Differentiation of Osteophytes and Disc Herniations in Spinal Radiculopathy Using Susceptibility-Weighted Magnetic Resonance Imaging.
The aim of this study was to evaluate the diagnostic performance of susceptibility-weighted magnetic resonance imaging (SW-MRI) for the differentiation of osteophytes and disc herniations of the spine compared with that of conventional spine MR sequences and radiography. This study was approved by the local ethics review board; written consent was obtained from all subjects. Eighty-one patients with suspected radiculopathy of the spine were included prospectively. Radiography, T1/T2, and SW-MRI of the cervical/lumbar spine were performed. As reference standard, 93 osteophytes (n = 48 patients) were identified on radiographs in combination with conventional T1/T2 images. One hundred fourteen posterior disc herniations (n = 60 patients) were identified on T1/T2 in combination with radiography excluding osteophytes. For this study, 2 observers independently assessed the presence of osteophytes and disc herniations on T1/T2 and SW-MRI, with radiographs excluded from the analysis. In a subgroup of patients (n = 19), additional computed tomography images were evaluated. Sensitivity, specificity, and interobserver agreement were calculated. Most osteophytes (n = 92 of 93) and disc herniations (n = 113 of 114) could be identified and differentiated on SW-MRI magnitude/phase images, if radiographs were excluded from analysis. Susceptibility-weighted magnetic resonance imaging achieved a sensitivity of 98.9% and specificity of 99.1% for the identification of osteophytes. Conventional T1/T2 spine MR sequences achieved a sensitivity and specificity of 68.6% and 86.5%, respectively, if radiographs were excluded from analysis. Regarding the size of osteophytes, SW-MRI showed a strong correlation with computed tomography (R = 0.96) and radiography (R = 0.95). In addition, SW-MRI achieved a higher interobserver agreement compared with conventional MR. Susceptibility-weighted magnetic resonance imaging enables the reliable differentiation of osteophytes and disc herniations in patients with spinal radiculopathy with a higher sensitivity and specificity compared with conventional T1/T2 MR sequences.


Inspiratory loading and limb locomotor and respiratory muscle deoxygenation during cycling exercise.
The aim of this study was to determine the effect of inspiratory loading on limb locomotor (LM) and respiratory muscle (RM) deoxygenation ([deoxy (Hb+Mb)]) using NIRS during constant-power cycling exercise. Sixteen, male cyclists completed three, 6-min trials. The intensity of the first 3-min of each trial was equivalent to ~80% V(O(2max)) (EX(80%)); during the final 3-min, subjects received an intervention consisting of either moderate inspiratory loading (Load(mod)), heavy inspiratory loading (Load(heavy)), or maximal exercise (Load(EX)). Load(heavy) significantly increased LM [deoxy(Hb+Mb)] from 12.2±9.0 μm during EX(80%) to 15.3±11.7 μm, and RM [deoxy(Hb+Mb)] from 5.9±3.6 μm to 9.5±6.6 μm. LM and RM [deoxy(Hb+Mb)] were significantly increased from EX(80%) to Load(EX); 12.8±9.1 μm to 16.4±10.3 μm and 5.9±2.9 μm to 11.0±6.4 μm, respectively. These data suggest an increase in respiratory muscle load increases muscle deoxy(Hb+Mb) and thus may indicate a reduction in oxygen delivery and/or increased oxygen extraction by the active muscles.


Prior episodic learning and the efficacy of retrieval practice.
In three experiments we investigated how the level of study-based, episodic knowledge influences the efficacy of subsequent retrieval practice (testing) as a learning event. Possibilities are that the efficacy of a test, relative to a restudy control, decreases, increases, or is independent of the degree of prior study-based learning. The degree of study-based learning was manipulated by varying the number of item repetitions in the initial study phase between one and eight. Predictions of the dual-memory model of test-enhanced learning for the case of one study-phase repetition were used as a reference. Results support the hypothesis that the advantage of testing over restudy is independent of the degree of prior episodic learning, and they suggest that educators can apply cued-recall testing with the expectation that its efficacy is similar across varying levels of prior content learning. Implications for testing effect theory are discussed.


Prevalence and Outcomes of Bicuspid Aortic Valve in Patients With Aneurysmal Sub-Arachnoid Hemorrhage: A Prospective Neurology Registry Report.
Background Intracranial aneurysms are reported in 6%-10% of patients with bicuspid aortic valve (BAV), and routine intracranial aneurysm surveillance has been advocated by some. We assessed the prevalence and features of the most important patient-outcome: aneurysmal sub-arachnoid hemorrhage (aSAH), as compared with controls without aSAH, and tricuspid aortic valve (TAV) with aSAH. Methods and Results Adult patients with accurate diagnosis of aSAH and at least one echocardiogram between 2000 and 2019 were identified from a consecutive prospectively maintained registry of aSAH admissions. Controls without a diagnosis of SAH were age- and sex-matched. BAV prevalence was confirmed echocardiographically. Severity of aSAH was categorized using modified Fisher and World Federation of Neurological Scale. Neurologic outcome was assessed using modified Rankin score. A total 488 aSAH cases and 990 controls were identified and BAV status was confirmed. Prevalence of BAV in patients with aSAH was 1.2% (6/488) versus 3.5% (35/990) in controls, P=0.01. BAV+aSAH were noted to be younger than TAV+aSAH (56±11 versus 68±14; P=0.03) with smaller aneurysms (5±2 versus 7±4; P=0.31). The severity of aSAH was lesser in BAV+aSAH than TAV (modified Fisher grade>2 50% versus 74%; P=0.19, World Federation of Neurological Scale grade>3 17% versus 36%; P=0.43). BAV+aSAH had less severe neurologic disability (modified Rankin score 3%-6 33% versus 49% in TAV; P=0.44) and comparable in-hospital mortality rates (P=0.93). BAV had lower odds for aSAH on multivariate analysis (odds ratio 0.23[CI 0.08-0.65]; P=0.01). Conclusions Prevalence of BAV was 3 times lower in the aSAH registry than in controls without aSAH. BAV+aSAH had clinically smaller aneurysms, clinically smaller bleeds, and better neurologic outcome as compared with TAV+aSAH, which needs to be confirmed in larger studies. These findings argue against routine surveillance for intracranial aneurysms in patients with BAV without aortic coarctation.


Revealing Dynamics of Accumulation of Systemically Injected Liposomes in the Skin by Intravital Microscopy.
Accumulation of intravenously injected cytotoxic liposomes in the skin induces serious toxicity. We used single time point and longitudinal intravital microscopy to understand skin accumulation dynamics of non-PEGylated and PEGylated liposomes after systemic injection into mice. Non-PEGylated egg phosphatidylcholine (PC) liposomes showed short circulation half-life (1.3 h) and immediate aggregation in the blood, with some aggregates lodging in skin microvasculature soon after the injection. At 24 h, and more prominently at 48 h postinjection, liposomes appeared in dermal and subdermal cells. PEGylated egg PC liposomes showed long circulation half-life (22 h) and no aggregation in the blood. PEGylated liposomes started to accumulate in the skin microvasculature as soon as 5 min after the injection. Within 3 h postinjection, PEGylated liposomes accumulated in extravascular cells in the dermis and subdermis. Liposomes were present in the skin for at least 7 days postinjection. A regulatory approved PEGylated liposomal doxorubicin (LipoDox) and empty liposomes of the same composition as LipoDox showed similar skin distribution as PEGylated egg PC liposomes, suggesting that this phenomenon is relevant to liposomes of different lipid composition. Decorating liposomes with shorter PEGs (350 or 700) in addition to PEG 2000 did not decrease the deposition. Outside the capillaries, liposomes partially colocalized with CD45-, F4/80+ cells. The accumulation of liposomes was not due to prior neutrophil/platelet binding and transport across endothelium. Moreover, our studies have excluded a role of complement in the skin accumulation of liposomes. Further understanding of mechanisms of this important phenomenon can improve the safety of liposomal nanocarriers.


Evidence synthesis for decision making 1: introduction.
We introduce the series of 7 tutorial papers on evidence synthesis methods for decision making, based on the Technical Support Documents in Evidence Synthesis prepared for the National Institute for Health and Clinical Excellence (NICE) Decision Support Unit. Although oriented to NICE's Technology Appraisal process, which examines new pharmaceutical products in a cost-effectiveness framework, the methods presented throughout the tutorials are equally relevant to clinical guideline development and to comparisons between medical devices, or public health interventions. Detailed guidance is given on how to use the other tutorials in the series, which propose a single evidence synthesis framework that covers fixed and random effects models, pairwise meta-analysis, indirect comparisons, and network meta-analysis, and where outcomes expressed in several different reporting formats can be analyzed without recourse to normal approximations. We describe the principles of evidence synthesis required by the 2008 revision of the NICE Guide to the Methods of Technology Appraisal and explain how the approach proposed in these tutorials was designed to conform to those requirements. We finish with some suggestions on how to present the evidence, the synthesis methods, and the results.


A critical review of electroencephalographic (EEG) correlates of higher cortical functions.
Since Berger's original observations of alpha intensity reductions accompanying mental activity, studies have attempted to elucidate spontaneous (background) EEG correlates of complex mental functions in normal adults. These efforts have failed to develop a model that successfully relates EEG patterns to aspects of cognition. Representative studies are criticized, and guidelines of minimal criteria for conducting research in this area are presented.


Assessing the Impact of Case Management on Medicaid Clients with Chronic Diseases.
State Medicaid programs increasingly use case management to manage enrollees with chronic conditions who may become cost intensive for the program. The authors examined the impact of Alabama Department of Public Health (ADPH)'s case management on care expenditures for Medicaid enrollees with various chronic diseases, over 2011 to 2014. The authors matched case-managed enrollees with three controls per case using health conditions and sociodemographics. Thereafter, the authors used a quasi-experimental approach to estimate how per-member-per-month costs changed following case management. Case -management appeared to result in program savings, driven largely by inpatient and emergency department cost-savings. There was noticeable variation in savings for members with different chronic conditions.


Interorgan crosstalk in pancreatic islet function and pathology.
Pancreatic β cells secrete insulin in response to glucose, a process that is regulated at multiple levels, including a network of input signals from other organ systems. Impaired islet function contributes to the pathogenesis of type 2 diabetes mellitus (T2DM), and targeting inter-organ communications, such as GLP-1 signalling, to enhance β-cell function has been proven to be a successful therapeutic strategy in the last decade. In this review, we will discuss recent advances in inter-organ communication from the metabolic, immune and neural system to pancreatic islets, their biological implication in normal pancreas endocrine function and their role in the (mal)adaptive responses of islet to nutrition-induced stress.


