Etiology of late free flap failures occurring after hospital discharge.
Vascular compromise of free flaps most commonly occurs in the immediate postoperative period in association with failure of the microvascular anastomosis. Rarely do flaps fail in the late postoperative period. It is not well understood why free flaps can fail after 7 postoperative days. We undertook a case review series to assess possible causes of late free flap failure. Retrospective review at two tertiary referral centers: Oregon Health Sciences University and University of Alabama at Birmingham. A review of 1,530 flaps performed in 1,592 patients between 1998 and 2006 were evaluated to identify late flap failure. Late flap failure was defined as failure occurring after postoperative day 7 or on follow-up visits after hospital discharge. A prospective database with the following variables was examined: age, medical comorbidities, postreconstructive complications (fistula or infection), hematoma, seroma, previous surgery, radiation therapy, intraoperative findings at the time of debridement, nutrition, and, possibly, etiologies. A total of 13 patients with late graft failure were identified in this study population of 1,530 (less than 1%) flaps; 6 radial forearm fasciocutaneous flaps, 2 rectus abdominis myocutaneous flaps, 4 fibular flaps, and 1 latissimus dorsi myocutaneous flap underwent late failure. The time to necrosis was a median of 21 (range, 7-90) days. Etiology was believed to possibly be pressure on the pedicle in the postoperative period in four patients (no sign of local wound issues at the pedicle), infection (abscess formation) in three patients, and regrowth of residual tumor in six patients. Loss occurring within 1 month was more common in radial forearm flaps and was presented in the context of a normal appearing wound at the anastomotic site, as opposed to loss occurring after 1 month, which happened more commonly in fibula flaps secondary to recurrence. Although late free flap failure is rare, local factors such as infection and possibly pressure on the pedicle can be contributing factors. Patients presenting with late flap failure should be evaluated for residual tumor growth.


Statin treatment before stroke reduces pro-inflammatory cytokine levels after stroke.
In this clinical case-control study, we investigated statin treatment in stroke patients on a range of inflammatory effectors in peripheral blood. We focus on RhoA GTPase and its downstream effectors as a future inflammatory target in stroke treatment. Data from 10 patients already on statins at stroke onset (Pre-S group) was compared with data from both 29 patients starting statin treatment right after stroke onset (Post-S group) and with 8 healthy controls. In T-cells isolated from stroke patients, we analyzed the activity of the main cytoskeletal regulator RhoA GTPase and its downstream effectors: rho-associated protein kinase (ROCK), myosin phosphatase targeting protein subunit 1 (pMYPT1), myosin light chain kinase (pMLC) and cofilin. In the blood samples, we further determined levels of 12 key plasma cytokines as well as C-reactive protein (CRP) and kallikrein. Compared to healthy controls, the Post-S group achieved significantly higher RhoA and ROCK activities, while the Pre-S did not differ from controls. Levels of pMYPT1, pMLC and cofilin did not differ from controls in the Pre-S and Post-S groups. At day 90 after stroke, interferon γ and IL-18 were significantly increased in the Post-S group compared to the Pre-S group. We found a positive correlation between CRP and NIHSS, whereas kallikrein levels showed no correlation with NIHSS at any of the days. Stroke induces changes in the RhoA-ROCK pathway in T-cells. CRP and NIHSS score correlated positively in the study. Statins may have an anti-inflammatory effect as statin treatment before stroke reduces post-stroke pro-inflammatory levels. RhoA GTPase and its downstream effectors are possibly the key to improve statin treatment in stroke.


No actual conflict over colony inheritance despite high potential conflict in the social wasp Polistes dominulus.
Social insect societies are outstanding examples of cooperation and conflict. Individuals work together, yet seek to increase their inclusive fitness at each others' expense. One such conflict is over colony inheritance, when a queen inherits the colony following the death of the previous queen. Colony inheritance is common in the social wasp Polistes dominulus, and it can have dramatic fitness consequences. The subordinate inheriting the colony is often unrelated to the initial foundress (alpha) and the workers, who therefore get zero inclusive fitness. Workers are capable of mating and reproducing, so that inheritance by a subordinate rather than by a related worker is surprising. Using patterns of egg-laying and egg destruction, we show in 32 laboratory colonies that, upon the removal of alpha, workers fully accepted a subordinate as the new breeder. This new alpha monopolized reproduction to the same extent as alpha, and there was no increase in reproduction by workers and other subordinates. Why workers accept a potentially unrelated subordinate as breeder rather than a full-sister worker is unclear. They may be constrained to do so, and they may seek fitness benefits by producing males later in the season or by absconding the nest.


Role of mitochondrial dysfunction and dopamine-dependent oxidative stress in amphetamine-induced toxicity.
To define the molecular mechanisms underlying amphetamine (AMPH) neurotoxicity, primary cultures of dopaminergic neurons were examined for drug-induced changes in dopamine (DA) distribution, oxidative stress, protein damage, and cell death. As in earlier studies, AMPH rapidly redistributed vesicular DA to the cytoplasm, where it underwent outward transport through the DA transporter. DA was concurrently oxidized to produce a threefold increase in free radicals, as measured by the redox-sensitive dye dihydroethidium. Intracellular DA depletion using the DA synthesis inhibitor alpha-methyl-p-tyrosine or the vesicular monoamine transport blocker reserpine prevented drug-induced free radical formation. Despite these AMPH-induced changes, neither protein oxidation nor cell death was observed until 1 and 4 days, respectively. AMPH also induced an early burst of free radicals in a CNS-derived dopaminergic cell line. However, AMPH-mediated attenuation of ATP production and mitochondrial function was not observed in these cells until 48 to 72 hours. Thus, neither metabolic dysfunction nor loss of viability was a direct consequence of AMPH neurotoxicity. In contrast, when primary cultures of dopaminergic neurons were exposed to AMPH in the presence of subtoxic doses of the mitochondrial complex I inhibitor rotenone, cell death was dramatically increased, mimicking the effects of a known parkinsonism-inducing toxin. Thus, metabolic stress may predispose dopaminergic neurons to injury by free radical-promoting insults such as AMPH.


Differential signaling and tumor necrosis factor receptor-associated factor (TRAF) degradation mediated by CD40 and the Epstein-Barr virus oncoprotein latent membrane protein 1 (LMP1).
Latent membrane protein 1 (LMP1) plays a critical role in B cell transformation by Epstein-Barr virus (EBV) and appears to mimic a constitutively active CD40 receptor. Intracellular tumor necrosis factor (TNF) receptor-associated factor (TRAF) adapter proteins, shown to contribute to signaling by both CD40 and LMP1, were recruited by both molecules to lipid-enriched membrane rafts. However, we found that TRAFs 2 and 3 were subsequently degraded after CD40- but not LMP1-induced signaling. This degradation was proteasome-dependent and required direct TRAF binding by CD40. Using a model system designed to directly compare the signaling potency of the cytoplasmic domains of LMP1 and CD40 in B lymphocytes, we found that LMP1 more potently activates c-Jun kinase and nuclear factor kappaB and induces higher levels of several B cell effector functions than does CD40. This suggests that LMP1 utilizes a modified CD40 signaling pathway. Failure to regulate TRAFs may contribute to the enhanced capacity of LMP1 to activate B cells as well as promote B cell transformation.


Mechanisms of Osteoclastogenesis in Orthodontic Tooth Movement and Orthodontically Induced Tooth Root Resorption.
Orthodontic tooth movement (OTM) is achieved by the simultaneous activation of bone resorption by osteoclasts and bone formation by osteoblasts. When orthodontic forces are applied, osteoclast-mediated bone resorption occurs in the alveolar bone on the compression side, creating space for tooth movement. Therefore, controlling osteoclastogenesis is the fundamental tenet of orthodontic treatment. Orthodontic forces are sensed by osteoblast lineage cells such as periodontal ligament (PDL) cells and osteocytes. Of several cytokines produced by these cells, the most important cytokine promoting osteoclastogenesis is the receptor activator of nuclear factor-κB ligand (RANKL), which is mainly supplied by osteoblasts. Additionally, osteocytes embedded within the bone matrix, T lymphocytes in inflammatory conditions, and PDL cells produce RANKL. Besides RANKL, inflammatory cytokines, such as interleukin-1, tumor necrosis factor-α, and prostaglandin E2 promote osteoclastogenesis under OTM. On the downside, excessive osteoclastogenesis activation triggers orthodontically-induced external root resorption (ERR) through pro-osteoclastic inflammatory cytokines. Therefore, understanding the mechanisms of osteoclastogenesis during OTM is essential in reducing the adverse effects of orthodontic treatment. Here, we review the current concepts of the mechanisms underlying osteoclastogenesis in OTM and orthodontically induced ERR.


