Following de novo triglyceride dynamics in ovaries of Aedes aegypti during the previtellogenic stage.
Understanding the molecular and biochemical basis of egg development is a central topic in mosquito reproductive biology. Lipids are a major source of energy and building blocks for the developing ovarian follicles. Ultra-High Resolution Mass Spectrometry (UHRMS) combined with in vivo metabolic labeling of follicle lipids with deuterated water (2H2O) can provide unequivocal identification of de novo lipid species during ovarian development. In the present study, we followed de novo triglyceride (TG) dynamics during the ovarian previtellogenic (PVG) stage (2-7 days post-eclosion) of female adult Aedes aegypti. The incorporation of stable isotopes from the diet was evaluated using liquid chromatography (LC) in tandem with the high accuracy (< 0.3 ppm) and high mass resolution (over 1 M) of a 14.5 T Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (14.5 T FT-ICR MS) equipped with hexapolar detection. LC-UHRMS provides effective lipid class separation and chemical formula identification based on the isotopic fine structure. The monitoring of stable isotope incorporation into de novo incorporated TGs suggests that ovarian lipids are consumed or recycled during the PVG stage, with variable time dynamics. These results provide further evidence of the complexity of the molecular mechanism of follicular lipid dynamics during oogenesis in mosquitoes.


CGBayesNets: conditional Gaussian Bayesian network learning and inference with mixed discrete and continuous data.
Bayesian Networks (BN) have been a popular predictive modeling formalism in bioinformatics, but their application in modern genomics has been slowed by an inability to cleanly handle domains with mixed discrete and continuous variables. Existing free BN software packages either discretize continuous variables, which can lead to information loss, or do not include inference routines, which makes prediction with the BN impossible. We present CGBayesNets, a BN package focused around prediction of a clinical phenotype from mixed discrete and continuous variables, which fills these gaps. CGBayesNets implements Bayesian likelihood and inference algorithms for the conditional Gaussian Bayesian network (CGBNs) formalism, one appropriate for predicting an outcome of interest from, e.g., multimodal genomic data. We provide four different network learning algorithms, each making a different tradeoff between computational cost and network likelihood. CGBayesNets provides a full suite of functions for model exploration and verification, including cross validation, bootstrapping, and AUC manipulation. We highlight several results obtained previously with CGBayesNets, including predictive models of wood properties from tree genomics, leukemia subtype classification from mixed genomic data, and robust prediction of intensive care unit mortality outcomes from metabolomic profiles. We also provide detailed example analysis on public metabolomic and gene expression datasets. CGBayesNets is implemented in MATLAB and available as MATLAB source code, under an Open Source license and anonymous download at http://www.cgbayesnets.com.


Preparation and evaluation of luteolin-loaded PLA-based shape memory gastroretentive drug delivery systems.
Luteolin, a natural flavonoid, is gaining growing attention for its potential in the treatment of gastric cancer. However, its clinical application is limited by factors such as poor aqueous solubility. This study aimed to develop a novel gastroretentive drug delivery system (GRDDS) to both enhance the oral bioavailability of luteolin and prolong its release and in vivo circulation time. Out of 10 luteolin-loaded PLA-based shape memory films prepared in this study, the LPC-PLA/PEG(7/3) formulation incorporated with PEG, HPMC, and NaHCO3 exhibited optimal properties in terms of drug release and inhibitory activity against SGC-7901 cells. Moreover, small-animal imaging revealed that LPC-PLA/PEG(7/3) exhibited a prolonged gastric retention time of approximately 8 h. Furthermore, the pharmacokinetic studies indicated a 354 % increase in the oral bioavailability of LPC-PLA/PEG(7/3) in rats compared to luteolin. In sum, a novel GRDDS was developed to enhance the relative bioavailability of luteolin, offering a potential strategy for practical oral administration.


A novel biosensor to study cAMP dynamics in cilia and flagella.
The cellular messenger cAMP regulates multiple cellular functions, including signaling in cilia and flagella. The cAMP dynamics in these subcellular compartments are ill-defined. We introduce a novel FRET-based cAMP biosensor with nanomolar sensitivity that is out of reach for other sensors. To measure cAMP dynamics in the sperm flagellum, we generated transgenic mice and reveal that the hitherto methods determining total cAMP levels do not reflect changes in free cAMP levels. Moreover, cAMP dynamics in the midpiece and principal piece of the flagellum are distinctively different. The sole cAMP source in the flagellum is the soluble adenylate cyclase (SACY). Although bicarbonate-dependent SACY activity requires Ca(2+), basal SACY activity is suppressed by Ca(2+). Finally, we also applied the sensor to primary cilia. Our new cAMP biosensor features unique characteristics that allow gaining new insights into cAMP signaling and unravel the molecular mechanisms underlying ciliary function in vitro and in vivo.


Recurrent Cesarean scar pregnancy: case series and literature review.
To determine the rate of recurrent Cesarean scar pregnancy (CSP) in our clinical practices and to evaluate whether the mode of treatment of a CSP is associated with the risk of recurrent CSP, as well as to review the published literature on recurrent CSP. We performed a retrospective search of our six obstetric and gynecological departmental ultrasound databases for all CSPs and recurrent CSPs between 2010 and 2019. We extracted various data, including number of CSPs with follow-up, number of cases attempting and number achieving pregnancy following treatment of CSP and number of recurrent CSPs, as well as details of the treatment of the original CSP. After analyzing the clinical data, we evaluated whether the mode of treatment terminating the previous CSP was associated with the risk of recurrent CSP. We also performed a PubMed search for: 'recurrent Cesarean scar pregnancy' and 'recurrent Cesarean scar ectopic pregnancy'. Articles were reviewed for year of publication, and extraction and analysis of the same data as those obtained from our departmental databases were performed. Our database search identified 252 cases of CSP. The overall rate of clinical follow-up ranged between 71.4% and 100%, according to treatment site (mean, 90.9%). Among these, 105 women had another pregnancy after treatment of the previous CSP. Of these, 36 (34.3%) pregnancies were recurrent CSP, with 27 women having a single recurrence and three women having multiple recurrences, one with two, one with three and one with four. We did not find any particular single or combination treatment mode terminating the previous CSP to be associated with recurrent CSP. The literature search identified 17 articles that yielded sufficient information for us to evaluate their reported prevalence of recurrent CSP. These reported 1743 primary diagnoses of CSP, of which 944 had reliable follow-up. Data were available for 489 cases that attempted to conceive again after treatment of a previous CSP, and on the 327 pregnancies achieved. Of these, 67 (20.5%) were recurrent CSP. On the basis of our pooled clinical data and review of the literature, recurrent CSP is apparently more common than was previously assumed based upon mostly single-case reports or series with few cases. This should be borne in mind when counseling patients undergoing treatment for CSP regarding their risk of recurrence. We found no obvious causal relationship or association between the type of treatment of the previous CSP and recurrence of CSP. Patients who become pregnant after treatment of a CSP should be encouraged to have an early (5-7-week) first-trimester transvaginal scan to determine the location of the gestation. © 2021 International Society of Ultrasound in Obstetrics and Gynecology.


Sodium channel gating currents. Origin of the rising phase.
There has been some uncertainty in the past as to the origin of the rising phase of the gating current. We present evidence here that proves that the gating current does not have a rising phase and that the observed rising phase is due to an uncompensated series resistance in the Frankenhaeuser-Hodgkin (F-H) space. When a squid giant axon is bathed in a solution that is 10-20% hyperosmotic with respect to the internal solution, the rising phase of the gating current is eliminated. In parallel with this, a component of the capacity transient (time constant, 20 microseconds) is reduced so that the capacity transient now appears to be closer to a single fast (5-10 microseconds) component. These changes in the capacity transient and gating current occur without altering the amount of charge moved in either. This indicates that the charge is simply redistributed in time. The gating current without a rising phase can still be immobilized by inactivation. Supporting evidence is provided by measuring the accumulation and washout of K+ from the F-H space. It was found that K+ washes out 35% faster when the axon is bathed in hyperosmotic solution. It was estimated that the F-H space thickness (theta) increased 2.5 +/- 0.4-fold (mean +/- SEM) in hyperosmotic solution. Similarly, K+ accumulation in the F-H space was decreased, leading to an estimate of a 5 +/- 1.4-fold increase in theta in hyperosmotic solution. These results are consistent with the simple structural model presented.


miRNAs, single nucleotide polymorphisms (SNPs) and age-related macular degeneration (AMD).
Advanced age-related macular degeneration (AAMD) is a complex sight-threating disease of public health significance. Micro RNAs (miRNAs) have been proposed as biomarkers for AAMD. The presence of certain single nucleotide polymorphisms (SNPs) may influence the explanatory value of these biomarkers. Here we present findings from an integrated approach used to determine whether AAMD-associated SNPs have the capacity to influence miRNA-mRNA pairing and, if so, to what extent such pairing may be manifested in a discrete AAMD transcriptome. Using a panel of 8854 SNPs associated with AAMD at p-values ≤5.0E-7 from a cohort of >30,000 elderly people, we identified SNPs in miRNA target-encoding constituents of: (1) regulator of complement activation (RCA) genes (rs390679, CFHR1, p≤2.14E-214 | rs12140421, CFHR3, p≤4.63E-29); (2) genes of major histocompatibility complex (MHC) loci (rs4151672, CFB, p≤8.91E-41 | rs115404146, HLA-C, p≤6.32E-12 | rs1055821, HLA-B, p≤1.93E-9 | rs1063355, HLA-DQB1, p≤6.82E-14); and (3) genes of the 10q26 AAMD locus (rs1045216, PLEKHA1, p≤4.17E-142 | rs2672603, ARMS2, p≤7.14E-46). We used these findings with existing data on AAMD-related retinal miRNA and transcript profiles for the purpose of making inferences on SNP-mRNA-miRNA-AAMD relationships. Four of 12 miRNAs significantly elevated in AAMD retina (hsa-miR-155-5p, hsa-let-7a-5p, hsa-let-7b-5p hsa-let-7d-5p) also showed strong pairing capacity (TarBase 7.1 context++ score <-0.2, miRanda 3.3 pairing score >150) with miRNA target transcripts encoded by AAMD-associated SNPs resident in HLA-DQB1 (rs1063355, hsa-miR-155-5p) and TGFBR1 (rs868, hsa-let-7). Three of the 12 miRNAs overexpressed in AAMD retina are inducible by NFkB and have high affinity targets in the complement factor H (CFH) mRNA 3' UTR. We used ENSEMBL to identify polymorphic regions in the CFH mRNA 3' UTR with the capacity to disrupt miRNA-mRNA pairing. Two variants (rs766666504 and rs459598) existed in DNA sequence encoding the seed region of hsa-miR-146a-5p in the CFH mRNA 3' UTR - as this miRNA is also elevated in both vitreous and serum of people with AAMD, it shows great value as a biomarker. Our findings suggest that knowledge on the nature of DNA sequence variation may increase the explanatory power of miRNA biomarkers in genetically diverse populations, while yielding information with which to develop: (1) mechanistic tests on processes implicated in AMD pathogenesis; and, (2) site-specific small molecules (synthetic mimetics or anti-miRNAs) with preventive or therapeutic efficacy for AAMD.


