Muscle-specific overexpression of lipoprotein lipase in transgenic mice results in increased alpha-tocopherol levels in skeletal muscle.
Lipoprotein lipase (LPL) has been implicated in the delivery of chylomicron-located alpha-tocopherol (alpha-TocH) to peripheral tissues. To investigate the role of LPL in the cellular uptake of alpha-TocH in peripheral tissue in vivo, three lines of transgenic mice [mouse creatine kinase- (MCK) L, MCK-M and MCK-H] expressing various amounts of human LPL were compared with regard to alpha-TocH levels in plasma, skeletal muscle, cardiac muscle, adipose tissue and brain. Depending on the copy number of the transgene, LPL activity was increased 3- to 27-fold in skeletal muscle and 1.3- to 3.7-fold in cardiac muscle. The intracellular levels of alpha-TocH in skeletal muscle were significantly increased in MCK-M and MCK-H animals and correlated highly with the tissue-specific LPL activity (r = 0.998). The highest levels were observed in MCK-H (21.4 nmol/g) followed by MCK-M (13.3 nmol/g) and MCK-L (8.2 nmol/g) animals when compared with control mice (7.3 nmol/g). Excellent correlation was also observed between intracellular alpha-TocH and non-esterified fatty acid (NEFA) levels (r = 0.998). Although LPL activities in cardiac muscle were also increased in the transgenic mouse lines, alpha-TocH concentrations in the heart remained unchanged. Similarly, alpha-TocH levels in plasma, adipose tissue and brain were unaffected by the tissue specific over-expression of LPL in muscle. The transgenic model presented in this report provides evidence that the uptake of alpha-TocH in muscle is directly dependent on the level of LPL expression in vivo. Increased intracellular alpha-TocH concentrations with increased triglyceride lipolysis and NEFA uptake might protect the myocyte from oxidative damage during increased beta-oxidation.


Simple PCR-based DNA microarray system to identify human pathogenic fungi in skin.
Fungal diseases in immunocompromised hosts pose significant threats to their prognoses. An accurate diagnosis and identification of the fungal pathogens causing the infection are critical to determine the proper therapeutic interventions, but these are often not achieved, due to difficulties with isolation and morphological identification. In an effort to ultimately carry out the simultaneous detection of all human pathogenic microbes, we developed a simple system to identify 26 clinically important fungi by using a combination of PCR amplification and DNA microarray assay (designated PCR-DM), in which PCR-amplified DNA from the internal transcribed spacer region of the rRNA gene was hybridized to a DNA microarray fabricated with species-specific probes sets using the Bubble Jet technology. PCR-DM reliably identified all 26 reference strains; hence, we applied it to cases of onychomycosis, taking advantage of the accessibility of tissue from skin. PCR-DM detected fungal DNA and identified pathogens in 92% of 106 microscopy-confirmed onychomycosis specimens. In contrast, culture was successful for only 36 specimens (34%), 3 of which had results inconsistent with the results of PCR-DM, but sequence analysis of the isolates proved that the PCR-DM result was correct. Thus, PCR-DM provides a powerful method to identify pathogenic fungi with high sensitivity and speed directly from tissue specimens, and this concept could be applied to other fungal or nonfungal infectious human diseases in less accessible anatomical sites.


Adult human neural crest-derived cells for articular cartilage repair.
In embryonic models and stem cell systems, mesenchymal cells derived from the neuroectoderm can be distinguished from mesoderm-derived cells by their Hox-negative profile--a phenotype associated with enhanced capacity of tissue regeneration. We investigated whether developmental origin and Hox negativity correlated with self-renewal and environmental plasticity also in differentiated cells from adults. Using hyaline cartilage as a model, we showed that adult human neuroectoderm-derived nasal chondrocytes (NCs) can be constitutively distinguished from mesoderm-derived articular chondrocytes (ACs) by lack of expression of specific HOX genes, including HOXC4 and HOXD8. In contrast to ACs, serially cloned NCs could be continuously reverted from differentiated to dedifferentiated states, conserving the ability to form cartilage tissue in vitro and in vivo. NCs could also be reprogrammed to stably express Hox genes typical of ACs upon implantation into goat articular cartilage defects, directly contributing to cartilage repair. Our findings identify previously unrecognized regenerative properties of HOX-negative differentiated neuroectoderm cells in adults, implying a role for NCs in the unmet clinical challenge of articular cartilage repair. An ongoing phase 1 clinical trial preliminarily indicated the safety and feasibility of autologous NC-based engineered tissues for the treatment of traumatic articular cartilage lesions.


Analysis of optical vortices with suppressed sidelobes using modified Bessel-like function and trapezoid annulus modulation structures.
Two amplitude modulation methods, including modified Bessel-like function modulation structure and trapezoid annulus structure, for suppressing sidelobes of optical vortices are studied. In the former approach, we propose that the order of the Bessel-like function can be an additional parameter to modulate diffraction patterns of optical vortices motivated by the idea of conventional annulus structures. Furthermore, new Bessel-like modulation functions are introduced to solve the problem of low diffraction efficiency of the original one. Trapezoid annulus structure is proposed as a compromise structure between the modified Bessel-like modulation structure and the conventional annulus one, and has advantages of both. It is demonstrated that these two approaches can achieve high-quality optical vortices with suppressed sidelobes effectively, and the relative structures behave as more flexible and applicable structures for producing optical vortices with large coverage of topological charges, which suggests great potential in simplifying the structure designing procedure. These reliable and generalized structures for generating high-quality optical vortices will help to promote the development of future optical communication and optical manipulation significantly.


Large-scale dewetting assembly of gold nanoparticles for plasmonic enhanced upconversion nanoparticles.
Plasmonic nanostructures have been broadly investigated for enhancing many photophysical properties of luminescent nanomaterials. Precisely controlling the distance between the plasmonic nanostructure and the luminescent material is challenging particularly for the large-scale production of individual nanoparticles. Here we report an easy and reliable method for the large-scale dewetting of plasmonic gold nanoparticles onto core-shell (CS) upconversion nanoparticles (UCNPs). A commensurate NaYF4 shell with a thickness between 5 nm and 15 nm is used as a tunable spacer to control the distance between the UCNP and the plasmonic gold nanoparticles. The upconversion emission intensity of single gold decorated core-inert shell (Au-CS) UCNPs is quantitatively characterized using a scanning confocal microscope. The results demonstrate the highest feasible enhancement of upconversion emission and a record reduction in lifetime for UCNPs fabricated in this manner. The Au-CS UCNPs are further investigated by simulation and synchrotron near edge X-ray absorption fine structure (NEXAFS) analysis.


Association of Widespread Adoption of the 39-Week Rule With Overall Mortality Due to Stillbirth and Infant Death.
To improve neonatal morbidity, efforts have been made to reduce elective deliveries prior to 39 weeks' gestation, also known as the 39-week rule. Prolonging pregnancies also prolongs exposure to the risk of stillbirth. The true association of a 39-week rule with mortality is unknown and studies to date have shown conflicting results. To determine if widespread adoption of a 39-week rule, limiting elective deliveries prior to 39 weeks' gestation, is associated with an increase or decrease in overall mortality when considering both stillbirths and infant deaths. This historical cohort study used birth certificate and infant death certificate data in the United States to compare years before and after the adoption of the 39-week rule. Births between 2008 and 2009 were considered to be in the preadoption period (n = 7 322 234), and those between 2011 and 2012 were considered to be in the postadoption period (n = 6 972 626). Included births were singleton, nonanomalous births between 37 0/7 weeks' and 42 6/7 weeks' gestation. Statistical analysis was performed from July 19, 2016, through June 27, 2019. The exposure of interest was the Joint Commission adoption of the 39-week rule as a quality measure. The primary outcomes of interest were stillbirth and infant death. A total of 7 322 234 births (49.0% girls and 51.0% boys) were included in the preadoption period and 6 972 626 births (49.1% girls and 50.9% boys) were included in the postadoption period. Compared with the preadoption period, there was a decrease in the proportion of deliveries at 37 weeks (-0.06%) and 38 weeks (-2.5%) and an increase in the proportion of deliveries at 39 weeks (6.8%) and 40 weeks (0.2%) in the postadoption period (P < .001). The stillbirth rate increased in the postadoption cohort compared with preadoption (0.09% vs 0.10%; P < .001). The infant death rate decreased in the postadoption period compared with preadoption (0.21% vs 0.20%; P < .001). An overall mortality rate of 0.31% was calculated for the preadoption period and 0.30% for the postadoption period (P = .06). Additional analysis in a counterfactual model suggests that up to 34.2% of the difference in mortality could be associated with the 39-week rule. Stable overall perinatal mortality rates were observed in the 2-year period immediately after adoption of the 39-week rule, despite an increase in stillbirth.


Causal pathways for incident lower-extremity ulcers in patients with diabetes from two settings.
To determine the frequency and constellations of anatomic, pathophysiologic, and environmental factors involved in the development of incident diabetic foot ulcers in patients with diabetes and no history of foot ulcers from Manchester, U.K., and Seattle, Washington, research settings. The Rothman model of causation was applied to the diabetic foot ulcer condition. The presence of structural deformities, peripheral neuropathy, ischemia, infection, edema, and callus formation was determined for diabetic individuals with incident foot ulcers in Manchester and Seattle. Demographic, health, diabetes, and ulcer data were ascertained for each patient. A multidisciplinary group of foot specialists blinded to patient identity independently reviewed detailed abstracts to determine component and sufficient causes present and contributing to the development of each patient's foot ulcer. A modified Delphi process assisted the group in reaching consensus on component causes for each patient. Estimates of the proportion of ulcers that could be ascribed to each component cause were computed. From among 92 study patients from Manchester and 56 from Seattle, 32 unique causal pathways were identified. A critical triad (neuropathy, minor foot trauma, foot deformity) was present in > 63% of patient's causal pathways to foot ulcers. The components edema and ischemia contributed to the development of 37 and 35% of foot ulcers, respectively. Callus formation was associated with ulcer development in 30% of the pathways. Two unitary causes of ulcer were identified, with trauma and edema accounting for 6 and < 1% of ulcers, respectively. The majority of the lesions were on the plantar toes, forefoot, and midfoot. The most frequent component causes for lower-extremity ulcers were trauma, neuropathy, and deformity, which were present in a majority of patients. Clinicians are encouraged to use proven strategies to prevent and decrease the impact of modifiable conditions leading to foot ulcers in patients with diabetes.