In Vitro Hemocompatibility Evaluation of Ventricular Assist Devices in Pediatric Flow Conditions: A Benchmark Study.
Development of pediatric ventricular assist devices (VADs) has significantly lagged behind that of adult devices. This frustrating reality is reflected by the fact that the Berlin Heart EXCOR VAD is currently the only approved pediatric-specific device in the USA. An alternative option is an off-label use of adult continuous-flow VADs, such as HeartMate II (HMII), which inevitably causes patient-device size mismatch in small children. We sought to conduct in vitro hemocompatibility testing in a pediatric flow condition, with a specific aim to provide benchmark values for future pediatric device development. Given the aforementioned fact that both pulsatile and continuous-flow devices are being used in the pediatric population, we opted to test both types of devices in the present study. The EXCOR and HMII blood pumps were tested using bovine blood under constant hemodynamic conditions (flow rate, Q = 2.5 ± 0.25L/min; differential pressure across the pump, ΔP = 68 ± 5mm Hg). Hemolysis was measured by Harboe assay. There was a steady increase in plasma free hemoglobin during in vitro testing, with a statistically significant difference between 5 and 360 min for both EXCOR (P < 0.0001) and HMII (P < 0.001). However, the degree of an increase in plasma free hemoglobin was more significant with HMII (P < 0.001). Normalized index of hemolysis for EXCOR and HMII were 0.003 ± 0.0026g/100 L and 0.085 ± 0.0119g/100 L, respectively. There was also a steady increase in platelet activation detected by CAPP2A antibody using flow cytometry, with a statistically significant difference between 5 and 360 min for both devices (P < 0.05). The degree of an increase in platelet activation was similar between the two devices (P = 0.218). High molecular weight von Willebrand factor (HMW vWF) multimer degradation measured by immunoblotting was evident for both devices, however, it was more pronounced with the EXCOR. EXCOR blood samples from all three time points (120, 240, and 360 min) were significantly different from the baseline (5 min), whereas only 360 min samples had a significant difference from the baseline with the HMII. In conclusion, we have observed similarities and differences in hemocompatibility profiles between the EXCOR and HMII, both of which are commonly used in the pediatric population. We anticipate the benchmark values in the present study will facilitate future pediatric VAD development.


Predictions and measurements of the stack effect on indoor airborne virus transmission in a high-rise hospital building.
As the viral diseases such as Severe Acute Respiratory Syndrome (SARS) and Influenza A (H1N1) occur in many countries recently, the epidemic of those influenza viruses causes many human casualties. Moreover, the second infection from infected patients particularly within general hospitals frequently takes places due to improperly hospitalized and/or quarantined patients. Accordingly, it becomes a great concern to accommodate safer ventilation system in general hospital wards against such airborne transmitted viruses. It is also a recent trend that many urban general hospitals are designed and constructed as high-rises. If a virus is transmitted through uncontrolled air movement within a hospital and then infected other patients or healthy visitors, it might be impossible to control the spread of the disease. Thus research has been preceded scrutinizing stack effect on the indoor airborne virus transmission in large hospitals by conducting both the field measurement and numerical analysis according to the outdoor temperature and the releasing vertical points of the tracer gas assumed as a viral contaminant. In the field measurement of a high-rise hospital, the indoor airflow was affected by the stack effect of vertical chute of the building. The numerical simulation was verified by comparing its prediction results and the field measurement data. In result, very high possibility has witnessed that the airborne contaminant emitted from the infected patients in the lower floors could be transported to the higher floors through the airflow driven by the stack effect.


Euplotespora binucleata n. gen., n. sp. (Protozoa: Microsporidia), a parasite infecting the hypotrichous ciliate Euplotes woodruffi, with observations on microsporidian infections in ciliophora.
A new microsporidian species, Euplotespora binucleata n. gen., n. sp., from the brackish-water ciliate Euplotes woodruffi is described and defined on the basis of life history characteristics, light and electron microscopic features, and small subunit (SSU) ribosomal DNA (rDNA) sequencing. The life cycle of E. binucleata n. sp. probably has rather short merogonic and relatively long sporogonic phases. Some uninuclear meronts and sporonts, along with diplokaryotic sporoblasts and spores, were found in experimentally infected host cells. Such a peculiar life cycle has been induced experimentally in Euplotes eurystomus and constitutively microsporidian-free stocks of E. woodruffi. Spores of E. binucleata n. sp. are monomorphic, ovoid-cylindrical in shape, 3.44+/-0.17 x 1.65+/-0.22 microm in size, and characterized by a diplokaryotic condition and a large posterior vacuole. The polar tube is isofilar, 4.5-5.5 microm in length when ejected, and lacking a distinctive coiled region (half-coiled). The polaroplast is divided into two regions: the anterior part has a few lamellae close to the anchoring disc; and the posterior part is a rounded body (sack), about one-quarter of the spore length. Spores do not appear to cluster together as a group. Each spore is surrounded by a sporophorous membrane closely adjacent to the exospore layer. A phylogenetic analysis of SSU rDNA sequences by different methods placed E. binucleata n. sp. in a clade with representatives of the microsporidian genera Cystosporogenes and Vittaforma. Observations of microsporidia in several other ciliates are discussed in view of the microsporidian infection frequency in the phylum Ciliophora.


Mechanism of the depressor effect of bromocriptine in the spontaneously hypertensive rat.
Bromocriptine, a dopamine (DA) agonist which passes the bloodbrain barrier, has been shown to have a depressor effect in the spontaneously hypertensive rat of the Okamoto strain. To elucidate the mechanism of this depressor effect, the responses of mean arterial blood pressure and plasma epinephrine (EP), norepinephrine, prolactin and renin activity to i.v. administration of bromocriptine (500 micrograms/kg), alone and after pretreatment with i.v. metoclopramide, a DA antagonist which crosses the blood-brain barrier, and domperidone, a DA antagonist which does not cross the blood-brain barrier, were examined in conscious unrestrained spontaneously hypertensive rats. Metoclopramide attenuated the depressor action of bromocriptine in a dose-related manner, but domperidone had no effect. Neither metoclopramide nor domperidone given alone altered mean arterial blood pressure. Bromocriptine given alone decreased plasma prolactin and increased plasma EP without altering plasma norepinephrine or plasma renin activity. Pretreatment with either metoclopramide or domperidone completely blocked the EP-stimulating effects of bromocriptine. Neither DA antagonist given alone had an effect on plasma EP, norepinephrine or plasma renin activity; both agents stimulated prolactin release. These results suggest that the depressor action of i.v. administered bromocriptine is mediated mainly through a central dopaminergic mechanism rather than by peripheral effects and that plasma EP responses to bromocriptine do not directly contribute to its depressor action.


Absolute Cluster Validity.
The application of clustering involves the interpretation of objects placed in multi-dimensional spaces. The task of clustering itself is inherently submitted to subjectivity, the optimal solution can be extremely costly to discover and sometimes even unreachable or nonexistent. This fact introduces a trade-off between accuracy and computational effort, moreover given that engineering applications usually work well with suboptimal solutions. In such applied scenarios, cluster validation is mandatory to refine algorithms and ensure that solutions are meaningful. Validity indices are commonly intended to benchmark diverse clustering setups, therefore they are coefficients with a relative nature, i.e., useful when compared to one another. In this paper, we propose a validation methodology that enables absolute evaluations of clustering results. Our method performs geometric measurements of the solution space and provides a coherent interpretation of the data structure by using indices based on inter- and intra-cluster distances, density, and multimodality within clusters. Conducted tests and comparisons with well-known indices show that our validation methodology improves the robustness of the clustering application for knowledge discovery. While clustering is often performed as a black box technique, our index is construable and therefore allows for the implementation of systems enriched with self-checking capabilities.


The low impact of fish traps on the seabed makes it an eco-friendly fishing technique.
Besides understanding the effects of fishing on harvested fish stocks, effects on non-target species, habitats and seafloor integrity also need to be considered. Static fishing gears have often been mentioned as a lower impact fishing alternative to towed gears, although studies examining their actual impact on the seafloor are scarce. In this study, we aimed to describe fish trap movements on the seafloor related to soaking time and trap retrieval. Impacts on the seafloor of lightweight rectangular traps and heavier circular traps were compared. We used 3D video cameras to estimate sweeping motion on the seabed and penetration into the sediment during soaking time. The area and distance swept by each type of trap during retrieval was determined by a camera set up facing the sea bottom. The potential rotation of the traps around the mainline was assessed using an Acoustic Doppler Current Profiler. Results showed that no penetration and almost no movements could be detected during soaking time for either lightweight or heavy commercial traps, even for high tidal coefficient (maximum 6 cm). No rotation could be observed when the tide turned. The swept area covered by a trap during retrieval was low (maximum 2.04 m2) compared to towed fishing gear and other static gear.


A novel microdeletion/microduplication syndrome of 19p13.13.
Whole genome interrogation by array-based comparative genomic hybridization has led to a rapidly increasing number of discoveries of novel microdeletion and/or microduplication syndromes. We here describe the clinical and cytogenomic correlates of a novel microdeletion/microduplication of 19p13.13. Among patients referred to the Cytogenetics laboratory for array-based comparative genomic hybridization analysis, we identified four with a deletion and one with a duplication within 19p13.13. Confirmatory fluorescence in situ hybridization and parental studies were performed. Detailed clinical findings and array profiles were reviewed and compared. Patients with deletions of 19p13.13 share a unique constellation of phenotypic abnormalities. In addition to developmental disabilities, the microdeletion manifested in overgrowth, macrocephaly, and ophthalmologic and gastrointestinal findings; in contrast, the single microduplication manifested in growth delay and microcephaly. The consistent constellation of clinical findings associated with copy number variation of this region warrants the designation of microdeletion/microduplication syndrome of 19p13.13. An approximately 311-340 Kb smallest region of overlap encompassing 16 genes was identified. Candidate genes include MAST1, NFIX, and CALR. Identification of this syndrome has led to recommendations for diagnostic work-up and follow-up of patients with this copy number variant. Integration of detailed clinical and array data is critical for advancing both patient care and human genomic research.


