-histidinaldehyde and then to L-histidine. This review will be focused on the biochemical and structural studies performed on HDH so far with the purpose to provide a structural background for the rational drug design of potent HDH inhibitors.


Methylmalonic Acid Levels and their Relation with Cobalamin Supplementation in Spanish Vegetarians.
Cobalamin deficiency represents a health issue for vegetarians, especially vegans, if supplements are not consumed. Vitamin B12 serum levels, traditionally used to assess the vitamin B12 status, can be normal under functional deficiency conditions. In this regard, methylmalonic acid (MMA) has proven to be a more specific marker to detect subclinical vitamin B12 deficiency. In this study, we present for the first time the cobalamin status of Spanish vegetarians using both vitamin B12 and MMA markers, and the effects of the plant-based diet and the intake of vitamin B12 supplements. Healthy adults were recruited (n = 103, 52% vegans). Dietary preferences and use of supplements were assessed by questionnaires and serum samples were collected and stored. Vitamin B12 was measured by chemiluminiscence and MMA by liquid chromatography tandem mass spectrometry (LC-MS/MS) using solid phase extraction for sample preparation. Obtained values, median (IQR), were: vitamin B12, 278.9 (160.2) pmol/l and MMA, 140.2 (78.9) nmol/l. No significant differences between lacto-ovo vegetarians and vegans were observed. Considering these two markers, 10% of the participants were mild vitamin B12 deficient. Supplementation (75% of the participants) was associated with higher vitamin B12 (p < 0.001) and lower MMA (p = 0.012). In conclusion, Spanish vegetarians have low risk of vitamin B12 deficiency due to vitamin B12 supplementation and the MMA determination is useful to detect mild deficiency.


Transforming Community Access Services through Client- and Family-Centred Homecare Transitions.
This article describes how one provincial health region adopted a client- and family-centred approach to improve access to community health services. Transition best practices and the "Triple Aim" supplied a framework for the transformation of transition of clients needing home healthcare services (Berwick et al. 2008). The need to improve the patient and family experience, establish and streamline professional practice standards, strengthen interprofessional collaborations, increase efficiency, create a critical mass of experts in the clinical domain of care transitions and program access, and evaluate customer experience were the organizational drivers for this transformation. The new framework identifies clients' needs and assigns a priority code. It also identifies which family member provides what support to the client and offers a one-stop service number staffed by individuals trained to provide client- and family-centred homecare services. This transformation of home healthcare transitions has improved the client and family experience, strengthened service provider satisfaction and generated efficiencies in prioritizing and delivering community healthcare services.


A Nationwide Survey of Hepatitis E Virus Infection and Chronic Hepatitis E in Liver Transplant Recipients in Japan.
Recently, chronic hepatitis E has been increasingly reported in organ transplant recipients in European countries. In Japan, the prevalence of hepatitis E virus (HEV) infection after transplantation remains unclear, so we conducted a nationwide cross-sectional study to clarify the prevalence of chronic HEV infection in Japanese liver transplant recipients. A total of 1893 liver transplant recipients in 17 university hospitals in Japan were examined for the presence of immunoglobulin G (IgG), IgM and IgA classes of anti-HEV antibodies, and HEV RNA in serum. The prevalence of anti-HEV IgG, IgM and IgA class antibodies was 2.9% (54/1893), 0.05% (1/1893) and 0% (0/1893), respectively. Of 1651 patients tested for HEV RNA, two patients (0.12%) were found to be positive and developed chronic infection after liver transplantation. In both cases, HEV RNA was also detected in one of the blood products transfused at the perioperative period. Analysis of the HEV genomes revealed that the HEV isolates obtained from the recipients and the transfused blood products were identical in both cases, indicating transfusion-transmitted HEV infection. The prevalence of HEV antibodies in liver transplant recipients was 2.9%, which is low compared with the healthy population in Japan and with organ transplant recipients in European countries; however, the present study found, for the first time, two Japanese patients with chronic HEV infection that was acquired via blood transfusion during or after liver transplantation.


Elevated antigen-driven IL-9 responses are prominent in peanut allergic humans.
Food allergies, and peanut allergy in particular, are leading causes of anaphylactic fatalities worldwide. The immune mechanisms that underlie food allergy remain ill-defined and controversial, in part because studies in humans typically focus on analysis of a limited number of prototypical Th1/Th2 cytokines. Here we determine the kinetics and prevalence of a broad panel of peanut-driven cytokine and chemokine responses in humans with current peanut allergy vs those with stable, naturally occurring clinical tolerance to peanut. Our primary focus is identification of novel indicators of immune dysregulation. Antigen-specific cytokine mRNA and protein responses were elicited in primary culture via peanut or irrelevant antigen (Leishmania extract, milk antigens) mediated stimulation of fresh peripheral blood cells from 40 individuals. Peanut extract exposure in vitro induced a broad panel of responses associated with Th2/Th9-like, Th1-like and Th17-like immunity. Peanut-dependent Type 2 cytokine responses were frequently found in both peanut allergic individuals and those who exhibit clinical tolerance to peanut ingestion. Among Th2/Th9-associated cytokines, IL-9 responses discriminated between allergic and clinically tolerant populations better than did commonly used IL-4, IL-5 or IL-13 responses. Comparison with responses evoked by unrelated control antigen-mediated stimulation showed that these differences are antigen-dependent and allergen-specific. Conversely, the intensity of IL-12, IL-17, IL-23 and IFN-γ production was indistinguishable in peanut allergic and peanut tolerant populations. In summary, the ability to generate and maintain cytokine responses to peanut is not inherently distinct between allergic and peanut tolerant humans. Quantitative differences in the intensity of cytokine production better reflects clinical phenotype, with optimally useful indicators being IL-9, IL-5, IL-13 and IL-4. Equivalent, and minimal, Ag-dependent pro-inflammatory cytokine levels in both healthy and peanut allergic volunteers argues against a key role for such cytokines in maintenance of clinical tolerance to food antigens in humans.


Rescuer fatigue: standard versus continuous chest-compression cardiopulmonary resuscitation.
Continuous chest-compression cardiopulmonary resuscitation (CCC-CPR) has been advocated as an alternative to standard CPR (STD-CPR). Studies have shown that CCC-CPR delivers substantially more chest compressions per minute and is easier to remember and perform than STD-CPR. One concern regarding CCC-CPR is that the rescuer may fatigue and be unable to maintain adequate compression rate or depth throughout an average emergency medical services response time. The specific aim of this study was to compare the effects of fatigue on the performance of CCC-CPR and STD-CPR on a manikin model. This was a prospective, randomized crossover study involving 53 medical students performing CCC-CPR and STD-CPR on a manikin model. Students were randomized to their initial CPR group and then performed the other type of CPR after a period of at least two days. Students were evaluated on their performance of 9 minutes of CPR for each method. The primary endpoint was the number of adequate chest compressions (at least 38 mm of compression depth) delivered per minute during each of the 9 minutes. The secondary endpoints were total compressions, compression rate, and the number of breaks taken for rest. The students' performance was evaluated on the basis of Skillreporter Resusci Anne (Laerdal, Wappingers Falls, NY) recordings. Primary and secondary endpoints were analyzed by using the generalized linear mixed model for counting data. In the first 2 minutes, participants delivered significantly more adequate compressions per minute with CCC-CPR than STD-CPR, (47 vs. 32, p = 0.004 in the 1st minute and 39 vs. 29, p = 0.04 in the 2nd minute). For minutes 3 through 9, the differences in number of adequate compressions between groups were not significant. Evaluating the 9 minutes of CPR as a whole, there were significantly more adequate compressions in CCC-CPR vs. STD-CPR (p = 0.0003). Although the number of adequate compressions per minute declined over time in both groups, the rate of decline was significantly greater in CCC-CPR compared with STD-CPR (p = 0.0003). The mean number of total compressions delivered in the first minute was significantly greater with CCC-CPR than STD-CPR (105 per minute vs. 58 per minute, p < 0.001) and did not change over 9 minutes in either group. There were no differences in compression rates or number of breaks between groups. CCC-CPR resulted in more adequate compressions per minute than STD-CPR for the first 2 minutes of CPR. However, the difference diminished after 3 minutes, presumably as a result of greater rescuer fatigue with CCC-CPR. Overall, CCC-CPR resulted in more total compressions per minute than STD-CPR during the entire 9 minutes of resuscitation.


Ischemia change in stable coronary artery disease is an independent predictor of death and myocardial infarction.
The aim of this study was to evaluate the independent prognostic significance of ischemia change in stable coronary artery disease (CAD). Recent randomized trials in stable CAD have suggested that revascularization does not improve outcomes compared with optimal medical therapy (MT). In contrast, the nuclear substudy of the COURAGE (Clinical Outcomes Utilizing Revascularization and Aggressive Drug Evaluation) trial found that revascularization led to greater ischemia reduction and suggested that this may be associated with improved unadjusted outcomes. Thus, the effects of MT versus revascularization on ischemia change and its independent prognostic significance requires further investigation. From the Duke Cardiovascular Disease and Nuclear Cardiology Databanks, 1,425 consecutive patients with angiographically documented CAD who underwent 2 serial myocardial perfusion single-photon emission computed tomography scans were identified. Ischemia change was calculated for patients undergoing MT alone, percutaneous coronary intervention, or coronary artery bypass grafting. Patients were followed for a median of 5.8 years after the second myocardial perfusion scan. Cox proportional hazards regression modeling was used to identify factors independently associated with the primary outcome of death or myocardial infarction (MI). Formal risk reclassification analyses were conducted to assess whether the addition of ischemia change to traditional predictors resulted in improved risk classification for death or MI. More MT patients (15.6%) developed ≥5% ischemia worsening compared with those undergoing percutaneous coronary intervention (6.2%) or coronary artery bypass grafting (6.7%) (p < 0.001). After adjustment for established predictors, ≥5% ischemia worsening remained a significant independent predictor of death or MI (hazard ratio: 1.634; p = 0.0019) irrespective of treatment arm. Inclusion of ≥5% ischemia worsening in this model resulted in significant improvement in risk classification (net reclassification improvement: 4.6%, p = 0.0056) and model discrimination (integrated discrimination improvement: 0.0062, p = 0.0057). In stable CAD, ischemia worsening is an independent predictor of death or MI, resulting in significantly improved risk reclassification when added to previously known predictors.


