Ancient hybridization and mitochondrial capture between two species of chipmunks.
Models that posit speciation in the face of gene flow are replacing classical views that hybridization is rare between animal species. We use a multilocus approach to examine the history of hybridization and gene flow between two species of chipmunks (Tamias ruficaudus and T. amoenus). Previous studies have shown that these species occupy different ecological niches and have distinct genital bone morphologies, yet appear to be incompletely isolated reproductively in multiple areas of sympatry. We compared data from four sequenced nuclear loci and from seven microsatellite loci to published cytochrome b sequences. Interspecific gene flow was primarily restricted to introgression of the T. ruficaudus mitochondrial genome into a sympatric subspecies of T. amoenus, T. a. canicaudus, with the four sequenced nuclear loci showing little to no interspecific allele sharing. Microsatellite data were consistent with high levels of differentiation between the species and also showed no current gene flow between broadly sympatric populations of T. a. canicaudus and T. ruficaudus. Coalescent analyses date the mtDNA introgression event from the mid-Pleistocene to late Pliocene. Overall, these data indicate that introgression has had a minimal impact on the nuclear genomes of T. amoenus and T. ruficaudus despite multiple independent hybridization events. Our findings challenge long-standing assumptions on patterns of reproductive isolation in chipmunks and suggest that there may be other examples of hybridization among the 23 species of Tamias that occur in western North America.


Oxytocin and Pain Perception: From Animal Models to Human Research.
An accumulating body of evidence suggests that the hypothalamic neuropeptide oxytocin (OT) has a modulatory effect on pain processing. Particularly strong evidence comes from animal models. Here, we review recent advances in animal research on the analgesic effects of OT and discuss possible target sites of OT within descending and ascending pain pathways in the brain. In addition to the spinal cord being a direct target of the neuropeptide, OT has also been shown to modulate the neuronal activity of limbic and cortical brain regions, which play a major role in the cognitive and emotional processing of pain. Human studies investigating the influence of OT on pain perception are less numerous and have revealed less consistent results. The human literature is therefore scanned thoroughly and different approaches to study the effects of OT on pain perception in humans are discussed. Moreover, we also address how OT might alleviate pain by influencing socio-emotional components in humans. We conclude that further investigating specific OT and OT-sensitive circuits, which modulate pain processing especially in primates, will improve our understanding of OT-analgesic effects. In human research, the increased use of neuroimaging and autonomic measures might help to bridge the gap to animal studies.


Functional and structural evaluation of the meibomian gland using a LipiView interferometer in thyroid eye disease.
The purpose of this study was to evaluate the structure and function of the meibomian gland and the incomplete blinking rate to understand the pathophysiology of dry eye in thyroid eye disease (TED) patients. Patients who were diagnosed with TED were enrolled between October 2015 and February 2016. Clinical measurements were performed in the following order: (i) external examination (Hertel exophthalmometer and palpebral fissure height), (ii) LipiView interferometer (lipid layer thickness [LLT], incomplete blinking rate, and meibography), (iii) slit-lamp biomicroscopy (corneal surface staining, tear breakup time, meibum expression, Marx line). Thirty eyes of 30 TED patients (male = 8; female = 22) were included in this study. The TED patient population had a mean age of 42.9 ± 11.8 years and a mean clinical activity score (CAS) of 2.33 ± 1.60. The meiboscore was 1.17 ± 0.90 in the upper eyelid and 0.70 ± 0.65 in the lower eyelid; scores were significantly higher in the upper eyelid (p < 0.001). The mean LLT was 82.43 ± 24.52 nm, and the mean incomplete blinking rate was 51.04 ± 33.62% (0-100%). CAS was the only variable that correlated with the meiboscore. There was no significant correlation between incomplete blinking and a degree of palpebral fissure height or proptosis. Tear film instability due to increased incomplete blinking can cause dry eye in TED. In addition to the increase in CAS, meibomian gland dysfunction may also be a cause of dry eye in TED. However, further comparative studies are needed to confirm these results.


Comparison of conventional, model-based quantitative planar, and quantitative SPECT image processing methods for organ activity estimation using In-111 agents.
Accurate quantification of organ radionuclide uptake is important for patient-specific dosimetry. The quantitative accuracy from conventional conjugate view methods is limited by overlap of projections from different organs and background activity, and attenuation and scatter. In this work, we propose and validate a quantitative planar (QPlanar) processing method based on maximum likelihood (ML) estimation of organ activities using 3D organ VOIs and a projector that models the image degrading effects. Both a physical phantom experiment and Monte Carlo simulation (MCS) studies were used to evaluate the new method. In these studies, the accuracies and precisions of organ activity estimates for the QPlanar method were compared with those from conventional planar (CPlanar) processing methods with various corrections for scatter, attenuation and organ overlap, and a quantitative SPECT (QSPECT) processing method. Experimental planar and SPECT projections and registered CT data from an RSD Torso phantom were obtained using a GE Millenium VH/Hawkeye system. The MCS data were obtained from the 3D NCAT phantom with organ activity distributions that modelled the uptake of (111)In ibritumomab tiuxetan. The simulations were performed using parameters appropriate for the same system used in the RSD torso phantom experiment. The organ activity estimates obtained from the CPlanar, QPlanar and QSPECT methods from both experiments were compared. From the results of the MCS experiment, even with ideal organ overlap correction and background subtraction, CPlanar methods provided limited quantitative accuracy. The QPlanar method with accurate modelling of the physical factors increased the quantitative accuracy at the cost of requiring estimates of the organ VOIs in 3D. The accuracy of QPlanar approached that of QSPECT, but required much less acquisition and computation time. Similar results were obtained from the physical phantom experiment. We conclude that the QPlanar method, based on 3D organ VOIs and accurate models of the projection process, provided a substantial increase in accuracy of organ activity estimates from planar images compared to CPlanar processing and had accuracy approaching that of QSPECT.


Performance of isolated rat hearts perfused with helium- or nitrogen-containing, gas-saturated solutions.
Effects of helium on the isolated perfused rat heart were studied employing the Langendorff technique. The perfusate consisted of Krebs-Henseleit solution saturated with one of three gas mixtures: 1) 95% O2-5% CO2, 2) 50% O2-45% He-5% CO2, and 3) 50% o2-45% N2-5% CO2. Contractile indices measured revealed the performance of hearts with the helium mixture to be equivalent to those perfused with the 95% O2-5% CO2 mixture. Those perfused with the nitrogen gas mixture exhibited contractile activity lower than that in the other two groups. It was concluded that helium exerts a direct effect on the coronary vasculature of the isolated rat heart by reducing its resistance to flow. A greater oxygen delivery to hearts perfused with the He-saturated solution compared to the N2-perfused hearts may account for the difference in performance.


Optical buffering technique under a space radiation environment.
We propose an optical buffering technique for a programmable gate array under a space radiation environment. The technique allows rapid recovery of a programmable device that is damaged by high-energy charged particles. The technique uses invalid configuration data, including error bits that have already been damaged by the particles, while the configuration data are transferred using wireless communications and are retained on electronically erasable, programmable read-only, or static random-access-memory devices.


Acute pancreatitis and hyperamylasaemia in pancreatic carcinoma.
To determine the incidence of acute pancreatitis in pancreatic carcinoma, a retrospective study was performed in 174 patients. Acute pancreatitis was found in 24 (13.8%), and hyperamylasaemia, but no clinical manifestation of acute pancreatitis, was found in 17 (9.8%) of these patients. The incidence of acute pancreatitis was higher when the papilla of Vater or the head of the pancreas was involved. Clinically, pancreatitis was slight to moderate, and the underlying disease was soon diagnosed with invasive procedures. Pancreatic carcinoma should be considered an etiological factor in patients with relapsing and/or unexplained pancreatitis of long duration.


Conditionally immortalized human proximal tubular epithelial cells isolated from the urine of a healthy subject express functional calcium-sensing receptor.
The calcium-sensing receptor (CaSR) is a G protein-coupled receptor, which plays an essential role in regulating Ca(2+) homeostasis. Here we show that conditionally immortalized proximal tubular epithelial cell line (ciPTEC) obtained by immortalizing and subcloning cells exfoliated in the urine of a healthy subject expresses functional endogenous CaSR. Immunolocalization studies of polarized ciPTEC revealed the apical localization of the receptor. By Western blotting of ciPTEC lysates, both monomeric and dimeric forms of CaSR at 130 and ∼250 kDa, respectively, were detected. Functional studies indicated that both external calcium and the positive CaSR allosteric modulator, NPS-R568, induced a significant increase in cytosolic calcium, proving a high sensitivity of the endogenous receptor to its agonists. Calcium depletion from the endoplasmic reticulum using cyclopiazonic acid abolished the increase in cytosolic calcium elicited by NPS-R568, confirming calcium exit from intracellular stores. Activation of CaSR by NPS-R significantly reduced the increase in cAMP elicited by forskolin (FK), a direct activator of adenylate cyclase, further confirming the functional expression of the receptor in this cell line. CaSR expressed in ciPTEC was found to interact with Gq as a downstream effector, which in turn can cause release of calcium from intracellular stores via phospholipase C activation. We conclude that human proximal tubular ciPTEC express functional CaSR and respond to its activation with a release of calcium from intracellular stores. These cell lines represent a valuable tool for research into the disorder associated with gain or loss of function of the CaSR by producing cell lines from patients.


Spectral dynamics of incoherent waves with a noninstantaneous nonlinear response.
We study the influence of a constant background noise on the dynamics of spectral incoherent solitons, which are incoherent structures sustained by a noninstantaneous (Raman-like) nonlinearity. As the level of the noise background increases, the incoherent wave enters a novel nonlinear regime characterized by oscillatory dynamics of the incoherent spectrum, which develop within a spectral cone during the propagation. In contrast to the conventional Raman-like spectral red shift, such incoherent spectral dynamics can be characterized by a significant spectral blue shift. On the basis of the kinetic wave theory, we derive explicit analytical expressions of these incoherent oscillatory spectral dynamics.


