Pain Modulation and Autonomic Function: The Effect of Clonidine.
The α2-agonist clonidine is an analgesic agent, whose yet uncertain action may involve either increase in pain modulation efficiency, change in autonomic function, and/or decrease in anxiety level. The present study aimed to examine the effect of oral clonidine on pain perception in healthy subjects in order to reveal its mode of action. Randomized, double-blind, placebo-controlled study. Forty healthy subjects. Subjects received either 0.15 mg oral clonidine or placebo. We measured pain parameters of heat pain thresholds, tonic heat stimulus, mechanical temporal summation, offset analgesia (OA) and conditioned pain modulation (CPM); autonomic parameters of deep breathing ratio and heart rate variability indices obtained before, during, and after tonic heat stimulus; and psychological parameters of anxiety and pain catastrophizing. Clonidine decreased systolic blood pressure (P = 0.022) and heart rate (P = 0.004) and increased rMSSD (P = 0.020), though no effect was observed on pain perception, pain modulation, and psychological parameters. Autonomic changes were correlated with pain modulation capacity; for OA, the separate slope model was significant (P = 0.008); in the clonidine group, more efficient OA was associated with lower heart rate (r = 0.633, P = 0.005), unlike in the placebo group. The change in autonomic function that was related to the increase in pain modulation capacity, and the lack of change in anxiety, suggest a combined modulatory-autonomic mode of analgesic action for clonidine.


Predictability of individual circadian phase during daily routine for medical applications of circadian clocks.
BACKGROUNDCircadian timing of treatments can largely improve tolerability and efficacy in patients. Thus, drug metabolism and cell cycle are controlled by molecular clocks in each cell and coordinated by the core body temperature 24-hour rhythm, which is generated by the hypothalamic pacemaker. Individual circadian phase is currently estimated with questionnaire-based chronotype, center-of-rest time, dim light melatonin onset (DLMO), or timing of core body temperature (CBT) maximum (acrophase) or minimum (bathyphase).METHODSWe aimed at circadian phase determination and readout during daily routines in volunteers stratified by sex and age. We measured (a) chronotype, (b) every minute (q1min) CBT using 2 electronic pills swallowed 24 hours apart, (c) DLMO through hourly salivary samples from 1800 hours to bedtime, and (d) q1min accelerations and surface temperature at anterior chest level for 7 days, using a teletransmitting sensor. Circadian phases were computed using cosinor and hidden Markov modeling. Multivariate regression identified the combination of biomarkers that best predicted core temperature circadian bathyphase.RESULTSAmong the 33 participants, individual circadian phases were spread over 5 hours, 10 minutes (DLMO); 7 hours (CBT bathyphase); and 9 hours, 10 minutes (surface temperature acrophase). CBT bathyphase was accurately predicted, i.e., with an error less than 1 hour for 78.8% of the subjects, using a new digital health algorithm (INTime), combining time-invariant sex and chronotype score with computed center-of-rest time and surface temperature bathyphase (adjusted R2 = 0.637).CONCLUSIONINTime provided a continuous and reliable circadian phase estimate in real time. This model helps integrate circadian clocks into precision medicine and will enable treatment timing personalization following further validation.FUNDINGMedical Research Council, United Kingdom; AP-HP Foundation; and INSERM.


[Modern methods of molecular targeted therapy for disseminated melanoma].
Significant changes occurred in drug therapy for disseminated melanoma during the last 5 years. New classes of pharmaceuticals appeared in daily clinical practice: inhibitors of MAPK pathway components (BRAF inhibitors vemurafenib and dabrafenib, MEK--inhibitors trametinib and cobimetinib) and immune checkpoint inhibitors that modulate immunologic synapse activity. This article presents information about MAPK pathway inhibitors, their mechanism of action and clinical trials experience including specific related adverse events. Relation of the therapies describes to the other methods in the field are also described. Algorithm of personalized use of current antineoplastic drugs in melanoma is presented. Modern therapeutic approaches in melanoma provide profound and long lasting effects and can even cure some patients. Rational consecutive and combined application of current methods, proper diagnostic and management of related AE can prolong life span of patients and meaningfully increase their quality of life.


Fluorescent electrospun PMMA microfiber mats with embedded NaYF4: Yb/Er upconverting nanoparticles.
Functional upconverting nanoparticles (UCNPs) can offer new possibilities in fluorescent applications as they exhibit desired characteristic properties like large shift between the fluorescent emission signal and the infrared excitation wavelength, multi- and narrow-band absorption and emission in visible and near infrared - Vis/NIR, together with excellent photostability and low toxicity as opposed to semiconducting quantum dots. The upconversion luminescence emission or quenching characteristics of UCNPs can be altered upon exposure to physical or chemical environmental factors providing thus a functionality that can be utilized for sensing or imaging. Furthermore their functionalization with suitable indicator dyes or recognition elements can extend the range of luminescence response and ratiometric sensing to specific analytes. Synergistically, electrospun nano- and microfibers offering large surface area can enhance the functionality of UCNPs by retaining the fluorescence efficiency and improving the overall responsivity due to dramatically increased surface. For the optimization of this hybrid material system the controllable incorporation of UCNPs is required especially at increased concentration conditions needed for high brightness. Herein, we report the fabrication, morphological and optical characterization of electrospun polymer-based nanocomposite fibers, consisting of poly(methyl methacrylate) (PMMA) and upconverting lanthanide doped nanoparticles of the type NaYF4 : 20% Yb3+/2% Er3+ @ NaYF4. Morphological studies regarding the uniformity and aggregation effects of the UCNP inclusion within the fibers have been implemented followed by upconversion emission characterization by pulsed near-infrared excitation. The study and optimization of such nanocomposite fibrous systems could provide useful insights for the development of efficient upconverting electrospun fiber mats for a number of imaging and sensing applications.


Layered Structure and Wear Mechanism of Concentrated Polymer Brushes.
Concentrated polymer brushes (CPBs), which are significantly denser and thicker than conventional semidilute polymer brushes, have received increasing attention in the field of tribology because of their superlow friction properties. However, despite numerous studies aimed at enhancing CPBs for mechanical applications, the relationship between the specific layered structure and lubrication mechanisms of CPBs is still not completely understood. In this study, to reveal the relationship, simultaneous time-resolved measurements of the interfacial gap, static mechanical response, and dynamic mechanical response of the CPB at the contact interface were conducted using optical interference and precise force measuring methods. Two types of tests (i.e., the "indentation" and "sliding" tests) were alternately performed on a glass substrate coated with the CPB against a steel ball immersed in an ionic liquid. The indentation tests measuring the time-resolved interfacial gap and changes in static and dynamic mechanical responses quantitatively confirmed the presence of dilute, middle, and concentrated layers in the CPB. In the sliding tests, the wear of the CPB was detected by observing a decrease in the interfacial gap at the contact interface. Moreover, the thickness of the dilute layer remained constant with sliding, whereas the thicknesses of the other layers decreased, indicating that the dilute layer was continuously formed due to sliding. Therefore, CPB wear occurs randomly at the friction interface alongside the formation of a dilute layer with low density and stiffness on the surface.


Early Bacillus anthracis-macrophage interactions: intracellular survival survival and escape.
This study describes early intracellular events occurring during the establishment phase of Bacillus anthracis infections. Anthrax infections are initiated by dormant endospores gaining access to the mammalian host and becoming engulfed by regional macrophages (Mphi). During systemic anthrax, late stage events include vegetative growth in the blood to very high titres and the synthesis of the anthrax exotoxin complex, which causes disease symptoms and death. Experiments focus on the early events occurring during the first few hours of the B. anthracis infectious cycle, from endospore germination up to and including release of the vegetative cell from phagocytes. We found that newly vegetative bacilli escape from the phagocytic vesicles of cultured Mphi and replicate within the cytoplasm of these cells. Release from the Mphi occurs 4-6 h after endospore phagocytosis, timing that correlates with anthrax infection of test animals. Genetic analysis from this study indicates that the toxin plasmid pXO1 is required for release from the Mphi, whereas the capsule plasmid pXO2 is not. The transactivator atxA, located on pXO1, is also found to be essential for release, but the toxin genes themselves are not required. This suggests that Mphi release of anthrax bacilli is atxA regulated. The putative 'escape' genes may be located on the chromosome and/or on pXO1.


In vivo invasion of modified chorioallantoic membrane by tumor cells: the role of cell surface-bound urokinase.
The ability of the chick embryo chorioallantoic membrane (CAM) to withstand invasion by tumor cells can be intentionally compromised by altering its morphological integrity. Using a newly developed quantitative assay of invasion we showed that intact CAMs were completely resistant to invasion by tumor cells, wounded CAMs did not pose a barrier to penetration, and CAMs that were wounded and then allowed to reseal displayed partial susceptibility to invasion. The invasion of resealed CAMs required catalytically active plasminogen activator (PA) of the urokinase type (uPA); the invasive efficiency of tumor cells was reduced by 75% when tumor uPA activity or tumor uPA production was inhibited. The invasive ability of human tumor cells, which have surface uPA receptors but which do not produce the enzyme, could be augmented by saturating their receptors with exogenous uPA. The mere stimulation of either uPA or tissue plasminogen activator production, in absence of binding to cell receptors, did not result in an enhancement of invasiveness. These findings suggest that the increased invasive potential of tumor cells is correlated with cell surface-associated proteolytic activity stemming from the interaction between uPA and its surface receptor.


Efficacy of an anti-cancer strategy targeting SET in canine osteosarcoma.
Osteosarcoma (OSA) is the most common bone tumor in dogs. Protein phosphatase 2A (PP2A), an evolutionary conserved serine/threonine protein phosphatase, is a crucial tumor suppressor. SET is a PP2A inhibitory protein that directly interacts with PP2A and suppresses its phosphatase activity. SET has been reported as a contributor of wide range of human and dog tumor malignancies. However, the role of SET in canine OSA (cOSA) remains unknown. In this study, we investigated the role of SET in cOSA by using 2 cOSA cell lines: POS (primary origin) and HM-POS (metastatic origin). Knockdown (KD) of SET expression was noted to slightly suppress POS cell proliferation only. Furthermore, SET KD effectively suppressed colony formation ability of both POS and HM-POS cells. SET KD was observed to repress ERK1/2, mTOR, E2F1, and NF-κB signaling in HM-POS cells, whereas it inhibited only ERK1/2 signaling in POS. Further, it was observed that SET-targeting drug, FTY720, exerted anti-cancer effects in both POS and HM-POS cells. Moreover, the drug also enhanced the anti-cancer effect of cisplatin. The data suggested that a combination therapy, based on SET targeting drugs and cisplatin, could be a potent strategy for cOSA.