A descriptive study of selected oocyte, blood and organ/tissue donation features among fertility patients in Ireland.
Anonymous oocyte donation and participation in organ and blood/tissue donation programmes were studied specifically among Irish fertility patients. An anonymous questionnaire measured patient perceptions of, and participation in, blood/organ/tissue donor programmes, and to record opinion on anonymous donor oocyte compensation. A total of 337 patents were sampled; 56.7% had no children. None had participated in a donor oocyte programme either as donor or recipient. At baseline, 19.6% had previous in vitro fertilisation experience, more than one-third (35.9%) had donated blood anonymously, 19.9% were organ/tissue donors and 52.2% indicated that anonymous oocyte donors should receive some compensation. We found patients with infertility for extended periods were more likely to view oocyte donation favourably, compared to those with infertility of shorter durations (p = 0.022, by Krusksal-Wallis Rank Sum test). Average recommended compensation for anonymous oocyte donor was euro 2177 (range euro 200-euro 9500), and most (77.2%) favoured confidential protections for recipient and donor identity. This is the first investigation of blood and organ/tissue donation features among fertility patients in Ireland; the rate of blood donation in this group was more than 10 times higher than in the general Irish population. Protection of anonymity for both donors and recipients was supported by most patients, even opponents of compensated anonymous donation. Further studies should clarify patient perceptions about oocyte donation as a function of involvement in organ/tissue procurement programmes and blood banks.


Detection and analysis of the bacterium, Xylella fastidiosa, in glassy-winged sharpshooter, Homalodisca vitripennis, populations in Texas.
The glassy-winged sharpshooter, Homalodisca vitripeninis Germar (Hemiptera: Cicadellidae), is a xylophagous insect that is an endemic pest of several economically important plants in Texas. H. vitripennis is the main vector of Xylella fastidiosa Wells (Xanthomonadales: Xanthomonadaceae), the bacterium that causes Pierce's disease of grapevine and can travel long distances putting much of Texas grape production at risk. Understanding the movement of H. vitripennis populations capable of transmitting X. fastidiosa into Pierce's-disease-free areas is critical for developing a management program for Pierce's disease. To that end, the USDA-APHIS has developed a program to sample vineyards across Texas to monitor populations of H. vitripennis. From this sampling, H vitripennis collected during 2005 and 2006 over the months of May, June, and July from eight vineyards in different regions of Texas were recovered from yellow sticky traps and tested for the presence of X. fastidiosa. The foregut contents were vacuum extracted and analyzed using RT-PCR to determine the percentage of H. vitripennis within each population that harbor X. fastidiosa and have the potential to transmit this pathogen. H. vitripennis from vineyards known to have Pierce's disease routinely tested positive for the presence of X. fastidiosa. While almost all H. vitripennis collected from vineyards with no history of Pierce's disease tested negative for the presence of the pathogen, three individual insects tested positive. Furthermore, all three insects were determined, by DNA sequencing, to be carrying a strain of X. fastidiosa homologous to known Pierce's disease strains, signifying them as a risk factor for new X. fastidiosa infections.


Enhanced responses of spinal dorsal horn neurons to heat and cold stimuli following mild freeze injury to the skin.
The effects of a mild freeze injury to the skin on responses of nociceptive dorsal horn neurons to cold and heat stimuli were examined in anesthetized rats. Electrophysiological recordings were obtained from 72 nociceptive spinal neurons located in the superficial and deep dorsal horn. All neurons had receptive fields (RFs) on the glabrous skin of the hindpaw, and neurons were functionally divided into wide dynamic range (WDR) and high-threshold (HT) neurons. Forty-four neurons (61%) were classified as WDR and responded to both innocuous and noxious mechanical stimuli (mean mechanical threshold of 12.8 +/- 1.6 mN). Twenty-eight neurons (39%) were classified as HT and were excited only by noxious mechanical stimuli (mean mechanical threshold of 154.2 +/- 18.3 mN). Neurons were characterized for their sensitivity heat (35 to 51 degrees C) and cold (28 to -12 degrees C) stimuli applied to their RF. Among WDR neurons, 86% were excited by both noxious heat and cold stimuli, while 14% responded only to heat. For HT neurons, 61% responded to heat and cold stimuli, 32% responded only to noxious heat, and 7% responded only to noxious cold. Effects of a mild freeze injury (-15 degrees C applied to the RF for 20 s) on responses to heat and cold stimuli were examined in 30 WDR and 22 HT neurons. Skin freezing was verified as an abrupt increase in skin temperature at the site of injury due to the exothermic reaction associated with crystallization. Freezing produced a decrease in response thresholds to heat and cold stimuli in most WDR and HT neurons. WDR and HT neurons exhibited a mean decrease in response threshold for cold of 9.0 +/- 1.3 degrees C and 10.0 +/- 1.6 degrees C, respectively. Mean response thresholds for heat decreased 4.0 +/- 0.4 degrees C and 4.3 +/- 1.3 degrees C in WDR and HT neurons, respectively. In addition, responses to suprathreshold cold and heat stimuli increased. WDR and HT neurons exhibited an 89% and a 192% increase in response across all cold stimuli, and a 93 and 92% increase in responses evoked across all heat stimuli, respectively. Our results demonstrate that many spinal neurons encode intensity of noxious cold as well as noxious heat over a broad range of stimulus temperatures. Enhanced responses of WDR and HT neurons to cold and heat stimuli after a mild freeze injury is likely to contribute to thermal hyperalgesia following a similar freeze injury in humans.


Natural and engineered isoforms of the inflammasome adaptor ASC form non-covalent, pH-responsive hydrogels.
The protein ASC polymerizes into intricate filament networks to assemble the inflammasome, a filamentous multiprotein complex that triggers the inflammatory response. ASC carries two Death Domains integrally involved in protein self-association for filament assembly. We have leveraged this behavior to create non-covalent, pH-responsive hydrogels of full-length, folded ASC by carefully controlling the pH as a critical factor in the polymerization process. We show that natural variants of ASC (ASC isoforms) involved in inflammasome regulation also undergo hydrogelation. To further demonstrate this general capability, we engineered proteins inspired in the ASC structure that successfully form hydrogels. We analyzed the structural network of the natural and engineered protein hydrogels using transmission and scanning electron microscopy, and studied their viscoelastic behavior by shear rheology. Our results reveal one of the very few examples of hydrogels created by the self-assembly of globular proteins and domains in their native conformation and show that Death Domains can be used alone or as building blocks to engineer bioinspired hydrogels.


Analysis of genetically modified red-fleshed apples reveals effects on growth and consumer attributes.
Consumers of whole foods, such as fruits, demand consistent high quality and seek varieties with enhanced health properties, convenience or novel taste. We have raised the polyphenolic content of apple by genetic engineering of the anthocyanin pathway using the apple transcription factor MYB10. These apples have very high concentrations of foliar, flower and fruit anthocyanins, especially in the fruit peel. Independent lines were examined for impacts on tree growth, photosynthesis and fruit characteristics. Fruit were analysed for changes in metabolite and transcript levels. Fruit were also used in taste trials to study the consumer perception of such a novel apple. No negative taste attributes were associated with the elevated anthocyanins. Modification with this one gene provides near isogenic material and allows us to examine the effects on an established cultivar, with a view to enhancing consumer appeal independently of other fruit qualities.


Prognostic significance of depth of invasion in cancer of the larynx.
To determine the prognostic significance of depth of invasion in laryngeal cancer, the depth of invasion of tumor was measured with an ocular micrometer on the laryngectomy specimens of 94 surgically treated patients with T1, T2, and T3 laryngeal cancer and was expressed in millimeters. There was a significant negative correlation between the depth of invasion and disease-free survival. The tumors with no clinical involvement of regional lymph nodes in neck (NO neck) had significantly less depth of invasion than those with involvement (N+ neck). The tumors with pathologically confirmed cervical lymph node metastasis had significantly more depth of invasion than those without metastasis. For tumors with a depth of invasion equal to or greater than 3.25 mm, the rate of cervical metastasis in this study has always been significantly higher than for those with a depth of invasion less than 3.25 mm (P < .05). The mean depths of invasion for cases with and without recurrence were not significantly different. According to the multivariate analysis, depth of invasion (P = .047) and patient age (P = .113) significantly affected the disease-free survival independently. The depth of invasion did not significantly affect the recurrence and the interval between surgery and the development of recurrence (P > .15). The depth of invasion should be measured in every laryngectomy specimen. The depth of invasion influences the cervical metastasis and disease-free survival significantly but does not affect the recurrence rate. The depth of invasion plays an independent role in determining the disease-free survival.