Missense mutation clustering in the survival motor neuron gene: a role for a conserved tyrosine and glycine rich region of the protein in RNA metabolism?
The Survival Motor Neuron (SMN) gene shows deletions in the majority of patients with Spinal Muscular Atrophy (SMA), a disease of motor neuron degeneration. To date only two missense mutations have been reported in SMN in patients with SMA. The fact that no SMN-homologues have been forthcoming from data-base searching has resulted in a lack of hypotheses concerning the structural and functional consequences of these mutations. Recently SMN has been shown to interact with heterogeneous nuclear ribonucleoproteins (hnRNPs) suggesting a role in mRNA metabolism. We describe a novel missense mutation and the subsequent identification of a triplicated tyrosine-glycine (Y-G) peptide sequence at the C-terminal of SMN which encompasses each of the three predicted amino acid sequence substitutions. We have identified apparent orthologues of SMN in Caenorhabditis elegans and Schizosaccharomyces pombe. These sequences retain the highly conserved Y-G motif and provide additional support for a role of SMN in mRNA metabolism.


Determinants of Replication-Fork Pausing at tRNA Genes in Saccharomyces cerevisiae.
Transfer RNA (tRNA) genes are widely studied sites of replication-fork pausing and genome instability in the budding yeast Saccharomyces cerevisiae tRNAs are extremely highly transcribed and serve as constitutive condensin binding sites. tRNA transcription by RNA polymerase III has previously been identified as stimulating replication-fork pausing at tRNA genes, but the nature of the block to replication has not been incontrovertibly demonstrated. Here, we describe a systematic, genome-wide analysis of the contributions of candidates to replication-fork progression at tDNAs in yeast: transcription factor binding, transcription, topoisomerase activity, condensin-mediated clustering, and Rad18-dependent DNA repair. We show that an asymmetric block to replication is maintained even when tRNA transcription is abolished by depletion of one or more subunits of RNA polymerase III. By contrast, analogous depletion of the essential transcription factor TFIIIB removes the obstacle to replication. Therefore, our data suggest that the RNA polymerase III transcription complex itself represents an asymmetric obstacle to replication even in the absence of RNA synthesis. We additionally demonstrate that replication-fork progression past tRNA genes is unaffected by the global depletion of condensin from the nucleus, and can be stimulated by the removal of topoisomerases or Rad18-dependent DNA repair pathways.


"Novel p.Tyr284Cys BEST1 genotype-phenotype correlations of Vitelliform Macular Dystrophy in a family with incomplete penetrance".
Purpose: Vitelliform Macular Dystrophy is an inherited autosomal dominant disease with variable expressivity, caused by a mutation in the BEST1 gene. We report a family with variable expressivity and incomplete penetrance in its members.Materials and Methods: A Mexican family was studied. It was comprised of six individuals (father, mother, and four children). A clinical history was taken, and a complete ophthalmological examination (distance best-corrected visual acuity, slit-lamp biomicroscopy, optical coherence tomography, fundus autofluorescence, optical coherence tomography angiography, and electrophysiological studies) was performed in each individual.Results: Two members presented low visual acuity and vitelliform lesions in different stages in the ocular fundus. The assessment suggested a diagnosis of Vitelliform Macular Dystrophy. Genetic analysis was performed by sequencing of exons 2, 4, 5, 7, 8, and 9 of the BEST1 gene. All patients were carriers of the A variant allele of SNP rs1109748 located in exon 2 (c.219 C > A; p.Ile73=). Also, a missense mutation was identified in exon 7 in the mother and two children (c.851A>G; p.Tyr284Cys). The mother has a normal visual acuity, no abnormal findings in the ophthalmological examination and an abnormal electrooculogram, exhibiting incomplete penetrance.Conclusion: This represents one of the few cases of Vitelliform Macular Dystrophy with incomplete penetrance, being the first in our country and Latin America, and with our reported mutation with this characteristic.


Diurnal variation in melatonin effect on adenosine triphosphate and serotonin release by human platelets.
The effect of the pineal hormone melatonin on adenosine diphosphate-induced human platelet aggregation and adenosine triphosphate release was assessed in platelet-rich plasma obtained from normal volunteers at 08.30 and 20.30 h. In 10(-7)-10(-5) mol/l concentrations melatonin inhibited ADP-induced platelet aggregation only in the evening (p less than 0.05). ADP-induced ATP release, an index of platelet secretory processes, showed a generally greater, dose-dependent inhibition after adding melatonin (10(-9)-10(-5)mol/l) at 20.30 h as compared with 08.30 h. The inhibitory activity of melatonin (10(-9)-10(-5) mol/l) on [3H]serotonin release elicited by thrombin in washed human platelets obtained from normal volunteers was dose-dependent; the effect was generally greater at 20.30 h. The activity of the potent platelet anti-aggregating agent prostacyclin did not exhibit diurnal differences with respect to impairing ADP-induced platelet-rich plasma aggregation. These results indicate the existence of a diurnal variation of sensitivity to melatonin in human platelets.


Genome-wide analysis of mutations in mutant lineages selected following fast-neutron irradiation mutagenesis of Arabidopsis thaliana.
Ionizing radiation has long been known to induce heritable mutagenic change in DNA sequence. However, the genome-wide effect of radiation is not well understood. Here we report the molecular properties and frequency of mutations in phenotypically selected mutant lines isolated following exposure of the genetic model flowering plant Arabidopsis thaliana to fast neutrons (FNs). Previous studies suggested that FNs predominantly induce deletions longer than a kilobase in A. thaliana. However, we found a higher frequency of single base substitution than deletion mutations. While the overall frequency and molecular spectrum of fast-neutron (FN)-induced single base substitutions differed substantially from those of "background" mutations arising spontaneously in laboratory-grown plants, G:C>A:T transitions were favored in both. We found that FN-induced G:C>A:T transitions were concentrated at pyrimidine dinucleotide sites, suggesting that FNs promote the formation of mutational covalent linkages between adjacent pyrimidine residues. In addition, we found that FNs induced more single base than large deletions, and that these single base deletions were possibly caused by replication slippage. Our observations provide an initial picture of the genome-wide molecular profile of mutations induced in A. thaliana by FN irradiation and are particularly informative of the nature and extent of genome-wide mutation in lines selected on the basis of mutant phenotypes from FN-mutagenized A. thaliana populations.


Giant synovial cell sarcoma of the thorax in a 46-year-old man: a case report.
Although synovial cell sarcoma is a common tumor of the extremities, its occurrence in the thorax has been less frequently documented. A 46-year-old Pakistani man presented with a 2 month history of progressively increasing cough and left lower chest pain. Initial evaluation was done using a chest x-ray; the patient was found to have a large mass involving the lower portion of the left chest. A computed tomography scan was performed next which showed a large mass involving the left chest wall with invasion into the pericardium and left hemidiaphragm. En bloc surgical resection of the tumor was undertaken. Final pathology showed synovial cell sarcoma of the thorax. At one-year follow-up, the patient has shown no recurrence of the disease. We have described a rare case of a large synovial cell sarcoma of the thorax. Surgical resection appears an appropriate modus operandi for managing giant synovial cell sarcomas of the thorax. However, there is a need to clearly define post-operative strategies for cases with extensive involvement of surrounding structures.


Spatiotemporal EEG analysis and distributed source estimation in presurgical epilepsy evaluation.
In the attempts to localize electric sources in the brain on the basis of multichannel EEG and/or MEG measurements, distributed source estimation procedures have become of increasing interest. Several commercial software packages offer such localization programs and results using these methods are seen more and more frequently in the literature. It is crucial that the users understand the similarities and differences of these methods and that they become aware of the advantages and limitations that are inherent to each approach. This review provides this information from a theoretical as well as from a practical point of view. The theoretical part gives the algorithmic basis of the electromagnetic inverse problem and shows how the different a priori assumptions are formally integrated in these equations. The authors restrict this formalism to the linear inverse solutions i.e., those solutions in which the inversion procedure can be represented as a matrix applied to the data. It will be shown that their properties can be best characterized by their resolution kernels and that methods with optimal resolution matrices can be designed. The authors also discuss the important problem of regularization strategies that are used to minimize the influence of noise. Finally, a new kind of inverse solution, termed ELECTRA (for ELECTRical Analysis), is presented that is based on constraining the source model on the basis of the currents that can actually be measured by the scalp recorded EEG. The practical part of the review illustrates the localization procedures with different clinical data sets. Three aspects become important when working with real data: 1) Clinical data is usually far from ideal (limited number of electrodes, noise, etc.). The behavior of inverse procedures in such unfortunate situations has to be evaluated. 2) The selection of the time points or time periods of interest is crucial, especially in the analysis of spontaneous EEG. 3) Additional information coming from other modalities is usually available and can be incorporated. The authors are illustrating these important points in the case of interictal and ictal epileptiform activity. Spike averaging, frequency domain source localization, and temporal segmentation based on electric field topographies will be discussed. Finally, the technique of EEG-triggered functional magnetic resonance imaging (fMRI) will be illustrated, where EEG is recorded in the magnet and is used to synchronize fMRI acquisition with interictal events. The analysis of both functional data, i.e. the EEG in terms of three-dimensional source localization and the EEG-triggered fMRI, combines the advantages of the two techniques: the temporal resolution of the EEG and the spatial resolution of the fMRI.


Large-scale pancreatic cancer detection via non-contrast CT and deep learning.
Pancreatic ductal adenocarcinoma (PDAC), the most deadly solid malignancy, is typically detected late and at an inoperable stage. Early or incidental detection is associated with prolonged survival, but screening asymptomatic individuals for PDAC using a single test remains unfeasible due to the low prevalence and potential harms of false positives. Non-contrast computed tomography (CT), routinely performed for clinical indications, offers the potential for large-scale screening, however, identification of PDAC using non-contrast CT has long been considered impossible. Here, we develop a deep learning approach, pancreatic cancer detection with artificial intelligence (PANDA), that can detect and classify pancreatic lesions with high accuracy via non-contrast CT. PANDA is trained on a dataset of 3,208 patients from a single center. PANDA achieves an area under the receiver operating characteristic curve (AUC) of 0.986-0.996 for lesion detection in a multicenter validation involving 6,239 patients across 10 centers, outperforms the mean radiologist performance by 34.1% in sensitivity and 6.3% in specificity for PDAC identification, and achieves a sensitivity of 92.9% and specificity of 99.9% for lesion detection in a real-world multi-scenario validation consisting of 20,530 consecutive patients. Notably, PANDA utilized with non-contrast CT shows non-inferiority to radiology reports (using contrast-enhanced CT) in the differentiation of common pancreatic lesion subtypes. PANDA could potentially serve as a new tool for large-scale pancreatic cancer screening.