Development of transgenic rats producing human beta-amyloid precursor protein as a model for Alzheimer's disease: transgene and endogenous APP genes are regulated tissue-specifically.
Alzheimer's disease (AD) is a devastating neurodegenerative disorder that affects a large and growing number of elderly individuals. In addition to idiopathic disease, AD is also associated with autosomal dominant inheritance, which causes a familial form of AD (FAD). Some instances of FAD have been linked to mutations in the beta-amyloid protein precursor (APP). Although there are numerous mouse AD models available, few rat AD models, which have several advantages over mice, have been generated. Fischer 344 rats expressing human APP driven by the ubiquitin-C promoter were generated via lentiviral vector infection of Fischer 344 zygotes. We generated two separate APP-transgenic rat lines, APP21 and APP31. Serum levels of human amyloid-beta (Abeta)40 were 298 pg/ml for hemizygous and 486 pg/ml for homozygous APP21 animals. Serum Abeta42 levels in APP21 homozygous rats were 135 pg/ml. Immunohistochemistry in brain showed that the human APP transgene was expressed in neurons, but not in glial cells. These findings were consistent with independent examination of enhanced green fluorescent protein (eGFP) in the brains of eGFP-transgenic rats. APP21 and APP31 rats expressed 7.5- and 3-times more APP mRNA, respectively, than did wild-type rats. Northern blots showed that the human APP transgene, driven by the ubiquitin-C promoter, is expressed significantly more in brain, kidney and lung compared to heart and liver. A similar expression pattern was also seen for the endogenous rat APP. The unexpected similarity in the tissue-specific expression patterns of endogenous rat APP and transgenic human APP mRNAs suggests regulatory elements within the cDNA sequence of APP. This manuscript describes the generation of APP-transgenic inbred Fischer 344 rats. These are the first human AD model rat lines generated by lentiviral infection. The APP21 rat line expresses high levels of human APP and could be a useful model for AD. Tissue-specific expression in the two transgenic rat lines and in wild-type rats contradicts our current understanding of APP gene regulation. Determination of the elements that are responsible for tissue-specific expression of APP may enable new treatment options for AD.


[Chemical composition of essential oil obtained from Romanian fennel fruits].
For therapeutical purposes, fennel (Foeniculum vulgare Mill.), an important aromatic plant, is used for its expectorant, antispasmodic, carminative and diuretic properties. The chemical composition, especially of the volatile fraction, depends on the origin of the plant, and the ecological and environmental conditions in which it grows. To determine the chemical composition of the volatile fractions obtained from fennel fruits (Foeniculum vulgare Mill.) available on the Romanian market. The essential oils obtained from dried fruits by hydrodistillation have been characterized by using gas chromatography and mass spectroscopy (GC-MS). In all oils, the main components were: trans-anethole (9.72-56.57%), fenchone (7.72-34.66%) and estragole (4.69-49.04%). The essential oil from fennel samples showed an inadequate chemical profile for aromatherapy purposes.


Tissue distribution of breast cancer-associated antigens defined by monoclonal antibodies.
Balb/c mice were immunized with membrane preparations of primary and metastatic breast carcinomas or with live breast cancer cell lines. Sixty-two fusions were performed between immune splenocytes and SP2/0 mouse myeloma cells, and 107 hybridomas were cloned that produced antibody reactive with breast cancer membrane extract, cell lines, and frozen sections, but not with normal tissue membrane extracts or a human fibroblast line. Ninety-four monoclonal antibodies were purified and tested for binding to 16 normal tissue frozen sections and five blood cell types. Thirty-five of the 94 antibodies were also tested on breast cancer sections from 21 patients, 14 breast cancer cell lines, and 11 nonbreast tumor sections. Two of 94 antibodies showed no reactivity to any of the 21 normal tissues or 11 nonbreast neoplasms studied. These two antibodies, 451B7 and 452F2, bound 60% of the breast cancer cell lines and 25% of the breast cancer tissue sections. Eight additional antibodies bound three or fewer of 21 normal tissues. These antibodies bound 25-85% of breast cancer sections, 0-75% of breast cancer cell lines, and many of the nonbreast cancers. A comparison of normal tissues and nonbreast tumors bound by the breast cancer-reactive antibodies indicated that most of the cross-reacting normal tissue structures were epithelial in origin, and that the most cross-reactive nonbreast cancers were those of secondary sex organs (uterus, prostate, ovary).


Fabrication of Eco-Friendly Solid-State Symmetric Ultracapacitor Device Based on Co-Doped PANI/GO Composite.
An eco-friendly solid-state symmetric ultracapacitor (Uc) device was fabricated using a polyaniline graphene oxide composite co-doped with sulfuric acid (H2SO4) and dodecyl benzene sulfonic acid (DBSA) or camphor sulfonic acid (CSA), as electrode material utilizing gold sheets as current collectors. The device showed specific capacitance value of 150 F/g at 1 A/g current density, with a capacitance retention value of 93.33% at higher current density (10 A/g), indicating a high rate capability. An energy density of 15.30 Whkg-1 with a power density of 1716 Wkg-1 was obtained at the current density of 1 A/g. The values of areal capacitance, power density, and energy density, achieved at the current density of 5 mAcm-2, were 97.38 mFcm-2, 9.93 mWhcm-2, and 1.1 Wcm-2, respectively. Additionally, the device showed very low solution and charge transfer resistance (0.885 Ω and 0.475 Ω, respectively). A device was also fabricated utilizing copper as current collector; however, a lower value of specific capacitance (82 F/g) was observed in this case.


Bond strength of fiber posts in different root thirds using resin cement.
The aim of the study was to assess the in vitro bond strength (BS) of glass fiber posts (GF) and carbon fiber posts (CF) in the cervical, middle, and apical thirds of root canals cemented with RelyX-Unicem (RX) and Cement-Post (CP). Forty maxillary canines were divided into 4 groups (n = 10) according to the cement and post used: group 1: GF and RX; group 2: CF and RX; group 3: GF and CP; group 4: CF and CP. The push-out test was applied in the cervical, middle and apical thirds of each specimen to assess bond strength of the cement/post complex to the root canal wall. The data obtained were submitted to ANOVA (Bonferroni test, p < 0.05), and fracture analysis was done with SEM. The GF posts presented the best results when cemented with RX and with CP (p < 0.05). RX presented the highest BS values for both GF and CF (p < 0.05). For all the groups, BS was higher in the cervical third, followed by the middle and apical thirds. Fracture analysis showed a predominance of cohesive fracture of posts for RX, and a predominance of adhesive fracture between dentin/cement, and mixed failure mode for CP. GF posts cemented with RX presented the highest BS values in all root thirds.


Recovery after Acute Kidney Injury.
Little is known about how acute kidney injury (AKI) resolves, and whether patterns of reversal of renal dysfunction differ among patients with respect to ultimate recovery. We sought to examine different patterns for AKI reversal that are found in patients and assess how they relate to postdischarge outcomes. We studied 16,968 critically ill patients with Kidney Disease Improving Global Outcomes stage 2 or 3 AKI, using an electronic database. Reversal of AKI was defined as alive and no longer meeting criteria for even stage 1. Recovery was defined as reversal at hospital discharge. We observed five patterns. The most common (4,508; 26.6%) was early reversal that was sustained through discharge, but almost as many patients (4,496; 26.5%) had no reversal at all. The remaining patients had late reversal after Day 7 (9.7%); early reversal with one or more relapses, but with ultimate recovery (22.5%); and relapsing without recovery (14.7%). Outcomes for patients with these phenotypes were quite different, with age-adjusted 1-year survival varying from more than 90% for early reversal to less than 40% for patients never reversing. Relapses are common (37.3%), especially in the first 72 hours after reversal, and are associated with a fivefold increased risk for death by 1 year compared with early sustained reversal. We have identified five distinct recovery phenotypes on the basis of the clinical course over the first week after AKI manifestation. These phenotypes may identify patients amenable to therapeutic intervention. Long-term outcomes are associated with recovery status at hospital discharge.


FGF-2 enhances TGF-beta1-induced periosteal chondrogenesis.
The use of periosteum as a cell source for the in vitro engineering of grafts for articular cartilage repair requires the development of methods to obtain high viable cell numbers in the early stages of culture. In this study, we demonstrate that the addition of a mitogen, fibroblast growth factor-2 (FGF-2), during the early stage of the in vitro culture of periosteum in the presence of transforming growth factor-beta1 (TGF-beta1), significantly enhances cell proliferation, which results in increased neo-cartilage formation at later stages. Periosteal explants were cultured in vitro within alginate or agarose based gels in the presence of either FGF-2 for the first week, TGF-beta1 for the first 2 weeks, FGF-2 and TGF-beta1 for the first week and first 2 weeks respectively, or no added factors. Consistent with previous studies, periosteum derived neo-chondrogenesis occurred only in the presence of TGF-beta1. The neo-cartilage was found to contain cartilage specific proteoglycans and Type-II collagen as determined by safranin-O and immunohistochemical staining respectively. Further medium supplementation with FGF-2 stimulated early cell proliferation (>3 fold higher total DNA content per explant at day 10). This resulted in a marked increase in the size of the cultured explants and in the total area of the explant staining positive for safranin-O (from around 50% to 85%, (p<0.05)) after 6 weeks culture. The ability to generate significant quantities of neo-cartilage within a biocompatible and biodegradable matrix such as alginate, which lacks the immunogenicity of agarose, could open new pathways to utilizing such constructs in articular cartilage tissue engineering applications.