Estimation of leaf nitrogen content from spectral characteristics of rice canopy.
Ground-based remotely sensed reflectance spectra of hyperspectral resolution were monitored during the growing period of rice under various nitrogen application rates. It was found that reflectance spectrum of rice canopy changed in both wavelength and reflectance as the plants developed. Fifteen characteristic wavebands were identified from the apparent peaks and valleys of spectral reflectance curves, in accordance with the results of the first-order differentiation, measured over the growing season of rice. The bandwidths and center wavelengths of these characteristic wavebands were different among nitrogen treatments. The simplified features by connecting these 15 characteristic wavelengths may be considered as spectral signatures of rice canopy, but spectral signatures varied with developmental age and nitrogen application rates. Among these characteristic wavebands, the changes of the wavelength in band 11 showed a positive linear relationship with application rates of nitrogen fertilizer, while it was a negative linear relationship in band 5. Mean reflectance of wavelengths in bands 1, 2, 3, 5, 11, and 15 was significantly correlated with application rates. Reflectance of these six wavelengths changed nonlinearly after transplanting and could be used in combination to distinguish rice plants subjected to different nitrogen application rates. From the correlation analyses, there are a variety of correlation coefficients for spectral reflectance to leaf nitrogen content in the range of 350-2400 nm. Reflectance of most wavelengths exhibited an inverse correlation with leaf nitrogen content, with the largest negative value (r = -0.581) located at about 1376 nm. Changes in reflectance at 1376 nm to leaf nitrogen content during the growing period were closely related and were best fitted to a nonlinear function. This relationship may be used to estimate and to monitor nitrogen content of rice leaves during rice growth. Reflectance of red light minimum and near-infrared peak and leaf nitrogen content were correlated nonlinearly.


LiDAR-Based GNSS Denied Localization for Autonomous Racing Cars.
Self driving vehicles promise to bring one of the greatest technological and social revolutions of the next decade for their potential to drastically change human mobility and goods transportation, in particular regarding efficiency and safety. Autonomous racing provides very similar technological issues while allowing for more extreme conditions in a safe human environment. While the software stack driving the racing car consists of several modules, in this paper we focus on the localization problem, which provides as output the estimated pose of the vehicle needed by the planning and control modules. When driving near the friction limits, localization accuracy is critical as small errors can induce large errors in control due to the nonlinearities of the vehicle's dynamic model. In this paper, we present a localization architecture for a racing car that does not rely on Global Navigation Satellite Systems (GNSS). It consists of two multi-rate Extended Kalman Filters and an extension of a state-of-the-art laser-based Monte Carlo localization approach that exploits some a priori knowledge of the environment and context. We first compare the proposed method with a solution based on a widely employed state-of-the-art implementation, outlining its strengths and limitations within our experimental scenario. The architecture is then tested both in simulation and experimentally on a full-scale autonomous electric racing car during an event of Roborace Season Alpha. The results show its robustness in avoiding the robot kidnapping problem typical of particle filters localization methods, while providing a smooth and high rate pose estimate. The pose error distribution depends on the car velocity, and spans on average from 0.1 m (at 60 km/h) to 1.48 m (at 200 km/h) laterally and from 1.9 m (at 100 km/h) to 4.92 m (at 200 km/h) longitudinally.


Efficacy of Dural Sealant System for Preventing Brain Shift and Improving Accuracy in Deep Brain Stimulation Surgery.
The success of deep brain stimulation (DBS) depends heavily on surgical accuracy, and brain shift is recognized as a significant factor influencing accuracy. We investigated the factors associated with surgical accuracy and showed the effectiveness of a dural sealant system for preventing brain shift in 32 consecutive cases receiving DBS. Thirty-two patients receiving DBS between March 2014 and May 2015 were included in this study. We employed conventional burr hole techniques for the first 18 cases (Group I) and a dural sealant system (DuraSeal) for the subsequent 14 cases (Group II). We measured gaps between the actual positions of electrodes and the predetermined target positions. We then retrospectively evaluated the factors involved in surgical accuracy. The average gap between an electrode's actual and target positions was 1.55 ± 0.83 mm in all cases. Postoperative subdural air volume e, the only factor associated with surgical accuracy (r = 0.536, P < 0.0001), was significantly smaller in Group II (Group I: 43.9 ± 27.7, Group II: 12.1 ± 12.5 ml, P = 0.0006). The average electrode position gap was also significantly smaller in Group II (Group I: 1.77 ± 0.91, Group II: 1.27 ± 0.59 mm, P = 0.035). Use of a dural sealant system could significantly reduce intracranial air volume, which should improve surgical accuracy.


Mineralogy at Gusev Crater from the Mössbauer spectrometer on the Spirit Rover.
Mössbauer spectra measured on Mars by the Spirit rover during the primary mission are characterized by two ferrous iron doublets (olivine and probably pyroxene) and a ferric iron doublet (tentatively associated to nanophase ferric iron oxide). Two sextets resulting from nonstoichiometric magnetite are also present, except for a coating on the rock Mazatzal, where a hematite-like sextet is present. Greater proportions of ferric-bearing phases are associated with undisturbed soils and rock surfaces as compared to fresh rock surfaces exposed by grinding. The ubiquitous presence of olivine in soil suggests that physical rather than chemical weathering processes currently dominate at Gusev crater.


Complex spatiotemporal haemodynamic response following sensory stimulation in the awake rat.
Detailed understanding of the haemodynamic changes that underlie non-invasive neuroimaging techniques such as blood oxygen level dependent functional magnetic resonance imaging is essential if we are to continue to extend the use of these methods for understanding brain function and dysfunction. The use of animal and in particular rodent research models has been central to these endeavours as they allow in-vivo experimental techniques that provide measurements of the haemodynamic response function at high temporal and spatial resolution. A limitation of most of this research is the use of anaesthetic agents which may disrupt or mask important features of neurovascular coupling or the haemodynamic response function. In this study we therefore measured spatiotemporal cortical haemodynamic responses to somatosensory stimulation in awake rats using optical imaging spectroscopy. Trained, restrained animals received non-noxious stimulation of the whisker pad via chronically implanted stimulating microwires whilst optical recordings were made from the contralateral somatosensory cortex through a thin cranial window. The responses we measure from un-anaesthetised animals are substantially different from those reported in previous studies which have used anaesthetised animals. These differences include biphasic response regions (initial increases in blood volume and oxygenation followed by subsequent decreases) as well as oscillations in the response time series of awake animals. These haemodynamic response features do not reflect concomitant changes in the underlying neuronal activity and therefore reflect neurovascular or cerebrovascular processes. These hitherto unreported hyperemic response dynamics may have important implications for the use of anaesthetised animal models for research into the haemodynamic response function.


miR-122 regulates hepatic lipid metabolism and tumor suppression.
In this issue of JCI, two independent groups describe the effects of germline and liver-specific deletion of Mir122a, the predominant liver miRNA. Their findings reveal a critical role for miR-122 in fat and cholesterol metabolism but suggest that other metabolic actions of the liver are independent of miR-122. Knockout mice also displayed hepatic inflammation, fibrosis, and a high incidence of hepatocellular carcinoma, suggesting that miR-122 has a tumor suppressor role in hepatocytes.


Impact of mother's education on full immunization of children aged 12-23 months in Eritrea: population and health survey 2010 data analysis.
Although vaccination coverage in Eritrea has improved in recent years, some children are still missing out, and it's important to identify risk factors for lower coverage in order to target campaigns and interventions. The objective of this study was to assess: (1) the impact of maternal education on full immunization of children aged 12-23 months, and (2) whether the association was confounded or modified by other factors. This study was a secondary data analysis of the Eritrean Population and Health Survey 2010 (EPHS 2010). In this analysis 1323 mothers of children aged 12-23 months were included. The outcome of the study was full immunization, defined as receiving all the WHO recommended basic vaccines: one dose of Bacillus Calmette-Gué rin (BCG), three doses of diphtheria-pertussis-tetanus(DPT), three doses of polio, and one dose of measles vaccine. The primary exposure was maternal education. Data on immunization coverage came from vaccination cards and from mothers' or caretakers' verbal reports. Bivariate and multivariable logistic regression analyses were performed. Full vaccination coverage among children aged 12-23 months was 83%. Most children received BCG (95%), DPT1 (97%), DPT2 (96%), DPT3 (93%), polio1 (97%), polio2 (97%), polio3 (91%) and measles (92%). In unadjusted analyses, children of mothers with primary (OR = 2.75, 95% CI 1.74-4.37), and middle or above (OR = 3.16, 95% CI 2.09-4.78) education were more likely to be fully immunised. However, after adjusting for wealth, region, ANC visit, and vaccination card ownership, only the effect for primary education remained significant (OR = 2.34, 95% CI 1.30-4.21). The result of this study suggested that children of mothers who attained primary level were more likely to be fully vaccinated than children of mothers with no education. The association was influenced by wealth index of household, mothers ANC visit, region, and possession of vaccination card. The Expanded Program on Immunization of the Ministry of Health should target strategies to enhance full immunization among children of mothers with no education.