Effects of mutagens on the immunogenicity of murine tumor cells: immunological and biochemical evidence for altered cell surface antigens.
Eb lymphoma cells were subjected to treatment in vitro with the alkylating mutagen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and then cloned by limiting dilution. When tested in vivo for tumorigenicity in groups of syngeneic DBA/2 mice, 6 from 18 clones were found to be strongly reduced (tum- phenotype). The other clones showed only moderate or no change in tumorigenicity compared to the untreated control. All clones were able to grow in 400-rad-irradiated mice. Mice in which MNNG clones had regressed were able to generate tumor-specific cytolytic T-lymphocytes in vitro. Limiting dilution analysis indicated that 3 of 4 MNNG clones analyzed in detail displayed additional antigenic determinants that were detected by cytolytic T-lymphocytes. These data thus provided evidence for increased immunogenicity of some of the MNNG clones. Membrane proteins of MNNG clones and original Eb cells were compared biochemically after metabolic labeling with [35S]methionine, TX114 solubilization, and electrophoretic separation. Two-dimensional gel maps revealed a general quantitative decrease in the expression of membrane proteins in MNNG clones. In addition, several proteins were only found in MNNG clones but not in untreated cells. Two membrane proteins of molecular weight 22,000 and 38,000 were greatly increased in expression in all MNNG clones but could be detected at a low level in the original Eb cells. MNNG is known to be a strong mutagenic agent, but it can also interfere with DNA methylation and cause transcriptional activation of genes. We suggest that amplified cell surface structures may be the consequence of such transcriptional activation and could be involved in altered immunogenicity.


Ginkgo biloba extract improves brain uptake of ginsenosides by increasing blood-brain barrier permeability via activating A1 adenosine receptor signaling pathway.
Ginkgo biloba leaves and Panax ginseng are Chinese medicine commonly used in combination for cerebral disease. To investigate the effect of standard extract of Ginkgo biloba leaves (EGb) on facilitating brain uptake of ginsenoside and its underlying mechanisms. The increasing uptake of ginsenosides in the brain of rats by EGb were detected by LC-MS/MS analysis. Evans blue and FITC-dextran leakage were determined to evaluate blood-brain barrier (BBB) permeability in vivo. Transendothelial electrical resistance (TEER) and Na-F penetration rate were measured with a co-culture of the human cerebral microvascular endothelial cell line (hCMEC/D3) and human normal glial cell line (HEB) in vitro BBB model. WB were used to analyzed the expression of BBB tight junctions (TJs) related protein (ZO-1, Occludin, Claudin-3, p-ERM, and p-MLC), ultrastructure of TJs was determined by transmission electron microscope. LC-MS/MS analysis demonstrated that EGb could improve brain uptake of ginsenoside Rg1, Re, Rd and Rb1. In vivo study showed that, BBB permeability was significantly increased after EGb administration, evidenced by the markedly increased penetration of FITC-dextran and Evans Blue into the mice brain parenchyma. In the in vitro BBB model, reduced TEER and increased Na-F penetration rate was observed in EGb group, which was associated with alteration of TJs ultrastructure. Furthermore, the expression of p-ERM and p-MLC in hCMEC/D3 as well as mice brain microvessels were significantly upregulated, but no significant change on the expression of TJs proteins (ZO-1, Occludin and Claudin-3). Moreover, the effect of EGb on in vitro BBB permeability and ERM, MLC phosphorylation was counteracted by DPCPX, an A1 adenosine receptor (A1R) antagonist. EGb might induce ERM/MLC phosphorylation and increase the cell-cell junction gaps to cause a reversible increase of the BBB permeability via A1R signaling pathway. Our results may contribute to better use of EGb in the treatment of brain diseases.


Searching the co-occurrence of pathogenic mutations for Leber's hereditary optic neuropathy and hearing loss in more than 26,000 whole mitochondrial genomes.
The co-occurrence of pathogenic or candidate mutations for Leber's hereditary optic neuropathy (LHON) and hearing loss has long been suggested to be a rare incident. The "rare" is probably caused by inadequate database searches. In this study, we created and released a comprehensive database with detailed information of haplogroup, variants, coding sites, and potential pathogenic mutations for more than 26,000 whole mitochondrial genomes. We found the co-occurrence in more than 200 individuals including not only LHON or hearing loss patients but also individuals sampled from general populations with various haplogroup backgrounds. The results highlighted the significant importance of adequate database searching in the genetic analysis of mitochondrial disorders.


Selective sensitization to Penicillin V with tolerance to other betalactams.
Penicillin G and V have the same betalactam ring. Penicillin V (phenoxymethylpenicillin) results from the substitution of the phenyl acetic acid of benzylpenicillin by the phenoxy methyl side chain. Our patient was a 34-year-old man who experienced generalized urticaria after ingestion of Penicillin V. We performed skin prick tests and intradermal tests with a battery of betalactams including Penicillin V. We also determined specific IgE against penicillin V, penicillin G, amoxicillin, and ampicillin and performed a single-blind oral challenge with Penicillin V, amoxicillin, cefuroxime, and ceftazidime. The results of skin prick and intradermal tests with the betalactams included were negative. Specific IgE with betalactams was < 0.10 IU/L. The result of a single-blind oral challenge with Penicillin V was positive: 40 minutes after receiving 125 mg of Penicillin V, the patient presented generalized pruritus with hives on his back and chest. He tolerated oral administration of amoxicillin, cefuroxime, and ceftazidime. We report an exceptional case of sensitization to Penicillin V with negative results in the allergy workup. Diagnosis was based on a positive single-blind oral challenge result. The patient tolerated other betalactams. We provide a brief summary of the most relevant recent patents.


Three new genera representing novel lineages of Sordariomycetidae (Sordariomycetes, Ascomycota) from tropical freshwater habitats in Costa Rica.
Three new genera are established in the Sordariomycetidae based on morphological and molecular data (SSU and LSU nrDNA) to accommodate five ascomycete species collected from submerged woody debris in freshwater habitats from Costa Rica. The genus Bullimyces contains three new species, B. communis, B. costaricensis and B. aurisporus. Bullimyces is characterized by globose to subglobose, membranous, black, ostiolate ascomata; deliquescent, hyaline, globose cells that fill the center of the centrum; unitunicate asci that deliquesce early in some species; and septate, thick-walled ascospores with or without gelatinous sheaths or appendages. Bullimyces species form a well supported clade with 100% bootstrap support, but the position of the genus in the Sordariomycetidae remains unclear. The second genus, Riomyces, is represented by a single species, R. rotundus. Riomyces is characterized by globose to subglobose, membranous, black, ostiolate ascomata, unitunicate, cylindrical asci, hyaline, globose cells that fill the hamathecium and septate, thick-walled ascospores with a gelatinous sheath. Although Riomyces is morphologically similar to Bullimyces, the two genera did not group together with support in any analysis. The third genus, Hydromelitis, is represented by a single species, H. pulchella. Hydromelitis is characterized by pyriform, membranous, black, ostiolate ascomata, unitunicate asci lacking an apical structure, simple, thin-walled, septate paraphyses and hyaline to golden yellow, multiseptate, thick-walled ascospores with a gelatinous sheath. Bullimyces, Riomyces and Hydromelitis were nested within an unsupported clade consisting of members of the Ophiostomatales, Magnaporthales and freshwater Annulatacaceae sensu lato and sensu stricto.


Near-infrared optogenetic pair for protein regulation and spectral multiplexing.
Multifunctional optogenetic systems are in high demand for use in basic and biomedical research. Near-infrared-light-inducible binding of bacterial phytochrome BphP1 to its natural PpsR2 partner is beneficial for simultaneous use with blue-light-activatable tools. However, applications of the BphP1-PpsR2 pair are limited by the large size, multidomain structure and oligomeric behavior of PpsR2. Here, we engineered a single-domain BphP1 binding partner, Q-PAS1, which is three-fold smaller and lacks oligomerization. We exploited a helix-PAS fold of Q-PAS1 to develop several near-infrared-light-controllable transcription regulation systems, enabling either 40-fold activation or inhibition. The light-induced BphP1-Q-PAS1 interaction allowed modification of the chromatin epigenetic state. Multiplexing the BphP1-Q-PAS1 pair with a blue-light-activatable LOV-domain-based system demonstrated their negligible spectral crosstalk. By integrating the Q-PAS1 and LOV domains in a single optogenetic tool, we achieved tridirectional protein targeting, independently controlled by near-infrared and blue light, thus demonstrating the superiority of Q-PAS1 for spectral multiplexing and engineering of multicomponent systems.


Corpus callosum abnormalities and their association with psychotic symptoms in patients with schizophrenia.
While the neuroanatomical underpinnings of the functional brain disconnectivity observed in patients with schizophrenia (SZ) remain elusive, white matter fiber bundles of the brain are a likely candidate, given that they represent the infrastructure for long-distance neural communication. This study investigated for diffusion abnormalities in 19 patients with chronic SZ, relative to 19 matched control subjects, across tractography-defined segments of the corpus callosum. Diffusion-weighted images were acquired with 51 noncollinear gradients on a 3T scanner (1.7 mm isotropic voxels). The corpus callosum was extracted by means of whole-brain tractography and automated fiber clustering and was parcelled into six segments on the basis of fiber trajectories. The diffusion indexes of fractional anisotropy (FA) and mode were calculated for each segment. Relative to the healthy control subjects, the SZ patients exhibited mode increases in the parietal fibers, suggesting a relative absence of crossing fibers. Schizophrenia patients also exhibited FA reductions in the frontal fibers, which were underpinned by increases in radial diffusivity, consistent with myelin abnormalities. Significant correlations were observed between patients' degree of reality distortion and their FA and radial diffusivity, such that the most severely psychotic patients were the least abnormal in terms of their frontal fiber diffusivity. The SZ patients exhibited a variety of diffusion abnormalities in the corpus callosum, which were related to the severity of their psychotic symptoms. To the extent that diffusion abnormalities influence axonal transmission velocities, these results provide support for those theories that emphasize neural timing abnormalities in the etiology of schizophrenia.