Using a Faculty-Developed Documentary-Style Film to Communicate Authentic Chemistry Research to a High School Audience.
Described is the creation, deployment, and evaluation of a video produced about the synthesis and applications of metal-organic frameworks (MOFs). The goal of this project was to gauge the impact of viewing the video on high school students' conceptions of authentic chemistry practices and applications. Additionally, comparisons were made between the use of the video and more traditional face-to-face presentations given by professional scientists. Observations, student surveys, and an interview with the high school chemistry teacher demonstrated the utility of such a video. Specifically, the students who viewed the video reported learning more about the nature of laboratory work in chemistry than other students who did not view the video. Students, regardless of whether they viewed the video or just received a presentation, reported growth in understandings of the applications of chemistry research and porous nanomaterial. Other research chemists are encouraged to consider ways that they could document on video the research that they are performing in order to introduce an untapped audience (high school students) to authentic chemistry research in a practically simple manner. During times of crisis, such as a pandemic, online videos could be a useful tool for high school chemistry teachers to use in collaboration with research faculty, particularly when schools are closed.


Integrated optical payload envelope detection and label recovery device for optical packet switching networks.
We demonstrate an integrated device for optical payload envelope detection and optical label recovery for optical packet switching. The device is designed to handle asynchronous serially labeled packets with variable length 40 Gbps payloads preceded by 10 Gbps labels. The device outputs two signals: 1. a payload envelope signal corresponding to the temporal location and duration of the optical payload and 2. an electrical label recovered from the optical label. The payload envelope signal has rise and fall times of 3 ns with 150 ps RMS jitter and is used to perform error free label erasure and rewriting. Error free label recovery is also demonstrated.


Crack Formation Following Root-End Preparation in Roots with the Butterfly Effect.
To investigate apical cracks in roots that exhibit the butterfly effect and that have undergone apical resection and ultrasonic root-end cavity preparation. The effect of the obturation material was also studied. Forty extracted single-rooted teeth were decoronated at the cemento-enamel junction. Roots were viewed under a light microscope and coded according to the presence or absence of the butterfly effect. Canals were prepared using ProTaper Next instruments to size X3 and assigned to two obturation groups (gutta-percha and AH Plus, and ProRoot MTA alone). Each contained twenty roots (10 with the butterfly effect and 10 without the butterfly effect). Roots were resected perpendicular to their long axis, 3 mm from the apex, and cavities were cut using ultrasonic retrotips. Resin replicas were used for crack imaging from scanning electron micrographs. Statistical analyses were performed using Stata 13.1 (StataCorp, College Station, TX, USA). Cracks occurred more frequently in teeth with the butterfly effect (80%), with this difference being significant (P=0.001). Most cracks (73%) ran buccolingually. Teeth obturated with MTA developed fewer cracks compared to those obturated with GP and sealer. Root-ends with the butterfly effect had a significantly higher number of buccolingual cracks following resection and ultrasonic root-end preparation. This might explain the development of some vertical root fractures, which usually run buccolingually. Canal obturation with MTA may be protective.


Induced pluripotent stem cell models from X-linked adrenoleukodystrophy patients.
Because of a lack of an appropriate animal model system and the inaccessibility of human oligodendrocytes in vivo, X-linked adrenoleukodystrophy (X-ALD)-induced pluripotent stem cells (iPSCs) would provide a unique cellular model for studying etiopathophysiology and development of therapeutics for X-ALD. We generated and characterized iPSCs of the 2 major types of X-ALD, childhood cerebral ALD (CCALD) and adrenomyeloneuropathy (AMN), and differentiated them into oligodendrocytes and neurons. We evaluated disease-relevant phenotypes by pharmacological and genetic approaches. We established iPSCs from the patients with CCALD and AMN. Both CCALD and AMN iPSCs normally differentiated into oligodendrocytes, the cell type primarily affected in the X-ALD brain, indicating no developmental defect due to the ABCD1 mutations. Although low in X-ALD iPSCs, very long chain fatty acid (VLCFA) level was significantly increased after oligodendrocyte differentiation. VLCFA accumulation was much higher in CCALD oligodendrocytes than AMN oligodendrocytes but was not significantly different between CCALD and AMN neurons, indicating that the severe clinical manifestations in CCALD might be associated with abnormal VLCFA accumulation in oligodendrocytes. Furthermore, the abnormal accumulation of VLCFA in the X-ALD oligodendrocytes can be reduced by the upregulated ABCD2 gene expression after treatment with lovastatin or 4-phenylbutyrate. X-ALD iPSC model recapitulates the key events of disease development (ie, VLCFA accumulation in oligodendrocytes), provides new clues for better understanding of the disease, and allows for early and accurate diagnosis of the disease subtypes. X-ALD oligodendrocytes can be a useful cell model system to develop new therapeutics for treating X-ALD.


Mating-type genes from asexual phytopathogenic ascomycetes Fusarium oxysporum and Alternaria alternata.
Mating-type (MAT) loci were cloned from two asexual (mitosporic) phytopathogenic ascomycetes, Fusarium oxysporum (a pyrenomycete) and Alternaria alternata (a loculoascomycete), by a polymerase chain reaction (PCR)-based strategy. The conserved high mobility group (HMG) box domain found in the MAT1-2-1 protein was used as a starting point for cloning and sequencing the entire MAT1-2 idiomorph plus flanking regions. Primer pairs designed to both flanking regions were used to amplify the opposite MAT1-1 idiomorph. The MAT1-1 and MAT1-2 idiomorphs were approximately 4.6 and 3.8 kb in F. oxysporum and approximately 1.9 and 2.2 kb in A. alternata, respectively. In both species, the MAT1-1 idiomorph contains at least one gene that encodes a protein with a putative alpha box domain and the MAT1-2 idiomorph contains one gene that encodes a protein with a putative HMG box domain. MAT-specific primers were used to assess the mating type of F. oxysporum and A. alternata field isolates by PCR. MAT genes from A. alternata were expressed. The A. alternata genes were confirmed to be functional in a close sexual relative, Cochliobolus heterostrophus, by heterologous expression.


Enabling Low Cost Biopharmaceuticals: A Systematic Approach to Delete Proteases from a Well-Known Protein Production Host Trichoderma reesei.
The filamentous fungus Trichoderma reesei has tremendous capability to secrete proteins. Therefore, it would be an excellent host for producing high levels of therapeutic proteins at low cost. Developing a filamentous fungus to produce sensitive therapeutic proteins requires that protease secretion is drastically reduced. We have identified 13 major secreted proteases that are related to degradation of therapeutic antibodies, interferon alpha 2b, and insulin like growth factor. The major proteases observed were aspartic, glutamic, subtilisin-like, and trypsin-like proteases. The seven most problematic proteases were sequentially removed from a strain to develop it for producing therapeutic proteins. After this the protease activity in the supernatant was dramatically reduced down to 4% of the original level based upon a casein substrate. When antibody was incubated in the six protease deletion strain supernatant, the heavy chain remained fully intact and no degradation products were observed. Interferon alpha 2b and insulin like growth factor were less stable in the same supernatant, but full length proteins remained when incubated overnight, in contrast to the original strain. As additional benefits, the multiple protease deletions have led to faster strain growth and higher levels of total protein in the culture supernatant.


Towards Personalized Closed-Loop Mechanical CPR: A Model Relating Carotid Blood Flow to Chest Compression Rate and Duration.
There is a growing interest in the personalization of chest compressions to increase blood flow during cardiopulmonary resuscitation (CPR), but there has been very little systematic work to test the feasibility of a closed loop mechanical CPR system. The purpose of this study is to determine if it is possible to model the response of the carotid blood flow to different chest compression waveforms as a function of time during resuscitation from cardiac arrest. This work tests several approaches to predict the carotid blood flow generated by the next chest compression based on knowledge of the duration of resuscitation, the chest compression rate, and the last compression's carotid blood flow. Using an existing physiological database from swine cardiac arrest studies, we computed the features of CPR epoch, compression index, compression rate, and the previous carotid blood flow and used them as the inputs to our model in order to predict carotid blood flow using a Random Forest algorithm. We tested animal specific (estimated with data from a single animal) and global (estimated with data from all but one animals) models for effectiveness. Animal specific models did not generalize when applied to the rest of the animals. The global model performed reasonably well when trained on six animals and tested on the 7th, resulting in errors of 40-160 μL per compression, compared to an average of approximately 400 μL net carotid blood flow per compression in early compressions. In addition, the global model highlighted the inter-animal variability in carotid blood flow generated by identical chest compression waveforms. Generation of probability distribution functions of carotid blood flows suggested at least three different distribution profiles in seven animals. A single physiological metric, carotid blood flow, combined with information about the duration of resuscitation and the compression rate was sufficient to model and predict carotid blood flow in the next compression. This demonstrates that the physiological response to chest compression can be predicted from a relatively modest data set. This suggests that closed loop mechanical CPR is a viable medical device target.


Combining magnetic resonance spectroscopy and molecular genomics offers better accuracy in brain tumor typing and prediction of survival than either methodology alone.
Recent advents in magnetic resonance spectroscopy (MRS) techniques permit subsequent microarray analysis over the entire human transcriptome in the same tissue biopsies. However, extracting information from such immense quantities of data is limited by difficulties in recognizing and evaluating the relevant patterns of apparent gene expression in the context of the existing knowledge of phenotypes by histopathology. Using a quantitative approach derived from a knowledge base of pathology findings, we present a novel methodology used to process genome-wide transcription and MRS data. This methodology was tested to examine metabolite and genome-wide profiles in MRS and RNA in 55 biopsies from human subjects with brain tumors with ~100% certainty. With the guidance of histopathology and clinical outcome, 15 genes with the assistance of 15 MRS metabolites were able to be distinguished by tumor categories and the prediction of survival was better than when either method was used alone. This new method, combining MRS, genomics, statistics and biological content, improves the typing and understanding of the complexity of human brain tumors, and assists in the search for novel tumor biomarkers. It is an important step for novel drug development, it generates testable hypotheses regarding neoplasia and promises to guide human brain tumor therapy provided improved in vivo methods for monitoring response to therapy are developed.