Patient education for neck pain with or without radiculopathy.
Neck disorders are common, disabling, and costly. The effectiveness of patient education strategies is unclear. To assess whether patient education strategies, either alone or in combination with other treatments, are of benefit for pain, function, global perceived effect, quality of life, or patient satisfaction, in adults with neck pain with and without radiculopathy. Computerized bibliographic databases were searched from their start up to May 31, 2008. Eligible studies were quasi or randomized trials (RCT) investigating the effectiveness of patient education strategies for neck disorder. Paired independent review authors carried out study selection, data abstraction, and methodological quality assessment. Relative risk and standardized mean differences (SMD) were calculated. The appropriateness of combining studies was assessed on clinical and statistical grounds. Because of differences in intervention type or disorder, no studies were considered appropriate to pool. Of the 10 selected trials, two (20%) were rated high quality. Advice was assessed as follows:Eight trials of advice focusing on activation compared to no treatment or to various active treatments, including therapeutic exercise, manual therapy and cognitive behavioural therapy, showed either inferiority or no difference for pain, spanning a full range of follow-up periods and disorder types. When compared to rest, two trials that assessed acute whiplash-associated disorders (WAD) showed moderate evidence of no difference for various forms of advice focusing on activation. Two trials studying advice focusing on pain & stress coping skills found moderate evidence of no benefit for pain in chronic mechanical neck disorder (MND) at intermediate/long-term follow-up. One trial compared the effects of ' traditional neck school ' to no treatment, yielding limited evidence of no benefit for pain at intermediate-term follow-up in mixed acute/subacute/chronic neck pain. This review has not shown effectiveness for educational interventions in various disorder types and follow-up periods, including advice to activate, advice on stress coping skills, and 'neck school'. In future research, further attention to methodological quality is necessary. Studies of multimodal interventions should consider study designs, such as factorial designs, that permit discrimination of the specific educational components.


Effects of oral adenosine-5'-triphosphate supplementation on athletic performance, skeletal muscle hypertrophy and recovery in resistance-trained men.
Currently, there is a lack of studies examining the effects of adenosine-5'-triphosphate (ATP) supplementation utilizing a long-term, periodized resistance-training program (RT) in resistance-trained populations. Therefore, we investigated the effects of 12 weeks of 400 mg per day of oral ATP on muscular adaptations in trained individuals. We also sought to determine the effects of ATP on muscle protein breakdown, cortisol, and performance during an overreaching cycle. The study was a 3-phase randomized, double-blind, and placebo- and diet-controlled intervention. Phase 1 was a periodized resistance-training program. Phase 2 consisted of a two week overreaching cycle in which volume and frequency were increased followed by a 2-week taper (Phase 3). Muscle mass, strength, and power were examined at weeks 0, 4, 8, and 12 to assess the chronic effects of ATP; assessment performance variables also occurred at the end of weeks 9 and 10, corresponding to the mid and endpoints of the overreaching cycle. There were time (p<0.001), and group x time effects for increased total body strength (+55.3 ± 6.0 kg ATP vs. + 22.4 ± 7.1 kg placebo, p<0.001); increased vertical jump power (+ 796 ± 75 ATP vs. 614 ± 52 watts placebo, p<0.001); and greater ultrasound determined muscle thickness (+4.9 ± 1.0 ATP vs. (2.5 ± 0.6 mm placebo, p<0.02) with ATP supplementation. During the overreaching cycle, there were group x time effects for strength and power, which decreased to a greater extent in the placebo group. Protein breakdown was also lower in the ATP group. Our results suggest oral ATP supplementation may enhance muscular adaptations following 12-weeks of resistance training, and prevent decrements in performance following overreaching. No statistically or clinically significant changes in blood chemistry or hematology were observed. ClinicalTrials.gov NCT01508338.


TRPA1 is a major oxidant sensor in murine airway sensory neurons.
Sensory neurons in the airways are finely tuned to respond to reactive chemicals threatening airway function and integrity. Nasal trigeminal nerve endings are particularly sensitive to oxidants formed in polluted air and during oxidative stress as well as to chlorine, which is frequently released in industrial and domestic accidents. Oxidant activation of airway neurons induces respiratory depression, nasal obstruction, sneezing, cough, and pain. While normally protective, chemosensory airway reflexes can provoke severe complications in patients affected by inflammatory airway conditions like rhinitis and asthma. Here, we showed that both hypochlorite, the oxidizing mediator of chlorine, and hydrogen peroxide, a reactive oxygen species, activated Ca(2+) influx and membrane currents in an oxidant-sensitive subpopulation of chemosensory neurons. These responses were absent in neurons from mice lacking TRPA1, an ion channel of the transient receptor potential (TRP) gene family. TRPA1 channels were strongly activated by hypochlorite and hydrogen peroxide in primary sensory neurons and heterologous cells. In tests of respiratory function, Trpa1(-/-) mice displayed profound deficiencies in hypochlorite- and hydrogen peroxide-induced respiratory depression as well as decreased oxidant-induced pain behavior. Our results indicate that TRPA1 is an oxidant sensor in sensory neurons, initiating neuronal excitation and subsequent physiological responses in vitro and in vivo.


Active site plasticity in D-amino acid oxidase: a crystallographic analysis.
D-Amino acid oxidase (DAAO) is the prototype of the flavin-containing oxidases. It catalyzes the oxidative deamination of various D-amino acids, ranging from D-Ala to D-Trp. We have carried out the X-ray analysis of reduced DAAO in complex with the reaction product imino tryptophan (iTrp) and of the covalent adduct generated by the photoinduced reaction of the flavin with 3-methyl-2-oxobutyric acid (kVal). These structures were solved by combination of 8-fold density averaging and least-squares refinement techniques. The FAD redox state of DAAO crystals was assessed by single-crystal polarized absorption microspectrophotometry. iTrp binds to the reduced enzyme with the N, C alpha, C, and C beta atoms positioned 3.8 A from the re side of the flavin. The indole side chain points away from the cofactor and is bound in the active site through a rotation of Tyr224. This residue plays a crucial role in that it adapts its conformation to the size of the active site ligand, providing the enzyme with the plasticity required for binding a broad range of substrates. The iTrp binding mode is fully consistent with the proposal, inferred from the analysis of the native DAAO structure, that substrate oxidation occurs via direct hydride transfer from the C alpha to the flavin N5 atom. In this regard, it is remarkable that, even in the presence of the bulky iTrp ligand, the active center is made solvent inaccessible by loop 216-228. This loop is thought to switch between the "closed" conformation observed in the crystal structures and an "open" state required for substrate binding and product release. Loop closure is likely to have a role in catalysis by increasing the hydrophobicity of the active site, thus making the hydride transfer reaction more effective. Binding of kVal leads to keto acid decarboxylation and formation of a covalent bond between the keto acid C alpha and the flavin N5 atoms. Formation of this acyl adduct results in a nonplanar flavin, characterized by a 22 degrees angle between the pyrimidine and benzene rings. Thus, in addition to an adaptable substrate binding site, DAAO has the ability to bind a highly distorted cofactor. This ability is relevant for the enzyme's function as a highly efficient oxidase.


Synthesis of Benzannulated Medium-ring Lactams via a Tandem Oxidative Dearomatization-Ring Expansion Reaction.
Medium-ring natural products exhibit diverse biological activities but such scaffolds are underrepresented in probe and drug discovery efforts due to the limitations of classical macrocyclization reactions. We report herein a tandem oxidative dearomatization-ring-expanding rearomatization (ODRE) reaction that generates benzannulated medium-ring lactams directly from simple bicyclic substrates. The reaction accommodates diverse aryl substrates (haloarenes, aryl ethers, aryl amides, heterocycles) and strategic incorporation of a bridgehead alcohol generates a versatile ketone moiety in the products amenable to downstream modifications. Cheminformatic analysis indicates that these medium rings access regions of chemical space that overlap with related natural products and are distinct from synthetic drugs, setting the stage for their use in discovery screening against novel biological targets.


Calcium phosphate transfection generates mammalian recombinant cell lines with higher specific productivity than polyfection.
Transfection with polyethylenimine (PEI) was evaluated as a method for the generation of recombinant Chinese hamster ovary (CHO DG44) cell lines by direct comparison with calcium phosphate-DNA coprecipitation (CaPO4) using both green fluorescent protein (GFP) and a monoclonal antibody as reporter proteins. Following transfection with a GFP expression vector, the proportion of GFP-positive cells as determined by flow cytometry was fourfold higher for the PEI transfection as compared to the CaPO4 transfection. However, the mean level of transient GFP expression for the cells with the highest level of fluorescence was twofold greater for the CaPO4 transfection. Fluorescence in situ hybridization on metaphase chromosomes from pools of cells grown under selective pressure demonstrated that plasmid integration always occurred at a single site regardless of the transfection method. Importantly, the copy number of integrated plasmids was measurably higher in cells transfected with CaPO4. The efficiency of recombinant cell line recovery under selective pressure was fivefold higher following PEI transfection, but the average specific productivity of a recombinant antibody was about twofold higher for the CaPO4-derived cell lines. Nevertheless, no difference between the two transfection methods was observed in terms of the stability of protein production. These results demonstrated the feasibility of generating recombinant CHO-derived cell lines by PEI transfection. However, this method appeared inferior to CaPO4 transfection with regard to the specific productivity of the recovered cell lines.