A national survey of American Pediatric Anesthesiologists: patient-controlled analgesia and other intravenous opioid therapies in pediatric acute pain management.
The influence of patient characteristics, institutional demographics, and published practice guidelines on the provision of IV opioid analgesia, particularly as delivered through a patient-controlled analgesia (PCA) delivery device, to pediatric patients is unknown. We sent a national, web-based, descriptive survey of pediatric pain management practice to select members of the Society for Pediatric Anesthesia to assess institutional demographics, availability and implementation of IVPCA and PCA by proxy, and recalled occurrence of serious and life-threatening opioid-related side effects. Data from respondents at 252 institutions throughout the United States were collected and analyzed. Sixty-nine percent of respondents practiced in a children's hospital or children's center within a general hospital, and 51% of institutions had a pediatric pain service. Virtually all pediatric pain services (91%) were administered by departments of anesthesiology. Pediatric pain service availability correlated with the number of pediatric beds. IVPCA was available to pediatric patients at 96% of institutions surveyed, whereas IVPCA by proxy was available at only 38%. Eleven percent of respondents reported that their hospital no longer provided IVPCA by proxy as a result of the 2004 Joint Commission on Accreditation of Hospitals Sentinel Event Warning. Instructional material concerning IVPCA was provided to patients or their families by 40% of institutions. IVPCA orders were handwritten by 55% of respondents, despite 39% having computerized provider order entry systems. Ninety percent of respondents reported using pulse oximetry monitoring when patients were administered IVPCA. Forty-two respondents recalled patients having received naloxone to counteract the cardiopulmonary side effects of opioids during the year before receipt of the survey. Eight respondents recalled patient deaths having occurred over the past 5 years in patients receiving IVPCA, IVPCA by proxy, and continuous non-IVPCA opioid infusions. Although IVPCA was available to pediatric patients at most institutions surveyed, prescribing practices and supervision of pediatric pain management were influenced by patient characteristics, institutional demographics, and published national guidelines. Recalled life-threatening events were reported in conjunction with all modes of opioid infusion therapy. Interventions that might diminish the incidence of adverse events but are not used to their fullest extent include improved education and implementation of systems designed to minimize human error involved in the prescribing of opioids. Providing a more accurate accounting of complications would require institutions to participate in a prospective data-collecting consortium designed to track both the incidence of therapy and associated complications.


Cannabimimetic fatty acid derivatives: the anandamide family and other endocannabinoids.
In agreement with the highly lipophilic nature of (-)-delta9-tetrahydrocannabinol, all the endogenous ligands of cannabinoid receptors identified so far are derivatives of long chain fatty acids. N- Arachidonoylethanolamine (anandamide) and some of its polyunsaturated congeners have been found in mammalian brain and shown to activate the CB1 and, with a lower efficacy, CB2 cannabinoid receptor subtypes. More recently, 2-arachidonoylglycerol (2-AG), a widespread intermediate in the metabolism of phosphoglycerides, diacylglycerols and triglycerides, was also found to activate the cannabinoid receptors. The capability of palmitoylethanolamide, an anti-inflammatory metabolite, to activate CB2-like receptors is still being debated. Here we review: 1) the metabolic pathways suggested so far to underlie the biosynthesis and inactivation of anandamide and 2-AG, and 2) the current knowledge of the chemical bases for the interactions of anandamide and 2-AG with proteins of the endogenous cannabinoid system characterized so far, i.e. the CB1 and CB2 receptor subtypes, the membrane anandamide carrier , which facilitates anandamide diffusion into cells, and the enzyme fatty acid amide hydrolase , which catalyzes anandamide and, to a certain extent, 2-AG hydrolysis in vivo.


Quantitative assessment of seminal vesicle and prostate drug concentrations by use of a noninvasive method.
The male genital tract is a complex collection of anatomically and biochemically distinct compartments that contribute to the ejaculate. Understanding the pharmacokinetics in these compartments should inform rational therapeutics involving these glands. Nineteen men were administered a single dose of 600 mg chloroquine (base) and 975 mg aspirin before providing a semen sample by masturbation with fractionation into a 5-compartment collection device. Fractions were assayed for fructose (unique seminal vesicle marker), prostate-specific antigen (unique prostate marker), salicylate, and chloroquine. Seminal vesicle and prostate concentrations of salicylate and chloroquine were estimated via a novel analytic method involving a multilevel latent-variable model implemented by use of Bayesian methods. The geometric mean chloroquine semen/blood ratio was 4.02 (95% confidence interval [CI], 2.36-6.86); for salicylate, the primary metabolite of aspirin, the semen/blood ratio was 0.10 (95% CI, 0.08-0.14). The estimated mean prostate/seminal vesicle ratio for salicylate, 0.38 (95% CI by Bayesian methods, 0.12-0.73), was consistent with our hypothesis that salicylate would achieve higher concentrations in the seminal vesicle than in the prostate. Chloroquine, however, did not demonstrate a statistically significant seminal vesicle/prostate difference (4.41; 95% CI by Bayesian methods, 0.14-30.52). We successfully demonstrated the quantitative, noninvasive estimation of drug concentrations in the prostate gland fluid distinct from the seminal vesicle fluid using our optimized method of split-ejaculate collection and a novel mixed-effects model with Bayesian estimation. Our methods can be applied to gland-specific quantitation of drugs and other substances of interest, thus enabling pharmacokinetic, pharmacodynamic, and pathophysiologic studies to inform rational therapeutics within different glands of the male genital tract.


Gic2p may link activated Cdc42p to components involved in actin polarization, including Bni1p and Bud6p (Aip3p).
Gic2p is a Cdc42p effector which functions during cytoskeletal organization at bud emergence and in response to pheromones, but it is not understood how Gic2p interacts with the actin cytoskeleton. Here we show that Gic2p displayed multiple genetic interactions with Bni1p, Bud6p (Aip3p), and Spa2p, suggesting that Gic2p may regulate their function in vivo. In support of this idea, Gic2p cofractionated with Bud6p and Spa2p and interacted with Bud6p by coimmunoprecipitation and two-hybrid analysis. Importantly, localization of Bni1p and Bud6p to the incipient bud site was dependent on active Cdc42p and the Gic proteins but did not require an intact actin cytoskeleton. We identified a conserved domain in Gic2p which was necessary for its polarization function but dispensable for binding to Cdc42p-GTP and its localization to the site of polarization. Expression of a mutant Gic2p harboring a single-amino-acid substitution in this domain (Gic2p(W23A)) interfered with polarized growth in a dominant-negative manner and prevented recruitment of Bni1p and Bud6p to the incipient bud site. We propose that at bud emergence, Gic2p functions as an adaptor which may link activated Cdc42p to components involved in actin organization and polarized growth, including Bni1p, Spa2p, and Bud6p.


Physical activity level of adult cats with varied feeding frequency.
The prevalence of feline obesity is influenced by numerous factors, including inactivity and overconsumption of food. The objective of the present study was to evaluate the effect of feeding frequency on physical activity in adult cats. A total of twelve healthy adult cats were used in a cross-over study consisting of 32 d. In each of the two periods, six cats were fed either two meals or four meals daily. Throughout the study, cats were fed the same diet at amounts to maintain body weight and body condition score. Cats were individually housed 4 h/d at each scheduled feeding time, while for the other 20 h, cats were group-housed to allow for voluntary physical activity in the room with a 16 h light-8 h dark cycle. Voluntary activity levels were evaluated using Actical activity collars for seven consecutive days in each period. Daily average activity level for two-meal-fed cats (20.04 (SEM 2.19), activity counts/epoch (15 s)) was not different from four-meal-fed cats (20.14 (SEM 2.15), activity counts/epoch (15 s); P>0.05). In conclusion, when group-housed cats are fed to maintain their body weight and body condition score, varied feeding frequency between twice and four times daily may not affect activity levels.


Martelella endophytica sp. nov., an antifungal bacterium associated with a halophyte.
A Gram-staining-negative, non-spore-forming endophytic bacterium, designated strain YC6887(T), was isolated from a root sample of a halophyte, Rosa rugosa, collected from a tidal flat area of Namhae Island, located at the southern end of Korea. Strain YC6887(T) was found to exhibit inhibitory activity against oomycete plant pathogens. The cells were non-motile and aerobic rods. The strain was able to grow at 4-40 °C (optimum 28-30 °C) and at pH 5.0-9.0 (optimum pH 7.0-8.5). Strain YC6887(T) was able to grow at NaCl concentrations of 0-9 % (w/v) with optimum growth at 4-5 % (w/v) NaCl, but NaCl is not essential for growth. Comparison of 16S rRNA gene sequences showed that the strain was a member of the genus Martelella, a member of order Rhizobiales, exhibiting highest similarity with Martelella mediterranea (98.6 %). The DNA-DNA relatedness between strain YC6887(T) and M. mediterranea MACL11(T) was 19.8 ± 6.8. Chemotaxonomically, strain YC6887(T) contained C19 : 0 cyclo ω8c (28.0 %) and C18 : 1ω7c (17.9 %) as predominant fatty acids, confirming the affiliation of strain YC6887(T) with the genus Martelella. The major respiratory quinone was Q-10 and the DNA G+C content was 62.1 mol%. On the basis of phylogenetic analysis, physiological and biochemical characterization and DNA-DNA hybridization data, strain YC6887(T) should be classified as representing a novel species of the genus Martelella, for which the name Martelella endophytica sp. nov. is proposed. The type strain is YC6887(T) ( = KCCM 43011(T) = NBRC 109149(T)).


Analysis of complex cardiovascular flow with three-component acceleration-encoded MRI.
Functional information regarding cardiac performance, pressure gradients, and local flow derangement are available from blood acceleration fields. Thus, this study examines a 2D and 3D phase contrast sequence optimized to efficiently encode three-directional, time-resolved acceleration in vitro and in vivo. Stenosis phantom acceleration measurements were compared to acceleration derived from standard velocity encoded phase contrast-magnetic resonance imaging (i.e., "velocity-derived acceleration"). For in vivo analysis, three-directional 2D acceleration maps were compared to velocity-derived acceleration using regions proximal and distal to the aortic valve in six healthy volunteers at 1.5 and 3.0 T (voxel size = 1.4 × 2.1 × 8 mm, temporal resolution = 16-20 ms). In addition, a 4D acceleration sequence was evaluated for feasibility in a healthy volunteer and postrepair biscuspid aortic valve patient with an ascending aortic aneurysm. The phantom magnetic resonance acceleration measurements were more accurate (nonturbulent root mean square error = 2.2 vs. 5.1 m/s(2) for phase contrast-magnetic resonance imaging) and 10 times less noisy (nonturbulent σ = 0.9 vs. 13.6 m/s(2) for phase contrast-magnetic resonance imaging) than velocity-derived acceleration. Acceleration mapping of the left ventricular outflow tract and aortic arch exhibited signal voids colocated with complex flow events such as vortex formation and high order motion. 4D acceleration data, visualized in combination with the velocity data, may provide new insight into complex flow phenomena.