Weight Gain, Executive Functioning, and Eating Behaviors Among Girls.
Executive functioning and excess weight have been associated in cross-sectional and prospective studies, but mechanisms explaining this relationship are unclear. The current study aimed to further explore the longitudinal relationship between executive functioning and changes in body weight and to determine whether binge eating behaviors mediate this relationship. Community-based girls (N = 2450) were assessed by using the behavioral measure of planning, Mazes subtest, and a parent-report measure of impulsivity at age 10; a self-report measure of binge eating at ages 10, 12, and 14; and investigator-measured BMI annually between ages 10 and 16. Regression and bootstrapping analyses explored the relations among age 10 impulsivity and planning, age 12 and age 14 binge eating frequency, and age 10 to 16 BMI changes. Age 10 impulsivity and planning each independently predicted age 10 to 16 BMI changes, after accounting for demographics, verbal comprehension, and BMI at age 10 (Ps < .001). Binge eating tendencies at age 12 mediated the relation between age 10 impulsivity and age 10 to 16 BMI changes, after controlling for demographics, verbal comprehension, binge eating frequency, and BMI at age 10 (indirect effect estimate = 0.0007; 95% confidence interval = 0.0001-0.0020). Results support the hypothesis that poorer executive functioning predicts weight gain from middle childhood through adolescence in girls, and that this effect may be partially explained by binge-eating behaviors in early adolescence.


Gambian children successfully treated with chloroquine can harbor and transmit Plasmodium falciparum gametocytes carrying resistance genes.
Polymorphisms in two genes of Plasmodium falciparum (P. falciparum multidrug resistance 1 [pfmdr1] and P. falciparum chloroquine [CQ] resistance transporter [pfcrt]) are associated with CQ treatment failure. We found significant linkage disequilibrium between these loci among isolates from symptomatic Gambian children (P = 0.026) and strong selection for the resistance-associated alleles pfmdr1-86Tyr and pfcrt-76Thr in children with persistent or re-emerging P. falciparum trophozoites during post-treatment follow-up (P = 1.9 x 10(-7)). Therefore, this genotype is characteristic of resistant infections among our study population. Since the long-term public health impact of parasites carrying such resistant genotypes depends upon their transmissibility, we examined the prevalence of pfmdr1-86Tyr and pfcrt-76Thr among Gambian children harboring sexual stage parasites during post-treatment follow-up. Gametocytes that emerged after successful treatment with CQ were significantly more likely to be of this genotype than were those emerging after other treatments (P = 4.83 x 10(-4)), and were infective to Anopheles mosquitoes. Therapeutic success may thus be accompanied by public health failure as cured children pass resistance genes on to mosquitoes at an enhanced rate.


Sleep disruption in breast cancer patients and survivors.
Sleep disruption is prevalent in patients and survivors of breast cancer. Most patients undergoing chemotherapy will experience transient sleep disruption, and nearly 60% will have chronic sleep problems. Numerous factors contribute to sleep disruption in women diagnosed with breast cancer. Sleep disruption is a consequence of several biological alterations, including circadian disruption and immune and metabolic deregulations. These systems also play significant roles in the control and progression of breast cancer. Sleep disruption is associated with many side effects and psychiatric and medical comorbidities. This article discusses the relationship between stress and posttraumatic stress disorder, depression and fatigue, and how sleep disturbance might be the cause or consequence of these disorders. Current evidence for management of sleep disturbance in breast cancer and high chronic use of hypnotic medication in this population is also discussed. Finally, the differences in management of sleep disturbance during acute cancer care and during the survivorship phase are discussed. More research is needed on accurate and timely assessment of sleep disturbance associated with breast cancer, and additional tailored approaches for the management of sleep problems in breast cancer should be developed.


Striatal and nigral pathology in a lentiviral rat model of Machado-Joseph disease.
Machado-Joseph disease (MJD) is a fatal, dominant neurodegenerative disorder. MJD results from polyglutamine repeat expansion in the MJD-1 gene, conferring a toxic gain of function to the ataxin-3 protein. In this study, we aimed at overexpressing ataxin-3 in the rat brain using lentiviral vectors (LV), to generate an in vivo MJD genetic model and, to study the disorder in defined brain regions: substantia nigra, an area affected in MJD, cortex and striatum, regions not previously reported to be affected in MJD. LV encoding mutant or wild-type human ataxin-3 was injected in the brain of adult rats and the animals were tested for behavioral deficits and neuropathological abnormalities. Striatal pathology was confirmed in transgenic mice and human tissue. In substantia nigra, unilateral overexpression of mutant ataxin-3 led to: apomorphine-induced turning behavior; formation of ubiquitinated ataxin-3 aggregates; alpha-synuclein immunoreactivity; and loss of dopaminergic markers (TH and VMAT2). No neuropathological changes were observed upon wild-type ataxin-3 overexpression. Mutant ataxin-3 expression in striatum and cortex, resulted in accumulation of misfolded ataxin-3, and within striatum, loss of neuronal markers. Striatal pathology was confirmed by observation in MJD transgenic mice of ataxin-3 aggregates and substantial reduction of DARPP-32 immunoreactivity and, in human striata, by ataxin-3 inclusions, immunoreactive for ubiquitin and alpha-synuclein. This study demonstrates the use of LV encoding mutant ataxin-3 to produce a model of MJD and brings evidence of striatal pathology, suggesting that this region may contribute to dystonia and chorea observed in some MJD patients and may represent a target for therapies.


Communications: Evaluation of the nondiabaticity of quantum molecular dynamics with the dephasing representation of quantum fidelity.
We propose an approximate method for evaluating the importance of non-Born-Oppenheimer effects on the quantum dynamics of nuclei. The method uses a generalization of the dephasing representation (DR) of quantum fidelity to several diabatic potential energy surfaces and its computational cost is the cost of dynamics of a classical phase space distribution. It can be implemented easily into any molecular dynamics program and also can utilize on-the-fly ab initio electronic structure information. We test the methodology on three model problems introduced by Tully and on the photodissociation of NaI. The results show that for dynamics close to the diabatic limit, the decay of fidelity due to nondiabatic effects is described accurately by the DR. In this regime, unlike the mixed quantum-classical methods such as surface hopping or Ehrenfest dynamics, the DR can capture more subtle quantum effects than the population transfer between potential energy surfaces. Hence we propose using the DR to estimate the dynamical importance of diabatic, spin-orbit, or other couplings between potential energy surfaces. The acquired information can help reduce the complexity of a studied system without affecting the accuracy of the quantum simulation.


An experimental and computational analysis of primary cilia deflection under fluid flow.
In this study we have developed a novel model of the deflection of primary cilia experiencing fluid flow accounting for phenomena not previously considered. Specifically, we developed a large rotation formulation that accounts for rotation at the base of the cilium, the initial shape of the cilium and fluid drag at high deflection angles. We utilised this model to analyse full 3D data-sets of primary cilia deflecting under fluid flow acquired with high-speed confocal microscopy. We found a wide variety of previously unreported bending shapes and behaviours. We also analysed post-flow relaxation patterns. Results from our combined experimental and theoretical approach suggest that the average flexural rigidity of primary cilia might be higher than previously reported (Schwartz et al. 1997, Am J Physiol. 272(1 Pt 2):F132-F138). In addition our findings indicate that the mechanics of primary cilia are richly varied and mechanisms may exist to alter their mechanical behaviour.


Patterns of calcium-binding proteins support parallel and hierarchical organization of human auditory areas.
The human primary auditory cortex (AI) is surrounded by several other auditory areas, which can be identified by cyto-, myelo- and chemoarchitectonic criteria. We report here on the pattern of calcium-binding protein immunoreactivity within these areas. The supratemporal regions of four normal human brains (eight hemispheres) were processed histologically, and serial sections were stained for parvalbumin, calretinin or calbindin. Each calcium-binding protein yielded a specific pattern of labelling, which differed between auditory areas. In AI, defined as area TC [see C. von Economo and L. Horn (1930) Z. Ges. Neurol. Psychiatr.,130, 678-757], parvalbumin labelling was dark in layer IV; several parvalbumin-positive multipolar neurons were distributed in layers III and IV. Calbindin yielded dark labelling in layers I-III and V; it revealed numerous multipolar and pyramidal neurons in layers II and III. Calretinin labelling was lighter than that of parvalbumin or calbindin in AI; calretinin-positive bipolar and bitufted neurons were present in supragranular layers. In non-primary auditory areas, the intensity of labelling tended to become progressively lighter while moving away from AI, with qualitative differences between the cytoarchitectonically defined areas. In analogy to non-human primates, our results suggest differences in intrinsic organization between auditory areas that are compatible with parallel and hierarchical processing of auditory information.


Prevention of breast cancer skeletal metastases with parathyroid hormone.
Advanced breast cancer is frequently associated with skeletal metastases and accelerated bone loss. Recombinant parathyroid hormone [teriparatide, PTH(1-34)] is the first anabolic agent approved in the US for treatment of osteoporosis. While signaling through the PTH receptor in the osteoblast lineage regulates bone marrow hematopoietic niches, the effects of anabolic PTH on the skeletal metastatic niche are unknown. Here, we demonstrate, using orthotopic and intratibial models of 4T1 murine and MDA-MB-231 human breast cancer tumors, that anabolic PTH decreases both tumor engraftment and the incidence of spontaneous skeletal metastasis in mice. Microcomputed tomography and histomorphometric analyses revealed that PTH increases bone volume and reduces tumor engraftment and volume. Transwell migration assays with murine and human breast cancer cells revealed that PTH alters the gene expression profile of the metastatic niche, in particular VCAM-1, to inhibit recruitment of cancer cells. While PTH did not affect growth or migration of the primary tumor, it elicited several changes in the tumor gene expression profile resulting in a less metastatic phenotype. In conclusion, PTH treatment in mice alters the bone microenvironment, resulting in decreased cancer cell engraftment, reduced incidence of metastases, preservation of bone microarchitecture and prolonged survival.