Influence of urban form on landscape pattern and connectivity in metropolitan regions: a comparative case study of Phoenix, AZ, USA, and Izmir, Turkey.
Although ecological connectivity conservation in urban areas has recently been recognized as an important issue, less is known about its relationship to urban form and landscape pattern. This study investigates how urban morphology influences regional ecosystem pattern and landscape connectivity. Two metropolitan landscapes, Phoenix, AZ, USA, and Izmir, Turkey, were compared, both of which are fast-growing regions in their national context. A wide range of variables were considered for identifying natural and urban properties. The natural characteristics include typology of urban ecosystems, urban to natural cover ratio, dominant habitat type, urban biodiversity, landscape context, and connectivity conservation efforts. Urban parameters examine urban form, urban extent, urban cover proportion, growth rate, populations, urban gradient, major drivers of urbanization, urban density, and mode/approach of urban development. Twelve landscape metrics were measured and compared across the natural patches. Results show that there is little difference in landscape connectivity in the rural zones of Phoenix and Izmir, although Phoenix has slightly higher connectivity values. The connectivity variance in urbanized areas, however, is significantly dependent on the region. For example, Phoenix urban zones have substantially lower connectivity than either urban or suburban zones in Izmir. Findings demonstrate that small and compact urban settlements with more dense populations are more likely to conserve landscape connectivity compared to multiple-concentric but amalgamated urban form spreading all over the landscape (aka urban sprawl).


Screening, prevention and treatment of viral hepatitis B reactivation in patients with haematological malignancies.
The prevalence of hepatitis B virus (HBV) infection in patients with haematological malignancies is increased compared with the general population worldwide. HBV reactivation is common following chemotherapy and is associated with a high mortality despite prompt anti-viral treatment. HBV reactivation may necessitate interruption of chemotherapy with adverse prognostic consequences for the haematological disease. Chemotherapy-induced immune suppression may lead to increased HBV replication. Immune reconstitution within the weeks and months following recovery from chemotherapy may be associated with a flare of hepatitis B manifested by hepatocellular injury. Risk factors associated with HBV reactivation include detectable hepatitis B surface antigen (HBsAg), HBV DNA, Hepatitis B e (HBeAg) antigen, antibodies to hepatitis B core antigen (anti-HBc), treatment with corticosteroids, young age and male gender. Lamivudine is effective during HBV reactivation due to immune suppression. Clinical trials have demonstrated that pre-emptive antiviral treatment with lamivudine is superior to deferred treatment. Current recommendations emphasise screening for HBV infection in all haematology patients, particularly prior to chemotherapy. Patients who are HBsAg positive or HBV DNA positive should receive pre-emptive treatment with lamivudine before chemotherapy. The duration of lamivudine treatment may be prolonged commensurate with the degree of immunosuppression. HBV naïve patients should be immunised against hepatitis B, as should haematopoietic stem cell donors. In summary, overt and occult HBV pose a serious, but preventable, threat. Pre-treatment screening of patients at risk should be practiced diligently by all clinicians that treat patients with malignancies.


A case of giant malignant phaeochromocytoma.
Malignant phaeochromocytoma is defined as the presence of tumour deposits at sites that are normally devoid of chromaffin cells. We report on a 63-year-old man who had a giant malignant phaeochromocytoma of the right adrenal gland that encased the inferior vena cava. The urinary excretion rates of catecholamines and their metabolites were normal, except for normetanephrine, which was excreted at a higher rate than normal. The tumour was surgically unresectable by laparotomy. Postoperatively, the patient was given a 4-month trial of subcutaneous octreotide and intravenous meta-iodobenzylguanidine I 131. Occult lung secondary tumours were first detected by meta-iodobenzylguanidine scintigraphy after 2 years, and the patient died of bone and lung metastases 1 year later. Because phaeochromocytoma is rare, local experience in managing this disease is limited. This report alerts physicians of the methods of diagnosing and managing surgically unresectable malignant phaeochromocytoma.


Fast high-precision distance metrology using a pair of modulator-generated dual-color frequency combs.
We demonstrate fast high-precision non-contact distance measurements to technical surfaces using a pair of dual-color electro-optic frequency combs for synthetic-wavelength interferometry (SWI). The dual-color combs are generated from continuous-wave (cw) lasers at 1300 nm and 1550 nm, which are jointly fed to a pair of high-speed dual-drive Mach-Zehnder modulators. The dual-color approach is used for continuous and dead-zone-free compensation of temperature-induced fiber drift. We achieve standard deviations below 2 µm at an acquisition time of 9.1 µs for measurements through 7 m of single-mode fiber. Despite the technical simplicity of our scheme, our concept can well compete with other comb-based distance metrology approaches, and it can maintain its accuracy even under industrial operating conditions. The viability of the concept is demonstrated by attaching the fiber-coupled sensor head to an industrial coordinate measuring machine for acquisition of surface profiles of various technical samples. Exploiting real-time signal processing along with continuous fiber drift compensation, we demonstrate the acquisition of point clouds of up to 5 million data points during continuous movement of the sensor head.


Modelling the movement of interacting cell populations: a moment dynamics approach.
Mathematical models describing the movement of multiple interacting subpopulations are relevant to many biological and ecological processes. Standard mean-field partial differential equation descriptions of these processes suffer from the limitation that they implicitly neglect to incorporate the impact of spatial correlations and clustering. To overcome this, we derive a moment dynamics description of a discrete stochastic process which describes the spreading of distinct interacting subpopulations. In particular, we motivate our model by mimicking the geometry of two typical cell biology experiments. Comparing the performance of the moment dynamics model with a traditional mean-field model confirms that the moment dynamics approach always outperforms the traditional mean-field approach. To provide more general insight we summarise the performance of the moment dynamics model and the traditional mean-field model over a wide range of parameter regimes. These results help distinguish between those situations where spatial correlation effects are sufficiently strong, such that a moment dynamics model is required, from other situations where spatial correlation effects are sufficiently weak, such that a traditional mean-field model is adequate.


Burkholderia pseudomallei capsular polysaccharide conjugates provide protection against acute melioidosis.
Burkholderia pseudomallei, the etiologic agent of melioidosis, is a CDC tier 1 select agent that causes severe disease in both humans and animals. Diagnosis and treatment of melioidosis can be challenging, and in the absence of optimal chemotherapeutic intervention, acute disease is frequently fatal. Melioidosis is an emerging infectious disease for which there are currently no licensed vaccines. Due to the potential malicious use of B. pseudomallei as well as its impact on public health in regions where the disease is endemic, there is significant interest in developing vaccines for immunization against this disease. In the present study, type A O-polysaccharide (OPS) and manno-heptose capsular polysaccharide (CPS) antigens were isolated from nonpathogenic, select-agent-excluded strains of B. pseudomallei and covalently linked to carrier proteins. By using these conjugates (OPS2B1 and CPS2B1, respectively), it was shown that although high-titer IgG responses against the OPS or CPS component of the glycoconjugates could be raised in BALB/c mice, only those animals immunized with CPS2B1 were protected against intraperitoneal challenge with B. pseudomallei. Extending upon these studies, it was also demonstrated that when the mice were immunized with a combination of CPS2B1 and recombinant B. pseudomallei LolC, rather than with CPS2B1 or LolC individually, they exhibited higher survival rates when challenged with a lethal dose of B. pseudomallei. Collectively, these results suggest that CPS-based glycoconjugates are promising candidates for the development of subunit vaccines for immunization against melioidosis.


[Psychotherapy in the era of evidence-based medicine].
Psychotherapy is a popular treatment for many psychiatric symptoms and disorders. Finding evidence for the effectiveness of therapies and bridging the gap with daily practice are a challenge. The current state of psychotherapy is explored from the perspective of evidence-based medicine. Hundreds of meta-analyses, based on thousands of studies, showed an average effect size of 0.60. Discussion in this paper focuses on the achievements and limitations of research into methods and specific disorders. Much is known about the specificity of psychotherapies and about other issues relevant to service delivery. If patient-based indication is to be a goal for the future, we will need to have a differentiated spectrum of evidence-based psychotherapies. This approach will require the integration of effective elements taken from a variety of methods, the development of multi-dimensional indicators for patient-based indication, and research into cost-effectiveness to underpin the relevance of psychotherapy for the community at large.


Mapping of protein domains of hepatitis A virus 3AB essential for interaction with 3CD and viral RNA.
The small hydrophobic protein 3AB of the picornaviruses, encompassing the replication primer 3B, has been suggested to anchor the viral replication complex to membranes. For hepatitis A virus (HAV) 3AB, we have previously demonstrated its ability to form stable homodimers, to bind to membranes, and to interact specifically with RNA, implicating its multiple involvement in viral replication. In the present report, we show that HAV 3AB additionally interacts with HAV protein 3CD, a feature also described for the corresponding polypeptide of poliovirus. By assessing the interactions of three deletion mutants, distinct domains of HAV 3AB were mapped. The hydrophobic domain and the 3B moiety were found to be essential for the 3AB interaction with 3CD. Both electrostatic and hydrophobic forces are involved in this interaction. The cluster of charged amino acid residues at the C terminus of 3A seems to determine the specificity of 3AB interaction with RNA structures formed at either terminus of the HAV genome. Furthermore, our data implicate that 3A can interact with HAV RNA. Compared with poliovirus 3AB, which by itself is a nonspecific RNA-binding protein, HAV 3AB specifically recognizes HAV RNA structures that might be of relevance for initiation of viral RNA replication.


Childhood cancer and parental occupation in Finland.
A case-control study was conducted of the occupations of parents of children under 15 with diagnosed malignancies. The total series contained all childhood cancers cases reported to the Finnish Cancer Registry during the period 1959-75. The parental occupations, recorded at the time of pregnancy, were collected from maternity welfare centres. The cases were analysed as a singly group or as subgroups according to the diagnoses-brain tumours, leukaemia, and all other malignancies. The maternal occupations found more frequently among cases than controls included farmers' wives (1959-68 only), pharmacists, saleswomen, bakers, and factory work of an vehicle driving, machine repair, painting, and the work of men who gave an academic degree as their occupation. Some of these occupations involve possible exposure to harmful chemicals, although chance correlations cannot be excluded.