Structural basis for effects of CpA modifications on C/EBPβ binding of DNA.
CCAAT/enhancer binding proteins (C/EBPs) regulate gene expression in a variety of cells/tissues/organs, during a range of developmental stages, under both physiological and pathological conditions. C/EBP-related transcription factors have a consensus binding specificity of 5'-TTG-CG-CAA-3', with a central CpG/CpG and two outer CpA/TpG dinucleotides. Methylation of the CpG and CpA sites generates a DNA element with every pyrimidine having a methyl group in the 5-carbon position (thymine or 5-methylcytosine (5mC)). To understand the effects of both CpG and CpA modification on a centrally-important transcription factor, we show that C/EBPβ binds the methylated 8-bp element with modestly-increased (2.4-fold) binding affinity relative to the unmodified cognate sequence, while cytosine hydroxymethylation (particularly at the CpA sites) substantially decreased binding affinity (36-fold). The structure of C/EBPβ DNA binding domain in complex with methylated DNA revealed that the methyl groups of the 5mCpA/TpG make van der Waals contacts with Val285 in C/EBPβ. Arg289 recognizes the central 5mCpG by forming a methyl-Arg-G triad, and its conformation is constrained by Val285 and the 5mCpG methyl group. We substituted Val285 with Ala (V285A) in an Ala-Val dipeptide, to mimic the conserved Ala-Ala in many members of the basic leucine-zipper family of transcription factors, important in gene regulation, cell proliferation and oncogenesis. The V285A variant demonstrated a 90-fold binding preference for methylated DNA (particularly 5mCpA methylation) over the unmodified sequence. The smaller side chain of Ala285 permits Arg289 to adopt two alternative conformations, to interact in a similar fashion with either the central 5mCpG or the TpG of the opposite strand. Significantly, the best-studied cis-regulatory elements in RNA polymerase II promoters and enhancers have variable sequences corresponding to the central CpG or reduced to a single G:C base pair, but retain a conserved outer CpA sequence. Our analyses suggest an important modification-dependent CpA recognition by basic leucine-zipper transcription factors.


mRNAs involved in copper homeostasis are regulated by the nonsense-mediated mRNA decay pathway depending on environmental conditions.
The nonsense-mediated mRNA decay pathway (NMD) is an mRNA degradation pathway that degrades mRNAs that prematurely terminate translation. These mRNAs include mRNAs with premature termination codons as well as many natural mRNAs. In Saccharomyces cerevisiae a number of features have been shown to target natural mRNAs to NMD. However, the extent to which natural mRNAs from the same functional group are regulated by NMD and how environmental conditions influence this regulation is not known. Here, we examined mRNAs involved in copper homeostasis and are predicted to be sensitive to NMD. We found that the majority of these mRNAs have long 3'-UTRs that could target them for degradation by NMD. Analysis of one of these mRNAs, COX19, found that the long 3'-UTR contributes to regulation of this mRNA by NMD. Furthermore, we examined an additional mRNA, MAC1 under low copper conditions. We found that low copper growth conditions affect NMD sensitivity of the MAC1 mRNA demonstrating that sensitivity to NMD can be altered by environmental conditions. MAC1 is a copper sensitive transcription factor that regulates genes involved with high affinity copper transport. Our results expand our understanding of how NMD regulates mRNAs from the same functional group and how the environment influences this regulation.


Efficacy and safety of bevacizumab in patients with malignant melanoma: a systematic review and PRISMA-compliant meta-analysis of randomized controlled trials and non-comparative clinical studies.
Background: Malignant melanoma is a highly aggressive cancer that spreads and metastasizes quickly. In recent years, the antiangiogenic drug bevacizumab has been trialed to treat malignant melanoma. We conducted the first meta-analysis to examine the efficacy and safety of bevacizumab combined with other drugs in malignant melanoma. Methods: We searched for randomized controlled trials (RCTs) and non-comparative clinical studies of bevacizumab combined with chemotherapy, targeted medicine, and interferon to treat malignant melanoma in PubMed, Embase, the Cochrane Library, and Web of Science. Meta-analysis of RCT was performed using Review Manager (version 5.4), and non-comparative meta-analysis was performed using R (version 4.0.3). The primary outcome was the objective response rate. Depending on the heterogeneity of the included studies, the pooled outcomes and 95% CI were calculated using either random-effects or fixed-effect models. Subgroup outcomes were calculated with possible relevant variables. Sensitivity analyses were carried out by excluding each study from the highly heterogeneous pooled results in turn. Funnel plot and Begg's test were used to test the included studies' potential publication bias. The level of significance was set at p < 0.05. Results: This meta-analysis included 20 trials: five RCTs and 15 non-comparative clinical studies with a total of 23 bevacizumab intervention arms. In 14 treatment arms, bevacizumab was combined with chemotherapy drugs such as fotemustine, dacarbazine, carboplatin/paclitaxel, and temozolomide. In six treatment arms, bevacizumab was combined with targeted medicines such as imatinib, everolimus, sorafenib, erlotinib, and temsirolimus. There were also six treatment arms that used bevacizumab in combination with interferon. The pooled objective response rate was 15.8% (95% CI, 11.4%-20.2%). Bevacizumab plus carboplatin/paclitaxel significantly increased the overall survival compared to carboplatin/paclitaxel (HR = 0.64, 95% CI, 0.49-0.85, p < 0.01). Fatigue, nausea, leukopenia, thrombocytopenia, and neutropenia were the most common adverse events. The pooled incidence of hypertension of all bevacizumab arms in malignant melanoma was 32.4% (95% CI, 24.5%-40.3%). Conclusion: This study showed that bevacizumab combined with chemotherapy might be effective and well-tolerated in patients with stage III or IV unresectable malignant melanoma. Systematic Review Registration: [https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=304625], identifier [CRD42022304625].


Hearing Beyond the Normal Enabled by Therapeutic Devices: The Role of the Recipient and the Hearing Profession.
The time is near where 'therapeutic' bodily assistive devices, developed to mimic species-typical body structures in order to enable normative body functioning, will allow the wearer to outperform the species-typical body in various functions. Although such devices are developed for people that are seen to exhibit sub species-typical abilities, many 'therapeutic enhancements' might also be desired and used by people that exhibit species-typical body abilities. This paper presents the views of members of the World Federation of the Deaf on potential beyond species-typical abilities enabling therapeutic assistive devices (i.e. related to hearing). Survey respondents showed support for the development and uptake of beyond normal hearing enabling devices. The views of survey respondents as clients affect hearing-enabling professions (such as audiologist and speech pathologists). The paper analyzes what guidance code of ethics of hearing enabling professions give in regards to beyond normal hearing enabling devices. This paper suggests that people labeled impaired and the professions that serve them should more involved in the enhancement discourse.


Tyrosine kinases modulate K+ channel gating in mouse Schwann cells.
1. The whole-cell configuration of the patch-clamp technique and immunoprecipitation experiments were used to investigate the effects of tyrosine kinases on voltage-dependent K+ channel gating in cultured mouse Schwann cells. 2. Genistein, a broad-spectrum tyrosine kinase inhibitor, markedly reduced the amplitude of a slowly inactivating delayed-rectifier current (IK) and, to a lesser extent, that of a transient K+ current (IA). Similar results were obtained on IK with another tyrosine kinase inhibitor, herbimycin A. Daidzein, the inactive analogue of genistein, was without effect. 3. Unlike herbimycin A, genistein produced additional effects on IA by profoundly affecting its gating properties. These changes consisted of slower activation kinetics with an increased time to peak, a positive shift in the voltage dependence of activation (by +30 mV), a decrease in the steepness of activation gating (9 mV per e-fold change) and an acceleration of channel deactivation. 4. The steepness of the steady-state inactivation was increased by genistein treatment, while the recovery from inactivation was not significantly altered. 5. The action of genistein on voltage-dependent K+ (Kv) currents was accompanied by a decrease in tyrosine phosphorylation of Kv1.4 as well as Kv1.5 and Kv2.1 encoding transient and slowly inactivating delayed-rectifier K+ channel alpha subunits, respectively. 6. In conclusion, the present study shows that tyrosine kinases markedly affect the amplitude of voltage-dependent K+ currents in Schwann cells and finely tune the gating properties of the transient K+ current component IA. These modulations may be functionally relevant in the control of K+ channel activity during Schwann cell development and peripheral myelinogenesis.


YB-1 represses AP1-dependent gene transactivation and interacts with an AP-1 DNA sequence.
Involvement of the AP-1 (activator protein-1) transcription factor has been demonstrated previously in the regulation of cell proliferation and cell-cycle progression, in the control of cell migration, invasion and metastasis, and in signal transduction, stress responsiveness, DNA replication and DNA repair. YB-1 (Y-box-binding protein-1) has also been implicated in many of these processes. However, the mechanism by which YB-1 mediates these processes is poorly understood. In the present study, we report that overexpression of a transfected gene encoding YB-1 in human HeLa cervical carcinoma cells significantly represses the transactivation of a minimal AP-1 reporter construct in response to the tumour promoter PMA. YB-1 also represses mRNA expression and PMA-induced promoter transactivation of the endogenous AP-1 target gene encoding matrix metalloproteinase-12 (metalloelastase). YB-1 transrepression of both the minimal and matrix metalloproteinase-12 promoter reporter constructs is dependent on the AP-1 sequence. To identify new nuclear proteins that bind specifically to the AP-1 DNA-binding site, we devised a DNA-affinity-chromatography-based assay termed NAPSTER (nucleotide-affinity preincubation specificity test of recognition) and discovered a 49 kDa protein from human cancer cells that binds in a sequence-specific manner to the AP-1 DNA sequence. By tandem MS fragmentation sequencing analyses we determined that p49 is a YB-1. Immunoblotting of the NAPSTER-purified p49 protein using anti-YB-1 antibodies confirmed YB-1 binding to the AP-1 DNA sequence, as did gel mobility-supershift assays using YB-1 antibodies. This is the first report of YB-1 transrepression and interaction at the AP-1 DNA-binding site.