Less Than Half of Patients Recover Within 2 Weeks of Injury After a Sports-Related Mild Traumatic Brain Injury: A 2-Year Prospective Study.
To describe clinical recovery time and factors that might impact on recovery after a sports-related mild traumatic brain injury (SR-mTBI; concussion). Prospective cohort study (level IV evidence). New Zealand Sports Concussion Clinic. Eight hundred twenty-two patients presenting within 14 days of a SR-mTBI/concussion over a 2-year period. Clinical recovery measured as number of days after injury. Participants were assessed and managed using a standardized protocol consisting of relative rest followed by controlled cognitive and physical loading. A reassessment was performed 14 days after injury with initiation of an active rehabilitation program consisting of a subsymptom threshold exercise program ± cervicovestibular rehabilitation (if required) for participants who remained symptomatic. Participants were then assessed every 2 weeks until clinical recovery. A total of 594 participants were eligible for analysis (mean age 20.2 ± 8.7 years, 77% males) and were grouped into 3 age cohorts: children (≤12 years), adolescents (13-18 years), and adults (≥19 years). Forty-five percent of participants showed clinical recovery within 14 days of injury, 77% by 4 weeks after injury, and 96% by 8 weeks after injury. There was no significant difference in recovery time between age groups. Prolonged recovery was more common in females (P = 0.001), participants with "concussion modifiers" (P = 0.001), and with increased time between injury and the initial appointment (P = 0.003). This study challenges current perceptions that most people with a SR-mTBI (concussion) recover within 10 to 14 days and that age is a determinant of recovery rate. Active rehabilitation results in high recovery rates after SR-mTBI.


Upregulation of PBP1B and LpoB in cysB Mutants Confers Mecillinam (Amdinocillin) Resistance in Escherichia coli.
Mecillinam (amdinocillin) is a β-lactam antibiotic that inhibits the essential penicillin-binding protein 2 (PBP2). In clinical isolates of Escherichia coli from urinary tract infections, inactivation of the cysB gene (which encodes the main regulator of cysteine biosynthesis, CysB) is the major cause of resistance. How a nonfunctional CysB protein confers resistance is unknown, however, and in this study we wanted to examine the mechanism of resistance. Results show that cysB mutations cause a gene regulatory response that changes the expression of ∼450 genes. Among the proteins that show increased levels are the PBP1B, LpoB, and FtsZ proteins, which are known to be involved in peptidoglycan biosynthesis. Artificial overexpression of either PBP1B or LpoB in a wild-type E. coli strain conferred mecillinam resistance; conversely, inactivation of either the mrcB gene (which encodes PBP1B) or the lpoB gene (which encodes the PBP1B activator LpoB) made cysB mutants susceptible. These results show that expression of the proteins PBP1B and LpoB is both necessary and sufficient to confer mecillinam resistance. The addition of reducing agents to a cysB mutant converted it to full susceptibility, with associated downregulation of PBP1B, LpoB, and FtsZ. We propose a model in which cysB mutants confer mecillinam resistance by inducing a response that causes upregulation of the PBP1B and LpoB proteins. The higher levels of these two proteins can then rescue cells with mecillinam-inhibited PBP2. Our results also show how resistance can be modulated by external conditions such as reducing agents.


Glassy dynamics in models of confluent tissue with mitosis and apoptosis.
Recent work on particle-based models of tissues has suggested that any finite rate of cell division and cell death is sufficient to fluidize an epithelial tissue. At the same time, experimental evidence has indicated the existence of glassy dynamics in some epithelial layers despite continued cell cycling. To address this discrepancy, we quantify the role of cell birth and death on glassy states in confluent tissues using simulations of an active vertex model that includes cell motility, cell division, and cell death. Our simulation data is consistent with a simple ansatz in which the rate of cell-life cycling and the rate of relaxation of the tissue in the absence of cell cycling contribute independently and additively to the overall rate of cell motion. Specifically, we find that a glass-like regime with caging behavior indicated by subdiffusive cell displacements can be achieved in systems with sufficiently low rates of cell cycling.


Speech dominance is a better predictor of functional brain asymmetry than handedness: a combined fMRI word generation and behavioral dichotic listening study.
An unresolved issue in behavioral studies of hemispheric asymmetry is why both left-handers and right-handers show a right ear advantage at the group level. In the present study we screened left-handers for left- versus right-hemisphere speech dominance with fMRI by comparing right versus left hemisphere frontal lobe activity (in Broca's area) in a silent word generation task. A left hemisphere dominant right-handed control group was included as well. All participants took part in a dichotic listening task with consonant-vowel syllables. The results showed that left-handers and right-handers with left-hemisphere speech dominance showed a right ear advantage. However, the left-handers with right hemisphere speech dominance had a left ear advantage. Thus, at the group level the direction of the ear advantage in dichotic listening was predicted by language dominance but not by hand preference. At the individual level, the dichotic task we used showed more variability than the fMRI results. Further research will have to indicate whether this is a feature inherent to dichotic listening, or whether the variability is due to alternative explanations such as a more bilateral representation of speech perception compared to speech production.


Distinct behavioral and EEG topographic correlates of loss of consciousness in absences.
To describe the behavioral and EEG topographic correlates of absences with 3-Hz generalized spike-waves and partitioned impairment of consciousness. Two adult women had so-called "phantom" absences, characterized by brief and mild impairments of consciousness that were previously inconspicuous to both patient and physician. Neuropsychological examination was performed under video-EEG monitoring during absence status. EEG topographic mapping of spike-wave discharges was obtained in the two cases. Only mild attentional and executive disturbances were observed during absence status despite prolonged discharges. Spike-wave bursts were associated with selective impairment in the initiation of response and self-generated action, whereas short-term storage of external information during discharges was fully preserved. This is consistent with a predominant involvement of frontomesial cortex demonstrated by topographic mapping of spike-wave discharges in the two cases. By contrast, in two other patients with typical absences and a complete lack of retention for information given during the discharges, topographic mapping found a more lateral frontal involvement by spike-wave activity. Different types of absence seizures may impair distinct components of conscious behavior. A predominant involvement of frontomesial thalamocortical circuitry may underlie an "inconspicuous" disorder of consciousness as seen in phantom absences with selective loss of initiation and goal-oriented behavior, whereas involvement of more lateral frontal areas in typical absences may additionally disrupt working memory processes.


Catechol/polyethyleneimine conversion coating with enhanced corrosion protection of magnesium alloys: potential applications for vascular implants.
Magnesium (Mg) alloys are promising biodegradable materials but challenges remain due to their rapid degradation, especially in the potential use of Mg alloys as vascular stents. Surface modification techniques are the most straightforward way to address both the desired biocompatibility and inhibit the corrosion of Mg alloys. In this work, inspired by the functional moieties (catechols) of mussel adhesive proteins, a mimetic approach to construct organic protective conversion coatings on magnesium-zinc-manganese (MgZnMn) alloys is investigated. Based on the cross-linking of CA (catechol) and PEI (polyethyleneimine), a CA/PEI conversion coating is developed on a MgZnMn alloy. The CA/PEI conversion coating showed enhanced corrosion resistance due to the strong binding and aggregation of units. Moreover, such coatings could also provide enough primary amine groups, catechols and quinones, which can be used to immobilize further molecules. Heparin was further grafted onto the CA/PEI, endowing the conversion coating with the desired functionality. Vascular cell behavior on such a coating is also studied. The improved hemocompatibility, favorable anti-inflammatory ability, suppressed smooth muscle cell proliferation and enhanced endothelialization indicate the potential of such organic conversion coatings on MgZnMn alloys as vascular implants.


Isolated multiple lumbar transverse process fractures with spinal instability: an uncommon yet serious association.
Isolated vertebral transverse process fractures of thoracolumbar spine without other vertebral injuries and neurological deficit are generally considered as minor injuries with no concern for associated spinal instability. This report describes a case of multiple lumbar transverse fractures associated with an unexpected yet clinically significant spinal instability. A young male presented with right flank pain following being pushed and trapped against the ground by a reversing truck. The neurological examination was normal, and computed tomography (CT) imaging revealed multiple fractures at right transverse processes from L1 to L5, a single left-sided transverse process fracture at L2 and subtle facet joint distraction without other spinal lesions or visceral injuries. The injury was initially deemed as stable requiring symptomatic treatment and in-patient observation. However, discharge upright X-rays taken in a brace showed marked subluxation of L2/L3 and L3/L4 levels. Magnetic resonance imaging revealed significant discoligamentous injuries involving anterior and posterior longitudinal ligaments, annulus fibrosus as well as posterior ligamentous complex. The patient underwent posterior spinal instrumentation and fusion of L1 to L5. This is the first case description of association of multisegmental lumbar transverse process fractures with notoriously unstable injuries of the major soft-tissue stabilizers of the spine presenting subtle changes on CT images. When a seemingly benign spinal injury is caused by high-energy trauma, careful scrutiny for associated instability is needed. In this case, the standing in-brace X-ray was able to avoid a misdiagnosis and potentially unfavourable outcome.


Nanobubble-embedded inorganic 808 nm excited upconversion nanocomposites for tumor multiple imaging and treatment.
Contrast reagents for ultrasound imaging are widely used in clinical medical diagnosis because ultrasound resolution is limited. Contrast agents must be utilized to enhance the image resolution. At present, microbubbles (MBs) and nanobubbles (NBs) are the main contrast reagent candidates for improving the signal resolution. Fluorescence upconversion nanoparticles provide high sensitivity and also function as nanocarriers. This can label tumor cells in a specific organ under irradiation of near-infrared (NIR) light. However, despite the use of NIR light, the penetration depth of NIR is only approximately 15 mm. Thus, we combine fluorescence with ultrasonic imaging to achieve the effect of multiple imaging and solve the low penetration depth of fluorescence imaging and the poor resolution of ultrasound imaging. The dual imaging modalities achieved higher resolution or signal to noise ratios. In this study, Nd3+-sensitized upconversion nanoparticles (UCNPs) are combined with graphitic carbon nitride quantum dots (CNs) and embedded in NBs (UCNP-CN@NBs). The UCNPs are excited by 808 nm light and emit visible and ultraviolet light. Then, the energy of the ultraviolet light is transferred to the CNs to produce reactive oxygen species (ROS) for photodynamic therapy. Ultrasonic waves are also used to promote NB bursting and the release of ROS molecules in photodynamic therapy, leading to cancer cell apoptosis.


The Novel Single-Stroke Kayak Test: Can It Discriminate Between 200-m and Longer-Distance (500- and 1000-m) Specialists in Canoe Sprint?
To test whether the force-velocity (F-V) relationship obtained during a specific single-stroke kayak test (SSKT) and during nonspecific traditional resistance-training exercises (bench press and prone bench pull) could discriminate between 200-m specialists and longer-distance (500- and 1000-m) specialists in canoe sprint. A total of 21 experienced male kayakers (seven 200-m specialists and 14 longer-distance specialists) participated in this study. After a familiarization session, kayakers came to the laboratory on 2 occasions separated by 48 to 96 hours. In a randomized order, kayakers performed the SSKT in one session and the bench press and bench pull tests in another session. Force and velocity outputs were recorded against 5 loads in each exercise to determine the F-V relationship and related parameters (maximum force, maximum velocity, F-V slope, and maximum power). The individual F-V relationships were highly linear for the SSKT (r = .990 [.908, .998]), bench press (r = .993 [.974, .999]), and prone bench pull (r = .998 [.992, 1.000]). The F-V relationship parameters (maximum force, maximum velocity, and maximum power) were significantly higher for 200-m specialists compared with longer-distance specialists (all Ps ≤ .047) with large effect sizes (≥0.94) revealing important practical differences. However, no significant differences were observed between 200-m specialists and longer-distance specialists in the F-V slope (P ≥ .477). The F-V relationship assessed during both specific (SSKT) and nonspecific upper-body tasks (bench press and bench pull) may distinguish between kayakers specialized in different distances.