Quantum fluctuations of a fullerene cage modulate its internal magnetic environment.
To investigate the effect of quantum fluctuations on the magnetic environment inside a C60 fullerene cage, we have calculated the nuclear magnetic shielding constant of protons in H2@C60 and HD@C60 systems by on-the-fly ab initio path integral simulation, including both thermal and nuclear quantum effects. The most dominant upfield from an isolated hydrogen molecule occurs due to the diamagnetic current of the C60 cage, which is partly cancelled by the paramagnetic current, where the paramagnetic contribution is enlarged by the zero-point vibrational fluctuation of the C60 carbon backbone structure via a widely distributed HOMO-LUMO gap. This quantum modulation mechanism of the nuclear magnetic shielding constant is newly proposed. Because this quantum effect is independent of the difference between H2 and HD, the H2/HD isotope shift occurs in spite of the C60 cage. The nuclear magnetic constants computed for H2@C60 and HD@C60 are 32.047 and 32.081 ppm, respectively, which are in reasonable agreement with the corresponding values of 32.19 and 32.23 ppm estimated from the experimental values of the chemical shifts.


Development of a decision analytic model to support decision making and risk communication about thrombolytic treatment.
Individualised prediction of outcomes can support clinical and shared decision making. This paper describes the building of such a model to predict outcomes with and without intravenous thrombolysis treatment following ischaemic stroke. A decision analytic model (DAM) was constructed to establish the likely balance of benefits and risks of treating acute ischaemic stroke with thrombolysis. Probability of independence, (modified Rankin score mRS ≤ 2), dependence (mRS 3 to 5) and death at three months post-stroke was based on a calibrated version of the Stroke-Thrombolytic Predictive Instrument using data from routinely treated stroke patients in the Safe Implementation of Treatments in Stroke (SITS-UK) registry. Predictions in untreated patients were validated using data from the Virtual International Stroke Trials Archive (VISTA). The probability of symptomatic intracerebral haemorrhage in treated patients was incorporated using a scoring model from Safe Implementation of Thrombolysis in Stroke-Monitoring Study (SITS-MOST) data. The model predicts probabilities of haemorrhage, death, independence and dependence at 3-months, with and without thrombolysis, as a function of 13 patient characteristics. Calibration (and inclusion of additional predictors) of the Stroke-Thrombolytic Predictive Instrument (S-TPI) addressed issues of under and over prediction. Validation with VISTA data confirmed that assumptions about treatment effect were just. The C-statistics for independence and death in treated patients in the DAM were 0.793 and 0.771 respectively, and 0.776 for independence in untreated patients from VISTA. We have produced a DAM that provides an estimation of the likely benefits and risks of thrombolysis for individual patients, which has subsequently been embedded in a computerised decision aid to support better decision-making and informed consent.


Genome wide analysis of protein production load in Trichoderma reesei.
The filamentous fungus Trichoderma reesei (teleomorph Hypocrea jecorina) is a widely used industrial host organism for protein production. In industrial cultivations, it can produce over 100 g/l of extracellular protein, mostly constituting of cellulases and hemicellulases. In order to improve protein production of T. reesei the transcriptional regulation of cellulases and secretory pathway factors have been extensively studied. However, the metabolism of T. reesei under protein production conditions has not received much attention. To understand the physiology and metabolism of T. reesei under protein production conditions we carried out a well-controlled bioreactor experiment with extensive analysis. We used minimal media to make the data amenable for modelling and three strain pairs to cover different protein production levels. With RNA-sequencing transcriptomics we detected the concentration of the carbon source as the most important determinant of the transcriptome. As the major transcriptional response concomitant to protein production we detected the induction of selected genes that were putatively regulated by xyr1 and were related to protein transport, amino acid metabolism and transcriptional regulation. We found novel metabolic responses such as production of glycerol and a cellotriose-like compound. We then used this cultivation data for flux balance analysis of T. reesei metabolism and demonstrate for the first time the use of genome wide stoichiometric metabolic modelling for T. reesei. We show that our model can predict protein production rate and provides novel insight into the metabolism of protein production. We also provide this unprecedented cultivation and transcriptomics data set for future modelling efforts. The use of stoichiometric modelling can open a novel path for the improvement of protein production in T. reesei. Based on this we propose sulphur assimilation as a major limiting factor of protein production. As an organism with exceptional protein production capabilities modelling of T. reesei can provide novel insight also to other less productive organisms.


Homo- and hetero-oligomerization of beta-arrestins in living cells.
Arrestins are important proteins, which regulate the function of serpentine heptahelical receptors and contribute to multiple signaling pathways downstream of receptors. The ubiquitous beta-arrestins are believed to function exclusively as monomers, although self-association is assumed to control the activity of visual arrestin in the retina, where this isoform is particularly abundant. Here the oligomerization status of beta-arrestins was investigated using different approaches, including co-immunoprecipitation of epitope-tagged beta-arrestins and resonance energy transfer (BRET and FRET) in living cells. At steady state and at physiological concentrations, beta-arrestins constitutively form both homo- and hetero-oligomers. Co-expression of beta-arrestin2 and beta-arrestin1 prevented beta-arrestin1 accumulation into the nucleus, suggesting that hetero-oligomerization may have functional consequences. Our data clearly indicate that beta-arrestins can exist as homo- and hetero-oligomers in living cells and raise the hypothesis that the oligomeric state may regulate their subcellular distribution and functions.


Brain-derived neurotrophic factor serum levels correlate with cognitive performance in Parkinson's disease patients with mild cognitive impairment.
Brain-derived neurotrophic factor (BDNF) is a trophic factor regulating cell survival and synaptic plasticity. Recent findings indicate that BDNF could be a potential regulatory factor for cognitive functioning in normal and/or neuropathological conditions. With regard to neurological disorders, recent data suggest that individuals with Parkinson's disease (PD) may be affected by cognitive deficits and that they have altered BDNF production. Therefore, the hypothesis can be advanced that BDNF levels are associated with the cognitive state of these patients. With this in mind, the present study was aimed at exploring the relationship between BDNF serum levels and cognitive functioning in PD patients with mild cognitive impairment (MCI). Thirteen PD patients with MCI were included in the study. They were administered an extensive neuropsychological test battery that investigated executive, episodic memory, attention, visual-spatial and language domains. A single score was obtained for each cognitive domain by averaging z-scores on tests belonging to that specific domain. BDNF serum levels were measured by enzyme-linked immunoassay (ELISA). Pearson's correlation analyses were performed between BDNF serum levels and cognitive performance. Results showed a significant positive correlation between BDNF serum levels and both attention (p < 0.05) and executive (p < 0.05) domains. Moreover, in the executive domain we found a significant correlation between BDNF levels and scores on tests assessing working memory and self-monitoring/inhibition. These preliminary data suggest that BDNF serum levels are associated with cognitive state in PD patients with MCI. Given the role of BDNF in regulating synaptic plasticity, the present findings give further support to the hypothesis that this trophic factor may be a potential biomarker for evaluating cognitive changes in PD and other neurological syndromes associated with cognitive decline.


Constitutive and regulated alpha-secretase cleavage of Alzheimer's amyloid precursor protein by a disintegrin metalloprotease.
Amyloid beta peptide (Abeta), the principal proteinaceous component of amyloid plaques in brains of Alzheimer's disease patients, is derived by proteolytic cleavage of the amyloid precursor protein (APP). Proteolytic cleavage of APP by a putative alpha-secretase within the Abeta sequence precludes the formation of the amyloidogenic peptides and leads to the release of soluble APPsalpha into the medium. By overexpression of a disintegrin and metalloprotease (ADAM), classified as ADAM 10, in HEK 293 cells, basal and protein kinase C-stimulated alpha-secretase activity was increased severalfold. The proteolytically activated form of ADAM 10 was localized by cell surface biotinylation in the plasma membrane, but the majority of the proenzyme was found in the Golgi. These results support the view that APP is cleaved both at the cell surface and along the secretory pathway. Endogenous alpha-secretase activity was inhibited by a dominant negative form of ADAM 10 with a point mutation in the zinc binding site. Studies with purified ADAM 10 and Abeta fragments confirm the correct alpha-secretase cleavage site and demonstrate a dependence on the substrate's conformation. Our results provide evidence that ADAM 10 has alpha-secretase activity and many properties expected for the proteolytic processing of APP. Increases of its expression and activity might be beneficial for the treatment of Alzheimer's disease.


Modelling the effect of SPION size in a stent assisted magnetic drug targeting system with interparticle interactions.
Cancer is a leading cause of death worldwide and it is caused by the interaction of genomic, environmental, and lifestyle factors. Although chemotherapy is one way of treating cancers, it also damages healthy cells and may cause severe side effects. Therefore, it is beneficial in drug delivery in the human body to increase the proportion of the drugs at the target site while limiting its exposure at the rest of body through Magnetic Drug Targeting (MDT). Superparamagnetic iron oxide nanoparticles (SPIONs) are derived from polyol methods and coated with oleic acid and can be used as magnetic drug carrier particles (MDCPs) in an MDT system. Here, we develop a mathematical model for studying the interactions between the MDCPs enriched with three different diameters of SPIONs (6.6, 11.6, and 17.8 nm) in the MDT system with an implanted magnetizable stent using different magnetic field strengths and blood velocities. Our computational analysis allows for the optimal design of the SPIONs enriched MDCPs to be used in clinical applications.