A cyclophilin-regulated PP2A-like protein phosphatase in thylakoid membranes of plant chloroplasts.
Dephosphorylation of central photosynthetic proteins regulates their turnover in plant thylakoid membranes. A membrane protein phosphatase from spinach thylakoids was purified 13000-fold using detergent-engaged FPLC. The purified enzyme exhibited characteristics typical of eukaryotic Ser/Thr phosphatases of the PP2A family in that it was inhibited by okadaic acid (IC(50) = 0.4 nM) and tautomycin (IC(50) = 25 nM), irreversibly bound to microcystin-agarose, and recognized by a polyclonal antibody raised against a recombinant catalytic subunit of human PP2A. Furthermore, the anti-PP2A antibody inhibited protein dephosphorylation in isolated thylakoids. The phosphatase copurified with TLP40, a cyclophilin-like peptidyl-prolyl isomerase located in the thylakoid lumen. TLP40 could be released from the phosphatase immobilized on microcystin-agarose by high-salt treatment. Binding of cyclosporin A (CsA) to TLP40 led to thylakoid phosphatase activation, while cyclophilin substrates, prolyl-containing oligopeptides, inhibited protein dephosphorylation. This dephosphorylation could be modulated by CsA or oligopeptides only after the thylakoids had been ruptured to expose the lumenal membrane surface where the TLP40 is located. Regulation of the PP2A-like phosphatase at the outer thylakoid surface is likely to operate via reversible binding of TLP40 to the inner membrane surface. This is a first example of transmembrane regulation in which the activity of phosphatase is altered by the binding of a cyclophilin to a site other than the active one. We propose that signaling from TLP40 to the protein phosphatase coordinates dephosphorylation and protein folding, two processes required for protein turnover during the repair of photoinhibited photosystem II reaction centers.


Preexisting opioid use disorder is associated with poor outcomes in hospitalized acute pancreatitis patients.
Although opioids are widely used for pain management in acute pancreatitis, the impact of opioid use disorder (OUD) on outcomes in patients with acute pancreatitis remains unknown. In the current study, we aimed to evaluate the impact of the OUD on outcomes in patients hospitalized with acute pancreatitis and delineate the trends associated with OUD and acute pancreatitis using a nationally representative sample. This is a retrospective cohort study of patients with acute pancreatitis using the combined releases of the year 2005-2014 of the National (Nationwide) Inpatient Sample (NIS) database. Patients over the age of 18 years with a principal diagnosis of acute pancreatitis were divided into cohorts of patients with opioid use disorders and those without. The primary measured outcome was in-hospital mortality and secondary outcomes were healthcare utilization measures, including length of stay (LOS) and hospitalization costs. A total of 2 593 831 hospitalizations of acute pancreatitis were included; of which, 37 849 (1.46%) had a secondary diagnosis of OUD. Total acute pancreatitis-related hospitalizations increased from 237 882 in 2005 to 274 006 in 2014. At the same time prevalence of OUD in acute pancreatitis patients also increased from 1 to 2.1%. Patients with OUD had significantly increased mortality as compared to patients without OUD (aOR: 1.4; P < 0.001). At the same time, acute pancreatitis patients with OUD were associated with 1.3 days longer LOS as compared to other acute pancreatitis patients (P < 0.001]. The mean adjusted difference in total hospitalization costs was $2353 (P < 0.001). OUD is associated with a significant increase in LOS, healthcare utilization cost and in-hospital mortality in patients admitted for acute pancreatitis. Therefore, clinicians should exercise caution in prescribing opioid medications to this high-risk patient population and other modalities such as nonopioid pain medications should be tried as alternatives to opioid analgesics.


Different microRNA profiles reveal the diverse outcomes induced by EV71 and CA16 infection in human umbilical vein endothelial cells using high-throughput sequencing.
Enterovirus 71 (EV71) and Coxsackievirus A16 (CA16) remain the predominant pathogens in hand, foot, and mouth disease (HFMD), but the factors underlying the pathogenesis of EV71 and CA16 infections have not been elucidated. Recently, the functions of microRNAs (miRNAs) in pathogen-host interactions have been highlighted. In the present study, we performed comprehensive miRNA profiling in EV71- and CA16-infected human umbilical vein endothelial cells (HUVECs) at multiple time points using high-throughput sequencing. The results showed that 135 known miRNAs exhibited remarkable differences in expression. Of these, 30 differentially expressed miRNAs presented opposite trends in EV71- and CA16-infected samples. Subsequently, we mainly focused on the 30 key differentially expressed miRNAs through further screening to predict targets. Gene ontology (GO) and pathway analysis of the predicted targets showed the enrichment of 14 biological processes, 9 molecular functions, 8 cellular components, and 85 pathways. The regulatory networks of these miRNAs with predicted targets, GOs, pathways, and co-expression genes were determined, suggesting that miRNAs display intricate regulatory mechanisms during the infection phase. Consequently, we specifically analyzed the hierarchical GO categories of the predicted targets involved in biological adhesion. The results indicated that the distinct changes induced by EV71 and CA16 infection may be partly linked to the function of the blood-brain barrier. Taken together, this is the first report describing miRNA expression profiles in HUVECs with EV71 and CA16 infections using high-throughput sequencing. Our data provide useful insights that may help to elucidate the different host-pathogen interactions following EV71 and CA16 infection and offer novel therapeutic targets for these infections.


Restoration of sinus rhythm by the Maze procedure halts progression of tricuspid regurgitation after mitral surgery.
Tricuspid regurgitation (TR) is common in patients undergoing mitral valve surgery, and atrial fibrillation (AF) can cause progression of TR. This study examined the hypothesis that correction of AF with the Maze procedure can prevent the late progression of TR after mitral valve surgery. We compared severity of TR in 33 patients (January 1, 1993, to January 1, 2003) who underwent concomitant Maze procedure plus mitral valve surgery and converted to normal sinus rhythm postoperatively with case-matched control patients who underwent mitral valve surgery alone and remained in AF postoperative. Matched variables were age, sex, diabetes mellitus, left ventricular ejection fraction, and hypertension. Preoperative TR grade was similar between groups (Maze 2.2 +/- 0.8 versus no-Maze 2.3 +/- 0.8, p = 0.67). Patients with permanent transvenous pacemakers, organic tricuspid valve disease, and prior tricuspid valve surgery were excluded from this comparison. Before hospital dismissal, average TR grade improved to 1.9 +/- 0.9 in both groups; TR improved in 42% of patients in the Maze group and 36% of patients with preoperative AF and no Maze. At last follow-up, average TR grade remained stable at 1.9 +/- 0.9 in the Maze group (p = 0.078 versus preoperative) with TR progression in only 9% of patients (3 of 33). In contrast, TR grade worsened to 2.7 +/- 0.9 in the no-Maze group (p = 0.04 versus preoperative, p < 0.001 versus postoperative, p < 0.001 versus groups), and TR worsened in 45% of patients (15 of 33). In a multivariable model, performance of a Maze procedure was protective against the progression of TR. Continued AF after mitral valve surgery can predispose a patient to progression of TR, and this progression is prevented in patients having successful concomitant Maze procedure.


ChIP-chip designs to interrogate the genome of Xenopus embryos for transcription factor binding and epigenetic regulation.
Chromatin immunoprecipitation combined with genome tile path microarrays or deep sequencing can be used to study genome-wide epigenetic profiles and the transcription factor binding repertoire. Although well studied in a variety of cell lines, these genome-wide profiles have so far been little explored in vertebrate embryos. Here we report on two genome tile path ChIP-chip designs for interrogating the Xenopus tropicalis genome. In particular, a whole-genome microarray design was used to identify active promoters by close proximity to histone H3 lysine 4 trimethylation. A second microarray design features these experimentally derived promoter regions in addition to currently annotated 5' ends of genes. These regions truly represent promoters as shown by binding of TBP, a key transcription initiation factor. A whole-genome and a promoter tile path microarray design was developed. Both designs can be used to study epigenetic phenomena and transcription factor binding in developing Xenopus embryos.