Enhanced microbial safety of channel catfish (Ictalurus punctatus) fillet using recently invented medium molecular weight water-soluble chitosan coating.
Chitosan with higher molecular weight exhibited higher antimicrobial efficacy against foodborne pathogens. However, the poor water solubility of higher or medium molecular weight chitosan limits its applications. To overcome the challenge, our research team searched for simple preparation procedure for fast-dissolving medium molecular weight chitosan in water. Throughout the process, we were able to obtain a higher concentration of medium molecular weight water-soluble (MMWWS) chitosan (400 kDa). The MMWWS chitosan showed physicochemical properties that are suitable for edible coating. Antibacterial activities of 400-kDa chitosan coating prepared in acetic acid (1% v/v) or aspartic acid (1% or 3% w/v) were examined. The surface of catfish cubes was inoculated with six foodborne pathogens and then coated with chitosan solutions. The survival of each pathogen was evaluated during shelf life storage. Compared with the control, 3% w/v chitosan coating in aspartic acid solution exhibited the most effective antibacterial activities among other coating treatments, completely inhibiting Vibrio parahaemolyticus on the surface of catfish. The study suggested that chitosan dissolved in aspartic acid has the potential for use as an alternative antimicrobial coating for catfish fillet.


Investigation of PC36 (BoCD45R).
Eight monoclonal antibodies (mAbs) clustered together in a statistical analysis of data submitted to the Third Workshop on Ruminant Leucocyte Antigens to form provisional cluster (PC) 36. PC36 included the CD45R workshop control mAb CC76. The mAbs were compared by two-colour immunofluorescence with mAbs against other leucocyte subpopulation antigens. The flow cytometry results indicated that all of the mAbs identified CD45R.


Multiple sclerosis genomic map implicates peripheral immune cells and microglia in susceptibility.
We analyzed genetic data of 47,429 multiple sclerosis (MS) and 68,374 control subjects and established a reference map of the genetic architecture of MS that includes 200 autosomal susceptibility variants outside the major histocompatibility complex (MHC), one chromosome X variant, and 32 variants within the extended MHC. We used an ensemble of methods to prioritize 551 putative susceptibility genes that implicate multiple innate and adaptive pathways distributed across the cellular components of the immune system. Using expression profiles from purified human microglia, we observed enrichment for MS genes in these brain-resident immune cells, suggesting that these may have a role in targeting an autoimmune process to the central nervous system, although MS is most likely initially triggered by perturbation of peripheral immune responses.


Neurodevelopment in perinatally HIV-infected children: a concern for adolescence.
Globally, an estimated 3.4 million children are living with HIV, yet little is known about the effects of HIV and antiretroviral treatment (ART) on the developing brain, and the neurodevelopmental and behavioural outcomes of perinatally HIV-infected (PHIV+) adolescents. We reviewed the literature on neurodevelopmental outcomes in PHIV+ children and adolescents, and summarized the current evidence on behaviour, general cognition, specific domains, hearing and language, school performance and physical disabilities due to neurological problems. Evidence suggests that PHIV+ children do not perform as well as controls on general cognitive tests, processing speed and visual-spatial tasks, and are at much higher risk for psychiatric and mental health problems. Children with AIDS-defining diagnoses are particularly at risk for poorer outcomes. A striking finding is the lack of published data specific to the adolescent age group (10-25 years), particularly from resource-constrained countries, which have the highest HIV prevalence. In addition, extreme heterogeneity in terms of timing and source of infection, and antiretroviral experience limits our ability to summarize findings of studies and generalize results to other settings. Due to the complex nature of the developing adolescent brain, environmental influences and variation in access to ART, there is an urgent need for research on the longitudinal trajectory of neurodevelopment among children and adolescents perinatally infected with HIV, especially in high burden resource-constrained settings.


Extracellular vesicles as drug delivery systems: lessons from the liposome field.
Extracellular vesicles (EVs) are membrane-derived particles surrounded by a (phospho)lipid bilayer that are released by cells in the human body. In addition to direct cell-to-cell contact and the secretion of soluble factors, EVs function as another mechanism of intercellular communication. These vesicles are able to efficiently deliver their parental cell-derived molecular cargo to recipient cells, which can result in structural changes at an RNA, protein, or even phenotypic level. For this reason, EVs have recently gained much interest for drug delivery purposes. In contrast to these 'natural delivery systems', synthetic (phospho)lipid vesicles, or liposomes, have been employed as drug carriers for decades, resulting in several approved liposomal nanomedicines used in the clinic. This review discusses the similarities and differences between EVs and liposomes with the focus on features that are relevant for drug delivery purposes such as circulation time, biodistribution, cellular interactions and cargo loading. By applying beneficial features of EVs to liposomes and vice versa, improved drug carriers can be developed which will advance the field of nanomedicines and ultimately improve patient outcomes. While the application of EVs for therapeutic drug delivery is still in its infancy, issues regarding the understanding of EV biogenesis, large-scale production and in vivo interactions need to be addressed in order to develop successful and cost-effective EV-based drug delivery systems.


Single-incision multiport laparoscopic cholecystectomy: things to overcome.
To report on our initial experience with single-incision multiport laparoscopic cholecystectomy, together with its clinical outcomes. Nonrandomized prospective study. University department of surgery. Sixty-four patients with gallstones and gallbladder polyps were enrolled after providing informed consent. Based on our experience, we excluded patients with acute cholecystitis, concomitant choledocholithiasis, a history of previous upper abdominal surgery, and a suspicion of gallbladder cancer. We analyzed the outcomes and complications, based on our experience, according to the clinicopathologic and operative factors. We also compared patients who underwent single-incision multiport laparoscopic cholecystectomy with those who were converted to conventional laparoscopic cholecystectomy. There were 2 bile duct injuries and 4 surgical site infections. We had difficulties in visualizing the Calot triangle in 22 patients. Higher levels of inflammatory markers, longer operation times, and more frequent bile juice spillage were significantly observed in those patients. Ten patients were converted to conventional laparoscopic cholecystectomy. The mean age of patients who underwent conversion surgery was significantly older than that of the no-conversion group. The more the body mass index increased, the more the conversion rate increased. Experienced laparoscopic surgeons can safely perform cholecystectomy using conventional and curved laparoscopic instruments in selected patients. We recommend that you consider performing conventional laparoscopic cholecystectomy or that you use additional retraction devices for patients with a higher body mass index or acute cholecystitis.


Exogenously administered secreted frizzled related protein 2 (Sfrp2) reduces fibrosis and improves cardiac function in a rat model of myocardial infarction.
Secreted frizzled related protein 2 (Sfrp2) is known as an inhibitor for the Wnt signaling. In recent studies, Sfrp2 has been reported to inhibit the activity of Xenopus homolog of mammalian Tolloid-like 1 metalloproteinase. Bone morphogenic protein 1 (Bmp1)/Tolloid-like metalloproteinase plays a key role in the regulation of collagen biosynthesis and maturation after tissue injury. Here, we showed both endogenous Sfrp2 and Bmp1 protein expressions were up-regulated in rat heart after myocardial infarction (MI). We hypothesize that Sfrp2 could inhibit mammalian Bmp1 activity and, hence, the exogenous administration of Sfrp2 after MI would inhibit the deposition of mature collagen and improve heart function. Using recombinant proteins, we demonstrated that Sfrp2, but not Sfrp1 or Sfrp3, inhibited Bmp1 activity in vitro as measured by a fluorogenic peptide based procollagen C-proteinase activity assay. We also demonstrated that Sfrp2 at high concentration inhibited human and rat type I procollagen processing by Bmp1 in vitro. We further showed that exogenously added Sfrp2 inhibited type I procollagen maturation in primary cardiac fibroblasts. Two days after direct injection into the rat infarcted myocardium, Sfrp2 inhibited MI-induced type I collagen deposition. As early as 2 wk after injection, Sfrp2 significantly reduced left ventricular (LV) fibrosis as shown by trichrome staining. Four weeks after injection, Sfrp2 prevented the anterior wall thinning and significantly improved cardiac function as revealed by histological analysis and echocardiographic measurement. Our study demonstrates Sfrp2 at therapeutic doses can inhibit fibrosis and improve LV function at a later stage after MI.


Effectiveness of physical therapy in treating atraumatic full-thickness rotator cuff tears: a multicenter prospective cohort study.
To assess the effectiveness of a specific nonoperative physical therapy program in treating atraumatic full-thickness rotator cuff tears using a multicenter prospective cohort study design. Patients with atraumatic full-thickness rotator cuff tears who consented to enroll provided data via questionnaire on demographics, symptom characteristics, comorbidities, willingness to undergo surgery, and patient-related outcome assessments (Short Form 12 score, American Shoulder and Elbow Surgeons score, Western Ontario Rotator Cuff score, Single Assessment Numeric Evaluation score, and Shoulder Activity Scale). Physicians recorded physical examination and imaging data. Patients began a physical therapy program developed from a systematic review of the literature and returned for evaluation at 6 and 12 weeks. At those visits, patients could choose 1 of 3 courses: (1) cured (no formal follow-up scheduled), (2) improved (continue therapy with scheduled reassessment in 6 weeks), or (3) no better (surgery offered). Patients were contacted by telephone at 1 and 2 years to determine whether they had undergone surgery since their last visit. A Wilcoxon signed rank test with continuity correction was used to compare initial, 6-week, and 12-week outcome scores. The cohort consists of 452 patients. Patient-reported outcomes improved significantly at 6 and 12 weeks. Patients elected to undergo surgery less than 25% of the time. Patients who decided to have surgery generally did so between 6 and 12 weeks, and few had surgery between 3 and 24 months. Nonoperative treatment using this physical therapy protocol is effective for treating atraumatic full-thickness rotator cuff tears in approximately 75% of patients followed up for 2 years.