Oxygen Restriction Generates Difficult-to-Culture P. aeruginosa.
Induction of a non-culturable state has been demonstrated for many bacteria, e.g., Escherichia coli and various Vibrio spp. In a clinical perspective, the lack of growth due to these non-culturable bacteria can have major consequences for the treatment of patients. Here, we show how anoxic conditioning (restriction of molecular oxygen, O2) generates difficult-to-culture (DTC) bacteria during biofilm growth. A significant subpopulation of Pseudomonas aeruginosa entered a DTC state after anoxic conditioning, ranging from 5 to 90% of the total culturable population, in both planktonic and biofilm models. Anoxic conditioning also generated DTC subpopulations of Staphylococcus aureus and Staphylococcus epidermidis (89 and 42% of the total culturable population, respectively). Growth of the DTC populations were achieved by substituting O2 with 10 mM NO3 - as an alternative electron acceptor for anaerobic respiration or, in the case of P. aeruginosa, by adding sodium pyruvate or catalase as scavengers against reactive oxygen species (ROS) during aerobic respiration. An increase in normoxic plating due to addition of catalase suggests the molecule hydrogen peroxide as a possible mechanism for induction of DTC P. aeruginosa. Anoxic conditioning also generated a true viable but non-culturable (VBNC) population of P. aeruginosa that was not resurrected by substituting O2 with NO3 - during anaerobic respiration. These results demonstrate that habituation to an anoxic micro-environment could complicate diagnostic culturing of bacteria, especially in the case of chronic infections where oxygen is restricted due to the host immune response.


Role of HIF-1 in Cancer Progression: Novel Insights. A Review.
Hypoxia is a condition always present in tumor environment owing to the fast growth of tumor cells not supported by adequate blood supply. There is increasing evidence that hypoxia plays an important role in cancer dormancy and cancer metabolism, increasing stemness activity and bringing about cancer initiation and progression. This condition may influence the production of hypoxia inducible factor (HIF) a helix transcription factor which is involved in carcinogenesis and tumor growth through the regulation of genes involved in angiogenesis, glycolytic metabolism and other biological mechanisms. In normoxia condition HIF is inactivated by prolyl hydroxylase enzymes (EGLN 1-3, also known as PHD 1-3) using oxygen as a substrate. Once hydroxilated it binds to a protein called Von Hippel Lindau protein (VHL) for its degradation, whereas in hypoxia condition stabilization and nuclear translocation occur, leading to oncogenes activation. It has got three isoforms HIF-1 HIF-2 and HIF-3. The most studied factor is HIF-1 which is a heterodimer consisting of two forms, the form α is expressed in manner oxygen dependent, the form β is expressed constitutively. Its presence in tumor microenvironment could foster among other the expression of VEGF, HGF, Met protoncogene which induces degradation of the extracellular matrix and TWIST gene, which is in turn involved in a mechanism of cancer cell metastasis called epithelial-mesenchimal transition(EMT). In this review, we summarize the most important findings in HIF action in different types of cancer focusing on its properties to induce tumor cell growth and highlighting its poor prognostic value in different cancers sites.


GPX1 Pro(198)Leu polymorphism, erythrocyte GPX activity, interaction with alcohol consumption and smoking, and risk of colorectal cancer.
GPX1 encoding the enzyme glutathione peroxidase 1 (GPX1) and hOGG1 encoding the 8-oxoguanine glycosylase 1 (OGG1) may counteract oxidative stress and resulting DNA damage associated with lifestyle-related exposures. We examined whether the polymorphisms GPX1 Pro(198)Leu and OGG1 Ser(326)Cys or low erythrocyte GPX enzyme activity in pre-diagnostic blood samples are associated with colorectal cancer risk, and assessed possible interactions between the polymorphisms or enzyme activity and various lifestyle factors in relation to colorectal cancer risk. Additionally, we studied whether the GPX1 Pro(198)Leu polymorphism and several lifestyle factors predict GPX activity in erythrocytes. The present study was nested within the prospective "Diet, Cancer and Health" study of 57,053 Danes including 375 colorectal cancer cases and a comparison group of 779 individuals matched on gender. Biomaterial was sampled and information on lifestyle factors was obtained from questionnaires filled in at enrolment in 1993-1997. GPX1 Pro(198)Leu, hOGG1 Ser(326)Cys and erythrocyte GPX enzyme activity were not associated with risk of colorectal cancer. We observed a higher risk associated with alcohol consumption and smoking among homozygous GPX1(198)Leu carriers, with incidence rate ratios for colorectal cancer of 1.45 (95% CI: 1.17-1.81, P=0.02) per 10g alcohol intake per day and 2.56 (95% CI: 0.99-6.61, P=0.02) among ever smokers compared with never smokers at enrolment. Erythrocyte GPX activity was influenced by the GPX1 Pro(198)Leu genotype, gender, smoking intensity, and intake of fruits and vegetables. Our results indicate that lifestyle-related oxidative stress may be a risk factor for colorectal cancer among subjects with a lowered defence.


High-intensity preoperative training improves physical and functional recovery in the early post-operative periods after total knee arthroplasty: a randomized controlled trial.
The benefits of preoperative training programmes compared with alternative treatment are unclear. The purpose of this study was to evaluate the effectiveness of a high-intensity preoperative resistance training programme in patients waiting for total knee arthroplasty (TKA). Forty-four subjects (7 men, 37 women) scheduled for unilateral TKA for osteoarthritis (OA) during 2014 participated in this randomized controlled trial. Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), the Physical Functioning Scale of the Short Form-36 questionnaire (SF-36), a 10-cm visual analogue scale (VAS), isometric knee flexion, isometric knee extension, isometric hip abduction, active knee range of motion and functional tasks (Timed Up and Go test and Stair ascent-descent test) were assessed at 8 weeks before surgery (T1), after 8 weeks of training (T2), 1 month after TKA (T3) and finally 3 months after TKA (T4). The intervention group completed an 8-week training programme 3 days per week prior to surgery. Isometric knee flexion, isometric hip abduction, VAS, WOMAC, ROM extension and flexion and all the functional assessments were greater for the intervention group at T2, T3 and T4, whereas isometric knee extension was greater for this group at T2 and T4 compared with control. The present study supports the use of preoperative training in end-stage OA patients to improve early postoperative outcomes. High-intensity strength training during the preoperative period reduces pain and improves lower limb muscle strength, ROM and functional task performance before surgery, resulting in a reduced length of stay at the hospital and a faster physical and functional recovery after TKA. The present training programme can be used by specialists to speed up recovery after TKA. I.


Impact of executive dysfunction on verbal memory performance in patients with Alzheimer's disease.
It is currently accepted that there is a subset of patients diagnosed with Alzheimer's disease (AD) who show executive functioning (EF) impairments even in the earlier stages. These patients have been shown to present distinct psychiatric, behavioral, occupational, and even histopathological profiles. We assessed thirty patients with AD on two tasks of verbal memory (Logical Memory - LM, and the Rey Auditory-Verbal Learning Task - RAVLT), as well as classical tests of EF. AD patients were classified into either a spared EF (SEF) group if they showed impaired performance (z < -1.5 SD) in none or only one of the executive tests, or into an impaired EF (IEF) group if they showed impaired performance on two or more tasks of EF. Their performance was compared with fourteen healthy controls. SEF showed significantly more years of education than IEF, but the groups did not differ significantly on age, gender, mood symptoms, or performance on general screening tests or attentional tasks. With education as a covariate, both AD groups differed from controls on all measures of memory, but a significant difference was found between SEF and IEF patients only on the recognition phases of both logical memory (p < 0.01) and RAVLT (p = 0.02). Recognition scores significantly correlated with performance on executive tasks. Early AD patients who preserve their EF seem to have an advantage in their ability to recognize information that has been previously presented over patients with impaired EF. Such advantage seems to be strongly associated with executive performance.


Inhibition of microsomal lipid peroxidation by glutathione and glutathione transferases B and AA. Role of endogenous phospholipase A2.
Lipid peroxidation in vitro in rat liver microsomes (microsomal fractions) initiated by ADP-Fe3+ and NADPH was inhibited by the rat liver soluble supernatant fraction. When this fraction was subjected to frontal-elution chromatography, most, if not all, of its inhibitory activity could be accounted for by the combined effects of two fractions, one containing Se-dependent glutathione (GSH) peroxidase activity and the other the GSH transferases. In the latter fraction, GSH transferases B and AA, but not GSH transferases A and C, possessed inhibitory activity. GSH transferase B replaced the soluble supernatant fraction as an effective inhibitor of lipid peroxidation in vitro. If the microsomes were pretreated with the phospholipase A2 inhibitor p-bromophenacyl bromide, neither the soluble supernatant fraction nor GSH transferase B inhibited lipid peroxidation in vitro. Similarly, if all microsomal enzymes were heat-inactivated and lipid peroxidation was initiated with FeCl3/sodium ascorbate neither the soluble supernatant fraction nor GSH transferase B caused inhibition, but in both cases inhibition could be restored by the addition of porcine pancreatic phospholipase A2 to the incubation. It is concluded that the inhibition of microsomal lipid peroxidation in vitro requires the consecutive action of phospholipase A2, which releases fatty acyl hydroperoxides from peroxidized phospholipids, and GSH peroxidases, which reduce them. The GSH peroxidases involved are the Se-dependent GSH peroxidase and the Se-independent GSH peroxidases GSH transferases B and AA.


Gain switch laser based on micro-structured Yb-doped active fiber.
In this paper a near infrared gain-switched fiber laser based on oscillator stage only design with high peak power is presented. Output pulses reached 2.3 kW of peak power and duration of less than 60 ns. The dependence of the laser pulse duration on operation parameters was measured and theoretically explained. As the setup is based on flexible micro structured single polarization fiber, the laser output exhibits high polarization extinction ratio. Due to the narrow output spectrum the setup is suitable for second harmonic generation.