Negative impacts of ultraviolet-A radiation on antioxidant and oxidative stress biomarkers of African catfish Clarias gariepinus.
The present study was carried out to evaluate the ultraviolet-A (UVA) effects on biochemical, oxidative stress and antioxidant changes using aquatic species. The destructive effects of ultraviolet-A radiation on the African Catfish, Clarias gariepinus was revealed in terms of the carbonyl protein (CP), lipid peroxidation (LPO), DNA damage, super oxide dismutase (SOD), catalase (CAT), glutathione peroxidase (Gpx), glutathione reductase (GR), glucose-6-phosphate dehydrogenase (G6PDH), lactate dehydrogenase (LDH), glutathione (GSH) and total antioxidant (TAO) in the gills, kidney, liver, muscles and skin. Also, glucose, total lipid, total protein and cortisol content of fish serums was detected. These parameters can be used as biomarkers to identify the negative effects of UVA radiation (20, 60 and 180 min per day) for 3 days in different tissues of Clarias gariepinus. There was a significant decrease in the activity of SOD, CAT, Gpx, GSH, G6PDH, LDH, and TAO in all of the examined tissues. The pattern of GR activity in UVA exposed groups showed no significant differences compared with the control group. However, CP, LPO and DNA damage were increased significantly with exposure periods in all of the examined tissues. The exposure to different doses of UVA caused hypoglycaemia, hypolipidimia and hypoproteinimia. Cortisol levels showed a significant increase after UV exposure when compared with the control group. In conclusion, UVA exposure with different time periods has been shown to have negative effects on the blood biochemistry, hormonal and antioxidant capacity of Clarias gariepinus tissues.


Comparison of standard CPR versus diffuse and stacked hand position interposed abdominal compression-CPR in a swine model.
Interposed abdominal compression cardiopulmonary resuscitation (IAC-CPR) is an innovative basic life support technique requiring no mechanical adjuncts. Optimizing its performance remains a challenge. Hand-position technique over the abdomen during interposed abdominal compression (IAC) may be important. The purpose of this study was to determine if there is a difference in efficacy depending on the type of abdominal hand-position used. Two different hand positions were studied: open hands, placed side by side, resulting in diffuse abdominal compression and stacked hands, with one on top of the other, producing a more focal compression of the abdomen. Thirty swine were cannulated with micromanometer-tipped pressure transducers in the ascending aorta (Ao) and right atrium (RA), and Millar Doppler-tipped catheters in the descending aorta and inferior vena cava (IVC) to determine flow patterns during cardiopulmonary resuscitation (CPR), During CPR there were no differences in aortic systolic or right atrial systolic pressures. Both forms of IAC-CPR produced greater aortic diastolic and right atrial diastolic pressures then standard CPR (STD-CPR) (P<0.05). Coronary perfusion pressures (CPP), however, were not different. Blood flow directions and velocity patterns showed that STD-CPR chest compressions produce caudally directed blood flow in both the descending aorta and the IVC, and that such flows reverse (becoming cranially directed) during the relaxation phase of chest compression. IAC-CPR produced similar blood flow patterns in the aorta and IVC, as seen with STD-CPR. There were no differences in blood flow patterns between the different forms of IAC-CPR. No CPR-produced trauma difference was found. Abdominal hand position (diffuse or stacked) did not affect blood flow in either the aorta or IVC or resuscitation success in this experimental model. There was a trend towards better outcomes with stacked hands IAC-CPR with 90 versus 70% survival with STD-CPR.


Tolerance to proton irradiation in the eutardigrade Richtersius coronifer--a nuclear microprobe study.
The tardigrade Richtersius coronifer has previously been shown to tolerate very high doses of low linear energy transfer (low-LET) radiation (gamma rays). The purpose of this study was to extend our knowledge on radiation tolerance in this species by investigating the dose-response to high-LET radiation in terms of protons. Dehydrated tardigrades of the species R. coronifer were irradiated with 2.55 MeV (megaelectronvolts) protons at doses ranging from 500 gray (Gy) to 15,000 Gy, to investigate the dose-viability relationship. In addition, a focused proton microbeam was utilised to determine the areal mass distribution, using the ion beam analytical technique STIM (Scanning Transmission Ion Microscopy). The experiment suggests that R. coronifer is unaffected by doses of proton irradiation up to 10,000 Gy, but shows very little viability at higher doses. The STIM analysis revealed that the thickness of the dehydrated tardigrades exceeds 150 microm, and that a fraction of the protons may not be fully absorbed. Our results are in line with previous studies of exposure to high-LET radiation in tardigrades, indicating that these animals are equally or even more tolerant to high-LET compared to low-LET gamma radiation. The physiological background to this remarkable result is currently unknown, but deserves investigation.


Pathogenesis of experimental shock. II. Absence of endotoxic activity in blood of rabbits subjected to graded hemorrhage.
A series of biological test reactions was used in order to establish the presence of bacterial endotoxins in the blood of rabbits during the progression of hemorrhagic shock. 1. When the shocked animal was used as the test object, it was not possible to induce either the generalized Shwartzman reaction or the dermal Shwartzman phenomenon with exogenous endotoxin (S. enteritidis or E. coli) as one of the two provocative factors. 2. Epinephrine instilled into the skin of rabbits either before, during, or after an episode of hemorrhagic shock did not result in the hemorrhagic skin reaction which occurs in the presence of as little as 1 microg of endotoxin intravenously. 3. Passive transfer from a donor in the irreversible phase of shock of 20 to 25 ml of blood into a primed recipient (B.P. at 40 mm Hg for 1 hour) was uniformly lethal. 4. Similar amounts of blood from such shocked donor failed upon intravenous injection to elicit a protective hemorrhagic reaction in skin sites which were infiltrated with 100 microg of epinephrine. In the same animals 1 microg of endotoxin added to the blood samples caused a positive dermal response. 5. Blood was taken from rabbits which had been pretreated with S. enteritidis endotoxin and then subjected to hemorrhagic shock (35 mm Hg for 2 hours). Such samples upon passive transfer produced positive skin reactions in epinephrine sites but were not lethal to the primed test recipient used in these studies. It is concluded that the contribution of bacterial endotoxemia to the genesis of hemorrhagic shock remains to be determined.


Uterine artery embolization in the treatment of symptomatic uterine fibroid tumors (EMMY trial): periprocedural results and complications.
Uterine artery embolization (UAE) is an emerging treatment for symptomatic uterine fibroid tumors. This study was performed to evaluate the periprocedural results of the UAE procedure and identify risk factors for technical failure, fever after UAE, pain, and other complications. As part of a multicenter, randomized trial to compare UAE versus hysterectomy in patients with symptomatic uterine fibroid tumors, 81 patients underwent UAE. Univariate and multivariate analyses were used to identify predictors for technical failure, postprocedural fever, complications as defined by the Society of Interventional Radiology (SIR), and pain scores. The technical failure rate according to SIR guidelines was 5.3% (95% CI, 2.3%-10.1%). The procedural failure rate was 17.3% (95% CI, 9.8%-27.3%). Bilateral failure occurred in four of 81 patients and unilateral failure occurred in 10 of 81 patients. Technical failure occurred mainly as a result of difficult anatomy (3.7%) or absence of the uterine artery (3.1%). The overall complication rates were 28.4% during the patients' hospital stay and 60.5% for the 6 weeks after discharge. The risk of technical failure was found to increase in the presence of a single fibroid tumor (odds ratio [OR], 6.21; 95% CI, 1.65-23.41; P = .007) and/or a small uterine volume (<500 cm(3); OR, 10.8; 95% CI, 1.25-93.36; P = .03). The amount of embolization material was associated with the onset of fever after UAE (OR, 2.05; 95% CI, 1.09-3.87; P = .027), major complications (OR, 5.68; 95% CI, 2.05-15.75; P = .001), and high pain scores (OR, 1.97; 95% CI, 1.08-3.58; P = .027). The procedural failure rate for UAE was higher than those reported by others, mainly as a result of difficult anatomy and absence of a uterine artery in some cases. The risk of procedural failure was increased for patients with single fibroid tumors and/or small uterine volumes. A clear dose-effect response was revealed between the amount of embolization material used and the risk for postprocedural fever, major complications, and severe pain.