Foam sclerotherapy for lower-limb varicose veins: impact on saphenous vein diameter.
To assess changes in the great saphenous vein (GSV) after foam sclerotherapy for varicose veins. This was a prospective study of 33 patients who were treated with polidocanol foam sclerotherapy after having had varicose veins with a clinical severity-etiology-anatomy-pathophysiology classification of C4-C6 for three months. The patients were evaluated by ultrasound before, during, and after the procedure (on post-procedure days 7, 15, 30, 60, and 90). The GSV diameter, the rate of venous occlusion, and the rate of reflux elimination were determined. Two patients were excluded for having a history of deep vein thrombosis history, and one was excluded for having bronchial asthma. Thirty patients (26 females and 4 males, with mean age of 62 years) completed the protocol. The mean pre-procedure GSV diameter was 6.0 ± 0.32 mm (range, 3.6-11.2 mm). During the sclerotherapy, the mean GSV diameter was reduced to 1.9 ± 0.15 mm (range, 0.6-3.8 mm). On post-procedure day 7, the mean GSV diameter increased to 6.3 ± 0.28 mm (range, 3.9-9.7 mm). On post-procedure day 90, the mean GSV diameter was 4.0 ± 0.22 mm (range, 1.9-8.2 mm). The rate of GSV reflux was significantly lower in the assessment performed on post-procedure day 90 than in the pre-procedure assessment (p < 0.0028). On the basis of our ultrasound analysis, we can conclude that foam sclerotherapy for varicose veins results in a significant reduction in GSV diameter, as well as in the elimination of GSV reflux.


Urinary Biomarkers of Carcinogenic Exposure among Cigarette, Waterpipe, and Smokeless Tobacco Users and Never Users of Tobacco in the Golestan Cohort Study.
How carcinogen exposure varies across users of different, particularly noncigarette, tobacco products remains poorly understood. We randomly selected 165 participants of the Golestan Cohort Study from northeastern Iran: 60 never users of any tobacco, 35 exclusive cigarette, 40 exclusive (78% daily) waterpipe, and 30 exclusive smokeless tobacco (nass) users. We measured concentrations of 39 biomarkers of exposure in 4 chemical classes in baseline urine samples: tobacco alkaloids, tobacco-specific nitrosamines (TSNA), polycyclic aromatic hydrocarbons (PAH), and volatile organic compounds (VOC). We also quantified the same biomarkers in a second urine sample, obtained 5 years later, among continuing cigarette smokers and never tobacco users. Nass users had the highest concentrations of tobacco alkaloids. All tobacco users had elevated TSNA concentrations, which correlated with nicotine dose. In both cigarette and waterpipe smokers, PAH and VOC biomarkers were higher than never tobacco users and nass users, and highly correlated with nicotine dose. PAH biomarkers of phenanthrene and pyrene and two VOC metabolites (phenylmercapturic acid and phenylglyoxylic acid) were higher in waterpipe smokers than in all other groups. PAH biomarkers among Golestan never tobacco users were comparable to those in U.S. cigarette smokers. All biomarkers had moderate to good correlations over 5 years, particularly in continuing cigarette smokers. We observed two patterns of exposure biomarkers that differentiated the use of the combustible products (cigarettes and waterpipe) from the smokeless product. Environmental exposure from nontobacco sources appeared to contribute to the presence of high levels of PAH metabolites in the Golestan Cohort. Most of these biomarkers would be useful for exposure assessment in a longitudinal study.


The case for open-source software in drug discovery.
Widespread adoption of open-source software for network infrastructure, web servers, code development, and operating systems leads one to ask how far it can go. Will "open source" spread broadly, or will it be restricted to niches frequented by hopeful hobbyists and midnight hackers? Here we identify reasons for the success of open-source software and predict how consumers in drug discovery will benefit from new open-source products that address their needs with increased flexibility and in ways complementary to proprietary options.


Atomic view of the histidine environment stabilizing higher-pH conformations of pH-dependent proteins.
External stimuli are powerful tools that naturally control protein assemblies and functions. For example, during viral entry and exit changes in pH are known to trigger large protein conformational changes. However, the molecular features stabilizing the higher pH structures remain unclear. Here we elucidate the conformational change of a self-assembling peptide that forms either small or large nanotubes dependent on the pH. The sub-angstrom high-pH peptide structure reveals a globular conformation stabilized through a strong histidine-serine H-bond and a tight histidine-aromatic packing. Lowering the pH induces histidine protonation, disrupts these interactions and triggers a large change to an extended β-sheet-based conformation. Re-visiting available structures of proteins with pH-dependent conformations reveals both histidine-containing aromatic pockets and histidine-serine proximity as key motifs in higher pH structures. The mechanism discovered in this study may thus be generally used by pH-dependent proteins and opens new prospects in the field of nanomaterials.


The role of methionine on metabolism, oxidative stress, and diseases.
Methionine is an aliphatic, sulfur-containing, essential amino acid, and a precursor of succinyl-CoA, homocysteine, cysteine, creatine, and carnitine. Recent research has demonstrated that methionine can regulate metabolic processes, the innate immune system, and digestive functioning in mammals. It also intervenes in lipid metabolism, activation of endogenous antioxidant enzymes such as methionine sulfoxide reductase A, and the biosynthesis of glutathione to counteract oxidative stress. In addition, methionine restriction prevents altered methionine/transmethylation metabolism, thereby decreasing DNA damage and carcinogenic processes and possibly preventing arterial, neuropsychiatric, and neurodegenerative diseases. This review focuses on the role of methionine in metabolism, oxidative stress, and related diseases.


Open source software for the analysis of microarray data.
DNA microarray assays represent the first widely used application that attempts to build upon the information provided by genome projects in the study of biological questions. One of the greatest challenges with working with microarrays is collecting, managing, and analyzing data. Although several commercial and noncommercial solutions exist, there is a growing body of freely available, open source software that allows users to analyze data using a host of existing techniques and to develop their own and integrate them within the system. Here we review three of the most widely used and comprehensive systems, the statistical analysis tools written in R through the Bioconductor project (http://www.bioconductor.org), the Java-based TM4 software system available from The Institute for Genomic Research (http://www.tigr.org/software), and BASE, the Web-based system developed at Lund University (http://base.thep.lu.se).


Evaluation of multi-task learning in deep learning-based positioning classification of mandibular third molars.
Pell and Gregory, and Winter's classifications are frequently implemented to classify the mandibular third molars and are crucial for safe tooth extraction. This study aimed to evaluate the classification accuracy of convolutional neural network (CNN) deep learning models using cropped panoramic radiographs based on these classifications. We compared the diagnostic accuracy of single-task and multi-task learning after labeling 1330 images of mandibular third molars from digital radiographs taken at the Department of Oral and Maxillofacial Surgery at a general hospital (2014-2021). The mandibular third molar classifications were analyzed using a VGG 16 model of a CNN. We statistically evaluated performance metrics [accuracy, precision, recall, F1 score, and area under the curve (AUC)] for each prediction. We found that single-task learning was superior to multi-task learning (all p < 0.05) for all metrics, with large effect sizes and low p-values. Recall and F1 scores for position classification showed medium effect sizes in single and multi-task learning. To our knowledge, this is the first deep learning study to examine single-task and multi-task learning for the classification of mandibular third molars. Our results demonstrated the efficacy of implementing Pell and Gregory, and Winter's classifications for specific respective tasks.


Evolution of enzymatic activities in the enolase superfamily: structure of a substrate-liganded complex of the L-Ala-D/L-Glu epimerase from Bacillus subtilis.
The members of the mechanistically diverse enolase superfamily share a bidomain structure formed from a (beta/alpha)7beta-barrel domain [a modified (beta/alpha)8- or TIM-barrel] and a capping domain formed from N- and C-terminal segments of the polypeptide. The active sites are located at the interface between the C-terminal ends of the beta-strands in the barrel domain and two flexible loops in the capping domain. Within this structure, the acid/base chemistry responsible for formation and stabilization of an enediolate intermediate derived from a carboxylate anion substrate and the processing of it to product is "hard-wired" by functional groups at the C-terminal ends of the beta-strands in the barrel domain; the identity of the substrate is determined in part by the identities of residues located at the end of the eighth beta-strand in the barrel domain and two mobile loops in the capping domain. On the basis of the identities of the acid/base functional groups at the ends of the beta-strands, the currently available structure-function relationships derived from functionally characterized members are often sufficient for "deciphering" the identity of the chemical reaction catalyzed by sequence-divergent members discovered in genome projects. However, insufficient structural information for liganded complexes for specifying the identity of the substrate is available. In this paper, the structure of the complex of L-Ala-L-Glu with the L-Ala-D/L-Glu epimerase from Bacillus subtilis is reported. As expected for the 1,1-proton transfer reaction catalyzed by this enzyme, the alpha-carbon of the substrate is located between Lys 162 and Lys 268 at the ends of the second and sixth beta-strands in the barrel domain. The alpha-ammonium group of the l-Ala moiety is hydrogen bonded to both Asp 321 and Asp 323 at the end of the eighth beta-strand, revealing a novel strategy for substrate recognition in the superfamily. The delta-carboxylate group of the Glu moiety is hydrogen bonded to Arg 24 in one of the flexible loops in the capping domain, thereby providing a structural explanation for the restricted substrate specificity of this epimerase [Schmidt, D. M., Hubbard, B. K., and Gerlt, J. A. (2001) Biochemistry 40, 15707-15715]. These studies provide important new information about the structural bases for substrate specificity in the enolase superfamily.


Rational material design for ultrafast rechargeable lithium-ion batteries.
Rechargeable lithium-ion batteries (LIBs) are important electrochemical energy storage devices for consumer electronics and emerging electrical/hybrid vehicles. However, one of the formidable challenges is to develop ultrafast charging LIBs with the rate capability at least one order of magnitude (>10 C) higher than that of the currently commercialized LIBs. This tutorial review presents the state-of-the-art developments in ultrafast charging LIBs by the rational design of materials. First of all, fundamental electrochemistry and related ionic/electronic conduction theories identify that the rate capability of LIBs is kinetically limited by the sluggish solid-state diffusion process in electrode materials. Then, several aspects of the intrinsic materials, materials engineering and processing, and electrode materials architecture design towards maximizing both ionic and electronic conductivity in the electrode with a short diffusion length are deliberated. Finally, the future trends and perspectives for the ultrafast rechargeable LIBs are discussed. Continuous rapid progress in this area is essential and urgent to endow LIBs with ultrafast charging capability to meet huge demands in the near future.