Gene cluster of Pseudomonas syringae pv. "phaseolicola" controls pathogenicity of bean plants and hypersensitivity of nonhost plants.
Loss of the ability of Pseudomonas syringae pv. "phaseolicola" NPS3121 to elicit a hypersensitive response on tobacco and other nonhost plants was associated with loss of pathogenicity on the susceptible host bean. Eight independent, prototrophic transposon Tn5 insertion mutants which had lost the ability to elicit a hypersensitive response on tobacco plants were identified. Six of these mutants no longer produced disease lesions on primary leaves of the susceptible bean cultivar Red Kidney and failed to elicit a hypersensitive response on the resistant bean cultivar Red Mexican and on the nonhost plants tomato, cowpea, and soybean. The two remaining mutants had reduced pathogenicity on Red Kidney bean and elicited variable hypersensitive responses on the other plants tested. Southern blot analysis indicated that each mutant carried a single independent Tn5 insertion in one of three EcoRI fragments of about 17, 7, and 5 kilobases. Marker exchange mutagenesis further supported the conclusion that the pleiotropic mutant phenotype was not associated with multiple Tn5 insertions. A genomic library of the wild-type strain was constructed in the cosmid vector pLAFR3. A recombinant plasmid, designated pPL6, that carried P. syringae pv. "phaseolicola" genomic sequences was identified by colony hybridization. This plasmid restored the wild-type phenotype to all but one mutant, suggesting that genes affected by the insertions were clustered. Structural analysis of pPL6 and the wild-type genome indicated that the 17- and 5-kilobase EcoRI fragments were contiguous in the strain NPS3121 genome.


The evolutionary dynamics of endogenous retroviruses.
Endogenous retroviruses (ERVs) are vertically transmitted intragenomic elements derived from integrated retroviruses. ERVs can proliferate within the genome of their host until they either acquire inactivating mutations or are lost by recombinational deletion. We present a model that unifies current knowledge of ERV biology into a single evolutionary framework. The model predicts the possible long-term outcomes of retroviral germline infection and can account for the variable patterns of observed ERV genetic diversity. We hope the model will provide a useful framework for understanding ERV evolution, enabling the testing of evolutionary hypotheses and the estimation of parameters governing ERV proliferation.


Polar nephelometer for light-scattering measurements of ice crystals.
We report on a small, lightweight polar nephelometer for the measurement of the light-scattering properties of cloud particles, specifically designed for use on a balloonborne platform in cirrus cloud conditions. The instrument consists of 33 fiber-optic light guides positioned in a two-dimensional plane from 5 degrees to 175 degrees that direct the scattered light to photodiode detectors-amplifier units. The system uses an onboard computer and data acquisition card to collect and store the measured signals. The instrument's calibration is tested by measurement of light scattered into a two-dimensional plane from small water droplets generated by an ultrasonic humidifier. Excellent comparisons between the measured water-droplet scattering properties and expectations generated by Mie calculation are shown. The measured scattering properties of ice crystals generated in a cold chamber also compare reasonably well with the theoretical results based on calculations from a unified theory of light scattering by ice crystals that use the particle size distribution measured in the chamber.


A Simple Effective Method for Frailty in Heart Failure with Impact on Clinical Outcomes in North Indian Population.
Frailty has been studied extensively in elderly population as a predictor and prognostic marker for morbidity and mortality. Frailty is being increasingly recognized as a distinct pathophysiological condition which plays a major role in outcomes of various disease states including heart failure. Our aim was to study the prevalence of frailty in heart failure and see its prognostic significance in such patients. This was a prospective study conducted in an out-patient HF clinic. All consecutive patients with HF, ≥25 years age, with LVEF<40% were included. All patients were asked a simple frailty questionnaire. Hand-dynamometer was used to assess handgrip strength in kilograms and were classified as frail, pre-frail or non-frail. The primary end point was cardiovascular mortality and hospitalization, and secondary end-point was composite of all cause mortality, hospitalization, device implantation and documented arrythmia. 210 patients were studied for clinical outcomes. Mean age was 60.59 ± 11.55 years with 15% patients aged less than 50 years. Mean LVEF was 30.24 ± 6.8%. Handgrip strength was poor in the frail vs non/pre-frail patients (p = 0.001) with a strength >16.95 kg having sensitivity of 72% and specificity of 63% for the prediction of survival. Frailty was an independent predictor of mortality with higher mortality and re-hospitalization in frail population (p = 0.001). Hazard for mortality or hospitalization was 4.7 fold in frail population. Frailty is associated with a significant morbidity and mortality in heart failure. A simple bedside hand-dynamometer may aid as a frailty screening tool in these patients and help in planning treatment strategies.


A new domain of conjugative relaxase TrwC responsible for efficient oriT-specific recombination on minimal target sequences.
We show that relaxase TrwC promotes recombination between two directly repeated oriTs while related relaxases TraI of F and pKM101 do not. Efficient recombination required also relaxosome accessory protein TrwA even after deletion of TrwA binding sites at oriT, suggesting that the effect of TrwA is mediated by protein-protein interactions. TrwC relaxase domain was necessary but not sufficient to catalyse recombination efficiently. Full recombinase activity was obtained with the N-terminal 600 residues of TrwC. The minimal target sequences required for recombination were different at each of the two involved oriTs: oriT1 could be reduced to the nic site and TrwC binding site, while oriT2 required an extended sequence including a set of iterons that are not required for conjugation. TrwC-mediated integration of a transferred DNA into a resident oriT copy required a complete oriT in the recipient. We observed dramatic changes in the efficiency of recombination between tandem oriTs linked to the direction of plasmid replication and transcription through oriT1. We propose that recombination is triggered by the generation of a single-stranded DNA at oriT1 that causes TrwC nicking. The resulting TrwC-DNA complex reacts with oriT2, excising the intervening DNA. This intermediate can be resolved by host-encoded replication functions.


Therapeutic preparations of normal polyspecific IgG (IVIg) induce apoptosis in human lymphocytes and monocytes: a novel mechanism of action of IVIg involving the Fas apoptotic pathway.
Therapeutic preparations of normal human IgG for i.v. use (i.v.Ig) exhibit a broad spectrum of immunoregulatory activities in vitro and in vivo. I.v.Ig has been shown to inhibit the proliferation of activated B and T lymphocytes and of several autonomously growing cell lines. In this study, we demonstrate that i.v.Ig induces apoptosis in leukemic cells of lymphocyte and monocyte lineage and in CD40-activated normal tonsillar B cells, involving, at least in part, Fas (CD95/APO-1) and activation of caspases. I.v.Ig-induced apoptosis was higher in Fas-sensitive HuT78 cells than in Fas-resistant HuT78.B1 mutant cells, and soluble Fas inhibited IVIg-induced apoptosis. I.v.Ig immunoprecipitated Fas from Fas-expressing transfectants and recognized purified Fas/glutathione-S-transferase fusion proteins upon immunoblotting. Affinity-purified anti-Fas Abs from i.v.Ig induced apoptosis of CEM T cells at a 120-fold lower concentration than unfractionated i.v.Ig. Inhibitors of cysteine proteases of the caspase family, caspase 1 (IL-1beta-converting enzyme) and caspase 3 (Yama/CPP32b), partially inhibited i.v.Ig-induced apoptosis of CEM cells. Furthermore, cleavage of poly(A)DP-ribose polymerase into an 85-kDa signature death fragment was observed in CEM cells following i.v.Ig treatment. Thus, normal IgG induces apoptosis in lymphocytes and monocytes. Our results provide evidence for a role of Fas, bring new insights into the mechanisms of action of i.v.Ig in autoimmune diseases, and suggest a role of normal Ig in controlling cell death and proliferation.


Characterization of CD4(+) CTLs ex vivo.
The cytotoxic potential of CD8(+) T cells and NK cells plays a crucial role in the immune response to pathogens. Although in vitro studies have reported that CD4(+) T cells are also able to mediate perforin-mediated killing, the in vivo existence and relevance of cytotoxic CD4(+) T cells have been the subject of debate. Here we show that a population of CD4(+) perforin(+) T cells is present in the circulation at low numbers in healthy donors and is markedly expanded in donors with chronic viral infections, in particular HIV infection, at all stages of the disease, including early primary infection. Ex vivo analysis shows that these cells have cytotoxic potential mediated through the release of perforin. In comparison with more classical CD4(+) T cells, this subset displays a distinct surface phenotype and functional profile most consistent with end-stage differentiated T cells and include Ag experienced CD4(+) T cells. The existence of CD4(+) cytotoxic T cells in vivo at relatively high levels in chronic viral infection suggests a role in the immune response.


The ROKS nomogram for predicting a second symptomatic stone episode.
Most patients with first-time kidney stones undergo limited evaluations, and few receive preventive therapy. A prediction tool for the risk of a second kidney stone episode is needed to optimize treatment strategies. We identified adult first-time symptomatic stone formers residing in Olmsted County, Minnesota, from 1984 to 2003 and manually reviewed their linked comprehensive medical records through the Rochester Epidemiology Project. Clinical characteristics in the medical record before or up to 90 days after the first stone episode were evaluated as predictors for symptomatic recurrence. A nomogram was developed from a multivariable model based on these characteristics. There were 2239 first-time adult kidney stone formers with evidence of a passed, obstructing, or infected stone causing pain or gross hematuria. Symptomatic recurrence occurred in 707 of these stone formers through 2012 (recurrence rates at 2, 5, 10, and 15 years were 11%, 20%, 31%, and 39%, respectively). A parsimonious model had the following risk factors for recurrence: younger age, male sex, white race, family history of stones, prior asymptomatic stone on imaging, prior suspected stone episode, gross hematuria, nonobstructing (asymptomatic) stone on imaging, symptomatic renal pelvic or lower-pole stone on imaging, no ureterovesicular junction stone on imaging, and uric acid stone composition. Ten-year recurrence rates varied from 12% to 56% between the first and fifth quintiles of nomogram score. The Recurrence of Kidney Stone nomogram identifies kidney stone formers at greatest risk for a second symptomatic episode. Such individuals may benefit from medical intervention and be good candidates for prevention trials.


A room temperature vulcanizing silicone rubber sport splint.
Athletic hand and wrist injuries may be treated by immobilizing the affected extremity in a rigid splint to protect the injury site for an extended time period. For athletes, this means a reduction in playing time, perhaps being out for an entire playing season. The use of a room temperature vulcanizing silicone rubber compound allows for a playing splint that complies with athletic regulations. Proper application of the room temperature vulcanizing silicone rubber provides an excellent, reliable protective playing splint allowing for safe, functional athletic performance for the injured player during the game. This article describes the fabrication process of a silicone rubber protective playing splint for athletic hand and wrist injuries. The effectiveness of the silicone rubber protective playing splint is illustrated via case studies.