Comparative efficacy and safety of tacrolimus, mycophenolate mofetil, azathioprine, and cyclophosphamide as maintenance therapy for lupus nephritis : A Bayesian network meta-analysis of randomized controlled trials.
This study aimed to assess the relative efficacy and safety of tacrolimus, mycophenolate mofetil (MMF), azathioprine (AZA), and cyclophosphamide (CYC) as maintenance therapy for lupus nephritis. Randomized controlled trials (RCTs) examining the efficacy and safety of tacrolimus, MMF, AZA, and CYC for maintenance therapy in lupus nephritis patients were included. We performed a Bayesian random-effects network meta-analysis to combine direct and indirect evidence from the RCTs. Five RCTs including 525 patients were included. Although the difference was not statistically significant, tacrolimus showed a trend toward a lower renal relapse rate than AZA or CYC. Similarly, MMF showed a trend toward a lower relapse rate than AZA or CYC. Renal relapse tended to be lower in the AZA group than in the CYC group. Ranking probability based on the surface under the cumulative ranking curve (SUCRA) indicated that tacrolimus had the highest probability of being the best treatment based on the renal relapse, followed by MMF, AZA, and CYC. Analysis of withdrawal due to adverse events showed the same pattern. The leukopenia incidence was significantly lower in the MMF group than in the AZA group. Similarly, it tended to be lower in the tacrolimus group than in the AZA group. Ranking probability based on SUCRA indicated that MMF had the highest probability of being the safest treatment based on leukopenia incidence, followed by tacrolimus and AZA. Lower renal relapse rates combined with a more favorable safety profile suggest that tacrolimus and MMF are superior to AZA and CYC as maintenance treatments in these patients.


Linking species habitat and past palaeoclimatic events to evolution of the teleost innate immune system.
Host-intrinsic factors as well as environmental changes are known to be strong evolutionary drivers defining the genetic foundation of immunity. Using a novel set of teleost genomes and a time-calibrated phylogeny, we here investigate the family of Toll-like receptor (TLR) genes and address the underlying evolutionary processes shaping the diversity of the first-line defence. Our findings reveal remarkable flexibility within the evolutionary design of teleost innate immunity characterized by prominent TLR gene losses and expansions. In the order of Gadiformes, expansions correlate with the loss of major histocompatibility complex class II (MHCII) and diversifying selection analyses support that this has fostered new immunological innovations in TLRs within this lineage. In teleosts overall, TLRs expansions correlate with species latitudinal distributions and maximum depth. By contrast, lineage-specific gene losses overlap with well-described changes in palaeoclimate (global ocean anoxia) and past Atlantic Ocean geography. In conclusion, we suggest that the evolvability of the teleost immune system has most likely played a prominent role in the survival and successful radiation of this lineage.


A primal role for the vestibular sense in the development of coordinated locomotion.
Mature locomotion requires that animal nervous systems coordinate distinct groups of muscles. The pressures that guide the development of coordination are not well understood. To understand how and why coordination might emerge, we measured the kinematics of spontaneous vertical locomotion across early development in zebrafish (Danio rerio) . We found that zebrafish used their pectoral fins and bodies synergistically during upwards swims. As larvae developed, they changed the way they coordinated fin and body movements, allowing them to climb with increasingly stable postures. This fin-body synergy was absent in vestibular mutants, suggesting sensed imbalance promotes coordinated movements. Similarly, synergies were systematically altered following cerebellar lesions, identifying a neural substrate regulating fin-body coordination. Together these findings link the vestibular sense to the maturation of coordinated locomotion. Developing zebrafish improve postural stability by changing fin-body coordination. We therefore propose that the development of coordinated locomotion is regulated by vestibular sensation.


The constant region contributes to the antigenic specificity and renal pathogenicity of murine anti-DNA antibodies.
Affinity for DNA and cross-reactivity with renal antigens are associated with enhanced renal pathogenicity of lupus autoantibodies. In addition, certain IgG subclasses are enriched in nephritic kidneys, suggesting that isotype may determine the outcome of antibody binding to renal antigens. To investigate if the isotype of DNA antibodies affects renal pathogenicity by influencing antigen binding, we derived IgM, IgG1, IgG2b and IgG2a forms of the PL9-11 antibody (IgG3 anti-DNA) by in vitro class switching or PCR cloning. The affinity and specificity of PL9-11 antibodies for nuclear and renal antigens were analyzed using ELISA, Western blotting, surface plasmon resonance (SPR), binding to mesangial cells, and glomerular proteome arrays. Renal deposition and pathogenicity were assayed in mice injected with PL9-11 hybridomas. We found that PL9-11 and its isotype-switched variants had differential binding to DNA and chromatin (IgG3>IgG2a>IgG1>IgG2b>IgM) by direct and competition ELISA, and SPR. In contrast, in binding to laminin and collagen IV the IgG2a isotype actually had the highest affinity. Differences in affinity of PL9-11 antibodies for renal antigens were mirrored in analysis of specificity for glomeruli, and were associated with significant differences in renal pathogenicity in vivo and survival. Our novel findings indicate that the constant region plays an important role in the nephritogenicity of antibodies to DNA by affecting immunoglobulin affinity and specificity. Increased binding to multiple glomerular and/or nuclear antigens may contribute to the renal pathogenicity of anti-DNA antibodies of the IgG2a and IgG3 isotype. Finally, class switch recombination may be another mechanism by which B cell autoreactivity is generated.


A detoxifying oxygen reductase in the anaerobic protozoan Entamoeba histolytica.
We report the characterization of a bacterial-type oxygen reductase abundant in the cytoplasm of the anaerobic protozoan parasite Entamoeba histolytica. Upon host infection, E. histolytica is confronted with various oxygen tensions in the host intestine, as well as increased reactive oxygen and nitrogen species at the site of local tissue inflammation. Resistance to oxygen-derived stress thus plays an important role in the pathogenic potential of E. histolytica. The genome of E. histolytica has four genes that encode flavodiiron proteins, which are bacterial-type oxygen or nitric oxide reductases and were likely acquired by lateral gene transfer from prokaryotes. The EhFdp1 gene has higher expression in virulent than in nonvirulent Entamoeba strains and species, hinting that the response to oxidative stress may be one correlate of virulence potential. We demonstrate that EhFdp1 is abundantly expressed in the cytoplasm of E. histolytica and that the protein levels are markedly increased (up to ~5-fold) upon oxygen exposure. Additionally, we produced fully functional recombinant EhFdp1 and demonstrated that this enzyme is a specific and robust oxygen reductase but has poor nitric oxide reductase activity. This observation represents a new mechanism of oxygen resistance in the anaerobic protozoan pathogen E. histolytica.


Multicomponent synthesis of 4-aminophthalazin-1(2H)-ones by palladium-catalyzed isocyanide insertion.
4-Aminophthalazin-1(2H)-ones (APOs) are underexplored heterocyclic compounds with promising and diverse biological activities. The classical synthesis of these compounds is tedious and does not allow the regioselective introduction of substituents. Here, we present our full studies on the Pd-catalyzed cross-coupling of substituted o-(pseudo)halobenzoates and hydrazines with isocyanide insertion allowing straightforward access to diversely substituted APOs. We illustrate the advantages of this method compared to other approaches and describe solutions for the limitations we encountered. In addition, we have developed efficient diversifications of this heterocyclic scaffold that allow access to more diverse APOs as well as novel heterocyclic scaffolds.