A double-blind, randomized study of olanzapine and olanzapine/fluoxetine combination for major depression with psychotic features.
The purpose of this study was to compare the efficacy and safety of olanzapine (OLZ) monotherapy and an olanzapine/fluoxetine combination (OFC) with placebo (PLA) for unipolar major depression with psychotic features. Under a single protocol, two 8-week, double-blind trials were conducted at 27 sites. Patients (n = 124 trial 1, n = 125 trial 2) were randomized to 1 of 3 treatment groups: OLZ (5 to 20 mg/d), PLA, or OFC (olanzapine 5 to 20 mg/d + fluoxetine 20 to 80 mg/d). The primary outcome measure was the 24-item Hamilton Depression Rating Scale total score. For trial 1, endpoint improvement for the OLZ group (-14.9) was not significantly different from the PLA or OFC groups. The OFC group had significantly greater endpoint improvement (-20.9) than the PLA group (-10.4, P = 0.001); this significant difference was present within 7 days of therapy and maintained at every subsequent visit. The OFC group also had significantly higher response rate (63.6%) than the PLA (28.0%, P = 0.004) or OLZ (34.9%, P = 0.027) groups. For trial 2, there were no significant differences among treatment groups on the 24-item Hamilton Depression Rating Scale total scores or response rates. The combination exhibited a comparable safety profile with OLZ monotherapy and no significant increases in extrapyramidal symptoms compared with placebo. Patients with major depression with psychotic features treated with OLZ monotherapy did not demonstrate significant depressive symptom improvement compared with placebo in either trial; however, an olanzapine/fluoxetine combination was associated with significant improvement compared with placebo in one trial and was well tolerated.


[Functional analysis of mutations on murine chromosome 17 using tertiary trisomy].
Analysis of the functional nature of mutations can be based on their manifestation in organisms with a deletion or a duplication of a particular chromosome segment. With the use of reciprocal translocation T(16;17)43H it is feasible to produce mice with tertiary trisomy for proximal region of chromosome 17. The mutations on chromosome 17 we tested included brachyury (T), hairpin tail (Thp), kinky (Fuki), quaking (qk), tufted (tf), as well as tct (t-complex tail interaction) and tcl (t complex lethal), that are specific for t haplotypes. The set of dominant and recessive mutations was assigned to two groups, one obligatory manifesting itself in the phenotype independently of the number of normal alleles in di- and trisomics, and the other facultative, phenotypically manifesting itself, depending upon the dosage of mutant alleles. A model was derived from analysis of the interaction of the T and Thp mutations with t haplotypes which is to explain the morphogenetic effects of the mutations observed in mice of different genotypes. The tir gene is postulated to reside on chromosome 17 within its framework. It is suggested that the gene dosage ratio at the tir and tct loci determines tail length.


The unique immune ecosystems in pediatric brain tumors: integrating single-cell and bulk RNA-sequencing.
The significant progress of immune therapy in non-central nervous system tumors has sparked interest in employing the same strategy for adult brain tumors. However, the advancement of immunotherapy in pediatric central nervous system (CNS) tumors is not yet on par. Currently, there is a lack of comprehensive comparative studies investigating the immune ecosystem in pediatric and adult CNS tumors at a high-resolution single-cell level. In this study, we comprehensively analyzed over 0.3 million cells from 171 samples, encompassing adult gliomas (IDH wild type and IDH mutation) as well as four major types of pediatric brain tumors (medulloblastoma (MB), ependymoma (EPN), H3K27M-mutation (DIPG), and pediatric IDH-mutation glioma (P-IDH-M)). Our approach involved integrating publicly available and newly generated single-cell datasets. We compared the immune landscapes in different brain tumors, as well as the detailed functional phenotypes of T-cell and myeloid subpopulations. Through single-cell analysis, we identified gene sets associated with major cell types in the tumor microenvironment (gene features from single-cell data, scFes) and compared them with existing gene sets such as GSEA and xCell. The CBTTC and external GEO cohort was used to analyze and validate the immune-stromal-tumor patterns in pediatric brain tumors which might potentially respond to the immunotherapy. From the perspective of single-cell analysis, it was observed that major pediatric brain tumors (MB, EPN, P-IDH-M, DIPG) exhibited lower immune contents compared with adult gliomas. Additionally, these pediatric brain tumors displayed diverse immunophenotypes, particularly in regard to myeloid cells. Notably, the presence of HLA-enriched myeloid cells in MB was found to be independently associated with prognosis. Moreover, the scFes, when compared with commonly used gene features, demonstrated superior performance in independent single-cell datasets across various tumor types. Furthermore, our study revealed the existence of heterogeneous immune ecosystems at the bulk-RNA sequencing level among different brain tumor types. In addition, we identified several immune-stromal-tumor patterns that could potentially exhibit significant responses to conventional immune checkpoint inhibitors. The single-cell technique provides a rational path to deeply understand the unique immune ecosystem of pediatric brain tumors. In spite of the traditional attitudes of "cold" tumor towards pediatric brain tumor, the immune-stroma-tumor patterns identified in this study suggest the feasibility of immune checkpoint inhibitors and pave the way for the upcoming tide of immunotherapy in pediatric brain tumors.


Recent knowledge gained from long-term observations in the field of prosthodontics.
Both cross-sectional and longitudinal studies have contributed data which provide a scientific basis to the prediction of outcome in prosthodontic treatment. 'Standard' cases have a more favourable and more predictable prognosis than those in which tooth loss is scattered about the mouth. However, the variability of patients' problems and the wide range of their abilities to adapt to denture wearing and the regular maintenance of their oral tissues significantly influence outcome. Long term success with partial prostheses lies in the time scale of 5-10 years. A prerequisite for this is a standard of oral hygiene better than practised before the loss of the dentition. Adverse factors are inadequate diagnosis and treatment planning, undetected errors in denture construction, failure of materials, thermal and abrasive effects of foods, behavioural changes in the patient and ageing. In planning the restoration of abutment teeth by inlays or crowns their suitability for attachments must be considered. As yet, there are no clear guidelines to determine when fixed or removable prostheses should be used. The design of partial dentures should be as simple as possible; they should be stable, easy to clean and, if required, easily modified. Success is most likely to attend the efforts of those dentists who use techniques with which they and their technicians are thoroughly familiar.


Understanding the electrical behavior of the action potential in terms of elementary electrical sources.
A concept of major importance in human electrophysiology studies is the process by which activation of an excitable cell results in a rapid rise and fall of the electrical membrane potential, the so-called action potential. Hodgkin and Huxley proposed a model to explain the ionic mechanisms underlying the formation of action potentials. However, this model is unsuitably complex for teaching purposes. In addition, the Hodgkin and Huxley approach describes the shape of the action potential only in terms of ionic currents, i.e., it is unable to explain the electrical significance of the action potential or describe the electrical field arising from this source using basic concepts of electromagnetic theory. The goal of the present report was to propose a new model to describe the electrical behaviour of the action potential in terms of elementary electrical sources (in particular, dipoles). The efficacy of this model was tested through a closed-book written exam. The proposed model increased the ability of students to appreciate the distributed character of the action potential and also to recognize that this source spreads out along the fiber as function of space. In addition, the new approach allowed students to realize that the amplitude and sign of the extracellular electrical potential arising from the action potential are determined by the spatial derivative of this intracellular source. The proposed model, which incorporates intuitive graphical representations, has improved students' understanding of the electrical potentials generated by bioelectrical sources and has heightened their interest in bioelectricity.


Endothelial function and germ-line ACE I/D, eNOS and PAI-1 gene profiles in patients with coronary slow flow in the Canakkale population: multiple thrombophilic gene profiles in coronary slow flow.
We examined the effects of ACE, PAI-1 and eNOS gene polymorphisms on endothelial function. The genes are related to atherosclerosis and endothelial dysfunction in coronary slow flow (CSF). Thirty-three patients with angiographically proven CSF and 48 subjects with normal coronary flow were enrolled in this study. Coronary flow patterns were determined by the thrombolysis in myocardial infarction (TIMI) frame count method. Endothelial function was assessed in the brachial artery by endothelium-dependent flow-mediated dilatation (FMD). PAI-1 4G/5G, eNOS T-786C and ACE I/D polymorphisms were determined by polymerase chain reaction (PCR) amplification. No difference was found between the groups regarding age, heart rate and blood pressure. Males were more prevalent among patients with CSF than control subjects (58.8 vs 29.8%, p = 0.009). Mean TIMI frame counts were significantly higher in CSF patients (24.2 ± 4.0 vs 13.1 ± 2.5 fpm, p = 0.001). FMD was significantly lower in CSF patients than in the controls (4.9 ± 6.6 vs 7.9 ± 5.6%, p = 0.029). TIMI frame count and FMD were found to be negatively correlated in a correlation analysis (r = -0.269, p = 0.015). PAI-1 4G/5G, eNOS T-786C and ACE I/D polymorphisms were similar in the two groups. This study showed that endothelial function was impaired in patients with CSF. PAI-1, ACE and eNOS polymorphisms were not related to CSF in our study population.


RYR3 gene polymorphisms and cardiovascular disease outcomes in the context of antihypertensive treatment.
Nearly one-third of adults in the United States have hypertension, which is associated with increased cardiovascular disease (CVD) morbidity and mortality. The goal of antihypertensive pharmacogenetic research is to enhance understanding of drug response based on the interaction of individual genetic architecture and antihypertensive therapy to improve blood pressure control and ultimately prevent CVD outcomes. In the context of the Genetics of Hypertension Associated Treatment study and using a case-only design, we examined whether single-nucleotide polymorphisms in RYR3 interact with four classes of antihypertensive drugs, particularly the calcium channel blocker amlodipine versus other classes, to modify the risk of coronary heart disease (CHD; fatal CHD and non-fatal myocardial infarction combined) and heart failure (HF) in high-risk hypertensive individuals. RYR3 mediates the mobilization of stored Ca(+2) in cardiac and skeletal muscle to initiate muscle contraction. There was suggestive evidence of pharmacogenetic effects on HF, the strongest of which was for rs877087, with the smallest P-value=0.0005 for the codominant model when comparing amlodipine versus all other treatments. There were no pharmacogenetic effects observed for CHD. The findings reported here for the case-only analysis of the antihypertensive pharmacogenetic effect of RYR3 among 3058 CHD cases and 1940 HF cases show that a hypertensive patient's genetic profile may help predict which medication(s) might better lower CVD risk.