The rheumatic mitral valve and repair techniques in children.
The mitral valve is the most commonly affected valve in acute and chronic rheumatic heart disease in the first and second decades of life. Pure or predominant mitral regurgitation with non-significant stenosis (mitral valve area > 1.5 cm(2) on echocardiography) is the most frequently encountered valvular dysfunction in children. In our experience, based on 428 children operated between 1993 and 2011 at our institution, functional classification based on leaflet motion assessed by echocardiography and reconfirmed peroperatively revealed pure annulus dilatation (type I) in 7% of patients, anterior leaflet prolapse (type IIa) in 33%, combination of anterior leaflet pseudoprolapse with restricted motion of the posterior leaflet (type pseudoIIa/IIIp) in 34%, and restricted anterior and posterior leaflet motion (type IIIa/p) in 26%. Patients with type III were older than those with type IIa and type pseudoIIa/IIIp. Different techniques can be used to repair rheumatic mitral valve lesions: prolapse of the anterior leaflet caused by chordal elongation or rupture can be treated by chordal shortening, chordal transfer, or artificial chordal replacement; restricted motion of the anterior and/or posterior leaflet can be treated by commissurotomy, splitting of the papillary muscles, resection of the secondary, or sometimes primary posterior chordae, posterior leaflet free edge suspension, leaflet thinning, and leaflet enlargement using autologous pericardium. Because mitral annulus dilatation is present in almost all patients with mitral regurgitation, concomitant ring annuloplasty offers more stability in valve repair, improving long-term outcome. The major causes for failure of rheumatic mitral valve repair are the presence of ongoing rheumatic inflammation at the time of surgery, use of inappropriate techniques, technical failures requiring early reoperation, lack of concomitant ring annuloplasty, and progression of leaflet and chordal disease further resulting in more leaflet retraction, thickening, and deformity. Freedom from reoperation depends on mitral regurgitation functional type, the type IIa and type pseudoIIa/IIIp having a better long-term outcome than type I and type III, in our series. In conclusion, mitral valve repair should be a preferred strategy in children with rheumatic heart disease whenever feasible, providing stable actuarial survival with fewer thromboembolic complications in a pediatric population noncompliant to anticoagulation.


Lateral Canthal Position Following Lateral Orbitotomy and the Influence of Ocular Proptosis.
Anecdotal evidence suggests that detachment of the lateral canthal tendon during lateral orbitotomy results in shortening of the horizontal palpebral aperture (HPA) and rounding of the lateral canthal angle (LCA). This study investigated the change in HPA and LCA following lateral orbitotomy and the effect of proptosis on the HPA. Retrospective, single surgeon, review of all cases undergoing lateral orbitotomy, comprising of a canthotomy and cantholysis and reconstruction of the lateral canthus using a single suture. The HPA and lateral canthal height (LCH) were measured on pre- and postoperative photographs using ImageJ software. Changes in the LCA and degree of proptosis were also investigated. Exclusion criteria included previous eyelid surgery, acute trauma, and less than 6 months' follow up. There were 41 cases of lateral orbitotomy in 27 patients, 67% female, average age 51 years. Twenty patients had thyroid eye disease of which 14 underwent bilateral surgery; the other 7 patients had a variety of orbital pathologies. Thirteen patients underwent unilateral orbitotomy with the contralateral eye serving as a control. Average follow up was 23 months. Overall, the HPA and proptosis were reduced by 0.6 mm (p = 0.143) and 3.4 mm (p < 0.001), respectively. A 0.2-mm reduction in HPA was observed for every 1-mm reduction in proptosis. A difference in post-orbitotomy HPA was observed between cases and their matched controls (p = 0.016). No rounding of the LCA or significant change in LCH was observed following lateral orbitotomy. The described technique of lateral orbitotomy and simple reconstruction do not appear to cause lateral canthal drift or rounding. There was no significant change in HPA or LCH. The degree of change of proptosis does appear to influence the change of HPA.


Glomerular Filtration Rate Measured Based on Iomeprol Clearance Assessed at CT Urography in Living Kidney Donor Candidates.
Background Estimating glomerular filtration rate (GFR) from serum creatinine can be inaccurate, and current procedures for measuring GFR are time-consuming and cumbersome. Purpose To develop a method for measuring GFR based on iomeprol clearance assessed at CT urography in kidney donor candidates and compare this with iohexol clearance (reference standard for measuring GFR). Materials and Methods This cross-sectional retrospective study included data from kidney donor candidates who underwent both iohexol clearance and CT urography between July 2016 and October 2022. CT-measured GFR was calculated as the iomeprol excretion rate in the urinary system between arterial and excretory phases (Hounsfield units times milliliters per minute) divided by a surrogate for serum iomeprol concentration in the aorta at the midpoint (in Hounsfield units). Performance of CT-measured GFR was assessed with use of mean bias (mean difference between CT-measured GFR and iohexol clearance), precision (the distance between quartile 1 and quartile 3 of the bias [quartile 3 minus quartile 1], with a small value indicating high precision), and accuracy (percentage of CT-measured GFR values falling within 10%, 20%, and 30% of iohexol clearance values). Intraobserver agreement was assessed for 30 randomly selected individuals with the Lin concordance correlation coefficient. Results A total of 75 kidney donor candidates were included (mean age, 51 years ± 13 [SD]; 45 female). The CT-measured GFR was unbiased (1.1 mL/min/1.73 m2 [95% CI: -1.9, 4.1]) and highly precise (16.2 mL/min/1.73 m2 [quartiles 1 to 3, -6.6 to 9.6]). The accuracy of CT-measured GFR within 10%, 20%, and 30% was 61.3% (95% CI: 50.3, 72.4), 88.0% (95% CI: 80.7, 95.4), and 100%, respectively. Concordance between CT-based GFR measurements taken 2 months apart was almost perfect (correlation coefficient, 0.99 [95% CI: 0.98, 0.99]). Conclusion In living kidney donors, GFR measured based on iomeprol clearance assessed at CT urography showed good agreement with GFR measured based on iohexol clearance. © RSNA, 2023 Supplemental material is available for this article. See also the editorial by Davenport in this issue.


Cellular origin of hexokinase in pancreatic islets.
Transgenic or tumoral pancreatic islet beta cells with enhanced expression of low K(m) hexokinases (HK) exhibit a leftward shift of the normal dose-response curve for glucose-induced insulin release. Furthermore, HK catalyzes roughly 50% of total glucose phosphorylation measured in extracts from freshly isolated rodent islets, suggesting that HK participates in the process of glucose sensing in beta cells. We previously observed that HK activity represents 20% of total glucose phosphorylation in purified rat beta cell preparations and that HK is not homogenously distributed over these cells. The present study provides several arguments for the idea that HK detected in freshly isolated rat islets or islet cell preparations originates mainly from contaminating exocrine cells. First, reverse transcriptase-polymerase chain reaction using isoform-specific primers allowed detection of hexokinase I and IV mRNA in rat beta cells, whereas the messenger levels encoding the hexokinase II and III isoforms were undetectably low. However, immunoblots indicated that hexokinase I protein was 10-fold more abundant in freshly isolated islets and flow-sorted exocrine cells than in purified rat beta cell preparations. Second, comparison of HK activity in the different pancreatic cell types resulted in 15-25-fold higher values in exocrine than in endocrine cells (acinar cells: 21 +/- 3 pmol of glucose 6-phosphate formed/h/ng of DNA; duct cells: 30 +/- 8 pmol/h/ng of DNA; islet beta cells: 1.2 +/- 0.2 pmol/h/ng DNA; alpha cells: 0.9 +/- 0.4 pmol/h/ng of DNA). Since freshly purified beta cell preparations contain 3 +/- 1% exocrine cells, at least 50% of their HK activity can be accounted for by exocrine contamination. Third, after 5 days of culture of purified islet beta cells, both HK activity and the proportion of exocrine cells decreased by more than 1 order of magnitude, while the ratio of glucokinase over hexokinase activity increased more than 10-fold. Finally, preincubating the cells with 50 mmol/liter 2-deoxyglucose did not affect glucose stimulation of insulin biosynthesis and release. In conclusion, the observation that pancreatic exocrine cells are responsible for a major part of HK activity in islet cell preparations cautions against the use of HK measurements in islet extracts in the study of these enzymes in glucose sensing by pancreatic beta cells.


TMEM45A is involved in renal fibrosis in rats by regulating Jagged1/Notch pathway.
Fibrosis, or the excess deposition of fibrous tissue, is a critical feature of chronic kidney disease. Here, using renal fibrotic rat as a model, which was established via 5/6 nephrectomy (Nx), the role of TMEM45A transmembrane protein in renal fibrosis was investigated. The results indicated that 5/6 Nx gradually led to histopathological abnormalities and loss of kidney function in rats, which correlated with upregulation of TMEM45A and Notch1. Interestingly, in NRK-49F renal cells, overexpression of TMEM45A resulted in up-regulation of extracellular matrix (ECM) components as well as induction of Notch-1 and Jagged-1. These effects were weakened by DAPT, an inhibitor of the Notch pathway, suggesting an important role of Notch signaling in mediating the functions of TMEM45A in NRK-49F cells Moreover, TMEM45A knockdown by TMEM45A siRNA in NRK-49F cells diminished TGF-b1-induced upregulation of ECM components, inflammatory cytokines, Notch-1 and Jagged-1. Correspondingly, TGF-beta 1 exhibited pro-fibrogenic like effect in NRK-49F cells and induced TMEM45A and Jagged1/Notch expression. Collectively, these results demonstrate that TMEM45A plays an important role in renal fibrosis by regulating ECM components and Jagged1/Notch pathway.


The accuracy of the emergency physician at diagnosing CVA/TIA in the acute care setting
OBJECTIVE: To determine the accuracy of the ED physician at diagnosing CVA/TIA in the acute setting. METHODS: We reviewed 246 patients admitted for acute CVA/TIA during 1997. We reviewed admitting and discharge diagnoses, CT, MRI, and MRA results. We also reviewed the medical histories of the patients. Patients with tumors and ICH diagnosed on the initial CT scan were excluded. RESULTS: A total of 241 patients were included for analysis. Of the patients admitted for CVA, 67% were discharged with the same diagnosis. Of the patients admitted for TIA, 82% were discharged with the same diagnosis. 10% of TIAs diagnosed at admission received the diagnoses at discharge. 22% of CVAs at admission were diagnosed as TIAs. 11% of CVA/TIAs at admission were given other diagnoses at discharge. Some of these diagnoses included hemiplegic migraines, Bell's palsy, lumbosacral spondylosis, giant cell arteritis, basilar artery aneurysm, and viral meningitis. In considering thrombolysis for CVA, one may then overtreat approximately 30% of patients and miss approximately 10% of patients who could be candidates for treatment. The NNT for CVA and thrombolysis is approximately 10. If 30% overtreatment rate is accurate, then the NNH is approximately 333 assuming an ICH rate of 1%. For 1,000 patients treated, 100 may benefit (improved function) and 3 would die who did not have the disease. CONCLUSIONS: The accuracy rate of the diagnosis of CVA at initial presentation is 67%. This is due to many initial neurologic changes being TIAs and some other diagnoses which can mask as CVA. Choosing thrombolysis for CVA treatment will involve treatment of many patients who do not have disease.