Adhesion to zirconia used for dental restorations: a systematic review and meta-analysis.
Currently, no consensus exists regarding the best adhesion protocol for zirconia used in dentistry; this is important particularly for restorations where mechanical retention is deficient. This systematic review analyzed the adhesion potential of resin-based and glass-ionomer luting cements to zirconia and aimed to highlight the possible dominant factors affecting the bond strength results to this substrate. Original scientific papers on adhesion to zirconia published in the MEDLINE (PubMed) database between 01/01/1995 and 01/06/2011 were included in this systematic review. The following MeSH terms, search terms, and their combinations were used: "Dental bonding", "Zirconium", "Zirconia", "Y-TZP", "Y-TZP ceramic", "Materials Testing/methods", "Test", "Cement", and "Resin bonding". Two reviewers performed screening and data abstraction. Descriptive statistics were performed and the frequencies of the studied parameters, means, standard deviations, confidence intervals (95% CI; uncorrected and corrected), median values, and interquartile ranges (IQR) were calculated for the bond strength data reported for different factor levels: surface conditioning methods (control, physicochemical, physical, chemical), cements (bis-GMA-, MDP-, and 4-META-based resin cements, self-adhesive cements, glass ionomer), aging with and without thermocycling (TC), and test methods (macroshear, microshear, macrotensile, and microtensile). The final search provided 177 titles with abstracts. Further abstract screening yielded 72 articles, out of which 54 were found potentially appropriate to be included. After full text evaluation, 2 of these were eliminated. The selection process resulted in the final sample of 52 studies. In total, 169 different surface conditioning methods, mainly combinations of air-abrasion protocols and adhesive promoters (primers or silanes), were investigated. Altogether, the use of 5 types of cements and 4 testing methods was reported. While 26 studies were performed without TC as aging, 26 of them employed thermocycling at varying number of cycles. This review highlighted that adhesion of the luting cements is significantly influenced by the surface conditioning method (p = 0.044), cement type (p = 0.018), test method (p = 0.017) and aging condition (p = 0.003). In nonconditioned control groups without thermocycling, mean bond strength values ranged between 1.15 (IQR = 3.54) and 8.93 (IQR = 9), and 6.9 (IQR = 0) and 8.73 (IQR = 13.93) MPa for macroshear and macrotensile tests, respectively. After physical conditioning method, MDP monomer based cement presented the highest bond values compared to those of other resin cements using either the macrotensile (no TC: 34.2; IQR = 24.18 MPa, TC: 42.35; IQR = 0 MPa) or microtensile (no TC: 37.2; IQR = 41.5 MPa, TC: 17.1; IQR = 31.15 MPa) test method. Based on the results of this systematic review, increased adhesion could be expected after physicochemical conditioning of zirconia. MDP-based resin cements tend to present higher results than those of other cements types when tested using macro- and microtensile tests. Adhesion studies on zirconia and reporting of data require more standardization.


Mice carrying a schizophrenia-associated mutation of the Arhgap10 gene are vulnerable to the effects of methamphetamine treatment on cognitive function: association with morphological abnormalities in striatal neurons.
We recently found a significant association between exonic copy-number variations in the Rho GTPase activating protein 10 (Arhgap10) gene and schizophrenia in Japanese patients. Special attention was paid to one patient carrying a missense variant (p.S490P) in exon 17, which overlapped with an exonic deletion in the other allele. Accordingly, we generated a mouse model (Arhgap10 S490P/NHEJ mice) carrying a missense variant and a coexisting frameshift mutation. We examined the spatiotemporal expression of Arhgap10 mRNA in the brain and found the highest expression levels in the cerebellum, striatum, and nucleus accumbens (NAc), followed by the frontal cortex in adolescent mice. The expression levels of phosphorylated myosin phosphatase-targeting subunit 1 and phosphorylated p21-activated kinases in the striatum and NAc were significantly increased in Arhgap10 S490P/NHEJ mice compared with wild-type littermates. Arhgap10 S490P/NHEJ mice exhibited a significant increase in neuronal complexity and spine density in the striatum and NAc. There was no difference in touchscreen-based visual discrimination learning between Arhgap10 S490P/NHEJ and wild-type mice, but a significant impairment of visual discrimination was evident in Arhgap10 S490P/NHEJ mice but not wild-type mice when they were treated with methamphetamine. The number of c-Fos-positive cells was significantly increased after methamphetamine treatment in the dorsomedial striatum and NAc core of Arhgap10 S490P/NHEJ mice. Taken together, these results suggested that schizophrenia-associated Arhgap10 gene mutations result in morphological abnormality of neurons in the striatum and NAc, which may be associated with vulnerability of cognition to methamphetamine treatment.


Pharmacological Profile of Viltolarsen for the Treatment of Duchenne Muscular Dystrophy: A Japanese Experience.
Duchenne muscular dystrophy (DMD) is a fatal, X-linked recessive disorder characterized by progressive muscle loss and cardiorespiratory complications. Mutations in the DMD gene that eliminate the production of dystrophin protein are the underlying causes of DMD. Viltolarsen is a drug of phosphorodiamidate morpholino oligomer (PMO) chemistry, designed to skip exon 53 of the DMD gene. It aims to produce truncated but partially functional dystrophin in DMD patients and restore muscle function. Based on a preclinical study showing the ability of antisense PMOs targeting the DMD gene to improve muscle function in a large animal model, viltolarsen was developed by Nippon Shinyaku and the National Center of Neurology and Psychiatry in Japan. Following clinical trials conducted in Japan, Canada, and the United States showing significant improvements in muscle function, viltolarsen was approved for medical use in Japan in March 2020 and the United States in August 2020, respectively. Viltolarsen is a mutation-specific drug and will work for 8% of the persons with DMD who carry mutations amenable to exon 53 skipping. This review summarizes the pharmacological profile of viltolarsen, important clinical trials, and challenges, focusing on the contribution of Japanese patients and researchers in its development.


[Gamma knife radiosurgery for large trigeminal schwannomas].
To evaluate the role of gamma knife (GK) radiosurgery in controlling large trigeminal schwannoma. The clinical data of 30 patients suffering from large trigeminal schwannoma with a maximum diameter of 39.0 mm (31.0 to 53.0 mm), who underwent GK radiosurgery, used as the primary treatment modality in 20 patients and used to cure the residuals of tumor after microsurgery in 10 patients, with the maximum irradiation dose to tumor of 25.8 Gy (20.0 to 33.0 Gy) and a tumor margin dose of 12.2 Gy (9.0 to 14.0 Gy), were analyzed respectively. The 30 patients had been followed up for 78 months (24 to 136 months). Twenty-nine of the 30 patients were alive, and one patient with malignant trigeminal schwannoma died of tumor progression 36 months after the GK radiosurgery. Neurological deficits were improved in 24 patients and 3 patients' condition remained stable. Two patients' symptoms worsened 5 and 30 months after the GK radiosurgery respectively because of tumor swelling and tumor cyst enlarging. Then these two patients underwent microsurgery and their tumors were subtotally removed. Disappearance of tumor occurred in 4 patients, a marked decrease in tumor volume was observed in 20 patients, three tumors kept unchanged or slightly regressed in volume, and 3 tumors, including that of the patient who died, progressed. The tumor-growth control rate was 90% (27/30). Two patients underwent craniotomy after GK radiosurgery and one of these 2 resected specimens underwent histopathological examination. The histopathological findings of this tumor resected 30 months after GK radiosurgery revealed that most of the tumor tissue necrosed and was full of microcysts, the residual tumor cells in peripheral areas underwent hyaline degeneration, and the nuclei showed pyknosis. GK may be a good alternative treatment for selective patients suffering from large trigeminal schwannoma with a maximum diameter <or= 40 mm.


Triiodothyronine increases myocardial function and pyruvate entry into the citric acid cycle after reperfusion in a model of infant cardiopulmonary bypass.
Triiodothyronine (T3) supplementation improves clinical outcomes in infants after cardiac surgery using cardiopulmonary bypass by unknown mechanisms. We utilized a translational model of infant cardiopulmonary bypass to test the hypothesis that T3 modulates pyruvate entry into the citric acid cycle (CAC), thereby providing the energy support for improved cardiac function after ischemia-reperfusion (I/R). Neonatal piglets received intracoronary [2-(13)Carbon((13)C)]pyruvate for 40 min (8 mM) during control aerobic conditions (control) or immediately after reperfusion (I/R) from global hypothermic ischemia. A third group (I/R-Tr) received T3 (1.2 μg/kg) during reperfusion. We assessed absolute CAC intermediate levels and flux parameters into the CAC through oxidative pyruvate decarboxylation (PDC) and anaplerotic carboxylation (PC) using [2-(13)C]pyruvate and isotopomer analysis by gas and liquid chromatography-mass spectrometry and (13)C-nuclear magnetic resonance spectroscopy. When compared with I/R, T3 (group I/R-Tr) increased cardiac power and oxygen consumption after I/R while elevating flux of both PDC and PC (∼4-fold). Although neither I/R nor I/R-Tr modified absolute CAC levels, T3 inhibited I/R-induced reductions in their molar percent enrichment. Furthermore, (13)C-labeling of CAC intermediates suggests that T3 may decrease entry of unlabeled carbons at the level of oxaloacetate through anaplerosis or exchange reaction with asparate. T3 markedly enhances PC and PDC fluxes, thereby providing potential substrate for elevated cardiac function after reperfusion. This T3-induced increase in pyruvate fluxes occurs with preservation of the CAC intermediate pool. Our labeling data raise the possibility that T3 reduces reliance on amino acids for anaplerosis after reperfusion.