Relationship between timing of emergency procedures and limb amputation in patients with open tibia fracture in the United States, 2003 to 2009.
The authors aimed to characterize patterns in the timing of initial emergency procedures for patients with open tibia fracture and examine the relationship between initial procedure timing and in-hospital amputation. Data were analyzed from the Nationwide Inpatient Sample, 2003 to 2009. Adult patients were included if they had a primary diagnosis code of open tibia fracture. Patients were excluded for the following reasons: they were transferred from or to another hospital, an immediate amputation was performed, more than one amputation was performed, no emergency procedure was documented, or they were treated at a facility that did not perform any amputations. The authors evaluated the association between timing of the first procedure and the outcome of amputation using multiple logistic regression, controlled for patient risk factors and hospital characteristics. Of 7560 patients included in the analysis, 1.3 percent (n=99 patients) underwent amputation on hospital day 2 or later. The majority (52.6 percent) underwent the first operative procedure on day 0 or 1. In adjusted analyses, timing of the first operative procedure beyond the day of admission was associated with more than three times greater odds of amputation (day 1, odds ratio, 3.81; 95 percent CI, 1.80 to 8.07). Delay of the first operative procedure beyond the day of admission appears to be associated with a significantly increased probability of amputation in patients with open tibia fracture. All practitioners involved in the management of these patients should seek a solution for any barrier, other than medical stability of the patient, to achieving early operative intervention. Risk, II.


Molecular properties of F9 embryoglycan recognized by a unique antibody in sera from patients with germ cell tumors.
Human antibody against an embryoglycan present on a mouse teratocarcinoma cell line F9 was found in sera from 16 of 29 patients with embryonal carcinoma, yolk sac tumor, immature teratoma, and choriocarcinoma of gonadal and extragonadal origins by Farr assay. In contrast, none of the sera from patients (77 cases) with dysgerminoma, seminoma, germinoma, and mature teratoma or from patients (118 cases) with nongerm cell types of ovarian tumors contained this antibody. The antigenic embryoglycan was of high molecular weight (Mr greater than 70,000) on Sephacryl S300 column chromatography and carried binding sites for Grifonia simplicifolia agglutinin-1. The antigenic embryoglycan was also found in F9 cell-cultured medium. Absorption of patients' sera with synthetic Blood Group B trisaccharides failed to remove the antibody against F9 embryoglycan. None of these patients' sera showed higher hemagglutination titer to rabbit erythrocytes than the normal range. In contrast, alpha-galactosyl carbohydrates obtained from Ehrlich ascites tumor cells effectively inhibited the binding of patients' sera with F9 embryoglycan. These results indicate that the human antibody against F9 embryoglycan recognizes alpha-galactosyl structures that are distinct from B blood group antigen, but are cross-reactive with alpha-galactosyl structures on Ehrlich ascites cells.


Simultaneous Golgi-Cox and immunofluorescence using confocal microscopy.
Visualization of neuronal elements is of fundamental importance in modern neuroscience. Golgi-Cox impregnation is a widely employed method that provides detailed information about morphological characteristics of neurons, but none regarding their neurochemical features. Immunocytochemical procedures, on the other hand, can provide a high degree of biochemical specificity but poorer morphological details, in particular if compared to Golgi-Cox impregnation. Hence, the combined use of these two approaches is highly desirable, especially for confocal microscopy that can exploit the advantages of both methods simultaneously. Here we show an innovative procedure of perfusion and fixation of brain tissue, that allows, by applying Golgi-Cox impregnation and immunofluorescence in the same histological section, to obtain high-quality histological material, with a very simple and inexpensive method. This procedure is based on three simple fixation steps: (1) a paraformaldehyde perfusion followed by a standard post-fixation to stabilize the subsequent immunofluorescence reaction; (2) the classical Golgi-Cox impregnation and (3) an immunofluorescence reaction in previously impregnated material. This combination allows simultaneous visualization of (a) the structural details (Golgi-Cox impregnated neurons), (b) the antigens' characterization, (c) the anatomical interactions between discrete neuronal elements and (d) the 3D reconstruction and modeling. The method is easy to perform and can be reproducibly applied by small laboratories and expanded through the use of different antibodies. Overall, the method presented in this study offers an innovative and powerful approach to study the nervous system, especially by using confocal microscopy.


Endovascular repair of traumatic descending thoracic aortic disruptions: should endovascular therapy become the gold standard?
Acute arterial disruptions of the thoracic aorta are rare and often lethal. They are most often due to blunt trauma and occur most commonly just distal to the left subclavian artery origin in the setting of multisystem injuries. The very proximal nature of the lesion in survivors makes open surgical repair hazardous, with mortality rates reaching over 20%. Endovascular therapy is a new and attractive option for the treatment of those challenging patients. Since March 2002, 3 patients with blunt and 1 patient with a stable iatrogenic descending aortic injury were successfully treated at this institution. Spiral computed tomography followed by angiography was used for diagnosis. No preexisting aortic pathology was present. Major associated injuries included unstable thoracic spinal fractures, abdominal solid organ injuries, and hip fractures, making the subjects poor surgical candidates. The Talent Endovascular Graft was deployed via open femoral or iliac artery access under fluoroscopic guidance. All 4 patients underwent successful exclusion of their thoracic pseudoaneurysm with use of the Talent endovascular graft. One patient required partial coverage of the left subclavian artery, and a second patient had an iliac artery stent deployed after traversal of an area of stenosis with the delivery system. There were no cardiac, neurologic, pulmonary, or peripheral vascular complications. Acute aortic disruption, with its associated high surgical morbidity and mortality, is an excellent indication for endovascular therapy to improve patient outcomes. More long-term data are needed on repair durability.


Management of nondiagnostic thyroid fine-needle aspiration biopsy: survey of endocrinologists.
To evaluate the approach of endocrinologists in the setting of nondiagnostic thyroid fine-needle aspiration (FNA) biopsies. In 2002, we surveyed physicians attending the national annual meetings of the American Association of Clinical Endocrinologists and the Endocrine Society of North America, using a 13-item questionnaire. The responses were tallied and analyzed. Of the 143 respondents, 139 were endocrinologists, with a male:female ratio of 2.5:1. Most respondents were involved in a medical practice in North America, but Europe, Asia, New Zealand, and Australia were also represented. Of those performing thyroid FNA biopsy, 31% used thyroid ultrasound guidance. Among the survey respondents, 16%, 49%, 20%, and 15% performed less than 2, 2 to 5, 6 to 10, and more than 10 thyroid FNA biopsies per month, respectively. Among the respondents, 13.5%, 44%, 28.5%, 10%, and 4% had non-diagnostic rates of less than 5%, 5 to 10%, 11 to 20%, 21 to 30%, and more than 30%, respectively. The approach of the respondents to an initially nondiagnostic FNA was repeated FNA biopsy in 87%, observation in 7%, levothyroxine suppression in 4%, and thyroid scintigraphy in 2%. Respondents believed that the most cost-effective approach in a patient with nondiagnostic FNA was repeated biopsy (82%), monitoring the size of the thyroid nodule (17%), and surgical referral (<1%). No one was willing to repeat the thyroid biopsy more than three times. On the basis of findings in our survey, most endocrinologists repeat thyroid FNA at least once when confronted with a nondiagnostic result. No published studies have demonstrated the cost-effectiveness of this approach versus proceeding to surgical intervention or observation. We hope that this survey will encourage further studies on this issue.


Biophysical properties of cadherin bonds do not predict cell sorting.
Differential binding between cadherin subtypes is widely believed to mediate cell sorting during embryogenesis. However, a fundamental unanswered question is whether cell sorting is dictated by the biophysical properties of cadherin bonds, or by broader, cadherin-dependent differences in intercellular adhesion or membrane tension. This report describes atomic force microscope measurements of the strengths and dissociation rates of homophilic and heterophilic cadherin (CAD) bonds. Measurements conducted with chicken N-CAD, canine E-CAD, and Xenopus C-CAD demonstrated that all three cadherins cross-react and form multiple, intermolecular bonds. The mechanical and kinetic properties of the heterophilic bonds are similar to the homophilic interactions. The thus quantified bond parameters, together with previously reported adhesion energies were further compared with in vitro cell aggregation and sorting assays, which are thought to mimic in vivo cell sorting. Trends in quantified biophysical properties of the different cadherin bonds do not correlate with sorting outcomes. These results suggest that cell sorting in vivo and in vitro is not governed solely by biophysical differences between cadherin subtypes.


Automated Liver Fat Quantification at Nonenhanced Abdominal CT for Population-based Steatosis Assessment.
Background Nonalcoholic fatty liver disease and its consequences are a growing public health concern requiring cross-sectional imaging for noninvasive diagnosis and quantification of liver fat. Purpose To investigate a deep learning-based automated liver fat quantification tool at nonenhanced CT for establishing the prevalence of steatosis in a large screening cohort. Materials and Methods In this retrospective study, a fully automated liver segmentation algorithm was applied to noncontrast abdominal CT examinations from consecutive asymptomatic adults by using three-dimensional convolutional neural networks, including a subcohort with follow-up scans. Automated volume-based liver attenuation was analyzed, including conversion to CT fat fraction, and compared with manual measurement in a large subset of scans. Results A total of 11 669 CT scans in 9552 adults (mean age ± standard deviation, 57.2 years ± 7.9; 5314 women and 4238 men; median body mass index [BMI], 27.8 kg/m2) were evaluated, including 2117 follow-up scans in 1862 adults (mean age, 59.2 years; 971 women and 891 men; mean interval, 5.5 years). Algorithm failure occurred in seven scans. Mean CT liver attenuation was 55 HU ± 10, corresponding to CT fat fraction of 6.4% (slightly fattier in men than in women [7.4% ± 6.0 vs 5.8% ± 5.7%; P < .001]). Mean liver Hounsfield unit varied little by age (<4 HU difference among all age groups) and only weak correlation was seen with BMI (r2 = 0.14). By category, 47.9% (5584 of 11 669) had negligible or no liver fat (CT fat fraction <5%), 42.4% (4948 of 11 669) had mild steatosis (CT fat fraction of 5%-14%), 8.8% (1025 of 11 669) had moderate steatosis (CT fat fraction of 14%-28%), and 1% (112 of 11 669) had severe steatosis (CT fat fraction >28%). Excellent agreement was seen between automated and manual measurements, with a mean difference of 2.7 HU (median, 3 HU) and r2 of 0.92. Among the subcohort with longitudinal follow-up, mean change was only -3 HU ± 9, but 43.3% (806 of 1861) of patients changed steatosis category between first and last scans. Conclusion This fully automated CT-based liver fat quantification tool allows for population-based assessment of hepatic steatosis and nonalcoholic fatty liver disease, with objective data that match well with manual measurement. The prevalence of at least mild steatosis was greater than 50% in this asymptomatic screening cohort. © RSNA, 2019.