Structure and multinuclear solid-state NMR of a highly birefringent lead-gold cyanide coordination polymer.
The coordination polymer Pb(H2O)[Au(CN)2]2 (1) was synthesized by the reaction of KAu(CN)2 and Pb(NO3)2. The structure contains 1-D chains of lead(II)-OH2 linked via Au(CN)2(-) moieties, generating a 2-D slab; weak aurophilic interactions of 3.506(2) and 3.4885(5) A occur within and between slabs. The geometry about each lead(II) is bicapped trigonal prismatic, having six N-bound cyanides at the prism vertices and waters at two of the faces. Dehydration at 175 degrees C yields microcrystalline Pb[Au(CN)2]2 (2), which, along with 1, was examined by 13C, 15N, 1H, and 207Pb solid-state NMR methods. Two 15N resonances are assigned to the mu2-bridging and hydrogen-bonding cyanides in 1. Upon dehydration, the 207Pb NMR spectrum becomes axially symmetric and yields a reduced shielding span, indicating higher site symmetry, while the 13C and 15N spectra reveal a single cyanide. Although no single-crystal X-ray structure of 2 could be obtained, a structure is proposed on the basis of the NMR and X-ray powder data, consisting of a lead(II) center in a distorted square-prismatic environment, with cyanides present at each corner. The birefringence of single crystals of 1 is found to be 7.0 x 10(-2) at room temperature. This value is large compared to that of most optical materials and can be attributed to the anisotropy of the 2-D slabs of 1, with all CN bonds aligned in the same direction by the polarizable lead(II) center.


HLA class II polymorphisms and susceptibility to recurrent respiratory papillomatosis.
Recurrent respiratory papillomatosis (RRP) is characterised by multiple laryngeal papillomas. Left untreated, the lesions enlarge, spread, and endanger the airway. Medical treatments are unsatisfactory, and repeated surgery remains the mainstay of therapy. RRP is caused by human papillomavirus (HPV) infection. However, since oral HPV infection is common and RRP is rare, other host and/or viral factors may contribute to pathogenesis. In an attempt to identify such factors, we have investigated 60 patients. The patients were HLA class I, II, and tumor necrosis factor TNF typed by sequence-specific primer PCR, and the results compared to those for 554 healthy controls by using Fisher's exact test. Peripheral blood mononuclear cell proliferative responses of 25 controls and 10 patients to HPV-11 L1 virus-like particles (VLP) were compared. Short-term VLP-specific T-cell lines were established, and recognition of L1 was analyzed. Finally, the L1 open reading frames of HPV isolates from four patients were sequenced. Susceptibility to RRP was associated with HLA DRB1*0301 (33 of 60 patients versus 136 of 554 controls, P < 0.0001). The three most severely affected patients were homozygous for this allele. A range of T-cell proliferative responses to HPV-11 VLP were observed in DRB1*0301-positive healthy donors which were comparable to those in DRB1*0301-negative controls. Individuals with juvenile-onset RRP also mounted a range of VLP responses, and their magnitude was negatively correlated with the clinical staging score (P = 0.012 by the Spearman rank correlation). DRB1*0301-positive patients who responded to L1 recognized the same epitope as did matched controls and produced similar cytokines. Sequencing of clinical isolates excluded the possibility that nonresponsiveness was the result of mutation(s) in L1.


OptiBIRTH: a cluster randomised trial of a complex intervention to increase vaginal birth after caesarean section.
Despite evidence supporting the safety of vaginal birth after caesarean section (VBAC), rates are low in many countries. OptiBIRTH investigated the effects of a woman-centred intervention designed to increase VBAC rates through an unblinded cluster randomised trial in 15 maternity units with VBAC rates < 35% in Germany, Ireland and Italy. Sites were matched in pairs or triplets based on annual birth numbers and VBAC rate, and randomised, 1:1 or 2:1, intervention versus control, following trial registration. The intervention involved evidence-based education of clinicians and women with one previous caesarean section (CS), appointment of opinion leaders, audit/peer review, and joint discussions by women and clinicians. Control sites provided usual care. Primary outcome was annual hospital-level VBAC rates before the trial (2012) versus final year of the trial (2016). Between April 2014 and October 2015, 2002 women were recruited (intervention 1195, control 807), with mode-of-birth data available for 1940 women. The OptiBIRTH intervention was feasible and safe across hospital settings in three countries. There was no statistically significant difference in the change in the proportion of women having a VBAC between intervention sites (25.6% in 2012 to 25.1% in 2016) and control sites (18.3 to 22.3%) (odds ratio adjusted for differences between intervention and control groups (2012) and for homogeneity in VBAC rates at sites in the countries: 0.87, 95% CI: 0.67, 1.14, p = 0.32 based on 5674 women (2012) and 5284 (2016) with outcome data. Among recruited women with birth data, 4/1147 perinatal deaths > 24 weeks gestation occurred in the intervention group (0.34%) and 4/782 in the control group (0.51%), and two uterine ruptures (one per group), a rate of 1:1000. Changing clinical practice takes time. As elective repeat CS is the most common reason for CS in multiparous women, interventions that are feasible and safe and that have been shown to lead to decreasing repeat CS, should be promoted. Continued research to refine the best way of promoting VBAC is essential. This may best be done using an implementation science approach that can modify evidence-based interventions in response to changing clinical circumstances. The OptiBIRTH trial was registered on 3/4/2013. Trial registration number ISRCTN10612254.


Ectopic thyroid carcinoma causing right ventricular outflow tract obstruction in a dog.
A 9-year-old Bouvier des Flandres was presented with coughing, lethargy, chylous pleural effusion, and a heart murmur. An echocardiogram revealed the presence of an intracardiac mass causing right ventricular outflow tract obstruction. The mass was successfully removed surgically, using total inflow occlusion. Histopathology and immunohistochemistry identified the tumor as an ectopic thyroid carcinoma. The dog was euthanized 11 months after diagnosis at the request of the owner because of nonresolving chylothorax.


Solid-state YVO4/Nd:YVO4/KTP green laser system for the generation of subnanosecond pulses with adjustable kilohertz repetition rate.
A solid-state green laser generating subnanosecond pulses with adjustable kilohertz repetition rate is presented. This pulse laser system is composed of a Q-switched and mode-locked YVO(4)/Nd:YVO(4)/KTP laser simultaneously modulated by an electro-optic (EO) modulator and a central semiconductor saturable absorption mirror. Because the repetition rate of the Q-switched envelope in this laser depends on the modulation frequency of the EO modulator, so long as the pulsewidth of the Q-switched envelope is shorter than the cavity roundtrip transmit time, i.e., the time interval of two neighboring mode-locking pulses, only one mode-locking pulse exists underneath a Q-switched envelope, resulting in the generation of subnanosecond pulses with kilohertz repetition rate. The experimental results show that the pulsewidth of subnanosecond pulses decreases with increasing pump power and the shortest pulse generated at 1 kHz was 450 ps with pulse energy as high as 252 μJ, corresponding to a peak power of 560 kW. In addition, this laser was confirmed to have high stability, and the pulse repetition rate could be freely adjusted from 1 to 4 kHz.


Potential virulence factors of Proteus bacilli.
The object of this review is the genus Proteus, which contains bacteria considered now to belong to the opportunistic pathogens. Widely distributed in nature (in soil, water, and sewage), Proteus species play a significant ecological role. When present in the niches of higher macroorganisms, these species are able to evoke pathological events in different regions of the human body. The invaders (Proteus mirabilis, P. vulgaris, and P. penneri) have numerous factors including fimbriae, flagella, outer membrane proteins, lipopolysaccharide, capsule antigen, urease, immunoglobulin A proteases, hemolysins, amino acid deaminases, and, finally, the most characteristic attribute of Proteus, swarming growth, enabling them to colonize and survive in higher organisms. All these features and factors are described and commented on in detail. The questions important for future investigation of these facultatively pathogenic microorganisms are also discussed.


Mutations in the hepatocyte nuclear factor-1alpha gene in maturity-onset diabetes of the young (MODY3)
The disease non-insulin-dependent (type 2) diabetes mellitus (NIDDM) is characterized by abnormally high blood glucose resulting from a relative deficiency of insulin. It affects about 2% of the world's population and treatment of diabetes and its complications are an increasing health-care burden. Genetic factors are important in the aetiology of NIDDM, and linkage studies are starting to localize some of the genes that influence the development of this disorder. Maturity-onset diabetes of the young (MODY), a single-gene disorder responsible for 2-5% of NIDDM, is characterized by autosomal dominant inheritance and an age of onset of 25 years or younger. MODY genes have been localized to chromosomes 7, 12 and 20 (refs 5, 7, 8) and clinical studies indicate that mutations in these genes are associated with abnormal patterns of glucose-stimulated insulin secretion. The gene on chromosome 7 (MODY2) encodes the glycolytic enzyme glucokinases which plays a key role in generating the metabolic signal for insulin secretion and in integrating hepatic glucose uptake. Here we show that subjects with the MODY3-form of NIDDM have mutations in the gene encoding hepatocyte nuclear factor-1alpha (HNF-1alpha, which is encoded by the gene TCF1). HNF-1alpha is a transcription factor that helps in the tissue-specific regulation of the expression of several liver genes and also functions as a weak transactivator of the rat insulin-I gene.