Effect of finishing and polishing procedures on the gap width between a denture base resin and two long-term, resilient denture liners.
The junction between a long-term, resilient denture liner and the denture base is difficult to finish and polish due to differences in the hardness of the materials. Gaps tend to form during finishing and polishing procedures. This study measured the junctional gap between 2 long-term, resilient denture liners and a denture base material after different finishing and polishing procedures were performed. The surface smoothness of the 2 liner materials also was evaluated. Molloplast-B and an experimental, heat-polymerized methyl siloxane-resin-based denture liner were processed (according to the manufacturer's instructions) against Lucitone 199 acrylic resin. Control specimens (n = 20) were 2 mm thick and flat. Experimental specimens (n = 64) were fabricated with a raised center section: a 3- x 5- x 15-mm half-cylindrical ridge with a junction at the top of the cylinder between the liner material and acrylic resin. The specimens were finished with 1 of 6 types of burs and polished with different combinations of rubber-impregnated acrylic polishers, pumice, and tin oxide. The finished specimens were examined and photographed with a scanning electron microscope, and the largest gap along the liner/denture base junction on each specimen was measured. Three-way analysis of variance without replication (P<.05) and post-hoc t tests were used to analyze the data and compare groups. RESULTS; Averaged across finishing and polishing techniques, a larger gap was recorded for the experimental liner material (22 microm) than for Molloplast-B (14 microm) (P<.00005). Qualitative evaluation suggested that the experimental liner material polished better than Molloplast-B. The smoothest surfaces were obtained when specimens were finished with fine-tooth cross-cut carbide burs and polished with both pumice and tin oxide. A comparison between polished and unpolished specimens, averaged over materials and finishing techniques, revealed that polishing reduced gap size (P=.015). Within the limitations of this study, the 2 denture liners tested behaved similarly. The results suggest that the size of the gap at the liner/denture base junction may be affected by finishing and polishing techniques and vary among materials.


The impact of the parent-physician relationship on parental vaccine safety perceptions.
In times of declining immunization rates among children in many countries and an increasing threat of potentially vaccine-preventable diseases, there is a strong need for new strategies to improve trust in vaccinations and acceptance of recommendations, especially in parents of infants and children. A survey to evaluate vaccination acceptance has been conducted in Vienna, Austria, based on a US CDC survey, applying a cross-sectional approach with districts and public as well as private kindergartens and preschools as selection base. The survey aimed to investigate the impact of parent satisfaction with, and overall trust in the physician on vaccine acceptance, as well as the impact of quality and completeness of safety information delivered during the vaccination consultation. Overall 1101 parents, predominantly (84.2%) mothers, participated in the survey. The majority (82.7%) of participants had a generally positive view concerning childhood vaccination. However, 25.1% refused at least one of the recommended vaccinations. In multivariate analysis, confidence in vaccinations was significantly influenced by education (lower confidence at higher levels of education), gender (higher confidence in females), and positively associated with trust in physician, smooth vaccination procedure, and information about vaccine risks. Similar results were obtained for compliance with recommended vaccinations with information about vaccine benefits being the most important predictor. This large survey indicates an important role of the physician in communicating balanced information about benefits and risks associated with childhood vaccinations. A trustworthy parent-physician relationship is crucial for vaccination decisions of parents.


Endocarditis presenting as acute abdomen.
Infective endocarditis remains an interesting and challenging disease in which the presenting features have been modified by medical progress. We report a case of a 63-year-old woman who presented with pyrexia, weight loss, night sweats and fatigue over a period of 7 weeks, at the end of which she developed severe right hypochondrial pain, mimicking acute abdomen. She was provisionally diagnosed with and treated for acute cholecystitis to no avail. An abdominal computed tomography scan was performed and revealed the presence of right kidney infarction, the source of which was later proven to originate from aortic valve endocarditis. The patient made a remarkable recovery following 6 weeks of treatment with intravenous antibiotics. This case demonstrates that endocarditis can present as an acute abdomen which is caused mainly by embolism in various organs (the kidney in our patient).


Arizona hinshawii infection of an atherosclerotic abdominal aorta.
A patient with Arizonae hinshawii infection of an atherosclerotic abdominal aortic aneurysm is described to emphasize the similarity of the nonenteric infections of man by Arizona and Salmonella organisms. These genera are similar bacteriologically, ecologically, and clinically. This case emphasizes the need for thoroughly considering the differential diagnosis when encountering cryptic bacteremia with either organism.


Two Synchronous Neonatal Tumors: An Extremely Rare Case.
We report a case of a newborn with two synchronous tumors-sialoblastoma and hepatoblastoma-diagnosed at 20 weeks of gestation by magnetic resonance imaging (MRI) and ultrasonography (US). The aim of this study was to describe the management of this case together with a review of the literature. Our patient had a large facial tumor associated with extremely high alpha-fetoprotein levels. Diagnosis of the tumors was made by surgical biopsy, showing typical features in both. Sialoblastoma is a potentially aggressive tumor. In our case, the Ki67 index in the sialoblastoma was between 20 and 30%, indicating a possibly unfavorable behavior. The infant underwent surgery and chemotherapy in different steps. Complete surgical resection with clean margins is considered to be the best treatment option for sialoblastoma. Only four similar cases were previously reported. Timely management by a multidisciplinary team is essential in these difficult cases. In our patient, outcome was good at the time of this report.


Systematic assessment of process analytical technologies for biologics.
The application of process analytical technology (PAT) for biotherapeutic development and manufacturing has been employed owing to technological, economic, and regulatory advantages across the industry. Typically, chromatographic, spectroscopic, and/or mass spectrometric sensors are integrated into upstream and downstream unit operations in in-line, on-line, or at-line fashion to enable real-time monitoring and control of the process. Despite the widespread utility of PAT technologies at various unit operations of the bioprocess, a holistic business value assessment of PAT has not been well addressed in biologics. Thus, in this study, we evaluated PAT technologies based on predefined criteria for their technological attributes such as enablement of better process understanding, control, and high-throughput capabilities; as well as for business attributes such as simplicity of implementation, lead time, and cost reduction. The study involved an industry-wide survey, where input from subject matter industry experts on various PAT tools were collected, assessed, and ranked. The survey results demonstrated on-line liquid Chromatography (LC), in-line Raman, and gas analysis techniques are of high business value especially at the production bioreactor unit operation of upstream processing. In-line variable path-length UV/VIS measurements (VPE), on-line LC, multiangle light scattering (MALS), and automated sampling are of high business value in Protein A purification and polishing steps of the downstream process. We also provide insights, based on our experience in clinical and commercial manufacturing of biologics, into the development and implementation of some of the PAT tools. The results presented in this study are intended to be helpful for the current practitioners of PAT as well as those new to the field to gauge, prioritize and steer their projects for success.


A systematic review of menopausal symptom management decision aid trials.
To systematically review the literature regarding the effects of menopausal symptom management decision aids. Using pre-designated inclusion and exclusion criteria, relevant articles were located using the PubMed.gov online search engine and reviewing reference lists of relevant articles. Full-text, English-language, peer-reviewed articles relevant to testing decision aids in uncontrolled trials (UCT) and randomized controlled trials (RCT) were reviewed. The 18 articles represented 15 trials focused on natural health products decision aids (1 UCT, 1 RCT) or hormone therapy decision aids (1 UCT, 12 RCT). Whereas the natural health products aid was intended for women deciding about menopausal symptom management strategies, decision aids for hormone therapy were intended for a broader group of menopausal women and included indications for symptom management, prevention of heart disease, and prevention of osteoporosis. Many trials occurred prior to two pivotal events: the 2002 announcement of the Women's Health Initiative findings and the 2006 publication of the International Patient Decision Aids Standards. Study limitations may help explain contradictory findings for outcomes such as decisional conflict, decisional confidence, decisional satisfaction, knowledge and values, and decisions. There is a relatively scant contemporary literature related to menopausal symptom management decision aids. Additional methodologically sound studies are needed to develop and subsequently test decision aids that are based on (a) contemporary knowledge regarding the wide array of available therapies and (b) international standards for decision aids that include consideration of women's values and preferences.