Contrasting migratory response of astrocytoma cells to tenascin mediated by different integrins.
Tenascin, an extracellular matrix protein, is expressed in human gliomas in vitro and in vivo. The distribution of tenascin at the invasive edge of these tumors, even surrounding solitary invading cells, suggests a role for this protein as a regulator of glioma cell migration. We tested whether purified tenascin, passively deposited on surfaces, influenced the adhesion or migration of a human gliomaderived cell line, SF-767. Adhesion of glioma cells to tenascin increased in a dose-dependent fashion up to a coating concentration of 10 micrograms/ml. Higher coating concentrations resulted in progressively fewer cells attaching. Cell adhesion could be blocked to basal levels using anti-beta 1 integrin antibodies. In contrast, when anti-alpha v antibodies were added to the medium of cells on tenascin, cell adhesion was enhanced slightly. Using a microliter scale migration assay, we found that cell motility on tenascin was dose dependently stimulated at coating concentrations of 1 and 3 micrograms/ml, but migration was inhibited below levels of non-specific motility when tested at coating concentrations of 30 and 100 micrograms/ml. Migration on permissive concentrations of tenascin could be reversibly inhibited with anti-beta 1, while treatment with anti-alpha v antibodies increased migration rates. We conclude that SF-767 glioma cells express two separate integrin receptors that mediate contrasting adhesive and migratory responses to tenascin.


Selective recruitment of breast cancer anti-estrogen resistance genes and relevance for breast cancer progression and tamoxifen therapy response.
Although endocrine treatment of breast cancer is effective and common practice, in advanced disease the development of resistance is nearly inevitable. To get more insight into individual genes that account for resistance against hormonal agents, we have executed functional genetic screens and subsequently evaluated the clinical relevance of several identified genes with respect to tumor aggressiveness and tamoxifen resistance in estrogen receptor-positive patients. Estrogen-dependent human breast cancer cells were transduced with different retroviral cDNA expression libraries and subjected to selective cultures with various anti-estrogens. From a total of 264 resistant cell clones, 132 different genes were recovered by PCR. By applying stringent selection criteria, we identified 15 breast cancer anti-estrogen resistance (BCAR) genes individually yielding resistance. BCAR genes were recovered with differential frequencies for the diverse culture conditions and anti-estrogen drugs. Analysis of the relation of BCAR genes (EIF1, FBXL10, HRAS, NRG1, PDGFRA, PDGFRB, RAD21, and RAF1) with tamoxifen treatment in patients with advanced disease showed significant association with clinical benefit and progression-free survival for EIF1 and PDGFRA mRNA levels. Furthermore, PDGFRA and HRAS mRNA levels were significantly associated with tumor aggressiveness in lymph node-negative patients who had not received adjuvant systemic therapy. In conclusion, our functional genetic screens showed that BCAR genes differ in their ability to confer resistance towards distinct anti-estrogens. Based on the clinical relevance of several BCAR genes, further studies are warranted to characterize the underlying mechanisms, which may ultimately lead to the development of novel treatments and more individualized management of breast cancer patients.


Diet: Cause or Consequence of the Microbial Profile of Cholelithiasis Disease?
Recent dietary habits and lifestyle could explain the shaping of the gut microbiota composition and, in consequence, the increasing prevalence of certain pathologies. However, little attention has been paid to the influence of diet on microbiotas, other than the gut microbiota. This is important in cholelithiasis, given that changes in the production of bile acids may affect gallbladder microbial communities. Our aim was to assess the association between regular dietary intake and gallbladder microbial composition. Fourteen adults with cholelithiasis and 14 controls, sex‒age-matched and without gastrointestinal pathology, were included. Diet was assessed through a food frequency questionnaire and quantification of gallbladder microbiota sequences by Illumina 16S rRNA gene-based analysis. The cholelithiasic patients showed greater intake of potatoes and lower consumption of vegetables, non-alcoholic drinks, and sauces, which resulted in a lower intake of energy, lipids, digestible polysaccharides, folate, calcium, magnesium, vitamin C, and some phenolic compounds. Regarding the altered bile microorganisms in cholelithiasic patients, dairy product intake was negatively associated with the proportions of Bacteroidaceae and Bacteroides, and several types of fiber, phenolics, and fatty acids were linked to the abundance of Bacteroidaceae, Chitinophagaceae, Propionibacteraceae, Bacteroides, and Escherichia‒Shigella. These results support a link between diet, biliary microbiota, and cholelithiasis.


[Predisposing factors in candidiasis. Animal experimental studies and clinical findings with special regard to immunity].
In the presence of predisposing factors which are considered as a local or general impairment of the host's defence mechanisms, the opportunistically pathogenic character of Candida albicans transforms into a parasitic one thus favoring the development of candidosis. The clinically reporting part of the paper deals with the predisposing factors and clinical features of candidosis. In animal experiments, we attempted to find out to what extent and whether exclusively the cellular immune system protects the host from Candida albicans infection. Our findings suggest that the cell-mediated immunity and phagocytosis are of fundamental importance in overcoming a candidosis and in resistance to Candida albicans. The humoral immune system plays a minor role, which may be limited to the stimulation of macrophages.


Do practical laboratory indices predict the outcomes of children with Henoch-Schönlein purpura?
Objectives: Henoch Schönlein Purpura (HSP) is the most common systemic vasculitis of childhood and often has a self-limiting course. We aimed to study whether practical laboratory parameters at the diagnosis predict disease course including recurrence and nephritis in addition to severe gastrointestinal involvement in children with HSP. Methods: This retrospective cohort study included 214 HSP patients, 43.5% (n = 93) female and 56.5% (n =121) male, who were diagnosed in our department. Laboratory parameters before treatment, including neutrophil, lymphocyte and platelet counts, mean platelet volume (MPV), neutrophil-to-lymphocyte (NLR), and platelet-to-lymphocyte ratios (PLR) were obtained retrospectively. Age at diagnosis, duration of follow-up, gender, preceding infections, medications, arthritis and arthralgia, abdominal pain, severe GI involvement, invagination, renal involvement and presence of nephritis, outcomes, and presence of recurrences were retrospectively recorded from medical files. Severe GI involvement was determined as severe colicky abdominal pain, bowel edema in ultrasonography or overt GI bleeding. A relapse was defined as a new flare of cutaneous lesions or other manifestations in a patient at least four asymptomatic weeks after the initial HSP episode. Results: Mean age at diagnosis was 7.6 ± 3.1 years. Biopsy-proven nephritis was found in 16 (7.5%) patients. Severe GI involvement was present in 77 (36%) patients, whereas only 12 (5.6%) patients were diagnosed with intussusception and in 29 (13.5%) patients, HSP recurred. Neutrophil count and NLR were found higher in HSP patients with severe gastrointestinal involvement and biopsy-proven nephritis. Additionally, only platelet count was lower and MPV was higher in patients with recurrent HSP. Conclusion: Elevated neutrophil count and NLR may be relevant markers for severe GI involvement and nephritis, whereas platelet count and MPV were the only laboratory parameters associated with disease recurrence.


Chromosome 1p36.22p36.21 duplications/triplication causes Setleis syndrome (focal facial dermal dysplasia type III).
Focal facial dermal dysplasias (FFDD) are characterized by congenital bitemporal or preauricular atrophic skin lesions, and either autosomal dominant or autosomal recessive inheritance. Setleis syndrome (SS), FFDD type III, is a severe form of FFDD with the ectodermal lesions plus other striking facial features. Autosomal recessive nonsense and frameshift mutations in TWIST2 have been found to cause SS in some but not all individuals. Here, we report on four unrelated individuals, one with an unclassified FFDD and the other three with classic SS. Chromosomal microarray analyses revealed unique copy number variants of 1p36 in two individuals with duplications at 1p36.22p36.21 and one with a triplication at 1p36.22p36.21. The fourth patient had normal chromosomes by microarray analysis. All four patients had normal TWIST2 exonic sequences. We propose that a dosage effect of one or more of the 30 genes in the 1.3 Mb 1p36.22p36.21 region of overlap is responsible for FFDD/SS manifestations in some individuals, and this mechanism would be inherited as an autosomal dominant trait. In patients with no duplication/triplication of the 1p36.22p36.21 region and no mutations in TWIST2, there are mutation(s) in one of the 30 genes in this region or mutations in other as yet unidentified genes at different locations that may affect the expressions of genes in this region or act independently to cause this developmental disease phenotype.


Early optimization in finger dexterity of skilled pianists: implication of transcranial stimulation.
It has been shown that non-invasive transcranial direct current stimulation (tDCS) facilitates motor functions in healthy adults and stroke patients. However, little is known about neuroplastic changes induced by tDCS in highly-trained individuals. Here we addressed this issue by assessing the effect of tDCS on dexterity of finger movements in healthy adult pianists. Twelve pianists practiced bimanual keystrokes in an in-phase manner while bilateral tDCS (left anodal/right cathodal or vice versa) of the primary motor cortex was performed. Before and after stimulation, each participant was asked to perform the trained successive keystrokes, and to repetitively strike a key with each of the fingers as fast and accurate as possible while keeping the remaining fingers immobilized voluntarily. In contrast to previous findings in untrained individuals, tDCS yielded overall no apparent improvement of fine control of finger movements in the professional pianists. However, for some movement features, pianists who commenced training at later age demonstrated larger improvements of fine motor control following tDCS. These findings, in combination with lack of any correlation between the age at which pianists commenced the training and motor improvements for sham stimulation conditions, supports the idea that selectively late-started players benefit from tDCS, which we interpret as early optimization of neuroplasticity of the motor system.