Medical Management of Recurrent Left Ventricular Assist Device Thrombosis in a Patient With Biventricular Assist Devices.
Ventricular assist device (VAD) pump thrombosis is a known complication and while the preferred standard treatment is surgical pump exchange this procedure is not without risk and for some patients the risks are prohibitive. This is a case of a 68-year-old female with bilateral HeartWare ventricular assist devices (HVAD) implanted as destination therapy who presented with signs of recurrent pump thrombosis. Surgical pump exchange was deemed to confer prohibitive risk due to her underlying medical co-morbidities and therefore not an option for treatment. After careful consideration of possible options for treatment, she received systemic thrombolysis (Alteplase 5 mg IV bolus followed by 3 mg/hour infusion for 10 hours through a central line) which was successful. This case highlights, not only the rarity of bilateral VADs as destination therapy, but also demonstrates the safety and efficacy of using systemic thrombolytics in patients with bilateral HVADs for treatment of pump thrombosis.


Minimum methionine requirement in adult cats as determined by indicator amino acid oxidation.
There is a lack of empirical data on the dietary Met requirement, in the presence of Cys or cystine, in adult cats. Thus, the aim of this study was to determine the Met requirement, in the presence of excess Cys, in adult cats at maintenance using the indicator amino acid oxidation (IAAO) technique. Six adult neutered male cats were initially selected and started the study. Cats were adapted to the basal diet sufficient in Met (0.24% dry matter, DM) for 14 d prior to being randomly allocated to one of eight dietary levels of Met (0.10, 0.13, 0.17, 0.22, 0.27, 0.33, 0.38 and 0.43% DM). Different dietary Met concentrations were achieved by supplementing the basal diet with Met solutions. Alanine was additionally included in the solutions to produce isonitrogenous and isoenergetic diets. Cats underwent a 2d adaptation period to each experimental diet prior to each IAAO study day. On IAAO study days, thirteen meals were offered corresponding to 75% of each cat's daily food allowance. The remaining 25% of their daily food intake was offered after each IAAO study. A bolus dose of NaH13CO3 (0.44 mg·kg-1) and L-[1-13C]-Phenylalanine (13C-Phe; 4.8 mg·kg-1) were provided in 5th and 6th meal, respectively, followed by a constant dose of 13C-Phe (1.04 mg·kg-1) in the next meals. Breath samples were collected and total production of 13CO2 was measured every 25 min through respiration calorimetry chambers. Steady state of 13CO2 achieved over at least three breath collections was used to calculate oxidation of 13C-Phe (F13CO2). Competing models were applied using the NLMIXED procedure in SAS to determine the effects of dietary Met on 13CO2. Two cats were removed from the study as they did not eat all meals, which is required to achieve isotopic steady. A breakpoint for the mean Met requirement, with excess of Cys, was identified at 0.24% DM (22.63 mg·kg-1) with an upper 95% confidence limit of 0.40% DM (37.71 mg·kg-1), on an energy density of 4,164 kcal of metabolizable energy / kg DM calculated using the modified Atwater factors. The estimated Met requirement, in the presence of excess of Cys, is higher than the current recommendations proposed by the National Research Council's Nutrient Requirement of Dogs and Cats, the Association of American Feed Control Officials and the European Pet Food Industry Federation.


Utilizing external ultrasonic energy to improve the results of tumescent liposculpture.
External ultrasound has been used safely for many years for therapy and diagnosis by multiple medical specialists. It has also been utilized safely to treat postoperative swelling after liposculpture. Recently, internal tumescent ultrasonic liposculpture has been introduced. Since internal and external ultrasonic devices utilize injected fluid to create an environment in the fat that efficiently transmits sound waves, it seemed logical that external ultrasound incorporated with tumescent liposculpture would be successful. This study was designed to compare tumescent liposculpture combined with external ultrasound with tumescent liposculpture alone. A side-by-side comparison of the results of tumescent liposculpture combined with external ultrasound versus tumescent liposculpture alone was performed. Thirty patients were infused with therapeutic amounts of tumescent solution to achieve anesthesia and vasoconstriction in the areas to be treated. External ultrasound was then applied to one-half of the areas to be treated in a randomized fashion. Either the left side or the right side of the body was treated with ultrasound. Liposculpture was then performed using a variety of cannulas. Evaluations were made of the temperature of the tissues, the ease of cannula movement by the surgeon, and the time of performing the technique in each area. Postoperatively, the amount of bruising and swelling were evaluated, as was the discomfort of the patient. Clinical assessment results revealed that external ultrasound combined with tumescent liposculpture was easier for the surgeon, requiring less physical effort with less operating time, and less bruising, swelling, and discomfort for the patient postoperatively. External ultrasound combined with tumescent liposculpture produced significant doctor and patient benefit both operatively and postoperatively. Ongoing studies are currently being performed to evaluate higher wattage ultrasonic units.


Value of EUS in Determining Curative Resectability in Reference to CT and FDG-PET: The Optimal Sequence in Preoperative Staging of Esophageal Cancer?
The separate value of endoscopic ultrasonography (EUS), multidetector computed tomography (CT), and 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) in the optimal sequence in staging esophageal cancer has not been investigated adequately. The staging records of 216 consecutive operable patients with esophageal cancer were reviewed blindly. Different staging strategies were analyzed, and the likelihood ratio (LR) of each module was calculated conditionally on individual patient characteristics. A logistic regression approach was used to determine the most favorable staging strategy. Initial EUS results were not significantly related to the LRs of initial CT and FDG-PET results. The positive LR (LR+) of EUS-fine-needle aspiration (FNA) was 4, irrespective of CT and FDG-PET outcomes. The LR+ of FDG-PET varied from 13 (negative CT) to 6 (positive CT). The LR+ of CT ranged from 3-4 (negative FDG-PET) to 2-3 (positive FDG-PET). Age, histology, and tumor length had no significant impact on the LRs of the three diagnostic tests. This study argues in favor of PET/CT rather than EUS as a predictor of curative resectability in esophageal cancer. EUS does not correspond with either CT or FDG-PET. LRs of FDG-PET were substantially different between subgroups of negative and positive CT results and vice versa.


Identification of Candida species associated with vulvovaginal candidiasis by multiplex PCR.
Vulvovaginal candidiasis is a common infection. The aim of this study was to identify the species of vaginal Candida isolates by using multiplex PCR technique. 191 isolates from patients admitted to Mahdieh hospital were identified. The vaginal swab specimens were cultured on Sabouraud Dextrose Agar. The ITS1 region between the 18S and 5.8S rRNA genes and a specific DNA fragment within the ITS2 region were amplified. The multiplex PCR products were separated by electrophoresis in 2% agarose gel, visualized by staining with ethidium bromide, and photographed. Descriptive statistics, Chi-square test, and Spearman correlation were used to summarize the findings. C. albicans and C. glabrata were the most common species isolated from the specimens. A mix of C. glabrata and C. albicans was the most common mixed infection isolated from the samples. The analysis revealed a significant positive association between older age and infection with C. glabrata isolates (Spearman's rho = 0.89, P = 0.015). Multiplex PCR is a fast, yet reliable method to identify Candida species. C. albicans and then C. glabrata are the two most common causes of vulvovaginal candidiasis. The number of mixed fungal infections is higher among Iranian population compared to international reports.


Exposure to Perfluoroalkyl Substances during Fetal Life and Pubertal Development in Boys and Girls from the Danish National Birth Cohort.
It remains unsettled whether prenatal exposure to perfluoroalkyl substances (PFASs) affects human reproductive health through potential endocrine disruption. We aimed to explore the associations between prenatal exposure to several PFASs and various aspects of pubertal development in boys and girls. We studied two samples ([Formula: see text] and 445) from the Puberty Cohort, nested within the Danish National Birth Cohort (DNBC), measuring PFAS in maternal plasma from early gestation. Data on pubertal development were collected biannually from the age of 11 y until full maturation, using web-based questionnaires. Outcomes were age at menarche, voice break, first ejaculation, and Tanner stages 2 to 5 for pubic hair, breast, genital development, and a combined puberty indicator. A regression model for censored data was used to estimate mean difference (months) in age at achieving the pubertal outcomes across tertiles of PFAS concentrations and with a doubling of PFAS concentrations (continuous). For perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS), a meta-analysis was used to provide a weighted average of the point estimates from samples 1 and 2. Overall, prenatal exposure to PFOS, perfluorohexane sulfonate (PFHxS), perfluoroheptane sulfonate (PFHpS), perfluorononanoic acid (PFNA), and perfluorodecanoic acid (PDFA) (girls) and PFHxS and PFHpS (boys) was associated with lower mean age at puberty marker onset. PFDA and PFNA exposure was associated with higher mean age at onset of puberty in boys. Nonmonotonic associations in girls (PFOS, PFHpS, PFDA) and boys (PFDA, PFNA) were observed, showing larger mean age differences for the combined puberty indicator in the middle tertile [girls: PFOS: [Formula: see text] mo, 95% confidence interval (CI): [Formula: see text], [Formula: see text]; PFHpS: [Formula: see text] mo, 95% CI: [Formula: see text], 1.85; PFDA: [Formula: see text] mo, 95% CI: [Formula: see text], 1.83; and boys: PFNA: 4.45 mo, 95% CI: [Formula: see text], 10.21; PFDA: 4.59 mo, 95% CI: [Formula: see text], 10.11] than in the highest tertile with the lowest as reference. Our population-based cohort study suggests sex-specific associations of altered pubertal development with prenatal exposure to PFASs. These findings are novel, and replication is needed. https://doi.org/10.1289/EHP3567.


Invited Review Article: Gas puff imaging diagnostics of edge plasma turbulence in magnetic fusion devices.
Gas puff imaging (GPI) is a diagnostic of plasma turbulence which uses a puff of neutral gas at the plasma edge to increase the local visible light emission for improved space-time resolution of plasma fluctuations. This paper reviews gas puff imaging diagnostics of edge plasma turbulence in magnetic fusion research, with a focus on the instrumentation, diagnostic cross-checks, and interpretation issues. The gas puff imaging hardware, optics, and detectors are described for about 10 GPI systems implemented over the past ∼15 years. Comparison of GPI results with other edge turbulence diagnostic results is described, and many common features are observed. Several issues in the interpretation of GPI measurements are discussed, and potential improvements in hardware and modeling are suggested.