CBFB-MYH11 fusion neoantigen enables T cell recognition and killing of acute myeloid leukemia.
Proteins created from recurrent fusion genes like CBFB-MYH11 are prevalent in acute myeloid leukemia (AML), often necessary for leukemogenesis, persistent throughout the disease course, and highly leukemia specific, making them attractive neoantigen targets for immunotherapy. A nonameric peptide derived from a prevalent CBFB-MYH11 fusion protein was found to be immunogenic in HLA-B*40:01+ donors. High-avidity CD8+ T cell clones isolated from healthy donors killed CBFB-MYH11+ HLA-B*40:01+ AML cell lines and primary human AML samples in vitro. CBFB-MYH11-specific T cells also controlled CBFB-MYH11+ HLA-B*40:01+ AML in vivo in a patient-derived murine xenograft model. High-avidity CBFB-MYH11 epitope-specific T cell receptors (TCRs) transduced into CD8+ T cells conferred antileukemic activity in vitro. Our data indicate that the CBFB-MYH11 fusion neoantigen is naturally presented on AML blasts and enables T cell recognition and killing of AML. We provide proof of principle for immunologically targeting AML-initiating fusions and demonstrate that targeting neoantigens has clinical relevance even in low-mutational frequency cancers like fusion-driven AML. This work also represents a first critical step toward the development of TCR T cell immunotherapy targeting fusion gene-driven AML.


Shearing interferometer using the grating as the beam splitter.
The theoretical interpretation of the shearing interferometer based on the moiré method using the fourier image of the grating is described. To obtain a pattern with good contrast, the observing plane must coincide with the normal fourier image plane of the grating or with the reversed fourier image plane. The information obtained by this method is the first partial derivative and under certain conditions the second partial derivative of the distortion from the reference wavefront, which is planar or spherical. Applications to measurement of the phase gradient and lens aberration are shown.


A role for NMDA receptors in posthypoxic frequency decline in the rat.
Posthypoxic frequency decline (PHFD) refers to the undershoot in respiratory frequency that follows brief hypoxic exposures. Lateral pontine neurons are required for PHFD. The neurotransmitters involved in the circuit that activate and/or are released by these pontine neurons regulating PHFD are unknown. We hypothesized that N-methyl-D-aspartate (NMDA) receptors are required for PHFD, because of the similarity in respiratory pattern after blocking lateral pontine activity or NMDA receptors. Furthermore, we hypothesized that the location of these NMDA receptors could be visualized by optimizing binding affinity with spermidine. In vagotomized, anesthetized rats (n = 16), cardiorespiratory responses to hypoxia (8% O2, 30-90 s) were recorded before and after dizocilpine (10 microg-1 mg/kg iv), and NMDA receptors were mapped with [3H]dizocilpine (n = 6). Dizocilpine elicited a dose-related effect on PHFD, blocking PHFD at high doses. Resting arterial blood pressure and breathing frequency decreased with high doses of dizocilpine, but the respiratory response to hypoxia remained intact. Our novel anatomical data indicate that NMDA receptors were widespread but distributed differentially in the brain stem. We conclude that NMDA receptors are located in pontine and medullary respiratory-related regions and that PHFD requires NMDA-receptor activation.


Priority setting for new systematic reviews: processes and lessons learned in three regions in Africa.
Priority setting to identify topical and context relevant questions for systematic reviews involves an explicit, iterative and inclusive process. In resource-constrained settings of low-income and middle-income countries, priority setting for health related research activities ensures efficient use of resources. In this paper, we critically reflect on the approaches and specific processes adopted across three regions of Africa, present some of the outcomes and share the lessons learnt while carrying out these activities. Priority setting for new systematic reviews was conducted between 2016 and 2018 across three regions in Africa. Different approaches were used: Multimodal approach (Central Africa), Modified Delphi approach (West Africa) and Multilevel stakeholder discussion (Southern-Eastern Africa). Several questions that can feed into systematic reviews have emerged from these activities. We have learnt that collaborative subregional efforts using an integrative approach can effectively lead to the identification of region specific priorities. Systematic review workshops including discussion about the role and value of reviews to inform policy and research agendas were a useful part of the engagements. This may also enable relevant stakeholders to contribute towards the priority setting process in meaningful ways. However, certain shared challenges were identified, including that emerging priorities may be overlooked due to differences in burden of disease data and differences in language can hinder effective participation by stakeholders. We found that face-to-face contact is crucial for success and follow-up engagement with stakeholders is critical in driving acceptance of the findings and planning future progress.


Antenatal Hypoxia Accelerates the Onset of Alzheimer's Disease Pathology in 5xFAD Mouse Model.
Alzheimer's disease (AD) is a chronic neurodegenerative disorder associated with cognitive impairment and later dementia among the elderly. Mounting evidence shows that adverse maternal environments during the fetal development increase the risk of diseases later in life including neurological disorders, and suggests an early origin in the development of AD-related dementia (ADRD) in utero. In the present study, we investigated the impact of antenatal hypoxia and fetal stress on the initiation of AD-related pathology in offspring of 5xFAD mice. We showed that fetal hypoxia significantly reduced brain and body weight in the fetal and the early postnatal period, which recovered in young adult mice. Using spontaneous Y-maze, novel object recognition (NOR), and open field (OF) tasks, we found that antenatal hypoxia exacerbated cognitive decline in offspring of 5xFAD compared with normoxia control. Of interest, fetal hypoxia did not alter intraneuronal soluble amyloid-β (Aβ) oligomer accumulation in the cortex and hippocampus in 5xFAD mouse offspring, indicating that antenatal hypoxia increased the vulnerability of the brain to synaptotoxic Aβ in the disease onset later in life. Consistent with the early occurrence of cognitive decline, we found synapse loss but not neuronal death in the cerebral cortex in 5xFAD but not wild-type (WT) offspring exposed to antenatal hypoxia. Furthermore, we also demonstrated that antenatal hypoxia significantly increased microglial number and activation, and reactive astrogliosis in the cerebral cortex in WT offspring. Moreover, antenatal hypoxia resulted in an exacerbated increase of microgliosis and astrogliosis in the early stage of AD in 5xFAD offspring. Together, our study reveals a causative link between fetal stress and the accelerated onset of AD-related pathology, and provides mechanistic insights into the developmental origin of aging-related neurodegenerative disorders.


The Significance of Endometrial Scratching for Clinical Pregnancy Rate in Long Agonist and Antagonist Protocols.
Background and Objectives: Assisted reproductive techniques today have helped many infertile couples achieve their desired pregnancy. However, unsuccessful implantation is usually the key failure in in vitro fertilization cycles. Many factors are now being studied to improve the implantation rate, one being endometrial artificial injury (endometrial scratching). The purpose of this study was to assess whether local endometrial artificial injury improved pregnancy rate, in long agonist and antagonist protocols. Materials and Methods: A retrospective analysis was carried out at the JSC Baltic American Clinic from January 1, 2016 to December 31, 2018. Women who were undergoing in vitro fertilization treatment were enrolled in the study. Medical data including demographic factors, menstrual history, and anamnesis of infertility were collected from medical records. Subjects were divided into intervention and control groups. The outcome of this study was clinical pregnancy rate (CPR). Results: A total of 137 women presenting with primary or secondary infertility were enrolled in the study. Clinical pregnancy was observed in 44/71 (61.9%) patients in the intervention group and 33/66 (50%) in the group without endometrial scratching (p value = 0.16). CPR in the intervention group was significantly higher in the patient group that had undergone ovarian stimulation with a long agonist protocol rather than the antagonist protocol (73.8% vs. 41.4%; p value = 0.006). In the multivariable logistic regression model, previously failed in vitro fertilization (IVF) (odds ratio (OR) = 0.07, 95% confidence interval (CI) 0.01-0.47), stimulation with the long agonist protocol (OR = 19.89, 95% CI 3.57-18.63), antagonist protocol (OR = 0.05, 95% CI 0.01-0.34), number of embryos (OR = 1.37, 95% CI 1.05-1.38), and number of blastocysts (OR = 2.96, 95% CI 1.43-8.36) remained important independent prognostic factors for clinical pregnancy. Conclusions: Our study findings indicate that randomized controlled studies are required to define patients for this procedure and to prove the effect and safety of local endometrial injury, before it is introduced to routine clinical practice.