Impact of small vessel disease in the brain on gait and balance.
Gait and balance impairment is highly prevalent in older people. We aimed to assess whether and how single markers of small vessel disease (SVD) or a combination thereof explain gait and balance function in the elderly. We analysed 678 community-dwelling healthy subjects from the Lothian Birth Cohort 1936 at the age of 71-74 years who had undergone comprehensive risk factor assessment, gait and balance assessment as well as brain MRI. We investigated the impact of individual SVD markers (white matter hyperintensity - WMH, microbleeds, lacunes, enlarged perivascular spaces, brain atrophy) as seen on structural brain MRI and of a global SVD score on the patients' performance. A regression model revealed that age, sex, and hypertension significantly explained gait speed. Among SVD markers white matter hyperintensity (WMH) score or volume were additional significant and independent predictors of gait speed in the regression model. A similar association was seen with the global SVD score. Our study confirms a negative impact of SVD-related morphologic brain changes on gait speed in addition to age, sex and hypertension independent from brain atrophy. The presence of WMH seems to be the major driving force for SVD on gait impairment in healthy elderly subjects.


Real-time particle tracking at 10,000 fps using optical fiber illumination.
We introduce optical fiber illumination for real-time tracking of optically trapped micrometer-sized particles with microsecond time resolution. Our light source is a high-radiance mercury arc lamp and a 600 μm optical fiber for short-distance illumination of the sample cell. Particle tracking is carried out with a software implemented cross-correlation algorithm following image acquisition from a CMOS camera. Our image data reveals that fiber illumination results in a signal-to-noise ratio usually one order of magnitude higher compared to standard Köhler illumination. We demonstrate position determination of a single optically trapped colloid with up to 10,000 frames per second over hours. We calibrate our optical tweezers and compare the results with quadrant photo diode measurements. Finally, we determine the positional accuracy of our setup to 2 nm by calculating the Allan variance. Our results show that neither illumination nor software algorithms limit the speed of real-time particle tracking with CMOS technology.


Molds in the homes of asthmatic patients in Isparta, Turkey.
This study was planned in order to determine the fungal spores in the air of inside the homes of asthmatic patients living in Isparta (from southwest region of Turkey). The seasonal properties of mold spores in the air of homes of 24 asthmatic and 14 control subjects living in the city of Isparta over a period of one year were investigated. Viable molds were recovered from all 38 houses. Twenty different molds were isolated and identified from the indoor air of the houses in which asthmatic patients and controls lived. The most common isolated genera were Penicillium spp. (27.9%), followed by Cladosporium spp. (26.3%), Aspergillus spp. (14.7%) and Alternaria spp. (13.1%) in the indoor air of the houses of asthmatic patients. No differences in colony numbers were observed between asthmatics and control groups. The percentage of molds was higher in kitchens than other parts of the houses such as living rooms and bedrooms (p < 0.05). A seasonal variety of the fungal flora in Isparta city region was observed. It is concluded that viable molds are common in houses in Isparta. Reducing indoor molds may improve the health of individuals with fungal-induced diseases like asthma.


Upregulation of extrinsic apoptotic pathway in curcumin-mediated antiproliferative effect on human pancreatic carcinogenesis.
Pancreatic cancer is one of the most lethal human cancers, with almost identical incidence and mortality rates. Curcumin, derived from the rhizome of Curcuma longa, has a long history of use as coloring agent and for a wide variety of disorders. Here, the antiproliferative activity of curcumin and its modulatory effect on gene expression of pancreatic cancer cell lines were investigated. The effect of curcumin on cellular proliferation and viability was monitored by sulphurhodamine B assay. Apoptotic effect was evaluated by flow cytometry and further confirmed by measuring amount of cytoplasmic histone-associated DNA fragments. Analysis of gene expression was performed with and without curcumin treatment using microarray expression profiling techniques. Array results were confirmed by real-time PCR. ingenuity pathway analysis (IPA) has been used to classify the list of differentially expressed genes and to indentify common biomarkergenes modulating the chemopreventive effect of curcumin. Results showed that curcumin induces growth arrest and apoptosis in pancreatic cancer cell lines. Its effect was more obvious on the highly COX-2 expressing cell line. Additionally, the expression of 366 and 356 cancer-related genes, involved in regulation of apoptosis, cell cycle, metastasis, was significantly altered after curcumin treatment in BxPC-3 and MiaPaCa-2 cells, respectively. Our results suggested that up-regulation of the extrinsic apoptotic pathway was among signaling pathways modulating the growth inhibitory effects of curcumin on pancreatic cancer cells. Curcumin effect was mediated through activation of TNFR, CASP 8, CASP3, BID, BAX, and down-regulation of NFκB, NDRG 1, and BCL2L10 genes.


Traumatic free wall and ventricular septal rupture - 'hybrid' management in a child.
A 8-year-old boy showed a traumatic ventricular septal rupture following a blunt chest trauma, and was scheduled for elective catheter closure. Two weeks later, a follow-up echocardiogram revealed a pseudoaneurysm of the anterior wall of the left ventricle. Because of the apical location of the VSD, it was decided to proceed with transcatheter occlusion. After successful VSD closure, the patient was taken to the operation room for surgical repair of the left ventricular pseudoaneurysm. Symptoms and signs seen in patients with ventricular pseudoaneurysms appear to be discrete and variable, and a high clinical index of suspicion with a very close echocardiographic follow-up is strongly recommended after occurrence of a blunt cardiac trauma. The combined 'hybrid' approach of transcatheter closure of the intraventricular rupture followed by surgical closure of the pseudoaneurysm allows for a less invasive and efficient management of this rare combination of post-traumatic ventricular free wall and septal rupture in a child.


Porphyria cutanea tarda, hepatitis C, and HFE gene mutations in North America.
In some, but not all countries, porphyria cutanea tarda (PCT) has been associated with chronic infection with the hepatitis C virus (HCV). Recently, PCT has also been associated with mutations in the HFE gene that are associated with HLA-linked hereditary hemochromatosis. Until now, few studies of these associations have been reported from North America. The aims of this study were: 1) to assess the prevalence of HCV infection and HFE mutations in North American patients with PCT; 2) to compare demographic and laboratory features between those who are HCV-positive and HCV-negative; and 3) to study urinary porphyrin excretions in American HCV-positive patients without clinically manifest PCT. Clinical and laboratory data, including tests for HCV and urinary porphyrins, were collected from 70 unselected patients with typical PCT. Urinary porphyrins were also measured in 110 non-PCT patients with chronic hepatitis C. Mutational analyses of the HFE gene were performed in 26 PCT patients. Thirty-nine of 70 (56%) of the PCT patients had evidence of HCV infection. Thirty-two of 39 PCT patients with HCV were men, all of whom used alcohol. In contrast, 22 of 31 PCT patients without HCV infection were women, 12 of whom had taken estrogens. The HCV-positive group was more likely to have used illicit intravenous drugs (45% vs. 0%; P = 0.01), to have had several (>4) sex partners (48% vs. 13%; P = 0.005), and less likely to have no known risk factors for HCV infection (33% vs. 78%; P = 0.004). Total urinary porphyrin excretion was the same in the two groups, but those with HCV infection had a significantly lower percentage of uroporphyrin and higher percentages of hepta-and hexa-carboxy porphyrins in urine. Sixteen of 110 (15%) HCV-positive subjects without PCT had increased urinary porphyrins, but, unlike PCT, these were mainly coproporphyrin. Forty-two percent of PCT patients carried the C282Y mutation of HFE (15% homozygous), and another 31% carried the H63D mutation (8% homozygous). Thus, 73% of PCT patients had one of these mutations. The prevalence of HCV infection (56%) and mutations in the HFE gene (73%) are high among North American patients with PCT. Alcohol and estrogen use are important additional risk factors. All PCT patients should be tested for HCV infection and for HFE gene mutations. Although HCV infection is a trigger for PCT, preclinical PCT is rare in chronic HCV hepatitis C in the United States.


[Oxygen tension in the skeletal muscles of rats adapted to the cold].
In control and cold-adapted rats the oxygen tension (pO2) was measured polarographically in the fast (gastrocnemic) and slow (soleus) muscles before and after blocking of beta-adrenoreceptors or the denervation of the muscle. The beta-adrenoreceptor blockade decreases the pO2 in both types of muscles in the control group of rats, whereas in cold-adapted rats the decrease of pO2 both types of muscles in the control group of rats, whereas in cold-adapted rats the decrease of pO2 is less pronounced in the coleus and remains unchanged in the gastrocnemic muscle. The sciatic nerve dissection augments the pO2 in both muscles, the increase being larger in cold-adapted rats as compared to the control group. It seems that the dependence of the oxygenation processes in muscles on nervous control and on the state of beta-adrenoreceptors increases during adaptation to cold mainly in the slow muscles.


Eigen-R2 for dissecting variation in high-dimensional studies.
We provide a new statistical algorithm and software package called 'eigen-R(2)' for dissecting the variation of a high-dimensional biological dataset with respect to other measured variables of interest. We apply eigen-R(2) to two real-life examples and compare it with simply averaging R(2) over many features. An R-package eigenR2 is available at http://www.genomine.org/eigenr2/ and will be made publicly available via Bioconductor.


Noncontact Cohesive Swimming of Bacteria in Two-Dimensional Liquid Films.
Bacterial swimming in confined two-dimensional environments is ubiquitous in nature and in clinical settings. Characterizing individual interactions between swimming bacteria in 2D confinement will help to understand diverse microbial processes, such as bacterial swarming and biofilm formation. Here we report a novel motion pattern displayed by flagellated bacteria in 2D confinement: When two nearby cells align their moving directions, they tend to engage in cohesive swimming without direct cell body contact, as a result of hydrodynamic interaction but not flagellar intertwining. We further found that cells in cohesive swimming move with higher directional persistence, which can increase the effective diffusivity of cells by ∼3 times as predicted by computational modeling. As a conserved behavior for peritrichously flagellated bacteria, cohesive swimming in 2D confinement may be key to collective motion and self-organization in bacterial swarms; it may also promote bacterial dispersal in unsaturated soils and in interstitial space during infections.


Ozone-induced acute pulmonary fibrosis in rats. Prevention of increased rates of collagen synthesis by methylprednisolone.
The net rate of collagen synthesis by lung minces prepared from rats exposed for 7 days to 0.8 to 1.5 ppm (1.6 to 3.0 mg/m3) of ozone was increased in a dose-dependent manner severalfold above the net rate obtained with lung minces prepared from rats that had breathed only filtered air. Concurrent administration of methylprednisolone (1 to 50 mg/kg/day) during the exposure to ozone prevented the increase in rate of collagen synthesis in a dose-dependent manner for each level of ozone tested. These results could be correlated with lower levels of inflammatory edema measured in the same steroid-treated rats as decreased wet weights of their right apical lung lobes. We conclude that acute pulmonary fibrosis caused by exposure of rats to high levels of inspired ozone can be partially or completely prevented, depending on the severity of the insult, by simultaneous administration of high levels of the anti-inflammatory steroid methylprednisolone.