Rational phonological lengthening in spoken Dutch.
Dutch allows optional schwa insertion between a liquid and obstruent in words like film/filəm ("film") and dorp/dorəp ("village"), lengthening the word by one syllable. This epenthesis is productive and widespread, and is understood to be a phonological rather than phonetic process. A corpus analysis shows that a speaker's choice between the variant forms is influenced by probability: words that are less frequent or less probable given their immediate or discourse context are more likely to be lengthened. This may reflect a rational communication strategy in which language is manipulated to efficiently transmit information. As these results unambiguously show that lengthening is probabilistically influenced, they are informative to the understanding of the production mechanisms underlying pronunciation variation.


Are friends really the family we choose? Local variations of hypothalamus activity when viewing personally known faces.
Sibling and friend relationships have significant impact on individuals' socio-emotional development. Hypothalamic supraoptic nucleus (SON) and paraventricular nucleus (PVN) synthesize and secrete neuropeptides, including oxytocin, associated with attachment behaviors. Here, using fMRI, we investigate the implication of these two hypothalamic nuclei in the visual processing of personally known faces. Faces of same-sex sibling, best friend, celebrity, and unknown person appear in the middle of the screen while participants perform a task requiring a button click each time a central white dot turns red. Ratings of familiarity (time spent together) and emotionality (feelings toward individual) toward the four individuals are recorded. Local activation within the hypothalamus is assessed via two complementary methods: (1) voxel-based analyses within inclusive mask of the hypothalamus; (2) region-of-interest (ROI) analysis of partial hypothalamic volumes using SON and PVN as center of mass coordinates, with percent signal change extracted and analyzed within these ROIs. Results suggest that the SON responds to all familiar individuals while the PVN has increased response to sibling compared to friend faces and is correlated to familiarity but not emotionality. These findings support differential involvement of local hypothalamic substructures SON and PVN in response to faces of individuals with different social relationships.


Towards in silico design of epitope-based vaccines.
Epitope-based vaccines (EVs) make use of immunogenic peptides (epitopes) to trigger an immune response. Due to their manifold advantages, EVs have recently been attracting growing interest. The success of an EV is determined by the choice of epitopes used as a basis. However, the experimental discovery of candidate epitopes is expensive in terms of time and money. Furthermore, for the final choice of epitopes various immunological requirements have to be considered. Numerous in silico approaches exist that can guide the design of EVs. In particular, computational methods for MHC binding prediction have already become standard tools in immunology. Apart from binding prediction and prediction of antigen processing, methods for epitope design and selection have been suggested. We review these in silico approaches for epitope discovery and selection along with their strengths and weaknesses. Finally, we discuss some of the obvious problems in the design of EVs. State-of-the-art in silico approaches to MHC binding prediction yield high accuracies. However, a more thorough understanding of the underlying biological processes and significant amounts of experimental data will be required for the validation and improvement of in silico approaches to the remaining aspects of EV design.


The German Multidimensional Attitude Scale Toward Persons With Disabilities (G-MAS): A factor analytical study among high-school students.
This study investigates the psychometric properties of the Multidimensional Attitudes Scale Toward Persons with Disabilities (MAS) among German 10th graders, to advance knowledge about the situation in German classrooms with regard to attitudes toward persons with disabilities. Four hundred twenty-one German 10th graders attending the most common regular schools of the German educational system completed the MAS. Confirmatory factor analyses, multigroup confirmatory factor analyses, multivariate analyses of variance and model based clustering were used to investigate: the factor structure, invariance of the factor structure across gender and school types, differences on the MAS between gender and school types, and the existence of groups of students with different attitudes. The authors reject previously proposed measurement models of the MAS, based on inadequate model fit. They identify a factor structure of the MAS with good model fit, the German MAS (G-MAS). The G-MAS, like 2 recently reported factor structures, assesses 4 dimensions: behavioral avoidance, positive cognitions, and 2 affective dimensions-calm and negative affect. The G-MAS is invariant across gender, but not across school types. The authors find no differences on these dimensions between students from different types of schools, but there are gender differences. Findings are discussed in light of the outcome that, of the studied sample, less than 1/3 of the students clearly expresses positive attitudes toward persons with disabilities. (PsycINFO Database Record


Effect of beta-cyclodextrin on pasting properties of wheat starch.
Changes of viscosity characteristics of genetically diverse hexaploid wheat starches during pasting in water, 1% NaCl solution, or at pH 4 or pH 10 were studied using a Rapid Visco-Analyzer. Peak viscosity (PV), hot paste viscosity (HPV) and cool paste viscosity (CPV) of all the wheat starches was little affected in pH 4 and pH 10 treatments. In 1% NaCl, all starches showed substantial increases in all three parameters relative to pasting in water. The use of 1% beta-cyclodextrin (beta-CD) (cycloheptaamylose) solution increased PV of high-swelling starches, but generally slightly decreased that of low swelling starches in all treatment conditions. HPV was always reduced by addition of 1% beta-CD, but CPV was increased in most treatments for Anza and Yecora Rojo (low swelling), but decreased for Klasic (high swelling). Bacterial alpha-amylase was added to starch or flour. The effect of beta-CD was shown to be independent of alpha-amylase inhibition in wheat starch, but beta-CD strongly inhibited alpha-amylase in wheat flour.


Loss of MicroRNA-21 Influences the Gut Microbiota, Causing Reduced Susceptibility in a Murine Model of Colitis.
microRNAs regulate gene expression and influence the pathogenesis of human diseases. The present study investigated the role of microRNA-21 [miR-21] in the pathogenesis of intestinal inflammation, because miR-21 is highly expressed in inflammatory bowel disease. Inflammatory bowel disease is associated with intestinal barrier dysfunction and an altered gut microbiota. Recent studies have demonstrated that host microRNAs can shape the microbiota. Thus, we determined the influence of miR-21 on the gut microbiota and observed the subsequent impact in a dextran sodium sulphate [DSS]-induced colitis model. The influence of miR-21 on the gut microbiota and inflammation was assessed in wild-type [WT] and miR-21-/- mice, in co-housed mice, following antibiotic depletion of the microbiota, or by colonization of germ-free [GF] mice with fecal homogenate, prior to DSS administration. We carried out 16S rRNA sequencing on WT and miR-21-/- mice to dissect potential differences in the gut microbiota. miR-21-/- mice have reduced susceptibility to DSS-induced colitis compared with WT mice. Co-housing conferred some protection to WT mice, while GF mice colonized with fecal homogenate from miR-21-/- were protected from DSS colitis compared with those colonized with WT homogenate. Further supporting a role for the microbiota in the observed phenotype, the protection afforded by miR-21 depletion was lost when mice were pre-treated with antibiotics. The 16S rRNA sequencing revealed significant differences in the composition of WT and miR-21-/- intestinal microbiota. These findings suggest that miR-21 influences the pathogenesis of intestinal inflammation by causing propagation of a disrupted gut microbiota.


Friction Durability of Extremely Thin Diamond-Like Carbon Films at High Temperature.
To clarify the friction durability, both during and after the high-temperature heating of nanometer-thick diamond-like carbon (DLC) films, deposited using filtered cathodic vacuum arc (FCVA) and plasma chemical vapor deposition (P-CVD) methods, the dependence of the friction coefficient on the load and sliding cycles of the DLC films, were evaluated. Cluster-I consisted of a low friction area in which the DLC film was effective, while cluster-II consisted of a high friction area in which the lubricating effect of the DLC film was lost. The friction durability of the films was evaluated by statistical cluster analysis. Extremely thin FCVA-DLC films exhibited an excellent wear resistance at room temperature, but their friction durability was decreased at high temperatures. In contrast, the durability of the P-CVD-DLC films was increased at high temperatures when compared with that observed at room temperature. This inverse dependence on temperature corresponded to the nano-friction results obtained by atomic force microscopy. The decrease in the friction durability of the FCVA-DLC films at high temperatures, was caused by a complex effect of temperature and friction. The tribochemical reaction produced by the P-CVD-DLC films reduced their friction coefficient, increasing their durability at high temperatures.


A phylogenomic approach reveals a low somatic mutation rate in a long-lived plant.
Somatic mutations can have important effects on the life history, ecology, and evolution of plants, but the rate at which they accumulate is poorly understood and difficult to measure directly. Here, we develop a method to measure somatic mutations in individual plants and use it to estimate the somatic mutation rate in a large, long-lived, phenotypically mosaic Eucalyptus melliodora tree. Despite being 100 times larger than Arabidopsis, this tree has a per-generation mutation rate only ten times greater, which suggests that this species may have evolved mechanisms to reduce the mutation rate per unit of growth. This adds to a growing body of evidence that illuminates the correlated evolutionary shifts in mutation rate and life history in plants.


Rice-based oral rehydration solution for treating diarrhoea.
Oral rehydration therapy is used to treat dehydration caused by diarrhoea. However the rehydration solution does not reduce stool loss or length of illness. A solution able to do this may lessen the use of ineffective diarrhoea treatments as well as improve morbidity and mortality related to diarrhoea. The objective of this review was to assess the effects of rice-based oral rehydration salts solution compared with glucose-based oral rehydration salts solution on reduction of stool output and duration of diarrhoea in patients with acute watery diarrhoea. We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase, Lilacs and the reference lists of relevant articles. We also contacted researchers in the field. Randomized trials comparing standard World Health Organization oral rehydration solution with an experimental oral rehydration salts solution in which glucose (20 grams per litre) was replaced by 50-80 grams per litre of rice powder, with the electrolytes remaining unchanged. Data were extracted independently by a statistician and a clinician. Twenty-two trials were included. Concealment of allocation was adequate in 15 of these trials. Irrespective of age, people with cholera who were given rice oral rehydration salts solution had substantially lower rates of stool loss than those given oral rehydration salts solution in the first 24 hours. Mean stool outputs in the first 24 hours were lower by 67 millilitres/kg of body weight (weighted mean difference -67.4, 95% confidence interval -94.3 to -41.0) in children, and by 51 millilitres/kg of body weight (weighted mean difference -51.1, 95% confidence interval -65.9 to -36.3) in adults. The rate of stool loss in infants and children with acute non-cholera diarrhoea was reduced by only four millilitres/kg of body weight (weighted mean difference -4.3, 95% confidence interval -9.3 to 0.8). Rice-based oral rehydration appears to be effective in reducing stool output in people with cholera. This effect was not apparent in infants and children with non-cholera diarrhoea.