Allopathic and Osteopathic Medicine Unify GME Accreditation: A Historic Convergence.
In 1968, the American Medical Association resolved to accept qualified graduates of osteopathic medical schools into its accredited Graduate Medical Education (GME) programs. An equally momentous decision was arrived at in 2014 when the Accreditation Council for Graduate Medical Education (ACGME), the American Osteopathic Association (AOA), and the American Association of Colleges of Osteopathic Medicine (AACOM) resolved to institute a single unified GME accreditation system by July 1, 2020. As envisioned, the unified accreditation system will all but assure system-wide consistency of purpose and practice in anticipation of the Next Accreditation System (NAS) of the ACGME. Governance integration replete with AOA and AACOM and osteopathic representation on the ACGME Board of Directors is now well underway. What is more, osteopathic representation on current Review Committees (RCs) and in a newly established one with an osteopathic focus has been instituted. Viewed broadly, the unification of the GME accreditation system goes a long way toward recognizing the overlapping characteristics in the training and practice of allopathic and osteopathic medicine. As such, this momentous development represents the latest, indeed boldest leap toward convergence between the two historic branches of American medicine. In this Health Policy Analysis we seek to place the impending unification of the GME accreditation process in its historical context, delineate its near-term impact, and discuss the potential long-term implications thereof.


Incidence of avian leukosis virus infection in broiler stocks and its effect on early growth.
Twenty-eight broiler breeder flocks were tested for avian leukosis virus (ALV) group-specific (gs) antigen shedding into the albumen. The rate of shedding ranged from 0 to 17%, rates substantially lower than those observed previously in egg production stocks. The flock shedding at the highest rate was trap nested and dams positive and negative for gs antigen shedding in the albumen produced progeny for a growth trial. Chicks produced by the positive and negative dams had 27 and 1% ALV isolations from their meconia, respectively. Progeny of positive dams or chicks having virus in their meconia weighed 1 to 3% less at 4 and 7 weeks than progeny of negative dams. More infected chicks died or were runted by 7 weeks. Although the proportion of infected chicks and the lower weight gain is very small, the previously reported loss in breeder flock productivity and livability suggest broiler breeders should consider reducing ALV shedding in higher incidence female lines.


Excited-state dynamics of charged dyes at alkane/water interfaces in the presence of salts and ionic surfactants.
The excited-state dynamics of the cationic dye malachite green (MG) and of the dianionic dye eosin B at the dodecane/water interface has been investigated using femtosecond time-resolved surface second harmonic generation (TR-SSHG). By using different probe wavelengths, the contributions of monomeric and aggregated MG to the signal could be spectroscopically distinguished. The effect of the addition of a small amount of surfactants was found to strongly depend on the relative charges of surfactant and dye. For surfactant/dye pairs with opposite charges, the TR-SSHG signal is dominated by the contribution from aggregates, whereas for pairs with the same charges, the signal intensity becomes vanishingly small. These effects are explained in terms of electrostatic interactions between surfactants and dyes that favor either attraction of the dye toward the interface or its repulsion toward the bulk. As a very similar behavior is observed with MG upon addition of NaSCN, we conclude that, in this case, this effect reflects the affinity of SCN¯ for the interface. On the other hand, the guanidinium cation was found to have a different effect than that of a positively charged surfactant on the SSHG signal of MG, indicating this cation does not accumulate in the interfacial region.


In vivo compatibility and degradation of crosslinked gelatin gels incorporated in knitted Dacron.
Gelatin gels were applied to porous Dacron meshes with the aim of using these gels for local drug delivery. In this article, the biocompatibility and degradation of gelatin gels with different crosslink densities applied in Dacron were studied in vivo by subcutaneous implantation in rats. Dacron discs were treated with carbon dioxide gas plasma to improve hydrophilicity, and subsequently impregnated with gelatin type B. The gelatin samples were crosslinked to different extents using various amounts of water-soluble carbodiimide (EDC) and N-hydroxysuccinimide (NHS). After 6 h, 2, 5, and 10 days, and 3, 6, and 10 weeks of postimplantation, the tissue reactions and biodegradation were studied by light microscopy. The early reaction of macrophages and polymorphonuclear cells to crosslinked gelatin was similar to or milder than Dacron. Giant cell formation was predominantly aimed at Dacron fibers and was markedly reduced in the presence of a crosslinked gelatin coating. At week 10 of implantation, the crosslinked gelatin gels were still present in the Dacron matrix. The gelatin degradation was less for samples with the highest crosslink density. The gelatin gel with the lowest crosslink density showed clear cellular ingrowth, starting after 6 weeks of implantation. The intermediate and high crosslinked gelatin gels showed little or no ingrowth. In these gels, giant cells were involved in the phagocytosis of gelatin parts at week 10. Application of carbodiimide crosslinked gelatin gels in Dacron is suitable for medical applications because of the good biocompatibility of the gels and the possibility of adapting the degradation rate of gelatin to a specific application.


Multifunctional Nucleus-targeting Nanoparticles with Ultra-high Gene Transfection Efficiency for In Vivo Gene Therapy.
Cancer stem cell-like cells (CSCL) are responsible for tumor recurrence associated with conventional therapy (e.g. surgery, radiation, and chemotherapy). Here, we developed a novel multifunctional nucleus-targeting nanoparticle-based gene delivery system which is capable of targeting and eradicating CSCL. These nanoparticles can facilitate efficient endosomal escape and spontaneously penetrate into nucleus without additional nuclear localization signal. They also induced extremely high gene transfection efficiency (>95%) even in culture medium containing 30% serum, which significantly surpassed that of some commercial transfection reagents, such as Lipofectamine 2000 and Lipofectamine 3000 etc. Especially, when loaded with the TRAIL gene, this system mediated remarkable depletion of CSCL. Upon systemic administration, the nanoparticles accumulated in tumor sites while sparing the non-cancer tissues and significantly inhibited the growth of tumors with no evident systemic toxicity. Taken together, our results suggest that these novel multifunctional, nucleus-targeting nanoparticles are a very promising in vivo gene delivery system capable of targeting CSCL and represent a new treatment candidate for improving the survival of cancer patients.


Predicting the short-term success of human influenza virus variants with machine learning.
Seasonal influenza viruses are constantly changing and produce a different set of circulating strains each season. Small genetic changes can accumulate over time and result in antigenically different viruses; this may prevent the body's immune system from recognizing those viruses. Due to rapid mutations, in particular, in the haemagglutinin (HA) gene, seasonal influenza vaccines must be updated frequently. This requires choosing strains to include in the updates to maximize the vaccines' benefits, according to estimates of which strains will be circulating in upcoming seasons. This is a challenging prediction task. In this paper, we use longitudinally sampled phylogenetic trees based on HA sequences from human influenza viruses, together with counts of epitope site polymorphisms in HA, to predict which influenza virus strains are likely to be successful. We extract small groups of taxa (subtrees) and use a suite of features of these subtrees as key inputs to the machine learning tools. Using a range of training and testing strategies, including training on H3N2 and testing on H1N1, we find that successful prediction of future expansion of small subtrees is possible from these data, with accuracies of 0.71-0.85 and a classifier 'area under the curve' 0.75-0.9.


Rapid and remarkable effectiveness of benralizumab for treating severe bronchial asthma with intractable eosinophilic rhinosinusitis and eosinophilic otitis media: A case report.
Severe bronchial asthma is a challenging disorder to treat and can impair quality of life (QOL) under conventional therapeutic modalities. We report the case of a 52-year-old woman with severe asthma associated with eosinophilic chronic rhinosinusitis (ECRS) and eosinophilic otitis media (EOM). Although the patient was treated with a full dose of inhaled corticosteroid, leukotriene receptor antagonist (LTRA), theophylline, burst use of oral corticosteroids (OCS), her asthmatic condition aggravated, disrupting her daily life. ECRS and EOM symptoms were also getting worse despite treatment with topical application of corticosteroids to the nose and ears, LTRA, and occasional use of OCS. In addition to asthmatic symptom, the patient always suffered from intractable nasal obstruction and hearing disturbance, which contributed to the heavily impaired QOL. However, the administration of benralizumab showed rapid and remarkable improvement not only in her asthmatic conditions but also in the symptoms of ECRS and EOM within a month. These results suggest that the use of benralizumab for the treatment of severe asthma with intractable ECRS and EOM should be considered when the patient's QOL is severely deteriorated.


Rapid Synthesis of Psychoplastogenic Tropane Alkaloids.
Tropane alkaloids are an important class of biologically active small molecules characterized by their 8-azabicyclo[3.2.1]octane core. Because of their numerous medicinal applications, microbial biosynthesis and a variety of chemical syntheses have been designed for individual family members. However, current approaches are not amenable to late-stage structural diversification at N8, C3, C6, or C7, positions that are critical for modulating the biological properties of these molecules. Here, we describe a general approach to the synthesis of tropane alkaloids and their analogues that relies on the construction of the 8-azabicyclo[3.2.1]octane core through aziridination of a cycloheptadiene intermediate, followed by vinyl aziridine rearrangement. Using this strategy, we synthesized six tropane alkaloids and several analogues in only 5-7 steps. Given that the tropane alkaloid scopolamine has been reported to promote structural neuroplasticity and produce antidepressant effects, we tested five tropane-containing compounds for their ability to promote dendritic spine growth in cultured cortical neurons. We found that the orientation of the C3 substituent may play a role in the psychoplastogenic effects of tropane alkaloids. Our work provides a robust platform for producing tropane analogs for future structure-activity relationship studies.


[The role of adrenocorticotropic hormone in inhibition of pain reaction in awake rats].
The effect of hormones of hypothalamic-pituitary-adrenocortical system on pain sensitivity were studied in experiments on awake Sprague-Dawley males rats. Pain sensitivity was tested by tail flick reaction induced by thermal stimuli. Systemic glucocorticoids and ACTH injection increased the tail flick latency. The ACTH-induced analgesic effect was unaffected by deficiency of glucocorticoids production in pretreatment with pharmacological dose of cortisol but was fully eliminated by pretreatment with opiate antagonist naltrexone. These findings suggest that ACTH-induced analgesic effect is mediated by opiate receptors but not by glucocorticoids released in response to ACTH injection.


Multiple DNA-protein interactions governing high-precision DNA transactions.
The precise association of DNA-binding proteins with localized regions of DNA is crucial for regulated replication and expression of the genome. For certain DNA transactions, the requirement for precision in localization and control is extremely high. High-precision events amenable to detailed biochemical analysis are the initiation of DNA replication and site-specific recombination by bacteriophage lambda and Escherichia coli. Recent experiments indicate that site-localization and control in these reactions involves the association of DNA-bound proteins to generate organized nucleoprotein structures in which the DNA is folded or wound. These specialized nucleoprotein structures are likely to provide the requisite accuracy for site localization and the necessary regulated reactivity to direct the DNA transaction. Multiple DNA-protein interactions are also required for controlled transcription of the eukaryotic genome. Distant upstream regulator and enhancer sequences may define protein-binding sites that form part of a reactive nucleoprotein structure capable of initiating transcription.