Wildfires in bamboo-dominated Amazonian forest: impacts on above-ground biomass and biodiversity.
Fire has become an increasingly important disturbance event in south-western Amazonia. We conducted the first assessment of the ecological impacts of these wildfires in 2008, sampling forest structure and biodiversity along twelve 500 m transects in the Chico Mendes Extractive Reserve, Acre, Brazil. Six transects were placed in unburned forests and six were in forests that burned during a series of forest fires that occurred from August to October 2005. Normalized Burn Ratio (NBR) calculations, based on Landsat reflectance data, indicate that all transects were similar prior to the fires. We sampled understorey and canopy vegetation, birds using both mist nets and point counts, coprophagous dung beetles and the leaf-litter ant fauna. Fire had limited influence upon either faunal or floral species richness or community structure responses, and stems <10 cm DBH were the only group to show highly significant (p = 0.001) community turnover in burned forests. Mean aboveground live biomass was statistically indistinguishable in the unburned and burned plots, although there was a significant increase in the total abundance of dead stems in burned plots. Comparisons with previous studies suggest that wildfires had much less effect upon forest structure and biodiversity in these south-western Amazonian forests than in central and eastern Amazonia, where most fire research has been undertaken to date. We discuss potential reasons for the apparent greater resilience of our study plots to wildfire, examining the role of fire intensity, bamboo dominance, background rates of disturbance, landscape and soil conditions.


Defining E3 ligase-substrate relationships through multiplex CRISPR screening.
Specificity within the ubiquitin-proteasome system is primarily achieved through E3 ubiquitin ligases, but for many E3s their substrates-and in particular the molecular features (degrons) that they recognize-remain largely unknown. Current approaches for assigning E3s to their cognate substrates are tedious and low throughput. Here we developed a multiplex CRISPR screening platform to assign E3 ligases to their cognate substrates at scale. A proof-of-principle multiplex screen successfully performed ~100 CRISPR screens in a single experiment, refining known C-degron pathways and identifying an additional pathway through which Cul2FEM1B targets C-terminal proline. Further, by identifying substrates for Cul1FBXO38, Cul2APPBP2, Cul3GAN, Cul3KLHL8, Cul3KLHL9/13 and Cul3KLHL15, we demonstrate that the approach is compatible with pools of full-length protein substrates of varying stabilities and, when combined with site-saturation mutagenesis, can assign E3 ligases to their cognate degron motifs. Thus, multiplex CRISPR screening will accelerate our understanding of how specificity is achieved within the ubiquitin-proteasome system.


Haematinic deficiency in pregnancy: another HELLP mimic.
Haematinic deficiency is not uncommon in pregnancy. Folate deficiency is more common than B12 deficiency because of the increased uptake of folate in pregnancy, and the fact that B12 stores take much longer to deplete. Described here are five cases of anaemia in pregnancy secondary to severe haematinic deficiency with subsequent management and pregnancy outcomes. In the majority of cases, the women were proteinuric, but systemically well and normotensive. Thrombotic thrombocytopenic purpura and HELLP were both considered, but the identification of very abnormal folate levels of less than 3 μg/L in all and low B12 deficiency in the majority made haematinic deficiency the most likely diagnosis. They all received high dose folic acid, parenteral vitamin B12 and oral iron and made good haematological recoveries. Adequate antenatal correction of vitamin deficiency like this avoids bone marrow suppression and helps minimise poor obstetric outcomes associated with pre-existing anaemia including post-partum haemorrhage.


Beetle horns are regulated by the Hox gene, Sex combs reduced, in a species- and sex-specific manner.
Discovering the mechanisms that underlie the origin of novel features represents a major frontier in developmental and evolutionary biology. Here we begin to characterize the role of the Hox gene Sex combs reduced (Scr) during the development and evolution of a morphologically novel trait: beetle horns. Beetle horns develop as epidermal outgrowths from the prothorax and/or head, and size and location vary dramatically across species and between sexes. Using both comparative gene expression and larval RNA interference in two species of the horned beetle genus Onthophagus, we show that Scr functions in patterning adult labial mouthpart identity and suppressing wing development in the prothorax. At the same time, however, our results illustrate that Scr has acquired, within its ancestral domain of expression, additional new functions including the regulation of prepupal growth and pupal remodeling of pronotal horn primordia. Furthermore, comparative analyses of our results across both Onthophagus species, which differ in location of horn development (thoracic horns vs. thoracic and head horns) as well as patterns of sexual dimorphism (traditional vs. reversed sexual dimorphism), reveal surprising differences in exactly when, where, and to what degree Scr regulates horn formation in different sexes. These observations suggest that the interactions between Scr and its targets in the regulation of horn development can diversify quickly over remarkably short phylogenetic distances. More generally, our results suggest that the Hox complex can play an integral role in the development and evolution of novel complex traits while maintaining traditional patterning responsibilities.


Mixed methods evaluation explains bypassing of vouchers in micronutrient powder trial in Mozambique.
Micronutrient powders (MNP) are recommended by the World Health Organization as an effective intervention to address anaemia in children. A formative process evaluation was conducted to assess the viability of a model using free vouchers in two districts of Mozambique to deliver MNP and motivate adherence to recommendations regarding its use. The evaluation consisted of (a) an examination of programme outcomes using a cross-sectional survey among caregivers of children 6-23 months (n = 1,028) and (b) an ethnographic study to investigate delivery experiences and MNP use from caregiver perspectives (n = 59), programme managers (n = 17), and programme implementers (n = 168). Using a mixed methods approach allowed exploration of unexpected programme outcomes and triangulation of findings. The survey revealed that receiving a voucher was the main implementation bottleneck. Although few caregivers received vouchers (11.5%, CI [9.7, 13.6]), one-fourth received MNP by bypassing the voucher system (26.3%, CI [23.6, 29.0]). Caregivers' narratives indicated that caregivers were motivated to redeem vouchers but encountered obstacles, including not knowing where or how to redeem them or finding MNP were not available at the shop. Observing these challenges, many programme implementers redeemed vouchers and distributed MNP to caregivers. Virtually, all caregivers who received MNP reported ever feeding it to their child. This study's findings are consistent with other studies across a range of contexts suggesting that although programmes are generally effective in motivating initial use, more attention is required to improve access to MNP and support continued use.


Treatment of bilateral mammary ptosis and pectus excavatum through the same incision in one surgical stage.
Congenital deformities of the anterior thoracic wall are characterized by unusual development of the costal cartilages. All these medical conditions are frequently associated with a variety of breast deformities. Several surgical techniques have been described for correcting them, going from sternochondroplasty to, nowadays, minimally invasive techniques and silicone prosthesis implantation. The present article reports the case of a young female patient who presented bilateral mammary ptosis and moderate pectus excavatum that caused a protrusion between the eighth and the tenth ribs and consequent esthetic disharmony. The proposed surgical treatment included not only subglandular breast implants of polyurethane, but also resection of part of the rib cartilage and a bone segment from the eighth, ninth and tenth ribs by means of a single submammary incision in order to make the scar minimally visible. Correction through a single incision benefited the patient and provided an excellent esthetic result. The techniques used to repair bilateral mammary ptosis and pectus excavatum by plastic and thoracic surgery teams, respectively, have been shown to be efficient for correcting both deformities. An excellent esthetic and functional result was obtained, with consequent reestablishment of the patient's self-esteem.


Contexts and data dependencies: a synthesis.
Two data-organization devices that have come out of AI re-search are data pools (``contexts'') and data dependencies. The latter are more flexible than the former, and have supplanted them. Data pools offer certain advantages of efficiency, however, so it is worth trying to make the two mechanisms compatible. Doing this requires generalizing the mark-and-sweep algorithms that maintain consistency in a data-dependency network, so that the labels passed around do not simply say whether a datum is IN or OUT, but say which data pools it is present in. The revised algorithm is essentially an algorithm for solving simultaneous Boolean equations. Other mechanisms are needed for per-forming useful chores like maintaining well-founded support links and orchestrating demon calls.


A Prototype High-Resolution Small-Animal PET Scanner Dedicated to Mouse Brain Imaging.
We developed a prototype small-animal PET scanner based on depth-encoding detectors using dual-ended readout of small scintillator elements to produce high and uniform spatial resolution suitable for imaging the mouse brain. The scanner consists of 16 tapered dual-ended-readout detectors arranged in a 61-mm-diameter ring. The axial field of view (FOV) is 7 mm, and the transaxial FOV is 30 mm. The scintillator arrays consist of 14 × 14 lutetium oxyorthosilicate elements, with a crystal size of 0.43 × 0.43 mm at the front end and 0.80 × 0.43 mm at the back end, and the crystal elements are 13 mm long. The arrays are read out by 8 × 8 mm and 13 × 8 mm position-sensitive avalanche photodiodes (PSAPDs) placed at opposite ends of the array. Standard nuclear-instrumentation-module electronics and a custom-designed multiplexer are used for signal processing. The detector performance was measured, and all but the crystals at the very edge could be clearly resolved. The average intrinsic spatial resolution in the axial direction was 0.61 mm. A depth-of-interaction resolution of 1.7 mm was achieved. The sensitivity of the scanner at the center of the FOV was 1.02% for a lower energy threshold of 150 keV and 0.68% for a lower energy threshold of 250 keV. The spatial resolution within a FOV that can accommodate the entire mouse brain was approximately 0.6 mm using a 3-dimensional maximum-likelihood expectation maximization reconstruction. Images of a hot-rod microphantom showed that rods with a diameter of as low as 0.5 mm could be resolved. The first in vivo studies were performed using (18)F-fluoride and confirmed that a 0.6-mm resolution can be achieved in the mouse head in vivo. Brain imaging studies with (18)F-FDG were also performed. We developed a prototype PET scanner that can achieve a spatial resolution approaching the physical limits of a small-bore PET scanner set by positron range and detector interaction. We plan to add more detector rings to extend the axial FOV of the scanner and increase sensitivity.