Assessing capability of semiconductors to split water using ionization potentials and electron affinities only.
We show in this article that the position of semiconductor band edges relative to the water reduction and oxidation levels can be reliably predicted from the ionization potentials (IP) and electron affinities (AE) only. Using a set of 17 materials, including transition metal compounds, we show that accurate surface dependent IPs and EAs of semiconductors can be computed by combining density functional theory and many-body GW calculations. From the extensive comparison of calculated IPs and EAs with available experimental data, both from photoemission and electrochemical measurements, we show that it is possible to sort candidate materials solely from IPs and EAs thereby eliminating explicit treatment of semiconductor/water interfaces. We find that at pH values corresponding to the point of zero charge there is on average a 0.5 eV shift of IPs and EAs closer to the vacuum due to the dipoles formed at material/water interfaces.


Sampling locality is more detectable than taxonomy or ecology in the gut microbiota of the brood-parasitic Brown-headed Cowbird (Molothrus ater).
Brown-headed Cowbirds (Molothrus ater) are the most widespread avian brood parasite in North America, laying their eggs in the nests of approximately 250 host species that raise the cowbird nestlings as their own. It is currently unknown how these heterospecific hosts influence the cowbird gut microbiota relative to other factors, such as the local environment and genetics. We test a Nature Hypothesis (positing the importance of cowbird genetics) and a Nurture Hypothesis (where the host parents are most influential to cowbird gut microbiota) using the V6 region of 16S rRNA as a microbial fingerprint of the gut from 32 cowbird samples and 16 potential hosts from nine species. We test additional hypotheses regarding the influence of the local environment and age of the birds. We found no evidence for the Nature Hypothesis and little support for the Nurture Hypothesis. Cowbird gut microbiota did not form a clade, but neither did members of the host species. Rather, the physical location, diet and age of the bird, whether cowbird or host, were the most significant categorical variables. Thus, passerine gut microbiota may be most strongly influenced by environmental factors. To put this variation in a broader context, we compared the bird data to a fecal microbiota dataset of 38 mammal species and 22 insect species. Insects were always the most variable; on some axes, we found more variation within cowbirds than across all mammals. Taken together, passerine gut microbiota may be more variable and environmentally determined than other taxonomic groups examined to date.


Desensitization and trafficking of μ-opioid receptors in locus ceruleus neurons: modulation by kinases.
The phosphorylation of μ-opioid receptors (MOPRs) by G protein-coupled receptor kinases (GRKs), followed by arrestin binding, is thought to be a key pathway leading to desensitization and internalization. The present study used the combination of intracellular and whole-cell recordings from rats and mice, as well as live cell imaging of Flag-tagged MOPRs from mouse locus ceruleus neurons, to examine the role of protein kinases in acute desensitization and receptor trafficking. Inhibition of GRKs by using heparin or GRK2-mutant mice did not block desensitization or alter the rate of recovery from desensitization. The nonselective kinase inhibitor staurosporine did not reduce the extent of [Met(5)]enkephalin (ME)-induced desensitization but increased the rate of recovery from desensitization. In the presence of staurosporine, ME-activated FlagMOPRs were internalized but did not traffic away from the plasma membrane. The increased rate of recovery from desensitization correlated with the enhancement in the recycling of receptors to the plasma membrane. ME-induced MOPR desensitization persisted and the trafficking of receptors was modified after inhibition of protein kinases. The results suggest that desensitization of MOPRs may be an early step after agonist binding that is modulated by but is not dependent on kinase activity.


Assisted reproductive technology surveillance--United States, 2006.
Assisted Reproductive Technology (ART) includes fertility treatments in which both eggs and sperm are handled in the laboratory (i.e., in vitro fertilization and related procedures).Patients who undergo ART procedures are more likely to deliver multiple-birth infants than women who conceive naturally. Multiple births are associated with increased risk for mothers and infants (e.g., pregnancy complications, premature delivery, low-birthweight infants, and long-term disability among infants). This report presents the most recent national data and state-specific results. 2006. In 1996, CDC initiated data collection regarding ART procedures performed in the United States, as mandated by the Fertility Clinic Success Rate and Certification Act of 1992 (FCSRCA) (Public Law 102-493 [October 24, 1992]). Beginning with 2004, CDC has contracted with Westat, Inc., a statistical survey research organization, to obtain data from ART medical centers in the United States. Westat, Inc., maintains the National ART Surveillance System (NASS), CDC's web-based data collection system. In 2006, a total of 138,198 ART procedures were reported to CDC. These procedures resulted in 41,343 live-birth deliveries, and 54,656 infants. ART procedures are categorized into four major procedure types: procedures that used embryos from freshly fertilized eggs (fresh embryos) from the patient's eggs (72%); procedures that used thawed embryos from the patient's eggs (16%); procedures that used fresh embryos from donor eggs (8%); and procedures that used thawed embryos from donor eggs (4%). Overall, 44% of ART transfer procedures resulted in a pregnancy, and 36% in a live-birth delivery (delivery of one or more live-born infants). Live-birth rates were generally higher among ART procedures that used fresh embryos from donor eggs (54%) than among other types. The highest numbers of ART procedures were performed among residents of California (18,886); New York (13,259); Illinois (9,594); New Jersey (9,237); and Massachusetts (8,305). All five states reported the highest number of live-birth deliveries as a result of ART. Of 54,656 infants born through ART, 48% were multiple-birth deliveries. The risk for a multiple-birth delivery was highest for women who underwent ART transfer procedures that used fresh embryos from either donor eggs (39%) or their own eggs (31%). Approximately 1% of U.S. infants born in 2006 were conceived through ART. Those infants accounted for 18% of multiple births nationwide. Approximately 9% of ART singletons, 57% of ART twins, and 96% of ART triplets or higher-order multiples were low birthweight. Similarly, 14% of ART singletons, 65% of ART twins, and 97% of ART triplets or higher-order multiples were born preterm. Whether an ART procedure resulted in a pregnancy and live-birth delivery varied, according to different patient and treatment factors. ART poses a major risk for multiple births associated with adverse maternal and infant outcomes (e.g., placenta previa, preterm delivery, cesarean delivery, low birthweight, and infant mortality). This risk varied according to the patient's age, the type of ART procedure performed, the number of embryos available for transfer to the uterus, the number transferred, and the day of transfer (day 3 or 5). ART-related multiple births represent a sizable proportion of all multiple births nationwide and in selected states. To minimize the adverse maternal and child health effects associated with multiple pregnancies, ongoing efforts to limit the number of embryos transferred in each ART procedure should be continued and strengthened. Adverse maternal and infant outcomes (e.g., low birthweight and preterm delivery) associated with ART treatment choices should be explained thoroughly when counseling patients considering ART.


Pathology of callosal damage in ALS: An ex-vivo, 7 T diffusion tensor MRI study.
The goal of this study was to better understand the changes in tissue microstructure that underlie white matter diffusion changes in ALS patients. Diffusion tensor imaging was carried out in postmortem brains of 4 ALS patients and two subjects without neurological disease on a 7 T MRI scanner using steady-state free precession sequences. Fractional anisotropy (FA) was measured in the genu, body, and splenium of the corpus callosum in formalin-fixed hemispheres. FA of the body and genu was expressed as ratio to FA of the splenium, a region unaffected in ALS. After imaging, tissue sections of the same segments of the callosum were stained for markers of different tissue components. Coded image fields were rated for pathological changes by blinded raters. The FA body/FA splenium ratio was reduced in ALS patients compared to controls. Patchy areas of myelin pallor and cells immunostained for CD68, a microglial-macrophage marker, were only observed in the body of the callosum of ALS patients. Blinded ratings showed increased CD68 + microglial cells in the body of the corpus callosum in ALS patients, especially those with C9orf72 mutations, and increased reactive astrocytes throughout the callosum. Reduced FA of the corpus callosum in ALS results from complex changes in tissue microstructure. Callosal segments with reduced FA had large numbers of microglia-macrophages in addition to loss of myelinated axons and astrogliosis. Microglial inflammation contributed to reduced FA in ALS, and may contribute to a pro-inflammatory state, but further work is needed to determine their role.


Inflammation Caused by Praziquantel Treatment Depends on the Location of the Taenia solium Cysticercus in Porcine Neurocysticercosis.
Neurocysticercosis (NCC), infection of the central nervous system by Taenia solium cysticerci, is a pleomorphic disease. Inflammation around cysticerci is the major cause of disease but is variably present. One factor modulating the inflammatory responses may be the location and characteristics of the brain tissue adjacent to cysticerci. We analyzed and compared the inflammatory responses to cysticerci located in the parenchyma to those in the meninges or cysticerci partially in contact with both the parenchyma and the meninges (corticomeningeal). Histological specimens of brain cysticerci (n = 196) from 11 pigs naturally infected with Taenia solium cysticerci were used. Four pigs were sacrificed after 2 days and four after 5 days of a single dose of praziquantel; 3 pigs did not receive treatment. All pigs were intravenously injected with Evans Blue to assess disruption of the blood-brain barrier. The degree of inflammation was estimated by use of a histological score (ISC) based on the extent of the inflammation in the pericystic areas as assessed in an image composed of several photomicrographs taken at 40X amplification. Parenchymal cysticerci provoked a significantly greater level of pericystic inflammation (higher ISC) after antiparasitic treatment compared to meningeal and corticomeningeal cysticerci. ISC of meningeal cysticerci was not significantly affected by treatment. In corticomeningeal cysticerci, the increase in ISC score was correlated to the extent of the cysticercus adjacent to the brain parenchyma. Disruption of the blood-brain barrier was associated with treatment only in parenchymal tissue. Inflammatory response to cysticerci located in the meninges was significantly decreased compared to parenchymal cysticerci. The suboptimal inflammatory response to cysticidal drugs may be the reason subarachnoid NCC is generally refractory to treatment compared to parenchymal NCC.