Prostasomes: Their Characterisation: Implications for Human Reproduction: Prostasomes and Human Reproduction.
The prostate is a principal accessory genital gland that is vital for normal fertility. Epithelial cells lining the prostate acini release in a defined fashion (exocytosis) organellar nanosized structures named prostasomes. They are involved in the protection of sperm cells against immune response in the female reproductive tract by modulating the complement system and by inhibiting monocyte and neutrophil phagocytosis and lymphocyte proliferation. The immunomodulatory function most probably involves small non-coding RNAs present in prostasomes. Prostasomes have also been proposed to regulate the timing of sperm cell capacitation and induction of the acrosome reaction, since they are rich in various transferable bioactive molecules (e.g. receptors and enzymes) that promote the fertilising ability of sperm cells. Antigenicity of sperm cells has been well documented and implicated in involuntary immunological infertility of human couples, and antisperm antibodies (ASA) occur in several body fluids. The propensity of sperm cells to carry attached prostasomes suggests that they are a new category of sperm antigens. Circulating human ASA recognise prostasomes, and among 12 identified prostasomal antigens, prolactin- inducible protein (95 %) and clusterin (85 %) were immunodominant at the expense of the other 10 that were sporadically occurring.


Phenotypic and Genetic Analysis of the Leopard Complex Spotting in Noriker Horses.
Genetic analyses of coat colors are frequently restricted to subjectively categorized phenotype information. The aim of this study was to develop a method to numerically quantify the variability of leopard complex (LP) spotting phenotypes introducing tools from image analysis. Generalized Procrustes analysis eliminates systematic errors due to imaging process. The binarization of normalized images and the application of principal component analysis (PCA) on the derived pixel matrices, transform pixel information into numerical data space. We applied these methods on 90 images to ascertain the specific leopard patterns within the Noriker breed. Furthermore, we genotyped a representative sample of 191 Noriker horses for the known LP spotting associated loci. Ninety-seven percentage of the genotyped leopard spotted horses were heterozygous for LP and had at least one copy of the PATN1 allele. However, the remaining pattern variation was great, indicating other genetic factors influencing the expression of LP spotting. Based upon this data, we estimated effect sizes of the modifier PATN1, and additional factors including sex, age, base color, and spotting phenotype of parents. The PCA of the pixel matrix resulted in 2 significant components accounting for 51% of the variation. Applying a linear model, we identified significant effects for age groups and base color on the first and second components, while for sex and parents' LP phenotype significant effects were found on 4 additional components.


Topical Review: Acute and Chronic Pain Experiences in Transgender and Gender-Diverse Youth.
To provide an overview of the existing literature on gender diversity in pediatric acute and chronic pain, propose an ecological systems model of understanding pain in transgender and gender-diverse (TGD) youth, and identify a direction for future work that will address the key knowledge gaps identified. Relevant literature on pain and gender diversity was reviewed, drawing from adult literature where there was insufficient evidence in pediatric populations. Existing relevant models for understanding minority stress, gender and pain, and pain experiences within marginalized groups were considered with the reviewed literature to develop a pain model in TGD youth. While there is an abundance of literature pointing to increased risk for pain experiences amongst TGD youth, there is comparably little empirical evidence of the rates of pain amongst TGD youth, prevalence of TGD identities in pain care settings, effective pain treatments for TGD youth and unique considerations for their care, and the role intersectional factors in understanding TGD youth identities and pain. Pediatric psychologists are well-positioned to advance the research on acute and chronic pain in TGD youth, make evidence-based adaptations to clinical care for TGD youth with pain, including pain related to gender affirmation, and support colleagues within the medical system to provide more inclusive care.


Deployment of spatial attention to a structural framework: exogenous (alerting) and endogenous (goal-directed) factors.
The main question examined in the present work was whether spatial attention can be deployed to an appropriate structural framework not only endogenously when the framework is displayed continuously, as in previous work, but also exogenously, when it is displayed transiently 100 ms before the target. The results of five experiments answered that question in the negative. We found that the onset transient triggered by a brief presentation of the structural framework did enhance the response to the upcoming target. That enhancement, however, was due not to the framework itself but to the alerting effect produced by its sudden onset, witness the finding that the same enhancement was produced by an onset transient triggered by a featureless stimulus (i.e., by a brief dimming of the entire screen, in the absence of a structural framework). We conclude that spatial attention can be deployed to the region demarcated by a structural framework when it is deployed endogenously but not when it is deployed exogenously. A theoretical account of the results is proposed in terms of the temporal dynamics of the locus cœruleus/norepinephrine neuromodulatory system.


Male to female and female to male transgender persons have different sexual risk behaviors yet similar rates of STDs and HIV.
The epidemiology of STDs and HIV among male-to-female (MTF) and female-to-male (FTM) transgender persons is limited, which makes prevention for transgender populations challenging. We examined data collected at visits for all self-identified MTF and FTM patients at the municipal STD clinic in San Francisco from January 1, 2006 to December 31, 2009. We compared demographic and socio-behavioral characteristics, as well as STD and HIV positivity and history of previous STD. Despite demographic and behavioral risk differences, there were no differences in STD positivity or HIV prevalence between MTF and FTM. A more complete understanding of the prevention needs for transgender persons is needed.


Targeted cancer gene therapy using a hypoxia inducible factor dependent oncolytic adenovirus armed with interleukin-4.
There is a need for novel therapies targeting hypoxic cells in tumors. These cells are associated with tumor resistance to therapy and express hypoxia inducible factor-1 (HIF-1), a transcription factor that mediates metabolic adaptation to hypoxia and activates tumor angiogenesis. We previously developed an oncolytic adenovirus (HYPR-Ad) for the specific killing of hypoxic/HIF-active tumor cells, which we now armed with an interleukin-4 gene (HYPR-Ad-IL4). We designed HYPR-Ad-IL4 by cloning the Ad E1A viral replication and IL-4 genes under the regulation of a bidirectional hypoxia/HIF-responsive promoter. The IL-4 cytokine was chosen for its ability to induce a strong host antitumor immune response and its potential antiangiogenic activity. HYPR-Ad-IL4 induced hypoxia-dependent IL-4 expression, viral replication, and conditional cytolysis of hypoxic, but not normoxic cells. The treatment of established human tumor xenografts with HYPR-Ad-IL4 resulted in rapid and maintained tumor regression with the same potency as that of wild-type dl309-Ad. HYPR-Ad-IL4-treated tumors displayed extensive necrosis, fibrosis, and widespread viral replication. Additionally, these tumors contained a distinctive leukocyte infiltrate and prominent hypoxia. The use of an oncolytic Ad that locally delivers IL-4 to tumors is novel, and we expect that HYPR-Ad-IL4 will have broad therapeutic use for all solid tumors that have hypoxia or active HIF, regardless of tissue origin or genetic alterations.


Effects of fatty acid intermediates on Na+-K+-ATPase activity of cardiac sarcolemma.
The effect of amphiphilic lipid intermediates on the Na+-stimulatable activity of the Na+-K+-ATPase of sarcolemma from adult canine cardiac myocytes was studied. Sarcolemma (mean Na+-stimulatable ATPase activity of 73 mumol.mg sarcolemmal protein-1.h-1) was preincubated (37 degrees C for 10 min at pH 7.2) or rapidly mixed at 0 degrees C with amphiphilic lipid intermediates prior to dilution and assay of enzyme activity. Their effects were dependent on temperature, initial concentration, and the ratio of bound amphiphile to sarcolemmal protein. In particular, pretreatment of freshly prepared sarcolemma at 0 degrees C with arachidonyl CoA (up to 0.25 mM) caused 110% stimulation above control activity; palmitoyl CoA or palmitoyl carnitine under the same conditions caused no significant effect. Despite strong binding to the sarcolemmal vesicles, palmitoyl carnitine (up to 0.4 mM or 5 mumol/mg protein) and palmitoyl CoA (0.1 mM or 1.0 mumol of membrane-bound palmitoyl CoA/mg protein) were ineffective even with preincubation. Palmitoyl CoA was inhibitory above this level. Preincubation (22 degrees C for 10 min) with lysophosphatidylcholine only produced inhibition (40% at 0.75 mM). Thus fatty acyl thioesters of CoA and lysophosphatidyl choline but not palmitoyl carnitine perturb sarcolemmal Na+-K+-ATPase activity.


Zebrafish CaV2.1 calcium channels are tailored for fast synchronous neuromuscular transmission.
The CaV2.2 (N-type) and CaV2.1 (P/Q-type) voltage-dependent calcium channels are prevalent throughout the nervous system where they mediate synaptic transmission, but the basis for the selective presence at individual synapses still remains an open question. The CaV2.1 channels have been proposed to respond more effectively to brief action potentials (APs), an idea supported by computational modeling. However, the side-by-side comparison of CaV2.1 and CaV2.2 kinetics in intact neurons failed to reveal differences. As an alternative means for direct functional comparison we expressed zebrafish CaV2.1 and CaV2.2 α-subunits, along with their accessory subunits, in HEK293 cells. HEK cells lack calcium currents, thereby circumventing the need for pharmacological inhibition of mixed calcium channel isoforms present in neurons. HEK cells also have a simplified morphology compared to neurons, which improves voltage control. Our measurements revealed faster kinetics and shallower voltage-dependence of activation and deactivation for CaV2.1. Additionally, recordings of calcium current in response to a command waveform based on the motorneuron AP show, directly, more effective activation of CaV2.1. Analysis of calcium currents associated with the AP waveform indicate an approximately fourfold greater open probability (PO) for CaV2.1. The efficient activation of CaV2.1 channels during APs may contribute to the highly reliable transmission at zebrafish neuromuscular junctions.


A computer program for estimating thresholds by means of Wetherill's Up-Down Transformed Response Rule (UDTR).
A Fortran IV computer program is presented which allows for automatic estimation of thresholds on an S-shaped psychometric curve by means of Wetherill's Up-Down Transformed Response Rule (UDTR). The efficiency and asymptotic properties of these rules are discussed and illustrated. The main program is in the form of a keyboard portable tutorial which calls the main decision-making subroutine. The subroutine can be detached from the tutorial. Only a single call with seven parameters is required to reach the subroutine. The program is particularly useful in forced-choice procedures.