Genetically determined alcohol consumption and cancer risk in Korea.
The purpose of this study was to determine the causal relationship between the genetically determined amount of alcohol consumption and the occurrence of major cancers. The data used in this study were from 129,324 people selected from the Korean Cancer Prevention Study-II, the participants of which visited 18 health examination centers between 2004 and 2013. Cancer incidence was confirmed as of 2020 using data from the National Cancer Center. A genome-wide association study (GWAS) on alcohol consumption was performed using PLINK 2.0, and sex, age, chip type, and principal components were adjusted. From the GWAS, a genetic risk score for alcohol consumption was calculated and genetically determined alcohol consumption (GDAC) was estimated. GDAC was divided into quintile groups and showed significant causal relationships with rectal cancer and liver cancer, but not with other cancers. For liver cancer, an association was shown in the hepatitis B surface antigen (HBsAg)-negative group, and a particularly strong association was found in the over-60-year-old HBsAg-negative group, in which, compared to the GDAC Q1 group, the Q4 group had a 2.35 times higher risk (95% confidence interval [CI], 1.05 to 5.23), and the Q5 group had a 2.40 times higher risk (95% CI, 1.09 to 5.30). The results of this study provided evidence that the amount of alcohol consumed is causally related to the occurrence of rectal cancer and liver cancer in HBsAg-negative individuals. Additional studies should be continued for other cancer types through long-term follow-up.


Characterization of Streptococcus pneumoniae clones from paediatric patients with cystic fibrosis.
The role of Streptococcus pneumoniae in cystic fibrosis (CF) is poorly understood. The pneumococcal population has changed over time after the introduction of the heptavalent conjugate vaccine (PCV7) and, more recently, the 13-valent conjugate vaccine (PCV13). Although serotypes and clones causing invasive pneumococcal disease or colonizing healthy children have been extensively analysed, little is known so far on the serotypes and clones of pneumococci in CF patients. The aim of this work was to investigate serotypes, antibiotic susceptibilities, genotypes and biofilm production of CF pneumococcal isolates. Overall, 44 S. pneumoniae strains collected from 32 paediatric CF patients from January 2010 to May 2012 in a large Italian CF Centre were tested for antimicrobial susceptibility testing by Etest, serotyped by the Quellung reaction and genotyped by a combination of different molecular typing methods, including pbp gene restriction profiling, pspA restriction profiling and sequencing, PFGE and multilocus sequence typing. Biofilm production by pneumococcal strains was also assessed. Penicillin non-susceptibility was 16 %. High resistance rates (>56 %) were observed for erythromycin, clindamycin and tetracycline. The most frequent serotype recovered was serotype 3 (31.8 %). The coverage of PCV7 and PCV13 was 6.8 and 47.7 %, respectively. More than 80 % of CF strains belonged to Pneumococcal Molecular Epidemiology Network (PMEN) reference clones, the most common being Netherlands(3)-ST180 (28.2 %), and Greece(21)-30/ST193 (15.4 %). All strains produced biofilm in vitro, although with large variability in biofilm formation efficiency. No correlation was found between biofilm levels and serotype, clone or antibiotic resistance. The high isolation rate of antibiotic-resistant serotype 3 pneumococci from CF patients suggests that PCV13 could increase protection from pneumococcal colonization and infection.


[Tissue oxygenation and tissue hypoxia in diabetes mellitus].
The-Polarographic method was applied to the study of free oxygen tension (PO2) in the subcutaneous cellular tissue and the muscle of the arm in 104 patients suffering from diabetes mellitus without any concomitant pathology of the lungs and in 25 healthy persons. At the period of decompensation of diabetes mellitus (in the patients without any concomitant diseases of the lungs) PO2 was decreased in the subcutaneous cellular tissue by the average of 42% in comparison with the norm, and in the skeletal muscle of the arm--by 38%. The most frequently (and more significant) was tissue hypoxia in complicated diabetes, particularly in ketacidosis. Effective treatment led to an elevation of PO2 in the tissues of the great majority of the patients; in some of the patients it became normal.


Interpreting the transmissibility of measles in two different post periods of supplementary immunization activities in Hubei, China.
Although evidence has shown that supplementary immunization activity (SIA) campaigns greatly reduce the incidence of measles, their effects on disease transmissibility have seldom been monitored. A great decrease in the number of cases may be a false signal of early success towards measles elimination to policy makers. By interpreting the transmissibility in two different post-SIA periods in Hubei, China, the current study showed sustained measles transmissions despite a reduced number of cases. Two population-based cross-sectional serological surveys of measles antibodies were conducted in Hubei province in mid-2010 and mid-2011 after the implementation of SIAs. Immunoglobulin G (IgG) antibodies against measles were measured by enzyme-linked immunosorbent assay (ELISA). Based on the estimated age-specific susceptibility levels, the effective reproduction number (R), a key indicator of disease transmissibility, was determined by the next generation matrix in transmission model. The results revealed an overall IgG seroprevalence of 88.0% (95% confidence interval [CI]: 85.6-90.4%) and 89.6% (95%CI: 88.0-91.2%), respectively, in the two different periods. Comparatively lower seroprevalence rates were observed among children less than 24months of age and young adults 15 to 19years of age in 2011. The Rs were 0.76 and 1.53 for the two study periods. In conclusion, even though the incidence was reduced to below 1/100,000 in both 2010 and 2011, the reproduction number in 2011 indicates a high risk for sustained measles transmission. This finding was potentially due to a lower seropositivity rate among young adults that had not been covered in the first SIA. Thus, implementation of SIA targeted to appropriate age groups is recommended. Regular monitoring of seroprevalence is also suggested to track disease transmissibility and to align SIA with the appropriate age groups.


Dynamical modeling of cascading failures in the Turkish power grid.
A reliable supply of electricity is critical for our modern society, and any large-scale disturbance of the electrical system causes substantial costs. In 2015, one overloaded transmission line caused a cascading failure in the Turkish power grid, affecting about 75×106 people. Here, we analyze the Turkish power grid and its dynamical and statistical properties. Specifically, we propose, for the first time, a model that incorporates the dynamical properties and the complex network topology of the Turkish power grid to investigate cascading failures. We find that the network damage depends on the load and generation distribution in the network with centralized generation being more susceptible to failures than a decentralized one. Furthermore, economic considerations on transmission line capacity are shown to conflict with stability.


Analysis of cell damage and proliferation in Helicobacter pylori-infected human gastric mucosa from patients with gastric adenocarcinoma.
Helicobacter pylori (HP) infection, a cause of multifocal atrophic gastritis, is considered an important factor related to the evolution of the human gastric mucosa from normal to intestinal-type adenocarcinoma. We examined cell proliferation and both double and single strand DNA damage in situ in 35 patients undergoing gastrectomy for adenocarcinoma with HP-infected gastric mucosa by immunolocalization of Ki-67, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling, and in situ nick translation. We also studied the distribution of intraepithelial neutrophils by elastase immunolocalization. HP infection was confirmed in all cases by serum anti-HP antibodies, ureas testing, and histopathological examination. HP-infected gastric mucosa was classified according to the degree of inflammation and intestinal metaplasia. Ki-67, terminal deoxynucleotidyl transferase-mediated labeling, in situ nick translation, and intraepithelial neutrophil indices all increased with the progression of gastritis and were highest in glands with incomplete intestinal metaplasia. All indices were lowest in gastric glands with complete intestinal metaplasia. Significant positive correlations were observed among these markers. Increased proliferative activity in HP-associated chronic gastritis in response to cell damage or injury was clearly demonstrated, suggesting that both HP-associated toxins and intraepithelial neutrophils are important in HP-related gastric epithelial injury. Increased cell turnover associated with incomplete intestinal metaplasia may result in DNA instability and subsequent development of intestinal-type gastric adenocarcinoma in HP-infected mucosa.


Age is the major determinant of recurrence in pediatric differentiated thyroid carcinoma.
A relationship between young age and increased risk of recurrence of pediatric differentiated thyroid carcinoma has been suggested; however, no attempts have been made to assess the prognostic factors or efficacy of treatment in very young children with this malignancy. The objectives of this study were to evaluate the association of age with outcome in pediatric differentiated thyroid carcinoma and to compare the clinical, pathologic, prognostic, and treatment variables between younger and older children with this disease. A retrospective review of all patients presenting to the British Columbia's Children's Hospital or British Columbia Cancer Agency <17 years of age at diagnosis with differentiated thyroid carcinoma between January, 1955, and December, 1996, was completed. Thirty-eight patients were identified, 12 of whom were </=10 years of age. The overall and relapse-free survivals at 20 years were 100% and 32.2%, respectively. Age at diagnosis was the only determinant of time to recurrence on univariate and multivariate regression analysis of prognostic factors (P = 0.022). The 20 year relapse-free survival for children < or =10 years of age was 10.1% vs. 48.3% for children >10 years. An association between young age and extrathyroidal tumor invasion was identified (P = 0.016); however, the latter factor did not independently predict outcome. There was a trend for suppressive doses of thyroid hormone to improve outcome, particularly with increasing age at diagnosis, but this was not statistically significant. Age is the major determinant of recurrence in pediatric differentiated thyroid carcinoma. The results suggest different tumor biology in young children requiring novel approaches to therapy to decrease recurrence rates.


Abnormal platelet trace amine profiles in migraine with and without aura.
Trace amines, including tyramine, octopamine and synephrine, are closely related to classic biogenic amines. In one study, where these substances were found elevated in plasma of migraineurs, it was hypothesized that trace amine metabolism is deranged in migraine. To confirm these findings, we studied, using a multichannel electrochemical high-performance liquid chromatography system, the concentrations of trace amines in platelets of migraine without aura (MoA) and migraine with aura (MA) patients in headache-free period, compared with controls. Platelet concentrations of trace amines, although elevated in both migraine types, showed a different profile in MoA and MA. Octopamine was significantly higher in MoA sufferers (0.69 +/- 0.43 ng/10(8) platelets) compared with both control subjects (0.22 +/- 0.16 ng/10(8) platelets) and MA patients (0.39 +/- 0.37 ng/10(8) platelets). Synephrine was significantly higher in MA patients (0.72 +/- 0.44 ng/10(8) platelets) with respect to both controls (0.33 +/- 0.25 ng/10(8) platelets) and MoA sufferers (0.37 +/- 0.29 ng/10(8) platelets). These results strengthen the hypothesis that tyrosine metabolism is deranged in migraine and may participate in its pathophysiology.