"It's Really Overwhelming": Patient Perspectives on Care Coordination.
Failures of care coordination among health care providers are known to lead to poorer health outcomes for patients with complex medical needs. However, there has been limited research into the perspectives of patients who receive care from a variety of health care providers. This qualitative study sought to characterize the factors leading to emergency department (ED) patient satisfaction and dissatisfaction with their care coordination. Semistructured telephone interviews were conducted with 25 adult patients following ED visits about their experiences with their care coordination. All patients interviewed had 2 or more ED visits and hospitalizations in the past year and/or health providers in more than one health system. Interview transcripts were coded and analyzed following a modified grounded theory approach. Four broad categories of themes emerged from the patient interviews covering the following: (1) perceptions of care coordination between their providers, (2) the role of electronic health records, (3) challenges with information exchange between health systems, and (4) sources of support for care coordination activities, emphasizing the important role of the primary care provider. Patients with multiple health care providers identified significant barriers to communication among providers and inadequate support with care coordination activities. Expansion of team-based models of primary care and prioritizing interoperable technology for sharing patient health information between providers will be critical to improving the patient experience and the safety of transitions of care.


Cell-type-specific signaling networks in heterocellular organoids.
Despite the widespread adoption of organoids as biomimetic tissue models, methods to comprehensively analyze cell-type-specific post-translational modification (PTM) signaling networks in organoids are absent. Here, we report multivariate single-cell analysis of such networks in organoids and organoid cocultures. Simultaneous analysis by mass cytometry of 28 PTMs in >1 million single cells derived from small intestinal organoids reveals cell-type- and cell-state-specific signaling networks in stem, Paneth, enteroendocrine, tuft and goblet cells, as well as enterocytes. Integrating single-cell PTM analysis with thiol-reactive organoid barcoding in situ (TOBis) enables high-throughput comparison of signaling networks between organoid cultures. Cell-type-specific PTM analysis of colorectal cancer organoid cocultures reveals that shApc, KrasG12D and Trp53R172H cell-autonomously mimic signaling states normally induced by stromal fibroblasts and macrophages. These results demonstrate how standard mass cytometry workflows can be modified to perform high-throughput multivariate cell-type-specific signaling analysis of healthy and cancerous organoids.


[Chronic recurrent multifocal osteomyelitis of the mandible: report of three cases].
To report three cases of chronic recurrent multifocal osteomyelitis of the mandible, an inflammatory disease affecting one or more bones with absence of isolated microorganisms in affected areas. The first case is a 13 year-old female presenting with pain and fever after dental treatment. The patient received antibiotic treatment for osteomyelitis, but developed progressive enlargement of the mandible and palmoplantar pustulosis. Bone scintigraphy showed intense and diffuse uptake in the mandible. The swelling decreased after indomethacin and hyperbaric oxygen therapy. Case 2 is a 9 year-old female patient with recurrent pain and edema of the right mandible for three years. The diagnosis of osteomyelitis was established and amoxicillin introduced. After three months, tomography showed diffuse mandible osteolysis. Indomethacin and hyperbaric oxygen therapy were introduced, however the patient presented a relapse and was treated with prednisone, rofecoxib and methotrexate. Patient 3, a 10 year-old male, had palmoplantar pustulosis and recurrent enlargement of the mandible. Tomography showed diffuse mandible osteolysis and scintigraphy revealed intense and diffuse uptake in the mandible. The patient was treated with prednisone. Rofecoxib was replaced after two relapses. Chronic recurrent multifocal osteomyelitis of the mandible is often associated with prolonged pain periods and periods of activity and remission of the inflammatory process. Its recognition is important to prevent the patient from being submitted to prolonged antibiotic therapy and unnecessary invasive procedures.


Association of impaired neuronal migration with cognitive deficits in extremely preterm infants.
Many extremely preterm infants (born before 28 gestational weeks [GWs]) develop cognitive impairment in later life, although the underlying pathogenesis is not yet completely understood. Our examinations of the developing human neocortex confirmed that neuronal migration continues beyond 23 GWs, the gestational week at which extremely preterm infants have live births. We observed larger numbers of ectopic neurons in the white matter of the neocortex in human extremely preterm infants with brain injury and hypothesized that altered neuronal migration may be associated with cognitive impairment in later life. To confirm whether preterm brain injury affects neuronal migration, we produced brain damage in mouse embryos by occluding the maternal uterine arteries. The mice showed delayed neuronal migration, ectopic neurons in the white matter, altered neuronal alignment, and abnormal corticocortical axonal wiring. Similar to human extremely preterm infants with brain injury, the surviving mice exhibited cognitive deficits. Activation of the affected medial prefrontal cortices of the surviving mice improved working memory deficits, indicating that decreased neuronal activity caused the cognitive deficits. These findings suggest that altered neuronal migration altered by brain injury might contribute to the subsequent development of cognitive impairment in extremely preterm infants.


The APOL1 gene and allograft survival after kidney transplantation.
Coding variants in the apolipoprotein L1 gene (APOL1) are strongly associated with nephropathy in African Americans (AAs). The effect of transplanting kidneys from AA donors with two APOL1 nephropathy risk variants is unknown. APOL1 risk variants were genotyped in 106 AA deceased organ donors and graft survival assessed in 136 resultant kidney transplants. Cox-proportional hazard models tested for association between time to graft failure and donor APOL1 genotypes. The mean follow-up was 26.4 ± 21.8 months. Twenty-two of 136 transplanted kidneys (16%) were from donors with two APOL1 nephropathy risk variants. Twenty-five grafts failed; eight (32%) had two APOL1 risk variants. A multivariate model accounting for donor APOL1 genotype, overall African ancestry, expanded criteria donation, recipient age and gender, HLA mismatch, CIT and PRA revealed that graft survival was significantly shorter in donor kidneys with two APOL1 risk variants (hazard ratio [HR] 3.84; p = 0.008) and higher HLA mismatch (HR 1.52; p = 0.03), but not for overall African ancestry excluding APOL1. Kidneys from AA deceased donors harboring two APOL1 risk variants failed more rapidly after renal transplantation than those with zero or one risk variants. If replicated, APOL1 genotyping could improve the donor selection process and maximize long-term renal allograft survival.


Interferon Lambda 4 Genotype Is Not Associated with Recurrence of Oral or Genital Herpes.
IFNL4-ΔG/TT (rs368234815) genotype is associated with hepatitis C virus clearance and may play a role in other infections. IFN-λ4 protein is generated only in individuals who carry the IFNL4-ΔG allele. The IFNL4 rs12979860-T allele, which is in strong linkage disequilibrium with IFNL4-ΔG, was recently reported to be associated with more frequent and severe oral herpes episodes. We investigated the association of IFNL4-ΔG/TT with herpes simplex virus (HSV)-related outcomes among 2,192 African American and European American participants in the Women's Interagency HIV Study (WIHS). WIHS is a prospective cohort study of human immunodeficiency virus (HIV)-infected and at-risk women that began in 1994. This report includes follow-up through 2013. Available data included: HSV-1 and HSV-2 antibodies at study entry; bi-annually ascertained episodes of (self-reported) oral herpes, (self-reported) genital sores and (clinician-observed) genital ulcers; HSV-2 DNA in cervicovaginal lavage (CVL) specimens. IFNL4-ΔG/TT genotyping was determined by TaqMan. We compared women with IFNL4-ΔG/ΔG or IFNL4-TT/ΔG genotypes (i.e., IFNL4-ΔG carriers) to those with the IFNL4-TT/TT genotype, adjusting for age, race and HIV status. For outcomes with repeated measurements, the adjusted odds ratio (aOR), 95% confidence interval [CI] and p-value were determined using a generalized estimating equations approach. Median participant age at enrollment was 36 years; 81% were African American, 74% were HIV-infected. Among 1,431 participants tested for antibodies, 72.8% were positive for HSV-1 and 79.0% were positive for HSV-2. We observed no association between IFNL4-ΔG/TT genotype and any outcome: HSV-1 or HSV-2 antibody prevalence (p>0.1, all comparisons); oral herpes (aOR, 1.2; p = 0.35); genital sores (aOR, 1.0; p = 0.71); genital ulcers (aOR, 1.1; p = 0.53); detectable HSV-2 DNA in CVL (N = 322; aOR, 0.71; p = 0.49); HSV-2 DNA level (p = 0.68). In this large prospective study, IFNL4-ΔG/TT genotype was not associated with HSV-related outcomes, including episodes of oral or genital herpes.


Pandoraviruses: amoeba viruses with genomes up to 2.5 Mb reaching that of parasitic eukaryotes.
Ten years ago, the discovery of Mimivirus, a virus infecting Acanthamoeba, initiated a reappraisal of the upper limits of the viral world, both in terms of particle size (>0.7 micrometers) and genome complexity (>1000 genes), dimensions typical of parasitic bacteria. The diversity of these giant viruses (the Megaviridae) was assessed by sampling a variety of aquatic environments and their associated sediments worldwide. We report the isolation of two giant viruses, one off the coast of central Chile, the other from a freshwater pond near Melbourne (Australia), without morphological or genomic resemblance to any previously defined virus families. Their micrometer-sized ovoid particles contain DNA genomes of at least 2.5 and 1.9 megabases, respectively. These viruses are the first members of the proposed "Pandoravirus" genus, a term reflecting their lack of similarity with previously described microorganisms and the surprises expected from their future study.


Immunoprotection against toxic biomarkers is retained during Parkinson's disease progression.
The aim was to ascertain any possible linkage between humoral immune responses to principal biomarkers (α-synuclein monomers, its toxic oligomers or fibrils, dopamine and S100B) and cellular immunity in Parkinson's disease development. There were elevated autoantibody titers to α-synuclein monomers, oligomers plus fibrils in 72%, 56%, and 17% of Parkinsonian patients respectively with a 5-year disease duration. Additionally, there were increased titers to dopamine and S100B (96% and 89%) in the 5-year patient group. All of these values subsided in 10-year sufferers. Furthermore, CD3+, CD4+, CD8+ T-lymphocyte and B-lymphocyte subsets declined in the patient cohort during Parkinsonism indicating disease associated reductions in these lymphocyte subsets.