Toward an interpretable Alzheimer's disease diagnostic model with regional abnormality representation via deep learning.
In this paper, we propose a novel method for magnetic resonance imaging based Alzheimer's disease (AD) or mild cognitive impairment (MCI) diagnosis that systematically integrates voxel-based, region-based, and patch-based approaches into a unified framework. Specifically, we parcellate the brain into predefined regions based on anatomical knowledge (i.e., templates) and derive complex nonlinear relationships among voxels, whose intensities denote volumetric measurements, within each region. Unlike existing methods that use cubical or rectangular shapes, we consider the anatomical shapes of regions as atypical patches. Using complex nonlinear relationships among voxels in each region learned by deep neural networks, we extract a "regional abnormality representation." We then make a final clinical decision by integrating the regional abnormality representations over the entire brain. It is noteworthy that the regional abnormality representations allow us to interpret and understand the symptomatic observations of a subject with AD or MCI by mapping and visualizing these observations in the brain space. On the baseline MRI dataset from the Alzheimer's Disease Neuroimaging Initiative (ADNI) cohort, our method achieves state-of-the-art performance for four binary classification tasks and one three-class classification task. Additionally, we conducted exhaustive experiments and analysis to validate the efficacy and potential of our method.


Cellulase production by Thermomonospora curvata isolated from municipal solid waste compost.
A cellulolytic, thermophilic actinomycete (previously isolated from municipal refuse compost samples) was identified as Thermomonospora curvata. A determination was made of the optimal conditions for cellulase production by T. curvata when grown at 55 C in a medium containing mineral salts, cellulose, and yeast extract. The pH and temperature optima (pH 6.0 and 65 C) for the cellulase produced by T. curvata were identical to those previously observed for the cellulase extracted from crude compost samples. Such similarities, together with the prevalence of T. curvata in compost samples and its ability to grow at composting temperatures, indicate that this actinomycete could possibly be considered as a major cellulose decomposer in the municipal refuse composting process.


Dual-energy micro-CT with a dual-layer, dual-color, single-crystal scintillator.
A wide range of X-ray imaging applications demand micrometer spatial resolution. In material science and biology especially, there is a great interest in material determination and material separation methods. Here we present a new detector design that allows the recording of a low- and a high-energy radiography image simultaneously with micrometer spatial resolution. The detector system is composed of a layered scintillator stack, two CCDs and an optical system to image the scintillator responses onto the CCDs. We used the detector system with a standard laboratory microfocus X-ray tube to prove the working principle of the system and derive important design characteristics. With the recorded and registered dual-energy data set, the material separation and determination could be shown at an X-ray tube peak energy of up to 160 keV with a spatial resolution of 12 μm. The detector design shows a great potential for further development and a wide range of possible applications.


Adaptive immune responses of legumin nanoparticles.
Legumin is one of the main storage proteins in the pea seeds (Pisum sativum L.) and the molecules of this protein have the capacity of binding together to form nanoparticles after aggregation and chemical cross-linkage with glutaraldehyde. The aim of this work was to study the adaptive immune response of legumin nanoparticles in rats. Following intradermal immunisation with the native protein legumin and legumin nanoparticles of about 250 nm, the humoral and cell-mediated immune responses were analysed in rats. The humoral responses against legumin and legumin nanoparticles were examined by western blot and ELISA analysis. Both techniques clearly showed that sera from rats immunised with legumin strongly expressed antibodies against this protein. On the contrary, serum samples from rats inoculated with legumin nanoparticles did not contain detectable amounts of antibodies. These results may be explained by a reduction on the antigenic epitopes of the protein induced by the glutaraldehyde used during the cross-linking step. Concerning the cell-mediated response, neither legumin nor legumin nanoparticles stimulated an immunogenic response. This absence of response of spleen lymphocytes for legumin and legumin nanoparticles may be explained by a cytostatic effect of legumin which was corroborated by the evaluation of the middle phase of cell apoptose. In fact, both legumin and legumin nanoparticles are potent inductors of a cytostatic phenomenon and showed a significant increase of the chromatin condensation (p < 0.05) as compared with control.


Assessment of the in vitro cytotoxicity and in vivo anti-tumor activity of the alcoholic stem bark extract/fractions of Mimusops elengi Linn.
Various parts of Mimusops elengi Linn. (Sapotaceae) have been used widely in traditional Indian medicine for the treatment of pain, inflammation and wounds. The study was conducted to explore the use of stem bark of M. elengi on pharmacological grounds and to evaluate the scientific basis of cytotoxic and anti-tumor activity. Extract/fractions were prepared and in vitro cytotoxicity was assessed using SRB assay. Most effective fractions were subjected to fluorescence microscopy based acridine orange/ethidium bromide (AO/EB) and Hoechst 33342 staining to determine apoptosis induction and DNA fragmentation assay. Comet and micronuclei assay were performed to assess genotoxicity. Cell cycle analysis was also performed. In vivo anti-tumor potential was evaluated by Ehrlich ascites carcinoma (EAC) model in mice. The alcoholic stem bark extract of M. elengi along with four fractions showed potential in vitro cytotoxicity in SRB assay. Of these, dichloromethane and ethyl acetate fractions were selected for further studies. The fractions revealed apoptosis inducing potential in AO/EB and Hoechst 33342 staining, which was further confirmed by DNA fragmentation assay. Genotoxic potential was revealed by comet and micronuclei assay. Fractions also exhibited specific cell cycle inhibition in G0/G1 phase. In EAC model, ethyl acetate fraction along with the standard (cisplatin) effectively reduced the increase in body weight compared to control and improved mean survival time. Both fractions were able to restore the altered hematological and biochemical parameters. Hence, M. elengi stem bark may be a possible therapeutic candidate having cytotoxic and anti-tumor potential.


Physical behaviors and fundamental movement skills in British and Iranian children: An isotemporal substitution analysis.
Although the relationship between fundamental movement skills (FMS) and physical behaviors has been established, differences between countries are scarcely explored. The impact of the whole physical behavior composition, in relation to FMS, has yet to be investigated in 9-11 y children. The aims were to investigate the associations of substitution of physical behaviors with FMS score and to compare traditional linear regression and compositional data analysis and compare between England and Iran. Measures included accelerometer-derived activity (sleep (SL), sedentary behavior (SB), light physical activity (LPA), and moderate-to-vigorous physical activity (MVPA) and FMS, using the TGMD-2, in 119 children (64 boys) from Iran (mean (±SD) age: 9.8 ± 0.3 y; BMI of 18.2 ± 3.3 kg/m2 ) and 139 (61 boys) children from England (mean (±SD) age: 9.5 ± 0.6 y; BMI of 17.7 ± 3.1 kg/m2 ). Isometric log-ratio multiple linear regression models were used to discern the association between FMS and the mean activity composition, and for new compositions, where fixed durations of time were reallocated from one behavior to another, while the remaining behaviors were unchanged. In physical behaviors as a composition, FMS was significantly associated in both ethnicities. English children responded significantly positively to adding 5 or more minutes LPA at the expense of SB (FMS unit change from 0.05 [0.01, 0.09] at 5 minutes to 0.72 [0.01, 1.34] at 60 minutes). Adding 10 minutes or more of SL, at the expense of SB, was associated with a significant, positive change in FMS in all children. Investigation is needed to understand the composition of SB and its potential influence on FMS development.


The mediating role of AKT/ERK/JNK signaling on the malignant phenotype of microcystin-LR in gastric adenocarcinoma cells.
Microcystin-leucine arginine (MC-LR), a widely distributed and highly toxic environmental pollutant, plays crucial roles in cancer malignancy by activating characteristically toxic signaling pathways. Traditional animal-based toxicity evaluation methods have proven insufficient for identifying the specific role of these signaling pathways. Therefore, this study aimed to uncover the regulatory relationship between the toxic pathways and the progression of gastric cancer (GC). The findings provide novel avenues for conducting in vitro toxicity tests based on the investigated pathways. We found that MC-LR promoted the migration and invasion of SGC-7901 cells while simultaneously inhibiting their apoptosis in a dose-dependent manner. This observed cytotoxicity was primarily mediated through the AKT, JNK, and ERK signaling pathways. By using a mediation analysis model, we determined that AKT and ERK exhibited competitive effects in MC-LR-treated GC malignancy, while AKT and JNK acted independently from one another. This study establishes an in vitro toxicity test model of MC-LR based on toxicity-related pathways and underscores the pivotal roles of AKT, ERK, and JNK signaling in MC-LR toxicity. The findings offer a novel, fundamental framework for conducting chemical toxicity risk assessment.


No deaths associated with patient refusal of transport after naloxone-reversed opioid overdose.
Naloxone is widely used in the treatment and reversal of opioid overdose. Most emergency medical services (EMS) systems administer naloxone by standing order, and titrate only to reverse respiratory depression without fully reversing sedation. Some EMS systems routinely administer sufficient naloxone to fully reverse the effects of opioid overdose. Frequently patients refuse further medical evaluation or intervention, including transport. The purpose of this study was to evaluate the safety of this practice and determine whether increased mortality is associated with full reversal of opioids. As a component of a comprehensive quality assurance initiative, we assessed mortality during the 48 hours after patients received naloxone to reverse opioid overdose followed by patient-initiated refusal of transportation. The setting was a large urban fire-based EMS system. Investigators provided the Bexar County Medical Examiner's Office (MEO) with a list of patients who were treated by the San Antonio Fire Department with naloxone, and not transported. Inclusion criteria were administration of naloxone and patient-initiated refusal. Patient dispositions also included aid only, referral to the MEO, or referral to law enforcement. The list was then compared with the MEO database. A chart review was completed on all patients treated and subsequently presented to the MEO within two days. A secondary time period of 30 days was also assessed. The list identified 592 patients treated with naloxone and not transported to the emergency department. Five-hundred fifty-two patients received naloxone and refused transport or were not transported. The remaining 40 patients all presented to EMS in cardiac arrest, naloxone was administered during the course of resuscitation, and subsequent efforts were terminated in the field. None of the patients receiving naloxone with a subsequent patient-initiated refusal were examined at the MEO within the two-day end point. The 30-day assessment revealed that nine individuals were treated with naloxone and subsequently died, but the shortest time interval between date of service and date of death was four days. The primary outcome was that no patients who were treated with naloxone for opioid overdose and then refused care were examined by the MEO within a 48-hour time frame.