Neurodevelopmental Disorders in the PICU Population.
Attention deficit hyperactivity disorder (ADHD), affecting 11% of children and adolescents, increases risk for injury and may predispose children to illness. However, the prevalence of ADHD and other developmental disorders in the pediatric intensive care unit (PICU) has not been previously studied. We performed a single-center, prospective cohort study of children aged 6 to 12 years who were hospitalized in the PICU from May through August 2016. Parents described their child's educational and neurodevelopmental history, and completed ADHD and emotional/behavioral disorder screening on enrollment and 1 month after discharge. Twenty-four children were enrolled. Ten patients (42%) had a prior neurodevelopmental diagnosis, and 7 (29%) met study criteria for ADHD. Children hospitalized for critical illness have a high prevalence of neurodevelopmental disabilities and are more susceptible to the impact of critical illness on development and behavior. More research is needed to better understand how to support this vulnerable population after critical illness.


Quality of life and functional outcomes 10 years after laparoscopic radical prostatectomy.
Minimally invasive laparoscopic radical prostatectomy (LRP) has proven equally effective as open surgery in terms of cancer control and peroperative complication rate with less bleeding and postoperative pain. However, long-term follow-up data after LRP are scarce, especially as related to quality of life (QoL). To compare QoL and functional outcomes at least 10 years after LRP with a population-based control group matched for age and region. Follow-up data were obtained by mailed questionnaires from patients who responded anonymously to five international questionnaires (EQ-5D, QLQ-C30, QLQ-PR25, IPSS, and IIEF). We collected self-reported outcome data directly from 49 patients who underwent LRP more than 10 years ago in our centre. The results of the patients' overall QoL and urinary continence rates were compared with 918 controls matched for region and age. Forty-two patients (86%) and 808 (88%) controls reported having no urinary leakage. Only 11 patients (24%) still had sexual activities 10 years after LRP, and three were without erectile dysfunction. There was no difference in four of five statements of the self-assessed QoL questionnaires between the LRP and control group. Anxiety level was higher in the LRP group (44%) than in the control group (23%). Patients reported high self-assessed QoL, although they also reported low sexual activity 10 years after LRP. Prevalence of urinary leakage was similar in both groups. However, anxiety was more common in LRP patients.


Olfm4 deletion enhances defense against Staphylococcus aureus in chronic granulomatous disease.
Chronic granulomatous disease (CGD) patients have recurrent life-threatening bacterial and fungal infections. Olfactomedin 4 (OLFM4) is a neutrophil granule protein that negatively regulates host defense against bacterial infection. The goal of this study was to evaluate the impact of Olfm4 deletion on host defense against Staphylococcus aureus and Aspergillus fumigatus in a murine X-linked gp91phox-deficiency CGD model. We found that intracellular killing and in vivo clearance of S. aureus, as well as resistance to S. aureus sepsis, were significantly increased in gp91phox and Olfm4 double-deficient mice compared with CGD mice. The activities of cathepsin C and its downstream proteases (neutrophil elastase and cathepsin G) and serum levels of IL-1β, IL-6, IL-12p40, CXCL2, G-CSF, and GM-CSF in Olfm4-deficient as well as gp91phox and Olfm4 double-deficient mice were significantly higher than those in WT and CGD mice after challenge with S. aureus. We did not observe enhanced defense against A. fumigatus in Olfm4-deficient mice using a lung infection model. These results show that Olfm4 deletion can successfully enhance immune defense against S. aureus, but not A. fumigatus, in CGD mice. These data suggest that OLFM4 may be an important target in CGD patients for the augmentation of host defense against bacterial infection.


[Study on the rule of acupoint selection in treatment of acute exacerbation of chronic obstructive pulmonary disease].
To explore the characteristics and rule of clinical acupoint selection in treatment of acute exacerbation of chronic obstructive pulmonary disease (AECOPD). The clinical articles of acupuncture in treatment of AECOPD were retrieved from the databases of PubMed, EMbase, Cochrane Library, Web of Science, CNKI, Wanfang, VIP and SinoMed, from the date of establishment to July 15, 2020. The articles were screened in accordance with the inclusion and exclusion criteria, the prescriptions of acupuncture and the relevant information of the acupoints and meridians were extracted to establish the database. The data mining methods i.e. Apriori association rule analysis and cluster analysis were used to analyse the using frequency, involving meridians, acupoint distributions, association rules and cluster of selected acupoints. A total of 54 articles were included, 67 acupuncture prescriptions were extracted, 69 acupoints were involved and the total using frequency was 475 times. The top 5 acupoints in frequency were Danzhong (CV 17), Feishu (BL 13), Zusanli (ST 36), Fenglong (ST 40) and Dingchuan (EX-B1). The main involved meridians were bladder meridian of foot-taiyang, conception vessel, stomach meridian of foot-yangming and lung meridian of hand-taiyin. The acupoints were mainly distributed in chest and abdomen, waist and dorsum. The mainly selected special points were five-shu points, front-mu points, convergent points and back-shu points. Twenty strong association rules were summarized by association rule analysis, including Fenglong (ST 40)Þ Zusanli (ST 36), Fengmen (BL 12)Þ Danzhong (CV 17), Tianshu (ST 25)Þ Zhongwan (CV 12), Dingchuan (EX-B1)+ Feishu (BL 13) Þ Danzhong (CV 17), Fenglong (ST 40)+ Feishu (BL 13) Þ Zusanli (ST 36), etc. Five cluster groups were summarized by cluster analysis, i.e. ①Feishu (BL 13), ② Dingchuan (EX-B1) and Danzhong (CV 17), ③ Chize (LU 5), Lieque (LU 7) and Fengmen (BL 12), ④Zusanli (ST 36) and Fenglong (ST 40), ⑤Tianshu (ST 25), Guanyuan (CV 4) and Zhongwan (CV 12). The local acupoints and acupoints along meridians are the main acupuncture prescriptions for AECOPD, and the special points are the predominated selection. The acupoint compatibility embodies the therapeutic principle of "strengthening vital qi to eliminate pathogenic factors and considering both the root cause and symptoms".


Kinetic Rate Constants of Gold Nanoparticle Deposition on Silicon.
We fabricated gold nanoparticles on nanoporous silicon microparticles using electroless deposition in a hydrofluoric acid solution containing gold chloride. The reaction was followed by UV spectrometer analysis of the absorbance of the solution (proportional to the nanoparticle concentration) for two temperatures (20 and 50 °C). The results indicate that the process is autocatalytic, described by a pseudo-first-order reaction, the apparent rate constant kobs of which was determined by utilizing UV spectrometer data. We found that the reaction rate constant at 20 °C is 7 × 10-3 s-1 and that at 50 °C is 2.9 × 10-2 s-1. Scanning electron microscope (SEM) analysis of samples and diffusion-limited aggregation (DLA) simulations were used to validate the results. This study aims to resolve the kinetics of the electroless deposition of gold on silicon at the nanoscale, in the present state of art missing a quantitative characterization, for certain conditions of growth and given values of temperature and concentration of the reagents. Results may have applications to the synthesis of gold nanoparticles and their use as nanosensors, drug delivery systems, or metal nanometamaterials with advanced optical properties.


Does bone mineral density predict fractures comparably in women and men?
Osteoporosis (OP) is a very common disease associated with increased morbidity, mortality and costs. For a 50-yr-old woman the lifetime risk of an osteoporotic fracture is 40%, while for a man of the same age the risk is 13%. Good evidence exists as to the correlation between bone mineral density (BMD) and fracture risk in post-menopausal women. The diagnosis of OP can be made when BMD is more >2.5 SD below the mean of normal young women (T-score < or = -2.5). In men it has not been possible, until now, to identify a definite T-score under which the diagnosis of OP can be made. Several studies produced conflicting results when they tried to answer the question as to whether males and females fracture at the same absolute BMD value. Men have a greater bone size than women even when this parameter is corrected for weight and body mass. As densitometric devices measure areal density, men appear to have a higher BMD than women. Some studies have shown that, for a given BMD, males and females have the same fracture risk, while other papers have demonstrated that fractured men have a higher BMD than fractured women. Another problem concerns the diagnosis of osteoporosis. In fact, when the T-score is calculated in men on the basis of a young female reference range the prevalence of osteoporosis can be underestimated. The official position of International Society for Clinical Densitometry (ISCD) may represent an "interim" answer in order to identify men at risk of fracture.


Cannabimimetic indoles, pyrroles and indenes.
In the course of efforts to develop new nonsteroidal antiinflammatory agents, it was discovered that 1-aminoalkyl-3-aroylindoles have affinity for the cannabinoid brain (CB1) receptor. This led to the synthesis of well over 100 cannabimimetic aminoalkylindoles by the group at Sterling Winthrop, and to the development of structure-activity relationships (SAR) for these compounds. These SAR require a heterocyclic aminoethyl group attached to the indole nitrogen, and a 1-naphthoyl group at C-3 for significant receptor affinity. Other workers subsequently demonstrated that an aminoalkyl group was not necessary for cannabinoid activity, but that an N-alkyl group of four to six carbons was sufficient. This led to the discovery that 1-propyl-3-(1-naphthoyl)indole is a selective ligand for the peripheral cannabinoid (CB2) receptor, and to the development of a series of cannabimimetic pyrroles. Comprehensive SAR for this group of cannabinoids have been developed. Two groups have described cannabimimetic indenes, which have been employed as rigid models for the receptor interactions of cannabimimetic indoles with the CB1 receptor. There is some evidence that the indoles interact at a somewhat different site on the receptor than traditional cannabinoids.


The effects of BungeeSkate training on measures of on-ice acceleration and speed.
Previous research has stated that dryland sled pulling trains first-step quickness in hockey players. Further research has demonstrated that off-ice horizontal training (sled pull, parachute, etc) relates well to on-ice acceleration and speed. However, there is limited literature pertaining to on-ice resistance training that aims to enhance speed and acceleration in hockey players. The purpose of the current study was to determine if on-ice BungeeSkate training would improve on-ice speed and acceleration in youth hockey players. Twenty-three Peewee and Bantam hockey players (age 11-14) were recruited, with 20 participants completing the study. Pretesting and posttesting consisted of an on-ice 44.8-m speed test, a 6.1-m acceleration test, and a 15.2-m full-speed test. The training protocol consisted of 8 sessions of 5 BungeeSkate training exercises per session, 2 times per week for a 4-wk period. The results of this study showed that speed and top speed were significantly increased (P < .05) by 4.2% and 4.3%, respectively. Acceleration was also slightly improved but not significantly. A 4-wk BungeeSkate training intervention can improve acceleration and speed in youth hockey players. This training method could be a valid adjunct to existing strategies to improve skill development in hockey and is shown to improve speed and acceleration in relatively short training sessions. This may be most advantageous for hockey coaches and players who are looking to maximize training benefits with limited ice time.