Peptide-Based Regulation of Metabolism as Related to Obesity.
Currently there are few, if any, approved and effective medicines for the attenuation of obesity, diabetes, and insulin resistance. This commentary addresses a communication describing the effect on glucose dynamics and obesity of a peptide monoclonal antibody for fibroblast growth factor (FGF) receptor 1c isoform (FGFR1c). The general lack of suitable, effective drugs is discussed, as is the treatment potential of FGF family receptors. The FGFR1c monoclonal antibody developed by the authors lowers body weight gain, blood glucose, and adipose tissue weight. It also enhances glucose uptake by fat cells [white adipose tissue (WAT) and 3T3-L1]. The robust weight loss, fat loss, and lower blood glucose were attributed to an observed potential futile cycle of continuous lipogenesis and lipolysis and adenosine triphosphate use in WAT.


Primary results of the SAVAL randomized trial of a paclitaxel-eluting nitinol stent versus percutaneous transluminal angioplasty in infrapopliteal arteries.
Effective and durable options for infrapopliteal artery revascularization for patients with chronic limb-threatening ischemia (CLTI) are limited. The SAVAL trial is a prospective, multicenter, randomized trial of patients with CLTI and infrapopliteal artery lesions with total lesion length ⩽ 140 mm, stenosis ⩾ 70%, and Rutherford category 4-5 assigned 2:1 to treatment with the SAVAL self-expandable paclitaxel drug-eluting stent (DES) or percutaneous transluminal angioplasty (PTA) with an uncoated balloon. The primary effectiveness endpoint was primary vessel patency (i.e., core lab-adjudicated duplex ultrasound-based flow at 12 months in the absence of clinically driven target lesion revascularization or surgical bypass of the target lesion). The primary safety endpoint was the 12-month major adverse event (MAE)-free rate; MAEs were defined as a composite of above-ankle index limb amputation, major reintervention, and 30-day mortality. The endpoints were prespecified for superiority (effectiveness) and noninferiority (safety) at a one-sided significance level of 2.5%. A total of 201 patients were enrolled and randomly assigned to treatment (N = 130 DES, N = 71 PTA). Target lesion length was 68.1 ± 35.2 mm for the DES group and 68.7 ± 49.2 mm for the PTA group, and 31.0% and 27.6% of patients, respectively, had occlusions. The 12-month primary patency rates were 68.0% for the DES group and 76.0% for the PTA group (Psuperiority = 0.8552). The MAE-free rates were 91.6% and 95.3%, respectively (Pnoninferiority = 0.0433). The SAVAL trial did not show benefit related to effectiveness and safety with the nitinol DES compared with PTA in infrapopliteal artery lesions up to 140 mm in length. Continued innovation to provide optimal treatments for CLTI is needed. (ClinicalTrials.gov Identifier: NCT03551496).


3D Vision Provides Shorter Operative Time and More Accurate Intraoperative Surgical Performance in Laparoscopic Hiatal Hernia Repair Compared With 2D Vision.
The aim of this study is to evaluate if 3-dimensional high-definition (3D) vision in laparoscopy can prompt advantages over conventional 2D high-definition vision in hiatal hernia (HH) repair. Between September 2012 and September 2015, we randomized 36 patients affected by symptomatic HH to undergo surgery; 17 patients underwent 2D laparoscopic HH repair, whereas 19 patients underwent the same operation in 3D vision. No conversion to open surgery occurred. Overall operative time was significantly reduced in the 3D laparoscopic group compared with the 2D one (69.9 vs 90.1 minutes, P = .006). Operative time to perform laparoscopic crura closure did not differ significantly between the 2 groups. We observed a tendency to a faster crura closure in the 3D group in the subgroup of patients with mesh positioning (7.5 vs 8.9 minutes, P = .09). Nissen fundoplication was faster in the 3D group without mesh positioning ( P = .07). 3D vision in laparoscopic HH repair helps surgeon's visualization and seems to lead to operative time reduction. Advantages can result from the enhanced spatial perception of narrow spaces. Less operative time and more accurate surgery translate to benefit for patients and cost savings, compensating the high costs of the 3D technology. However, more data from larger series are needed to firmly assess the advantages of 3D over 2D vision in laparoscopic HH repair.


Hereditary hydronephrosis in C57BL/KsJ mice.
We sought to determine if the incidence of renal hydronephrosis in male C57BL/KsJ mice increased with age and if grossly normal kidneys would develop hydronephrosis over time. Spontaneous hydronephrosis was found incidentally in 32% of 234 male C57BL/KsJ mice killed as pancreas donors for islet transplantation experiments. The incidence of hydronephrosis increased with age; the incidence was 15% in 6- to 8-week-old mice, 52% in 8- to 10-week-old mice and 63% in 11- to 15-week-old mice (P less than 0.001). Additional mice received islet isografts beneath the renal capsule. Only mice with grossly normal kidneys received islet grafts. These same kidneys were then re-examined when the graft recipients were killed at the end of the experiment and the incidence of hydronephrosis was determined. The conversion of normal kidneys to hydronephrotic kidneys increased with the time since islet transplantation. Kidneys re-examined less than 4 weeks since transplantation had only 5.8% new hydronephrosis, while those re-examined later than 4 weeks after transplantation had a new hydronephrosis incidence rate of 40% (P less than 0.001). Our findings suggest that hydronephrosis is hereditary but not congenital, that it develops rapidly, and that it can complicate experiments using this strain. This may also represent a useful new animal model of progressive hydronephrosis.


The mediator role of psychological morbidity in patients with chronic low back pain in differentiated treatments.
This study analyzed the mediating role of psychological morbidity and the variables that discriminated low versus high disability, in patients receiving physiotherapy and acupuncture. A total of 203 patients answered measures of illness and medication representations, coping, depression, anxiety, quality of life, and functional disability. Morbidity was a mediator between functional disability and quality of life. Treatment consequences and quality of life, in the acupuncture group, and emotional representations, quality of life, depression, anxiety, and active strategies for pain relief, in the physiotherapy group, discriminated patients with low versus high disability. These results have important implications for identifying high-risk patients.


The enzymatic defects in porphyria cutanea tarda and variegate porphyria.
Although enzymatic defects have been identified in porphyria cutanea tarda and variegate porphyria several important controversial points remain unresolved. Hepatic uroporphyrinogen decarboxylase activity is subnormal in all patients with porphyria cutanea tarda and is presumably responsible for the biochemical derangement which characterizes the disease. Several groups have found subnormal erythrocyte uroporphyrinogen decarboxylase activity as well and have demonstrated that the defect is inherited as an autosomal dominant trait. In other studies, erythrocyte enzyme activity has been normal and no evidence of an inherited factor has been identified. These two types of patients have been called "familial" and "sporadic" cases of porphyria cutanea tarda. Whether or not the hepatic enzyme defect is inherited in all cases remains to be determined. Two groups of investigators have identified two different enzymatic defects in variegate porphyria. One group has reported subnormal activity of heme synthase (ferrochelatase) to be the defect responsible for the disease and the other has reported subnormal activity of protoporphyrinogen oxidase to be the causative factor. Which of the two defects is responsible for the biochemical abnormalities in variegate porphyria remains to be resolved.


Gene-by-Environment Interactions on Alcohol Use Among Asian American College Freshmen.
Among northeast Asians, the variant aldehyde dehydrogenase allele, ALDH2*2 (rs671, A/G, minor/major), has been inversely associated with alcohol dependence. The strength of the associations between ALDH2*2 and drinking behaviors depends on the developmental stage, the phenotype studied, and other moderating variables. This study examined ALDH2 gene status as a moderator of the associations between parental drinking, peer drinking, and acculturation with alcohol use among 222 Chinese American and Korean American college freshmen. Negative binomial regressions were used to test the main and interactive effects of ALDH2 with contextual factors on alcohol frequency (drinking days) and quantity (drinks per drinking day) in the past 3 months. ALDH2*2 was associated with more subjective flushing symptoms and longer length of flushing but was unrelated to both alcohol frequency and quantity. Peer drinking was positively associated with both alcohol frequency and quantity, but neither was moderated by ALDH2. We observed a nonsignificant trend for the interaction between parental drinking and ALDH2 on alcohol frequency, where parental drinking was positively associated with alcohol frequency only among participants with ALDH2*2. We found a significant interaction between acculturation and ALDH2 on alcohol frequency, where acculturation was positively associated with alcohol frequency only among those with ALDH2*2. Exploratory analyses stratified by Asian ethnic subgroup indicated that this interaction was driven primarily by the Korean subsample. Parental drinking and acculturation may facilitate more frequent drinking among those who have more intense reactions to alcohol (i.e., those with ALDH2*2) during the transition from high school to college.


[Rectal cancer: current status of multimodal therapy--when and how?].
Preoperative 5-fluorouracil-based radiochemotherapy (RCT) followed by quality assessed total mesorectal excision (TME surgery) are the two most important elements of multimodal treatment for patients with locally advanced rectal cancer (UICC stages II and III). The optimum sequence of these neoadjuvant modalities complemented by adjuvant (postoperative) chemotherapy, has been addressed in several randomised trials. Especially within the trials of the German Rectal Cancer Study Group (GRCSG), preoperative RCT has been shown to be superior to postoperative treatment for a variety of endpoints (pathologically confirmed complete tumour remission (pCR), RCT-induced tumour regression, R0 resection rates (including circumferential resection margins) and long-term locoregional control). This neoadjuvant multimodal strategy has decreased the 5-year and 10-year local recurrence rates below 10%, and the development of distant metastases (e.g., 35% to 45% liver metastases) remains the predominant reason for failure. Furthermore, approximately 25% of patients do not receive adjuvant chemotherapy, mainly due to surgical complications, patients' refusal or the investigator's discretion. Thus, today, integrating more effective systemic therapy into (preoperative) multimodal regimens is the most accepted challenge! But from the clinical point of view this demand is also a dilemma. The question to be addressed is how and when to apply intensified systemic therapy with adequate dosage and intensity as well as acceptable treatment-associated toxicity. The increase of therapeutic options requires valid predictive biomarkers that may help to stratify patients into regimens associated with low toxicity (5-FU monotherapy alone) or into more intensified treatment for better long-term outcome. In summary, the use of biomarkers for individualised risk-adapted treatment is one of the most promising areas of clinical investigations, not only in rectal cancer. The assessment of individual tumour response, toxicity, and prognosis during multimodal treatment of rectal cancer as a model of a very common solid tumour offers radiooncologists, surgeons, pathologists, gastroenterologists as well as oncologists immense insights into the under-standing of tumour biology.