Crystal structure of the T877A human androgen receptor ligand-binding domain complexed to cyproterone acetate provides insight for ligand-induced conformational changes and structure-based drug design.
Cyproterone acetate (CPA) is a steroidal antiandrogen used clinically in the treatment of prostate cancer. Compared with steroidal agonists for the androgen receptor (AR) (e.g. dihydrotestosterone, R1881), CPA is bulkier in structure and therefore seemingly incompatible with the binding pockets observed in currently available x-ray crystal structures of the AR ligand-binding domain (LBD). We solved the x-ray crystal structure of the human AR LBD bound to CPA at 1.8A in the T877A variant, a mutation known to increase the agonist activity of CPA and therefore facilitate purification and crystal formation of the receptor.drug complex. The structure demonstrates that bulk from the 17alpha-acetate group of CPA induces movement of the Leu-701 side chain, which results in partial unfolding of the C-terminal end of helix 11 and displacement of the loop between helices 11 and 12 in comparison to all other AR LBD crystal structures published to date. This structural alteration leads to an expansion of the AR binding cavity to include an additional pocket bordered by Leu-701, Leu-704, Ser-778, Met-780, Phe-876, and Leu-880. Further, we found that CPA invokes transcriptional activation in the L701A AR at low nanomolar concentrations similar to the T877A mutant. Analogous mutations in the glucocorticoid receptor (GR) and progesterone receptor were constructed, and we found that CPA was also converted into a potent agonist in the M560A GR. Altogether, these data offer information for structure-based drug design, elucidate flexible regions of the AR LBD, and provide insight as to how CPA antagonizes the AR and GR.


Comparative removal of As(V) and Sb(V) from aqueous solution by sulfide-modified α-FeOOH.
Efficient elimination of As(V) and Sb(V) from wastewater streams has long been a major challenge. Herein, sulfide-modified α-FeOOH adsorbent was fabricated via a simple sulfidation reaction for removing As(V) and Sb(V) from aqueous media. Compared with the pristine α-FeOOH, sulfide-modified α-FeOOH increased the adsorption of As(V) from 153.8 to 384.6 mg/g, and Sb(V) adsorption from 277.8 to 1111.1 mg/g. The enhanced adsorption of both As(V) and Sb(V) was maintained at the pH range from 2 to 11, and was not interfered by various coexisting anions such as Cl-, SO42-, NO3-, SiO32- and PO43-. The adsorption affinity increased from 0.0047 to 0.0915 and 0.0053 to 0.4091 for As(V) and Sb(V), respectively. X-ray photoelectron spectroscopic investigation demonstrated a reductive conversion of As(V) to As(III) during the adsorption process with sulfide-modified α-FeOOH, but with no obvious variation of Sb(V) speciation. While the removal mechanism for As(V) was reduction followed by adsorption via hydroxyl groups, mainly surface complexation was involved in the removal of Sb(V). This study presented a simple strategy to enhance the adsorption capacity and adsorption affinity of α-FeOOH toward As(V)/Sb(V) via sulfide-modification.


Characterization of drug authenticity using thin-layer chromatography imaging with a mobile phone.
Thin-layer chromatography (TLC) has a myriad of separation applications in chemistry, biology, and pharmacology due to its simplicity and low cost. While benchtop laboratory sample application and detection systems for TLC provide accurate quantitation of TLC spot positions and densities, there are many applications where inexpensive and portable instruments would greatly expand the applicability of the technology. In this work, we demonstrate identity verification and concentration determination of pharmaceutical compounds via TLC using a custom 3D-printed cradle that interfaces with an ordinary mobile phone. The cradle holds the mobile phone's internal, rear-facing camera in a fixed position relative to a UV lamp and a TLC plate that includes a phosphor in the stationary phase. Analysis of photographs thus reveals the locations and intensities of principal spots of UV--absorbing drugs. Automated image analysis software determines the center location and density of dark spots, which, using integrated calibration spots of known drug compounds and concentrations, can be used to determine if a drug has been diluted or substituted. Two independent image processing approaches have been developed that may be selected based upon the processing capabilities of the smartphone. Each approach is able to discern 5% drug concentration differences. Using single-component solutions of nevirapine, amodiaquine, and paracetamol that have been manually applied, the mobile phone-based detection instrument provides measurements that are equivalent to those obtained with a commercially available lab-based desktop TLC densitometer.


Macroprudential Policy in a Heterogeneous Environment-An Application of Agent-Based Approach in Systemic Risk Modelling.
Assessment of welfare effects of macroprudential policy seems the most important application of the Dynamic Stochastic General Equilibrium (DSGE) framework of macro-modelling. In particular, the DSGE-3D model, with three layers of default (3D), was developed and used by the European Systemic Risk Board and European Central Bank as a reference tool to formally model the financial cycle as well as to analyze effects of macroprudential policies. Despite the extreme importance of incorporating financial constraints in Real Business Cycle (RBC) models, the resulting DSGE-3D construct still embraces the representative agent idea, making serious analyses of diversity of economic entities impossible. In this paper, we present an alternative to DSGE modelling that seriously departs from the assumption of the representativeness of agents. Within an Agent Based Modelling (ABM) framework, we build an environment suitable for performing counterfactual simulations of the impact of macroprudential policy on the economy, financial system and society. We contribute to the existing literature by presenting an ABM model with broad insight into heterogeneity of agents. We show the stabilizing effects of macroprudential policies in the case of economic or financial distress.


Development of Core Outcome Measures sets for paediatric and adult Severe Asthma (COMSA).
Effectiveness studies with biological therapies for asthma lack standardised outcome measures. The COMSA (Core Outcome Measures sets for paediatric and adult Severe Asthma) Working Group sought to develop Core Outcome Measures (COM) sets to facilitate better synthesis of data and appraisal of biologics in paediatric and adult asthma clinical studies. COMSA utilised a multi-stakeholder consensus process among patients with severe asthma, adult and paediatric clinicians, pharmaceutical representatives, and health regulators from across Europe. Evidence included a systematic review of development, validity and reliability of selected outcome measures plus a narrative review and a pan-European survey to better understand patients' and carers' views about outcome measures. It was discussed using a modified GRADE (Grading of Recommendations Assessment, Development and Evaluation) Evidence to Decision framework. Anonymous voting was conducted using predefined consensus criteria. Both adult and paediatric COM sets include forced expiratory volume in 1 s (FEV1) as z-scores, annual frequency of severe exacerbations and maintenance oral corticosteroid use. Additionally, the paediatric COM set includes the Paediatric Asthma Quality of Life Questionnaire and Asthma Control Test or Childhood Asthma Control Test, while the adult COM set includes the Severe Asthma Questionnaire and Asthma Control Questionnaire-6 (symptoms and rescue medication use reported separately). This patient-centred collaboration has produced two COM sets for paediatric and adult severe asthma. It is expected that they will inform the methodology of future clinical trials, enhance comparability of efficacy and effectiveness of biological therapies, and help assess their socioeconomic value. COMSA will inform definitions of non-response and response to biological therapy for severe asthma.


Watermarking of three-dimensional objects by digital holography.
We present an optical method for information watermarking of three-dimensional (3D) objects by digital holography. A hidden image is embedded by double phase encoding in a phase-shift digital hologram of the 3D object. We decode the watermarked hologram to reconstruct the hidden image and the 3D object. We use either the entire hologram or a part of it to decode the hidden image. Experiments are presented to illustrate the ability to recover both the 3D object and the decoded hidden image. Digital holograms of the 3D object are obtained by optical experiments. The watermarking process, 3D object reconstruction, and hidden image recovery are performed digitally. To the best of our knowledge, this is the first report of 3D object watermarking by use of a phase encoding technique and digital holography.


Scotland's first iron lung.
The history of artificial ventilation and the development of the iron lung in the USA by Drinker and his colleagues is discussed. The building and use of an iron lung by Dr R G Henderson in Aberdeen in 1933 is described. The development of other types of ventilator in the UK is recorded and the circumstances whereby positive pressure ventilation was introduced in Denmark in 1952 is outlined.


Food insecurity is associated with muscle dysmorphia symptomatology among a sample of Canadian adolescents and young adults.
Prior research has documented the association between food insecurity and eating disorders, disordered eating behaviors, and body dissatisfaction. No known research has investigated whether food insecurity is associated with muscle dysmorphia symptomatology, which was the aim of this study. Data from 912 adolescents and young adults in Canada were analyzed. Linear regression analyses were used to determine the association between experiencing past year food insecurity and current muscle dysmorphia symptomatology. Among the sample, 15.7% reported experiencing any food insecurity. In regression analyses, food insecurity was significantly associated with greater overall muscle dysmorphia symptomatology and symptoms of Functional Impairment and Appearance Intolerance. Nearly one in five (18.5%) participants who reported food insecurity were at clinical risk for muscle-dysmorphia. Findings add to the growing literature on the adverse correlates of food insecurity and underscore the need for more research and intervention efforts to address the relationship between food insecurity and muscle dysmorphia symptomatology.


Proteasome inhibitors in the treatment of multiple myeloma.
Proteasome inhibition has been shown to be an effective strategy for the treatment of multiple myeloma, as demonstrated by the clinical activity of the first-in-class agent bortezomib. Recently, the second-generation proteasome inhibitor carfilzomib has been approved in the USA in the relapsed and refractory setting, and several other investigational agents are in clinical development, including MLN9708, marizomib, oprozomib and delanzomib. Here, the authors provide a comprehensive review of the key role of proteasome inhibitors in the myeloma treatment pathway, and highlight the similarities and differences in pharmacology, routes of administration, and efficacy and safety profiles between bortezomib, carfilzomib and investigational agents. The authors also evaluate the potential for further improving myeloma treatment through the ongoing development of novel proteasome inhibitors.