Unique honey bee (Apis mellifera) hive component-based communities as detected by a hybrid of phospholipid fatty-acid and fatty-acid methyl ester analyses.
Microbial communities (microbiomes) are associated with almost all metazoans, including the honey bee Apis mellifera. Honey bees are social insects, maintaining complex hive systems composed of a variety of integral components including bees, comb, propolis, honey, and stored pollen. Given that the different components within hives can be physically separated and are nutritionally variable, we hypothesize that unique microbial communities may occur within the different microenvironments of honey bee colonies. To explore this hypothesis and to provide further insights into the microbiome of honey bees, we use a hybrid of fatty acid methyl ester (FAME) and phospholipid-derived fatty acid (PLFA) analysis to produce broad, lipid-based microbial community profiles of stored pollen, adults, pupae, honey, empty comb, and propolis for 11 honey bee hives. Averaging component lipid profiles by hive, we show that, in decreasing order, lipid markers representing fungi, Gram-negative bacteria, and Gram-positive bacteria have the highest relative abundances within honey bee colonies. Our lipid profiles reveal the presence of viable microbial communities in each of the six hive components sampled, with overall microbial community richness varying from lowest to highest in honey, comb, pupae, pollen, adults and propolis, respectively. Finally, microbial community lipid profiles were more similar when compared by component than by hive, location, or sampling year. Specifically, we found that individual hive components typically exhibited several dominant lipids and that these dominant lipids differ between components. Principal component and two-way clustering analyses both support significant grouping of lipids by hive component. Our findings indicate that in addition to the microbial communities present in individual workers, honey bee hives have resident microbial communities associated with different colony components.


No one accelerometer-based physical activity data collection protocol can fit all research questions.
Measuring physical activity and sedentary behavior accurately remains a challenge. When describing the uncertainty of mean values or when making group comparisons, minimising Standard Error of the Mean (SEM) is important. The sample size and the number of repeated observations within each subject influence the size of the SEM. In this study we have investigated how different combinations of sample sizes and repeated observations influence the magnitude of the SEM. A convenience sample were asked to wear an accelerometer for 28 consecutive days. Based on the within and between subject variances the SEM for the different combinations of sample sizes and number of monitored days was calculated. Fifty subjects (67% women, mean ± SD age 41 ± 19 years) were included. The analyses showed, independent of which intensity level of physical activity or how measurement protocol was designed, that the largest reductions in SEM was seen as the sample size were increased. The same magnitude in reductions to SEM was not seen for increasing the number of repeated measurement days within each subject. The most effective way of reducing the SEM is to have a large sample size rather than a long observation period within each individual. Even though the importance of reducing the SEM to increase the power of detecting differences between groups is well-known it is seldom considered when developing appropriate protocols for accelerometer based research. Therefore the results presented herein serves to highlight this fact and have the potential to stimulate debate and challenge current best practice recommendations of accelerometer based physical activity research.


Breast Cancer Screening: Knowledge, Attitudes, and Practices among Female University Students in The Gambia.
Breast cancer is the second most prevalent form of cancer in The Gambia, with an incidence rate of approximately 15% and a mortality rate exceeding 50% in 2020. The all-age prevalence stands at 11.25 per 100,000 population. In light of this, we conducted a study to assess the knowledge, attitude, and practice of breast cancer screening among female university students. We conducted an institutional cross-sectional study involving 361 randomly sampled female university students. Data collection was done using a pretested, self-administered questionnaire. We utilized descriptive statistics to describe the prevalence and burden of breast cancer screening among the participants. Our study revealed good knowledge regarding breast cancer screening among female university students, yet 82.8% had a negative attitude about the disease. More than three-quarters (76.6%) of the respondents had never practiced any form of breast cancer screening. Notably, there was a significant association between knowledge of breast cancer screening and attitude (p = 0.027), and factors such as level of study (p = 0.041), ethnicity (p = 0.026), parity (p = 0.018), and faculty of study (p = <0.001) influenced the participants' knowledge. It is crucial to implement comprehensive awareness campaigns to address the negative attitude and poor screening practices among female university students regarding breast cancer. Additionally, providing free and widespread breast cancer screening services to students should be considered as a means to combat this disease.


Light and electron microscopic distribution of nerve growth factor receptor-like immunoreactivity in the skin of the rat lower lip.
Nerve growth factor receptor distribution in the skin of the adult rat was studied by immunocytochemistry with the use of the monoclonal antibody 192-IgG. Immunoreaction occurred in a patchy pattern in the epidermis and outer root sheaths of hair follicles, as well as in nerves and in capsulated and non-capsulated sensory receptors. Electron microscopic analysis revealed that the areas of patchy epithelial immunostaining corresponded to the plasma membrane of groups of keratinocytes, which were always associated with intra-epithelial nerve fibres. Immunostaining was also associated with Merkel cells, hair follicle receptors, and the capsular cells of capsulated receptors. In the nerves, immunostaining was detected in perineurial cells and axons. After sensory denervation of the skin, immunostaining decreased considerably in the epithelia, as well as in the small nerves, although short segments of increased staining were observed in the latter. In larger nerves, immunostaining was markedly increased and mainly associated with newly formed Schwann cell processes. After sympathetic denervation, the pattern of immunostaining was identical to controls, except for augmented immunoreactivity in short segments of degenerating nerves around blood vessels and smooth muscle fibres. From these observations, it is suggested that the epithelial and perineurial cells synthesize and express nerve growth factor receptors, while receptors present in nerve fibres originate from the nerve cell bodies in the gasserian ganglion.


Alteration of human right bundle branch refractoriness by changes in duration of the atrial drive train.
We studied the effect of drive train duration on human right bundle branch refractoriness. Seven patients with a QRS duration of 0.10 sec or less and no preexcitation had functional right bundle branch block induced reproducibly with premature atrial stimulation. Refractoriness of the right bundle branch was defined as the longest H1H2 interval resulting in right bundle branch block and was determined in each patient at a constant pacing cycle length but at five to seven different atrial drive train durations varying from four to 99 complexes. In all seven patients, right bundle branch refractoriness decreased with increasing drive train duration (mean 15 msec). One-third of this decrease (5.0 msec) occurred between drive train durations of four and eight complexes, and close to two-thirds (9.3 msec) occurred between drive train durations of eight and 64 complexes. Very little further decrease (0.7 msec) occurred between 64 and 99 complexes. We conclude that right bundle branch refractoriness shortens progressively as the preceding drive train duration increases. This phenomenon may in part explain the disappearance of functional right bundle branch block during supraventricular tachycardia after a variable number of complexes without a change in cycle length of tachycardia.


The kinetoplast DNA of Trypanosoma equiperdum.
We have analyzed the kinetoplast DNA for Trypanosoma equiperdum (American Type Culture Collection 30019) and two dyskinetoplastic strains derived from it. The DNA networks from the kinetoplastic strain are made up of catenated mini-circles and maxi-circles, like the networks from the closely-related Trypanosoma brucei. The mini-circles of T. equiperdum lack the pronounced sequence heterogeneity of T. brucei mini-circles, as shown by the fragment distribution of restriction digests and by the predominance of well-matched duplexes in electron micrographs of renatured DNA. The electrophoretic analysis of kinetoplast DNA digested with various restriction endonucleases shows the maxi-circle of T. equiperdum to consist of circular DNA molecules of 8.4 x 10(6) daltons, without size or sequence heterogeneity or repetitious segments. A comparison of the sequence by restriction endonuclease fragmentation and hybridization shows extensive sequence homology. The size difference between both maxi-circles is due to the deletion of one continuous segment of 5.10(6) daltons. In the two dyskinetoplastic strains, we cannot detect DNA sequences that hybridize with kinetoplast DNA from T. brucei or from the kinetoplastic strain of T. equiperdum. In one of these strains, a 'low-density' DNA fraction contained a simple sequence DNA, cleaved by restriction endonuclease HindIII into fragments of 180 base-pairs and multimers of this. The relation of this DNA to kinetoplast DNA, if any, is unknown.


Rapid depletion of B lymphocytes by ultra-low-dose rituximab delivered intrathecally.
We are conducting an open-label phase 1b study on the efficacy of intrathecal (IT) administration of rituximab, provided via an Ommaya reservoir, for the treatment of progressive multiple sclerosis (PMS). The objective of this initial study was to monitor B lymphocytes in peripheral blood (PB) and CSF from the first 10 patients 1 year posttreatment. Dose titration was performed with daily escalation from 1 mg to 25 mg IT rituximab (n = 3). Lymphocyte subpopulations were monitored daily during dose escalation in PB by flow cytometry and subsequently every 3 months for 1 year, after a total dose of 3 × 25 mg. PB B-lymphocyte subpopulations for the remaining patients (n = 7) were monitored at regular intervals. CSF lymphocyte subpopulations for all patients were monitored by flow cytometry every 2-3 months. The PB B-lymphocyte count dropped rapidly after the first 2 injections (total dose of 3.5 mg IT rituximab) to undetectable levels. Three 25-mg doses given once per week depleted peripheral B lymphocytes entirely for the following 3-6 month period. Monoclonal antibodies seem to rapidly redistribute to the peripheral compartment following IT injection. Ultra-low doses of rituximab given IT are sufficient to cause complete depletion of peripheral B lymphocytes, indicating that low-dose IT treatment has the potential to be effective in both the CNS and systemic compartments. This study provides Class IV evidence that for patients with PMS, rituximab provided via an Ommaya reservoir depletes peripheral blood B lymphocytes.


Jail-based medication for opioid use disorder and patterns of reincarceration and acute care use after release: A sequence analysis.
Treatment with methadone and buprenorphine medications for opioid use disorder (MOUD) during incarceration may lead to better community re-entry, but evidence on these relationships have been mixed. We aimed to identify community re-entry patterns and examine the association between in-jail MOUD and a pattern of successful reentry defined by rare occurrence of reincarceration and preventable healthcare utilization. Data came from a retrospective, observational cohort study of 6066 adults with opioid use disorder who were incarcerated in New York City jails and released to the community during 2011-14. An outcome was community re-entry patterns identified by sequence analysis of 3-year post-release reincarceration, emergency department visits, and hospitalizations. An exposure was receipt of in-jail MOUD versus out-of-treatment (42 % vs. 58 %) for the last 3 days before discharge. The study accounted for differences in baseline demographic, clinical, behavioral, housing, and criminal legal characteristics between in-jail MOUD and out-of-treatment groups via propensity score matching. This study identified five re-entry patterns: stability (64 %), hospitalization (23 %), delayed reincarceration (7 %), immediate reincarceration (4 %), and continuous incarceration (2 %). After addressing confounding, 64 % and 57 % followed the stability pattern among MOUD and out-of-treatment groups who were released from jail in 2011, respectively. In 2012-14, the prevalence of following the stability pattern increased year-by-year while a consistently higher prevalence was observed among those with in-jail MOUD. Sequence analysis helped define post-release stability based on health and criminal legal system involvement. Receipt of in-jail MOUD was associated with a marker of successful community re-entry.