Acoustic impedance matching of piezoelectric transducers to the air.
The purpose of this work is threefold: to investigate material requirements to produce impedance matching layers for air-coupled piezoelectric transducers, to identify materials that meet these requirements, and to propose the best solution to produce air-coupled piezoelectric transducers for the low megahertz frequency range. Toward this end, design criteria for the matching layers and possible configurations are reviewed. Among the several factors that affect the efficiency of the matching layer, the importance of attenuation is pointed out. A standard characterization procedure is applied to a wide collection of candidate materials to produce matching layers. In particular, some types of filtration membranes are studied. From these results, the best materials are identified, and the better matching configuration is proposed. Four pairs of air-coupled piezoelectric transducers also are produced to illustrate the performance of the proposed solution. The lowest two-way insertion loss figure is -24 dB obtained at 0.45 MHz. This increases for higher frequency transducers up to -42 dB at 1.8 MHz and -50 at 2.25 MHz. Typical bandwidth is about 15-20%.


Transport Metabolons and Acid/Base Balance in Tumor Cells.
Solid tumors are metabolically highly active tissues, which produce large amounts of acid. The acid/base balance in tumor cells is regulated by the concerted interplay between a variety of membrane transporters and carbonic anhydrases (CAs), which cooperate to produce an alkaline intracellular, and an acidic extracellular, environment, in which cancer cells can outcompete their adjacent host cells. Many acid/base transporters form a structural and functional complex with CAs, coined "transport metabolon". Transport metabolons with bicarbonate transporters require the binding of CA to the transporter and CA enzymatic activity. In cancer cells, these bicarbonate transport metabolons have been attributed a role in pH regulation and cell migration. Another type of transport metabolon is formed between CAs and monocarboxylate transporters, which mediate proton-coupled lactate transport across the cell membrane. In this complex, CAs function as "proton antenna" for the transporter, which mediate the rapid exchange of protons between the transporter and the surroundings. These transport metabolons do not require CA catalytic activity, and support the rapid efflux of lactate and protons from hypoxic cancer cells to allow sustained glycolytic activity and cell proliferation. Due to their prominent role in tumor acid/base regulation and metabolism, transport metabolons might be promising drug targets for new approaches in cancer therapy.


Wilms' tumor suppressor gene (WT1) is expressed in primary breast tumors despite tumor-specific promoter methylation.
We analyzed Wilms' tumor suppressor 1 (WT1) expression and its regulation by promoter methylation in a panel of normal breast epithelial samples and primary carcinomas. Contrary to previous reports, WT1 protein was strongly expressed in primary carcinomas (27 of 31 tumors) but not in normal breast epithelium (1 of 20 samples). Additionally, the WT1 promoter was methylated in 6 of 19 (32%) primary tumors, which nevertheless expressed WT1. The promoter is not methylated in normal epithelium. Thus, although tumor-specific methylation of WT1 is established in primary breast cancer at a low frequency, other transcriptional regulatory mechanisms appear to supercede its effects in these tumors. Our results demonstrate expression of WT1 in mammary neoplasia, and that WT1 may not have a tumor suppressor role in breast cancer.


Psychometric Properties of Flourishing Scales From a Comprehensive Well-Being Assessment.
In this article, we develop a measure of complete well-being. The framework is derived from the theoretical model of human flourishing understood as a state in which all aspects of a human life are favorable. The approach extends beyond psychological well-being and reflects the World Health Organization definition of health that not only considers the health of body and mind but also embraces the wholeness of the person. The Well-Being Assessment (WBA) is a comprehensive instrument designed to assess holistic well-being in six domains: emotional health, physical health, meaning and purpose, character strengths, social connectedness, and financial security. Although each of these domains is distinct, all of them are nearly universally desired, and all but financial security constitute ends in themselves. Data were collected from a representative sample of working adults. A sample of 276 employees participated in the pilot, 2,370 participated in the first wave and 1,209 in the second wave of the survey. The WBA showed a good fitting (40 items, six factors), satisfactory reliability, test-retest correlation, and convergent/discriminant validity in relation to stability over time and relevant health measures, as well as a good fit to the data that were invariant over time, gender, age, education, and marital status. The instrument can be of use for scientists, practitioners, clinicians, public health officials, and patients. Adoption of more holistic measures of well-being that go beyond psychological well-being may help to shift the focus from health deficiencies to health and well-being promotion.


Generation of mature T cells from human hematopoietic stem and progenitor cells in artificial thymic organoids.
Studies of human T cell development require robust model systems that recapitulate the full span of thymopoiesis, from hematopoietic stem and progenitor cells (HSPCs) through to mature T cells. Existing in vitro models induce T cell commitment from human HSPCs; however, differentiation into mature CD3+TCR-αβ+ single-positive CD8+ or CD4+ cells is limited. We describe here a serum-free, artificial thymic organoid (ATO) system that supports efficient and reproducible in vitro differentiation and positive selection of conventional human T cells from all sources of HSPCs. ATO-derived T cells exhibited mature naive phenotypes, a diverse T cell receptor (TCR) repertoire and TCR-dependent function. ATOs initiated with TCR-engineered HSPCs produced T cells with antigen-specific cytotoxicity and near-complete lack of endogenous TCR Vβ expression, consistent with allelic exclusion of Vβ-encoding loci. ATOs provide a robust tool for studying human T cell differentiation and for the future development of stem-cell-based engineered T cell therapies.


Role of pro- and anti-inflammatory cytokines during inflammation: experimental and clinical findings.
There is an emerging concept that the net biological response of pro and anti-inflammatory cytokines affects the outcome of certain diseases. In inflammatory diseases, interleukin-1 (IL-1) and tumor necrosis factor (TNF) are produced and function primarily as pro-inflammatory cytokines. In addition, other cytokines such as IFN gamma, IL-12 and IL-18 are also produced and affect the production and cellular response to IL-1 and TNF. Biologically, IL-1 and TNF are closely related, although the respective structures and the specific receptors for IL-1 and TNF are clearly distinct. IL-1 and TNF are truly pleiotropic cytokines and are active in the low pM and fM range. Because of their multiple pro-inflammatory properties, these cytokines contribute to disease. Most of our knowledge on IL-1 and TNF are derived from experiments in which either humans or animals have been injected with the cytokine or added to cells in vitro. However, in models of inflammation where several cytokines are produced, specific blockade of either IL-1 or TNF or both results is a reduction in the severity of the inflammation. These latter data therefore implicate IL-1 and TNF as primary cytokines involved with inflammation. Moreover, IL-1 and TNF act synergistically in nearly every in vitro and in vivo model of inflammation. This review will focus on IL-1 and TNF as cytokines of strategic importance to the initiation and progression of inflammation. In addition, this review examines the effects of anti-inflammatory cytokines which non-specifically reduce the production and activity of IL-1 and TNF.


The tyranny of distance: maternity waiting homes and access to birthing facilities in rural Timor-Leste.
To examine the impact of maternity waiting homes on the use of facility-based birthing services for women in two remote districts of Timor-Leste. A before-and-after study design was used to compare the number of facility-based births in women who lived at different distances (0-5, 6-25, 26-50 and > 50 km) from the health centre before and after implementation of maternity waiting homes. Routine data were collected from health centre records at the end of 2007; they included 249 births in Same, Manufahi district, and 1986 births in Lospalos, Lautem district. Population data were used to estimate the percentage of women in each distance category who were accessing facility-based care. Most facility-based births in Same (80%) and Lospalos (62%) were among women who lived within 5 km of the health centre. There was no significant increase in the number of facility-based births among women in more remote areas following implementation of the maternity waiting homes. The percentage of births in the population that occurred in a health facility was low for both Manufahi district (9%) and Lautem district (17%), and use decreased markedly as distance between a woman's residence and the health facilities increased. The maternity waiting homes in Timor-Leste did not improve access to facility-based delivery for women in remote areas. The methods for distance analysis presented in this paper provide a framework that could be used by other countries seeking to evaluate maternity waiting homes.


Kinetics of delta antigen and delta antibody in acute delta hepatitis: evaluation with different enzyme immunoassays.
The kinetics of delta antigen (HDAg) and anti-delta antibody (anti-HD) was analyzed in 22 acute delta hepatitis infections (11 coinfections and 11 superinfections), with an enzyme immunoassay developed by Organon Teknika and with two commercially available assays: Deltassay, a test for HDAg from Noctech and Abbott anti-delta enzyme immunoassay, a test for anti-HD from Abbott Laboratories. In seven cases, HDAg was detected with the Organon assay but not with Deltassay. These discrepancies were not related to the type of hepatitis delta virus infection. All samples from these patients taken beyond week 4 of illness were found anti-HD positive with both the Organon and Abbott anti-HD assays. These data reflect the lack of sensitivity of Deltassay. In no instance were HDAg and anti-HD present simultaneously when tested with the Organon assays. On the contrary, 10 sera among the 28 that were HDAg positive with the Organon assay also were found anti-HD positive with the Abbott test. We suspected that the test procedure recommended by Abbott (a one-step competitive assay) may have yielded false-positive anti-HD results when HDAg present in the sera reacted with peroxidase-labeled anti-HD of the kit. To determine the specificity of the simultaneous presence of HDAg and anti-HD, these 10 sera were retested with the Abbott anti-HD assay, but by a modified two-step procedure that avoided contact between sera and labeled antibody. For six sera a negative result was obtained with the second procedure, suggesting that a false anti-HD reaction occurred with the standard test procedure. For four sera a positive result with both procedures was indicative of the simultaneous presence of HDAg and anti-HD. In conclusion, assay for HDAg was found very convenient for the early diagnosis of acute hepatitis delta virus infections. Seroconversion to anti-HD could be used for a late diagnosis 2 to 5 weeks after the beginning of illness. However, anti-HD results obtained with one-step competitive assays have to be interpreted carefully in HDAg-positive sera.