The Mex-Cog cognitive assessment battery: discriminant analysis of the cognitive performance profile in older adults.
To analyze the cognitive profile of a clinical sample using the Mex-Cog cognitive battery and establish which cognitive measures and domains contribute most to group separation. A group of 145 older adults previously diagnosed with dementia (n= 47), mild cognitive impairment MCI (n= 47), or as cognitively normal (n= 51) were assessed with the Mex-Cog cognitive battery. Six linear discriminant analyses (LDA) were estimated to compare dementia vs. cognitively normal, MCI vs. cognitively normal, and MCI vs. dementia, using ten individual measures and six cognitive domains. We used a leave-one-out cross-validation procedure to evaluate the predictive capacity of LDA models. Discriminant functions using individual measures and domains distinguished correctly 100% of dementia and cognitively normal groups showing a memory and executive function profile. The predictive group membership for MCI versus cognitively normal varied between 82 and 85%, with a cognitive profile associated with attention-executive function followed by memory. Group separation between MCI and dementia was between 80 and 87%, characterized by orientation, memory, and visuospatial abilities. The Mex-Cog cognitive battery is useful for identifying cognitive impairment in older adults.


A junctional PACSIN2/EHD4/MICAL-L1 complex coordinates VE-cadherin trafficking for endothelial migration and angiogenesis.
Angiogenic sprouting relies on collective migration and coordinated rearrangements of endothelial leader and follower cells. VE-cadherin-based adherens junctions have emerged as key cell-cell contacts that transmit forces between cells and trigger signals during collective cell migration in angiogenesis. However, the underlying molecular mechanisms that govern these processes and their functional importance for vascular development still remain unknown. We previously showed that the F-BAR protein PACSIN2 is recruited to tensile asymmetric adherens junctions between leader and follower cells. Here we report that PACSIN2 mediates the formation of endothelial sprouts during angiogenesis by coordinating collective migration. We show that PACSIN2 recruits the trafficking regulators EHD4 and MICAL-L1 to the rear end of asymmetric adherens junctions to form a recycling endosome-like tubular structure. The junctional PACSIN2/EHD4/MICAL-L1 complex controls local VE-cadherin trafficking and thereby coordinates polarized endothelial migration and angiogenesis. Our findings reveal a molecular event at force-dependent asymmetric adherens junctions that occurs during the tug-of-war between endothelial leader and follower cells, and allows for junction-based guidance during collective migration in angiogenesis.


Plasma parasitemia as assessed by quantitative PCR in relation to clinical disease severity in African adults with falciparum malaria with and without HIV co-infection.
When considering malaria disease severity, estimation of parasitemia in erythrocytes is important, but sometimes misleading, since the infected erythrocytes may be sequestered in peripheral capillaries. In African children and Asian adults with falciparum malaria, parasitemia as assessed by quantitative PCR (qPCR) in plasma seems to be a valuable indicator of disease severity, but data on African adults as well as the impact of co-infection with HIV is lacking. In 131 patients with falciparum malaria in a public tertiary teaching hospital in Mozambique, plasma malaria parasitemia as assessed by qPCR, compared to qualitative malaria PCR in blood cell fraction, was related to malaria disease severity and HIV co-infection. Of the 131 patients with falciparum malaria, based on positive qualitative PCR in the blood cell fraction, 93 patients (72%) had positive malaria qPCR in plasma. Patients with severe malaria as defined by the WHO criteria had higher malaria quantitative plasma parasitemia (median 143 genomes/µL) compared to those with uncomplicated malaria (median 55 genomes/µL, p = 0.037) in univariate analysis, but this difference was attenuated after adjusting for age, sex and HIV co-infection (p = 0.055). A quarter of the patients with severe malaria had negative qPCR in plasma. This study of adult African in-patients with falciparum malaria with and without HIV co-infection, neither confirms nor rejects previous studies of malaria qPCR in plasma as an indicator of disease severity in patients with falciparum malaria. There is a need for further and larger studies to clarify if parasitemia as assessed malaria qPCR in plasma could be a surrogate marker of disease severity in falciparum malaria.


Refining Markov Clustering for protein complex prediction by incorporating core-attachment structure.
Protein complexes are responsible for most of vital biological processes within the cell. Understanding the machinery behind these biological processes requires detection and analysis of complexes and their constituent proteins. A wealth of computational approaches towards detection of complexes deal with clustering of protein-protein interaction (PPI) networks. Among these clustering approaches, the Markov Clustering (MCL) algorithm has proved to be reasonably successful, mainly due to its scalability and robustness. However, MCL produces many noisy clusters, which either do not represent any known complexes or have additional proteins (noise) that reduce the accuracies of correctly predicted complexes. Consequently, the accuracies of these clusters when matched with known complexes are quite low. Refinement of these clusters to improve the accuracy requires deeper understanding of the organization of complexes. Recently, experiments on yeast by Gavin et al. (2006) revealed that proteins within a complex are organized in two parts: core and attachment. Based on these insights, we propose our method (MCL-CA), which couples core-attachment based refinement steps to refine the clusters produced by MCL. We evaluated the effectiveness of our approach on two different datasets and compared the quality of our predicted complexes with that produced by MCL. The results show that our approach significantly improves the accuracies of predicted complexes when matched with known complexes. A direct result of this is that MCL-CA is able to cover larger number of known complexes than MCL. Further, we also compare our method with two very recently proposed methods CORE and COACH, which also capitalize on the core-attachment structure. We also discuss several instances to show that our predicted complexes clearly adhere to the core-attachment structure as revealed by Gavin et al.


Pembrolizumab in lung cancer: current evidence and future perspectives.
Pembrolizumab is a humanized monoclonal antibody against PD-1 capable of enhancing antitumor immune activity. The KEYNOTE-001 study showed that pembrolizumab has activity in advanced non-small-cell lung cancer patients and identified programmed death ligand 1 (PD-L1) as a companion test to select patients most likely to benefit from pembrolizumab. Five randomized clinical trials showed the efficacy of pembrolizumab in non-small-cell lung cancer: in second-line setting PD-L1 ≥1% (KEYNOTE-010), in first-line setting PD-L1 ≥50% (KEYNOTE-024 and KEYNOTE-042) and in first-line setting in combination with platinum doublets, any expression of PD-L1 (KEYNOTE-189 and KEYNOTE-407). Future challenges are the identification of the role of pembrolizumab in adjuvant, neoadjuvant, locally advanced disease or oncogene-addicted patients, in combination with radiotherapy or other biological agents.


Antiviral activity and safety of aplaviroc, a CCR5 antagonist, in combination with lopinavir/ritonavir in HIV-infected, therapy-naïve patients: results of the EPIC study (CCR100136).
This phase IIb study explored the antiviral activity and safety of the investigational CC chemokine receptor 5 (CCR5) antagonist aplaviroc (APL) in antiretroviral-naïve patients harbouring R5- or R5X4-tropic virus. A total of 191 patients were randomized 2:2:2:1 to one of three APL dosing regimens or to lamivudine (3TC)/zidovudine (ZDV) twice daily (bid), each in combination with lopinavir/ritonavir (LPV/r) 400 mg/100 mg bid. Efficacy, safety and pharmacokinetic parameters were assessed. This study was terminated prematurely because of APL-associated idiosyncratic hepatotoxicity. A total of 141 patients initiated treatment early enough to have been able to complete 12 weeks on treatment [modified intent-to-treat (M-ITT) population]; of these, 133 completed the 12-week treatment phase. The proportion of subjects in the M-ITT population with HIV-1 RNA <400 copies/mL at week 12 was 50, 48, 54 and 75% in the APL 200 mg bid, APL 400 mg bid, APL 800 mg once a day (qd) and 3TC/ZDV arms, respectively. Similar responses were seen in the few subjects harbouring R5X4-tropic virus (n=17). Common clinical adverse events (AEs) were diarrhoea, nausea, fatigue and headache. APL demonstrated nonlinear pharmacokinetics with high interpatient variability. While target plasma concentrations of APL were achieved, the antiviral activity of APL+LPV/r did not appear to be comparable to that of 3TC/ZDV+LPV/r.


Altered Skeletal Muscle Mitochondrial Proteome As the Basis of Disruption of Mitochondrial Function in Diabetic Mice.
Insulin plays pivotal role in cellular fuel metabolism in skeletal muscle. Despite being the primary site of energy metabolism, the underlying mechanism on how insulin deficiency deranges skeletal muscle mitochondrial physiology remains to be fully understood. Here we report an important link between altered skeletal muscle proteome homeostasis and mitochondrial physiology during insulin deficiency. Deprivation of insulin in streptozotocin-induced diabetic mice decreased mitochondrial ATP production, reduced coupling and phosphorylation efficiency, and increased oxidant emission in skeletal muscle. Proteomic survey revealed that the mitochondrial derangements during insulin deficiency were related to increased mitochondrial protein degradation and decreased protein synthesis, resulting in reduced abundance of proteins involved in mitochondrial respiration and β-oxidation. However, a paradoxical upregulation of proteins involved in cellular uptake of fatty acids triggered an accumulation of incomplete fatty acid oxidation products in skeletal muscle. These data implicate a mismatch of β-oxidation and fatty acid uptake as a mechanism leading to increased oxidative stress in diabetes. This notion was supported by elevated oxidative stress in cultured myotubes exposed to palmitate in the presence of a β-oxidation inhibitor. Together, these results indicate that insulin deficiency alters the balance of proteins involved in fatty acid transport and oxidation in skeletal muscle, leading to impaired mitochondrial function and increased oxidative stress.


The Effect of Amino-Phosphate Interactions on the Biosensing Performance of Enzymatic Graphene Field-Effect Transistors.
The interaction between polyamines and phosphate species is found in a wide range of biological and abiotic systems, yielding crucial consequences that range from the formation of supramolecular colloids to structure determination. In this work, the occurrence of phosphate-amino interactions is evidenced from changes in the electronic response of graphene field effect transistors (gFETs). First, the surface of the transistors is modified with poly(allylamine), and the effect of phosphate binding on the transfer characteristics is interpreted in terms of its impact on the surface charge density. The electronic response of the polyamine-functionalized gFETs is shown to be sensitive to the presence of different phosphate anions, such as orthophosphate, adenosine triphosphate, and tripolyphosphate, and a simple binding model is developed to explain the dependence of the shift of the Dirac point potential on the phosphate species concentration. Afterward, the impact of phosphate-amino interactions on the immobilization of enzymes to polyamine-modified graphene surfaces is investigated, and a decrease in the amount of anchored enzyme as the phosphate concentration increases is found. Finally, multilayer polyamine-urease biosensors are fabricated while increasing the phosphate concentration in the enzyme solution, and the sensing properties of the gFETs toward urea are evaluated. It is found that the presence of simple phosphate anions alters the nanoarchitecture of the polyelectrolyte-urease assemblies, with direct implications on urea sensing.