Actin filament alignment causes mechanical hysteresis in cross-linked networks.
Cells dynamically control their material properties through remodeling of the actin cytoskeleton, an assembly of cross-linked networks and bundles formed from the biopolymer actin. We recently found that cross-linked networks of actin filaments reconstituted in vitro can exhibit adaptive behavior and thus serve as a model system to understand the underlying mechanisms of mechanical adaptation of the cytoskeleton. In these networks, training, in the form of applied shear stress, can induce asymmetry in the nonlinear elasticity. Here, we explore control over this mechanical hysteresis by tuning the concentration and mechanical properties of cross-linking proteins in both experimental and simulated networks. We find that this effect depends on two conditions: the initial network must exhibit nonlinear strain stiffening, and filaments in the network must be able to reorient during training. Hysteresis depends strongly and non-monotonically on cross-linker concentration, with a peak at moderate concentrations. In contrast, at low concentrations, where the network does not strain stiffen, or at high concentrations, where filaments are less able to rearrange, there is little response to training. Additionally, we investigate the effect of changing cross-linker properties and find that longer or more flexible cross-linkers enhance hysteresis. Remarkably plotting hysteresis against alignment after training yields a single curve regardless of the physical properties or concentration of the cross-linkers.


[Cutaneo-viscero-hemolytic loxoscelism with acute renal failure].
The Loxoscelism is caused by the bite of spider Loxosceles laeta gender, of worldwide distribution. The poisoning can cause lesions dermonecrotic and less frequently a systemic illness that can be fatal. The mechanism of venom action is multifactorial. The characteristic dermonecrotic lesion results from the direct effects of the venom on the celular and basal membrane components, as well as the extracelular matrix. The initial interaction between the poison and tissues, causes complement activation, migration of polymorphic neutrophils, liberation of proteolytic enzymes, cytoquines, aggregation platelet, and blood flow alterations that result in edema and ischemia, with development of necrosis. There is no a definitive treatment for loxoscelism. However, the value of specific antivenom, to decrease lesion size and limit systemic illness even when such administration is delayed. We present a case of cutaneous-visceral loxoscelismo with unfavorable evolution.


Rapid and reciprocal regulation of tenascin-C and tenascin-Y expression by loading of skeletal muscle.
Tenascin-C and tenascin-Y are two structurally related extracellular matrix glycoproteins that in many tissues show a complementary expression pattern. Tenascin-C and the fibril-associated minor collagen XII are expressed in tissues bearing high tensile stress and are located in normal skeletal muscle, predominantly at the myotendinous junction that links muscle fibers to tendon. In contrast, tenascin-Y is strongly expressed in the endomysium surrounding single myofibers, and in the perimysial sheath around fiber bundles. We previously showed that tenascin-C and collagen XII expression in primary fibroblasts is regulated by changes in tensile stress. Here we have tested the hypothesis that the expression of tenascin-C, tenascin-Y and collagen XII in skeletal muscle connective tissue is differentially modulated by mechanical stress in vivo. Chicken anterior latissimus dorsi muscle (ALD) was mechanically stressed by applying a load to the left wing. Within 36 hours of loading, expression of tenascin-C protein was ectopically induced in the endomysium along the surface of single muscle fibers throughout the ALD, whereas tenascin-Y protein expression was barely affected. Expression of tenascin-C protein stayed elevated after 7 days of loading whereas tenascin-Y protein was reduced. Northern blot analysis revealed that tenascin-C mRNA was induced in ALD within 4 hours of loading while tenascin-Y mRNA was reduced within the same period. In situ hybridization indicated that tenascin-C mRNA induction after 4 hours of loading was uniform throughout the ALD muscle in endomysial fibroblasts. In contrast, the level of tenascin-Y mRNA expression in endomysium appeared reduced within 4 hours of loading. Tenascin-C mRNA and protein induction after 4-10 hours of loading did not correlate with signs of macrophage infiltration. Tenascin-C protein decreased again with removal of the load and nearly disappeared after 5 days. Furthermore, loading was also found to induce expression of collagen XII mRNA and protein, but to a markedly lower level, with slower kinetics and only partial reversibility. The results suggest that mechanical loading directly and reciprocally controls the expression of extracellular matrix proteins of the tenascin family in skeletal muscle.


Simulation of bilateral movement training through mirror reflection: a case report demonstrating an occupational therapy technique for hemiparesis.
In rehabilitation for hemiparesis, one of the goals of an occupational therapist is to practice upper extremity tasks with the recovering individual. The practice is intended to strengthen muscles and refine movements. It also provides examples for the recovering body and brain as they attempt to reestablish the now delicate cognitive and neural connections mediating voluntary behavior. However, the paresis significantly limits the movement sequence possibilities that may be physically practiced. We outline a method for using simulation of movement, which is intended to provide a means for experiencing a range of smooth and controlled movements completed by a paretic limb. The simulation provides a compelling perceptual experience of bilateral motion beyond the current capabilities of the affected limb. The benefits of this technique after a 3-week course of the simulation practice are exemplified by the presented case study that reveals improved function as demonstrated by increases in Fugl-Meyer scores and faster movement speeds as demonstrated by decreased movement times for the Jebsen test of hand function.


Mitral valve surgery with concomitant treatment of atrial fibrillation.
Recovery of sinus rhythm after mitral valve surgery in patients with chronic atrial fibrillation lowers thromboembolic risk and improves survival and quality of life. This article reviews the principal surgical procedures devised in the 1980s and 1990s to treat atrial fibrillation during mitral valve operations. Advantages and drawbacks of the different techniques are discussed. Traditional atrial fibrillation surgery is technically demanding and increases operative morbidity. Simplified techniques, mostly limited to the left atrium, have been developed to reduce operation time and procedure-related complications. Intraoperative radiofrequency ablation has recently proven extremely effective in atrial fibrillation surgery, allowing a further simplification of the procedures. We report the results of an original technique for atrial fibrillation treatment during mitral valve surgery through epicardial radiofrequency ablation. Based on recently reported results of atrial fibrillation surgery and on prognostic considerations, specific treatment of both chronic and paroxysmal atrial fibrillation is indicated in virtually all affected patients undergoing mitral valve surgery.


Chronic lung allograft dysfunction phenotypes and treatment.
Chronic lung allograft dysfunction (CLAD) remains a major hurdle limiting long-term survival post lung transplantation. Given the clinical heterogeneity of CLAD, recently two phenotypes of CLAD have been defined [bronchiolitis obliterans syndrome (BOS) vs. restrictive allograft syndrome (RAS) or restrictive CLAD (rCLAD)]. BOS is characterized by an obstructive pulmonary function, air trapping on CT and obliterative bronchiolitis (OB) on histopathology, while RAS/rCLAD patients show a restrictive pulmonary function, persistent pleuro-parenchymal infiltrates on CT and pleuroparenchymal fibro-elastosis on biopsies. Importantly, the patients with RAS/rCLAD have a severely limited survival post diagnosis of 6-18 months compared to 3-5 years after BOS diagnosis. In this review, we will review historical evidence for this heterogeneity and we will highlight the clinical, radiological, histopathological characteristics of both phenotypes, as well as their risk factors. Treatment of CLAD remains troublesome, nevertheless, we will give an overview of different treatment strategies that have been tried with some success. Adequate phenotyping remains difficult but is clearly needed for both clinical and scientific purposes.


An Image Classification Approach to Pre-Determine Extent of Development of Post-Operative Necrosis in Skin Flaps.
Purpose - Clinicians and care workers currently cannot determine the final extent of necrosis once it begins. If the area is relatively small, it may heal like a normal wound, whereas re-operation may be the only option if it spreads. This paper represents work toward a predictive algorithm using full-thickness random rat skin flap photographs to determine whether the tissue will develop irretrievable necrosis.Methods - Using post-surgery images taken over a series of days of ischemic flaps, features were extracted, selected, and input into a classification algorithm to see if it could provide information on the future condition of the flaps. We split our data into two groups: flaps that underwent normal healing and slow healing. When consulting with a specialist, it was observed that the resulting dermal damage was not severe when a flap had ≤ 40% necrosis over its length on the final day. Three classifiers were implemented: K-Nearest Neighbor (KNN), Random Forest, and Support Vector Machine (SVM).Results - The trained KNN was able to correctly determine whether a flap developed a necrotic area larger or less than 40% of its length with an accuracy of 91% using only images one day post-surgery. Under leave-one-out cross-validation, the Random Forest and SVM achieved accuracies of 82% and 79.5%, respectively, using images spanning ten days post-surgery.Conclusion - We have shown that a classifier can accurately determine whether ischemic skin flaps will develop severe necrotic tissue.Clinical Relevance- An algorithm that assesses early timepoint images and predicts necrosis spread is not only a vital tool for patients and clinicians but is also an extremely important tool to accelerate research into necrosis reduction strategies that ultimately may find application for more life-threatening necrosis-related conditions.


Liver microsomal drug-metabolizing enzyme system: functional components and their properties.
The liver microsomal drug-metabolizing enzyme system consists of two protein components, cytochrome P-450 and NADPH-cytochrome c reductase, and a lipid, phosphatidylcholine. Cytochrome P-450 serves as the binding site for oxygen and substrate while the reductase acts as an electron carrier shuttling electrons from NADPH to cytochrome P-450. The phospholipid facilitates the transfer of electrons from NADPH-cytochrome c reductase to cytochrome P-450 but itself is not an electron carrier. Different cytochromes P-450 and P-448 have been purified; the spectral, catalytic, and immunological properties as well as the molecular weight (determined by SDS-gel electrophoresis) of all these hemeproteins differ from one another. The presence of multiple cytochrome P-450s may explain the species, strain, age, tissue, and sex differences as well as the effect of inducers and nutritional status in mammlian drug metabolism.


Indirect alloreactivity and chronic rejection.
The relative contribution of the direct versus indirect pathway of T-lymphocyte alloreactivity to the development of chronic rejection is incompletely understood. Utilizing a murine model of cardiac allograft vasculopathy (CAV) and a recipient strain with markedly reduced capacity for indirect alloreactivity, we sought to define the importance of indirect allorecognition in CAV. The cells from H2-M mutant mice are unable to present intact protein antigens via class II molecules and have a markedly reduced capacity to present exogenous peptides. B6C.H-2(bm12) strain donor hearts were transplanted into either C57Bl/6 wild-type (WT) or H2-M mutant mice (on C57Bl/6 background). Recipients were killed on day 24. T lymphocyte and macrophage infiltration were graded immunohistochemically. Intimal lesions were measured morphometrically. Donor hearts in WT recipients developed significant intimal lesions, as expected (50+/-7%). Moreover, the donor hearts in H2-M mutant mice also developed comparable intimal lesions (52+/-9%, P=NS vs. WT). Furthermore, the extent of T lymphocyte and macrophage infiltration was similar in both groups. This study demonstrates that a markedly reduced capacity for indirect alloreactivity does not affect the severity of intimal lesions in this model of CAV. The findings of this study question the role of indirect alloreactivity as the sole pathway of allorecognition leading to chronic rejection.