Prevention and self-treatment of traveler's diarrhea.
Of the millions who travel from the industrialized world to developing countries every year, between 20% and 50% will develop at least one episode of diarrhea, making it the most common medical ailment afflicting travelers. Although usually a mild illness, traveler's diarrhea can result in significant morbidity and hardship overseas. Precautions can be taken to minimize the risk of developing traveler's diarrhea, either through avoidance of potentially contaminated food or drink or through various prophylactic measures, including both nonpharmacological and antimicrobial strategies. If diarrhea does develop despite the precautions taken, effective treatment-usually a combination of an antibiotic and an antimotility agent-can be brought by the traveler and initiated as soon as symptoms develop. In the future, vaccines-several of which are in the advanced stages of clinical testing-may be added to the list of prophylactic measures.


Look youse guys and gals, dat just ain't right.
When I invite authors to submit a punctum to Autophagy, my e-mail includes the following: "Note for international authors: I would like to point out that I personally edit all the puncta for accuracy, but also for English grammar and spelling. I make this point to all international authors as I do not want you to worry extensively about the writing. As a native English speaker, it is easy for me to make small changes of this nature." I do not claim to be an expert in English grammar; however, I am indeed a native English speaker, I read a lot, and I am even fond of using the dictionary (both hard copy and online). Also, I do a lot of editing. Thus, I thought I would share some common mistakes to help reduce the required edits for papers that are submitted to Autophagy.


Global processing fosters donations toward charity appeals framed in an approach orientation.
People are often influenced by how persuasive appeals are framed. While decisions and preferences seem dependent on the effects of a fit between one's regulatory focus and the motivational orientation of a message, specific cognitive mechanisms involved are not yet clear. This study investigated how perceptual processing styles (global vs local) linked with the scope of attention (broad vs narrow) influence decisions depending on motivation-dependent framing (approach vs avoidance). We found that a global processing style fits approach-oriented message appeals and fosters monetary allocation toward charities framed in eager motivational terms. We discuss implications of the findings on processing styles in relation to affective versus deliberate modes of processing and the need to address in detail the role of attentional scope-dependent processing styles in decision making.


Humoral immunodeficiencies: conferred risk of infections and benefits of immunoglobulin replacement therapy.
Primary immunodeficiency (PID) diseases result from genetic defects of the immune system that increase a patient's susceptibility to infections. The types of infections that occur in patients with PID diseases are dictated largely by the nature of the immunodeficiency, which can be defined by dysfunction of cellular or humoral defenses. An increasing number of PID diseases, including those with both cellular and humoral defects, have antibody deficiency as a major feature, and as a result can benefit from immunoglobulin replacement therapy. In fact, the most common PID diseases worldwide are antibody deficiencies and include common variable immunodeficiency, congenital agammaglobulinemia, hyper-IgM syndrome, specific antibody deficiency, and Good syndrome. Although immunoglobulin replacement therapy is the cornerstone of treatment for the majority of these conditions, a thorough understanding of the specific infections for which these patients are at increased risk can hasten diagnosis and guide additional therapies. Moreover, the infection trends in some patients with PID disease who have profound defects of cellular immunity, such as autosomal-dominant hyper-IgE syndrome (Job/Buckley syndrome) or dedicator of cytokinesis 8 (DOCK8) deficiency, suggest that select patients might benefit from immunoglobulin replacement therapy even if their immunodeficiency is not limited to antibody defects. In this review, we provide an overview of the predisposition to infections seen in PID disease that may benefit from immunoglobulin replacement therapy.


Deoxynucleoside Salvage in Fission Yeast Allows Rescue of Ribonucleotide Reductase Deficiency but Not Spd1-Mediated Inhibition of Replication.
In fission yeast, the small, intrinsically disordered protein S-phase delaying protein 1 (Spd1) blocks DNA replication and causes checkpoint activation at least in part, by inhibiting the enzyme ribonucleotide reductase, which is responsible for the synthesis of DNA. The CRL4Cdt2 E3 ubiquitin ligase mediates degradation of Spd1 and the related protein Spd2 at S phase of the cell cycle. We have generated a conditional allele of CRL4Cdt2, by expressing the highly unstable substrate-recruiting protein Cdt2 from a repressible promoter. Unlike Spd1, Spd2 does not regulate deoxynucleotide triphosphate (dNTP) pools; yet we find that Spd1 and Spd2 together inhibit DNA replication upon Cdt2 depletion. To directly test whether this block of replication was solely due to insufficient dNTP levels, we established a deoxy-nucleotide salvage pathway in fission yeast by expressing the human nucleoside transporter human equilibrative nucleoside transporter 1 (hENT1) and the Drosophila deoxynucleoside kinase. We present evidence that this salvage pathway is functional, as 2 µM of deoxynucleosides in the culture medium is able to rescue the growth of two different temperature-sensitive alleles controlling ribonucleotide reductase. However, salvage completely failed to rescue S phase delay, checkpoint activation, and damage sensitivity, which was caused by CRL4Cdt2 inactivation, suggesting that Spd1-in addition to repressing dNTP synthesis-together with Spd2, can inhibit other replication functions. We propose that this inhibition works at the point of the replication clamp proliferating cell nuclear antigen, a co-factor for DNA replication.


Extracorporeal Photopheresis for Second-Line Treatment of Chronic Graft-versus-Host Diseases: Results from a Health Technology Assessment in Italy.
To develop a comparative, cost-effectiveness, and budget impact analysis of Therakos online extracorporeal photopheresis (ECP) compared with the main alternatives used for the treatment of steroid-refractory/resistant chronic graft-versus-host disease (cGvHD) in Italy. The current therapeutic pathway was identified by searching medical databases and from the results of a survey of practice in Italian clinical reference centers. A systematic review was performed to evaluate the efficacy and safety of second-line alternatives. Budget impact and cost-effectiveness analyses were performed from the Italian National Health Service perspective over a 7-year time horizon through the adaption of a Markov model. The following health states were considered: complete and partial response, stable disease, and progression. A discount rate of 3% was applied to costs and outcomes. The most common alternatives used in Italy for the management of steroid-refractory/resistant cGvHD were ECP, mycophenolate, pentostatin, and imatinib. The literature review highlighted that complete and partial responses are higher with ECP than with the alternatives while serious adverse events are less common. The economic analysis showed that Therakos online ECP represents the dominating alternative, in that it delivers greater benefit at a lower cost. In fact, according to the alternatives considered, cost saving ranged from €3237.09 to €19,903.51 per patient with 0.04 to 0.21 quality-adjusted life-year gained. Therakos online ECP should be considered an effective, safe, and cost-effective alternative in steroid-refractory/resistant cGvHD. There is inequality in access, and a dedicated reimbursement tariff, however, should be introduced to overcome these barriers.


Systemic sclerosis: a prototypic multisystem fibrotic disorder.
A unique feature of systemic sclerosis (SSc) that distinguishes it from other fibrotic disorders is that autoimmunity and vasculopathy characteristically precede fibrosis. Moreover, fibrosis in SSc is not restricted to a single organ, but rather affects many organs and accounts for much of the morbidity and mortality associated with this disease. Although immunomodulatory drugs have been used extensively in the treatment of SSc, no therapy to date has been able to reverse or slow the progression of tissue fibrosis or substantially modify the natural progression of the disease. In this Review, we highlight recent studies that shed light on the cellular and molecular mechanisms underlying the fibrotic process in SSc and that identify cellular processes and intra- and extracellular proteins as potential novel targets for therapy in this prototypic multisystemic fibrotic disease.


Vasopressin secretion in normotensive black and white men and women on normal and low sodium diets.
The plasma concentration and the 24-h urinary excretion of vasopressin were studied in normal black and white men and women on a normal sodium (150 mmol/day) diet for 3 days and a low sodium (9 mmol/day; furosemide, 1 mg/kg on first day) diet for 4 days. During the normal sodium diet, the 24-h urinary excretion of vasopressin was significantly (P less than 0.05) higher in men than in women and higher (P less than 0.05) in black than in white subjects. Corresponding differences in plasma vasopressin concentrations (P less than 0.05 to 0.01) were observed, with the exception that the difference between white men and women was not statistically significant. These data suggest that under basal conditions the secretion of vasopressin is higher in men and black subjects than in women and white subjects, although differences in the metabolic clearance of vasopressin cannot be ruled out. Reduction in sodium intake resulted in a significantly (P less than 0.01) decreased excretion of vasopressin in all groups except black women, but had no effect on plasma vasopressin concentrations. Significant (P less than 0.01) differences with respect to sex and race persisted for vasopressin excretion, but not for plasma vasopressin concentrations. Sex- and race-related differences in vasopressin could not be attributed to differences in blood pressure, plasma volume or plasma sodium concentration. The physiological consequences of these differences in vasopressin remain to be determined.


Diminished PKC activity and decreased binding of transcription factors are involved in the impaired production of nitric oxide by macrophages from tumor-bearing mice.
In previous studies we have shown that peritoneal macrophages (PEM) from mammary tumor-bearing BALB/c mice (T-PEM) display a diminished ability to lyse tumor cells upon stimulation with LPS, a phenomenon that is associated to a lower production of nitric oxide, and that is reverted upon costimulation with IFN-gamma. The reduced lytic activity and NO production displayed by T-PEM upon LPS activation were earlier shown by us to be due to a diminished transcription of the inducible nitric oxide synthase (iNOS) gene. In the present study, we have investigated the participation of possible signaling molecules and transcription factors - PKC, NF-kappaB, C/EBP and IRF-1 - in the downregulation of NO production in LPS-activated T-PEM. It was found that PKC activity was greatly reduced in T-PEM as compared to normal macrophages, and did not respond to activation. Interestingly, the different PKC isozyme levels were not significantly altered in T-PEM, with the exception of PKC delta. Alterations in the binding activity of the transcription factors NF-kappaB and C/EBP appeared to be involved in the reduced transcription of iNOS previously observed in T-PEM after LPS activation. These results provide evidence that reductions in iNOS transcription secondary to alterations in cell signaling may be responsible for the diminished capacity of macrophages of LPS-activated tumor-bearers to produce NO and lyse tumor targets.