The Combination of Electrochemistry and Microfluidic Technology in Drug Metabolism Studies.
Drugs are metabolized within the liver (pH 7.4) by phase I and phase II metabolism. During the process, reactive metabolites can be formed that react covalently with biomolecules and induce toxicity. Identifying and detecting reactive metabolites is an important part of drug development. Preclinical and clinical investigations are conducted to assess the toxicity and safety of a new drug candidate. Electrochemistry coupled to mass spectrometry is an ideal complementary technique to the current preclinical studies, a pure instrumental approach without any purification steps and tedious protocols. The combination of microfluidics with electrochemistry towards the mimicry of drug metabolism offers portability, low volume of reagents and faster reaction times. This review explores the development of microfluidic electrochemical cells for mimicking drug metabolism.


α-Synuclein transfer between neurons and astrocytes indicates that astrocytes play a role in degradation rather than in spreading.
Recent evidence suggests that disease progression in Parkinson's disease (PD) could occur by the spreading of α-synuclein (α-syn) aggregates between neurons. Here we studied the role of astrocytes in the intercellular transfer and fate of α-syn fibrils, using in vitro and ex vivo models. α-Syn fibrils can be transferred to neighboring cells; however, the transfer efficiency changes depending on the cell types. We found that α-syn is efficiently transferred from astrocytes to astrocytes and from neurons to astrocytes, but less efficiently from astrocytes to neurons. Interestingly, α-syn puncta are mainly found inside the lysosomal compartments of the recipient cells. However, differently from neurons, astrocytes are able to efficiently degrade fibrillar α-syn, suggesting an active role for these cells in clearing α-syn deposits. Astrocytes co-cultured with organotypic brain slices are able to take up α-syn fibrils from the slices. Altogether our data support a role for astrocytes in trapping and clearing α-syn pathological deposits in PD.


Activation of a calcium-permeable cation channel by insulin-like growth factor II in BALB/c 3T3 cells.
Insulin-like growth factor II (IGF II) is a member of somatomedin family and is a potent mitogen in various types of mammalian cells. We have recently reported that IGF II stimulates calcium influx in competent BALB/c 3T3 cells primed with epidermal growth factor (J. Biol. Chem. 262: 12120-12126, 1987). Using patch-clamp technique, we show here an IGF II-sensitive cation channel in BALB/c 3T3 cell plasma membrane. Calcium is permeable to this cation channel and its opening behavior is independent of membrane potential. IGF II increases the opening probability of the identical channel in cells pretreated sequentially with platelet-derived growth factor and epidermal growth factor, whereas IGF II does not affect the opening of the channel in G0-arrested cells. This cation channel activity is observed only when IGF II is included in the patch pipette, indicating direct regulation of the channel by IGF II. We suggest that IGF II stimulates calcium influx by opening this cation channel and that IGF II activates the channel in a unique cell cycle-dependent manner.


Temporal analysis of the antigenic composition of Borrelia burgdorferi during infection in rabbit skin.
The numbers of host-adapted Borrelia burgdorferi (HAB) organisms in rabbit skin were assessed by real-time PCR over the first 3 weeks of infection. Maximal numbers were found at day 11, while spirochete numbers decreased by more than 30-fold by day 21. The antigenic composition of HAB in skin biopsy samples was determined by use of a procedure termed hydrophobic antigen tissue Triton extraction. Immune sera from rabbits, sera from chronically infected mice, and monospecific antiserum to the antigenic variation protein, VlsE, were used to probe parallel two-dimensional immunoblots representing each time point. Individual proteins were identified using either specific antisera or by matching protein spots to mass spectrometry-identified protein spots from in vitro-cultivated Borrelia. There were significant changes in the relative expression of a variety of known and previously unrecognized HAB antigens during the 21-day period. OspC and the outer membrane proteins OspA and OspB were prominent at the earliest time point, day 5, when the antigenic variation protein VlsE was barely detected. OspA and OspB were not detected after day 5. OspC was not detected after day 9. VlsE was the most prominent antigen from day 7 through day 21. BmpA, ErpN, ErpP, LA7, OppA-2, DbpA, and an unidentified 15-kDa protein were also detected from day 7 through day 21. Immunoblot analysis using monospecific anti-VlsE revealed the presence of prominent distinct VlsE lower forms in HAB at days 9, 11, and 14; however, these lower forms were no longer detected at day 21. This marked diminution in VlsE lower forms paralleled the clearance of the spirochete from skin.


Mutations in the N- and D-helices of the N-domain of troponin C affect the C-domain and regulatory function.
Troponin C contains a 14-residue alpha-helix at the amino terminus, the N-helix, that calmodulin lacks. Deletion of the first 11-14 residues of troponin C alters function. In the present investigation a mutant lacking residues 1-7 of the N-helix has normal conformation, Ca2+ binding, and regulatory function. Thus, residues 8-14 of the N-helix are generally sufficient for troponin C function. In the x-ray structures of troponin C there is a salt bridge between Arg 11 in the N-helix and Glu 76 in the D-helix. Destroying the salt bridge by individually mutating the residues to Cys has no effect on function. However, mutation of both residues to Cys reduces troponin C's affinity for the troponin complex on the thin filament, reduces the stability of the N-domain in the absence of divalent cations, increases the Ca2+ affinity and reduces the cooperativity of the Ca2+Mg2+ sites in the C-domain, and alters the conformational change that takes place upon Ca2+ binding (but not Mg2+ binding) to the C-domain. Cross-linking with bis-(maleimidomethylether) partially restores function. The Ca2+-specific sites in the N-domain, those closest to the sites of the mutations, are unaffected in the assays employed. These results show that the N-helix is a critical structural element for interaction with and activation of the thin filament. Moreover, mutations in the N-helix affect the C-terminal domain, consistent with recent structural studies showing that the N-helix and C-terminal domain are physically close.


Object-position binding in visual memory for natural scenes and object arrays.
Nine experiments examined the means by which visual memory for individual objects is structured into a larger representation of a scene. Participants viewed images of natural scenes or object arrays in a change detection task requiring memory for the visual form of a single target object. In the test image, 2 properties of the stimulus were independently manipulated: the position of the target object and the spatial properties of the larger scene or array context. Memory performance was higher when the target object position remained the same from study to test. This same-position advantage was reduced or eliminated following contextual changes that disrupted the relative spatial relationships among contextual objects (context deletion, scrambling, and binding change) but was preserved following contextual change that did not disrupt relative spatial relationships (translation). Thus, episodic scene representations are formed through the binding of objects to scene locations, and object position is defined relative to a larger spatial representation coding the relative locations of contextual objects.


Endocrinological responses to 5-HT.
There is increasing evidence that neuroendocrine and temperature responses in humans can be employed to study the functional sensitivity of different 5-HT receptor subtypes. The evidence suggests that the PRL response to LTP is mediated by 5-HT1 receptors, perhaps the 5-HT1A subtype, though further studies are needed to confirm this effect. It is uncertain whether the PRL responses to other 'presynaptic' challenges of 5-HT function, for example, fenfluramine, are mediated by the same post-synaptic 5-HT receptor subtype as that for LTP. Conversely it seems likely that agonists which stimulate 5-HT2/1C receptors increase both plasma PRL and ACTH in humans. There is also evidence that 5-HT1A receptors can increase ACTH secretion. This suggests that in humans as in animals both the 5-HT1A and 5HT2/1C receptors can facilitate ACTH release, though the significance of this dual control is not understood. It is also possible that both 5-HT1A and 5-HT2/1C receptors stimulate PRL release, but 5-HT1A receptors may have a more prominent role in GH secretion. In both human and animal studies 5-HT1A and 5-HT2 receptor agonists may produce opposite effects on body temperature. These recent developments in 5-HT neuroendocrinology have been of great interest, but much is still uncertain. Progress in this field will be considerably advanced by the availability of new selective 5-HT receptor ligands, particularly selective receptor antagonists.


Tracking fungal species-level responses in soil environments exposed to long-term warming and associated drying.
Climate change is affecting fungal communities and their function in terrestrial ecosystems. Despite making progress in the understanding of how the fungal community responds to global change drivers in natural ecosystems, little is known on how fungi respond at the species level. Understanding how fungal species respond to global change drivers, such as warming, is critical, as it could reveal adaptation pathways to help us to better understand ecosystem functioning in response to global change. Here, we present a model study to track species-level responses of fungi to warming -and associated drying- in a decade-long global change field experiment; we focused on two free-living saprotrophic fungi which were found in high abundance in our site, Mortierella and Penicillium. Using microbiological isolation techniques, combined with whole genome sequencing of fungal isolates, and community level metatranscriptomics, we investigated transcription-level differences of functional categories and specific genes involved in catabolic processes, cell homeostasis, cell morphogenesis, DNA regulation and organization, and protein biosynthesis. We found that transcription-level responses were mostly species-specific but that under warming, both fungi consistently invested in the transcription of critical genes involved in catabolic processes, cell morphogenesis, and protein biosynthesis, likely allowing them to withstand a decade of chronic stress. Overall, our work supports the idea that fungi that invest in maintaining their catabolic rates and processes while growing and protecting their cells may survive under global climate change.


Thirtieth anniversary of the UN Convention on the Rights of the Child: advancing a child rights-based approach to child health and well-being.
Global challenges to children's health are rooted in social and environmental determinants. The UN Convention on the Rights of the Child (CRC) articulates the rights required to address these civil-political, social, economic and cultural determinants of child well-being. The principles of child rights-universality, interdependence and accountability-define the tenets of social justice and health equity required to ensure all rights accrue to all children, and the accountability of individuals and organisations (duty-bearers) to ensure these rights are fulfilled. Together, the CRC and child rights principles establish the structure and function of a child rights-based approach (CRBA) to child health and well-being-that provides the strategies and tools to transform child health practice into a rights, justice and equity-based paradigm. The 30th anniversary of the CRC is an opportune time to translate a CRBA to health and well-being into a global practice of paediatrics and child health.