Recent origin and phylogenetic utility of divergent ITS putative pseudogenes: a case study from Naucleeae (Rubiaceae).
The internal transcribed spacer (ITS) of nuclear ribosomal DNA has been widely used by systematists for reconstructing phylogenies of closely related taxa. Although the occurrence of ITS putative pseudogenes is well documented for many groups of animals and plants, the potential utility of these pseudogenes in phylogenetic analyses has often been underestimated or even ignored in part because of deletions that make unambiguous alignment difficult. In addition, long branches often can lead to spurious relationships, particularly in parsimony analyses. We have discovered unusually high levels of ITS polymorphism (up to 30%, 40%, and 14%, respectively) in three tropical tree species of the coffee family (Rubiaceae), Adinauclea fagifolia, Haldina cordifolia, and Mitragyna rubrostipulata. Both secondary structure stability and patterns of nucleotide substitutions in a highly conserved region (5.8S gene) were used for distinguishing presumed functional sequences from putative pseudogenes. The combination of both criteria was the most powerful approach. The sequences from A. fagifolia appear to be a mix of functional genes and highly distinct putative pseudogenes, whereas those from H. cordifolia and M. rubrostipulata were identified as putative pseudogenes. We explored the potential utility of the identified putative pseudogenes in the phylogenetic analyses of Naucleeae sensu lato. Both Bayesian and parsimony trees identified the same monophyletic groups and indicated that the polymorphisms do not transcend species boundaries, implying that they do not predate the divergence of these three species. The resulting trees are similar to those produced by previous analyses of chloroplast genes. In contrast to results of previous studies therefore, divergent putative pseudogenes can be useful for phylogenetic analyses, especially when no sequences of their functional counterparts are available. Our studies clearly show that ITS polymorphism may not necessarily mislead phylogenetic inference. Despite using many different PCR conditions (different primers, higher denaturing temperatures, and absence or presence of DMSO and BSA-TMACl), we recovered only a few functional ITS copies from A. fagifolia and none from H. cordifolia and M. rubrostipulata, which suggests that PCR selection is occurring and/or the presumed functional alleles are located at minor loci (with few ribosomal DNA copies).


Dietary tomato inhibits angiogenesis in TRAMP prostate cancer but is not protective with a Western-style diet in this pilot study.
Prostate cancer (PCa) remains the second most diagnosed cancer worldwide. Higher body weight is associated with chronic inflammation, increased angiogenesis, and treatment-resistant tumor phenotypes. Dietary tomato reduces PCa risk, which may be due to tomato inhibition of angiogenesis and disruption of androgen signaling. This pilot study investigated the interplay between tomato powder (TP), incorporated into control (CON) and obesogenic (OB) diets, and PCa tumor growth and blood perfusion over time in a transgenic model of PCa (TRAMP). Ultrasound microvessel imaging (UMI) results showed good agreement with gold-standard immunohistochemistry quantification of endothelial cell density, indicating that this technique can be applied to non-invasively monitor tumor blood perfusion in vivo. Greater body weight was positively associated with tumor growth. We also found that TP significantly inhibited prostate tumor angiogenesis but that this inhibition differentially affected measured outcomes depending on CON or OB diets. TP led to reduced tumor growth, intratumoral inflammation, and intratumoral androgen-regulated gene expression (srd5a1, srd5a2) when incorporated with the CON diet but greater tumor growth and intratumoral gene expression when incorporated with the OB diet. Results from this study show that protective benefits from dietary tomato are lost, or may become deleterious, when combined with a Western-style diet.


Discovering Sexual Health Conversations between Adolescents and Youth Development Professionals.
Youth development professionals (YDPs), working at community-based organizations are in a unique position to interact with the adolescents as they are neither parents/guardians nor teachers. The objectives of this study were to explore qualitatively what sexual health issues adolescents discuss with YDPs and to describe those issues using the framework of the Sexuality Information and Education Council of the United States (SIECUS) comprehensive sexuality education guidelines. YDPs reported conversations with adolescents that included topics related to the SIECUS key concepts of human development, relationships, personal skills, sexual behavior, and sexual health.


Feasibility and preliminary efficacy of a telerehabilitation approach to group adapted tango instruction for people with Parkinson disease.
People with Parkinson disease (PD) demonstrate improvements in motor function following group tango classes, but report long commutes as a barrier to participation. To increase access, we investigated a telerehabilitation approach to group tango instruction. Twenty-six people with mild-to-moderate PD were assigned based on commute distance to either the telerehabilitation group (Telerehab) or an in-person instruction group (In-person). Both groups followed the same twice-weekly, 12-week curriculum with the same instructor. Feasibility metrics were participant retention, attendance and adverse events. Outcomes assessed were balance, PD motor sign severity and gait. Participant retention was 85% in both groups. Attendance was 87% in the Telerehab group and 84% in the In-person group. No adverse events occurred. Balance and motor sign severity improved significantly over time ( p < 0.001) in both groups, with no significant group × time effects. Gait did not significantly change. Since a priori feasibility criteria were met or exceeded, and there were no notable outcome differences between the two instruction approaches, this pilot study suggests a telerehabilitation approach to group tango class for people with PD is feasible and may have similar outcomes to in-person instruction.


Etanercept for the Treatment of Chronic Arthritis Related to Chronic Granulomatous Disease: A Case.
Introduction: Chronic granulomatous disease (CGD) is a rare primary immunodeficiency, in which infections are the main presenting symptoms. Moreover, patients may also experience inflammatory and autoimmune manifestations. With proper management of infections, patients now survive to adulthood, and inflammatory manifestations have become more prominent problems. Treatment of the inflammatory manifestations in CGD is challenging and requires a multidisciplinary approach. Since tumor necrosis factor (TNF)-α has been cited as having a possible role on inflammatory conditions in CGD, etanercept, an anti-TNF agent, may represent a major advance in the management of inflammatory manifestations. Case Presentation: In this report, we described a 15-year-old boy, suffering concurrently both from human leukocyte antigen (HLA)-B27-positive chronic arthritis and CGD, whose arthritis did not respond to treatment with methotrexate and ibuprofen. Remission was achieved 6 months after etanercept initiation and during the next 18 months on medication, we did not encounter any signs of severe infections. Conclusion: Treatment of inflammatory conditions in CGD patients is still challenging in view of the lack of evidence-based therapeutic options. In this study, we report the first pediatric CGD case, in which chronic arthritis was successfully treated with etanercept.


Ichthyophonus sp. Infection in Opaleye (Girella nigricans).
Over a 3-year-period, 17 wild-caught opaleye (Girella nigricans) housed in a public display aquarium were found dead without premonitory signs. Grossly, 4 animals had pinpoint brown or black foci on coelomic adipose tissue. Histologically, liver, spleen, heart, and posterior kidney had mesomycetozoan granulomas in all cases; other organs were less commonly infected. Four opaleye had goiter; additional substantial lesions were not identified. Granulomas surrounded melanized debris, leukocytes, and mesomycetozoa represented by folded membranes (collapsed schizont walls), intact schizonts (50- to >200 µm in diameter with a multilaminate membrane), plasmodia (budding from schizonts or free in tissue), or rarely germinal tubes (budding from schizonts). Ichthyophonus was grown from fresh tissues in tissue explant broth cultures of the heart, liver, and/or spleen. Polymerase chain reaction using 18S ribosomal DNA primers amplified a 1730-bp region, and the DNA sequence was most similar to Ichthyophonus hoferi, which is often associated with freshwater aquaculture fish.


Rosuvastatin Failed to Improve Arteriovenous Fistula Patency for Hemodialysis in Diabetic Patients - A Randomized Clinical Trial.
Very limited therapeutic strategies exist to prevent the primary failure of arteriovenous (AV) fistulas in patients with diabetes. To investigate whether rosuvastatin could improve the primary patency of AV fistulas in diabetic patients with stage 5 chronic kidney disease (CKD). This was a double-blind randomized clinical trial. From July 2012 to September 2018, patients aged between 18 and 65 years with type 2 diabetes and stage 5 CKD were randomized to receive placebo or rosuvastatin (5 mg/day) for 7 days prior to the creation of an AV fistula on the forearm until the 21st day after surgery. Patients were followed up for 180 days after the operation. The primary composite endpoint was the development of fistula immaturity or stenosis. The secondary endpoints were changes in inflammatory markers, oxidative stress, and occurrence of postoperative complications. A total of 60 patients were enrolled in the study. Rosuvastatin resulted in a 20% reduction in total cholesterol from postoperative day 0 to 28 (p = .0006). The overall rate of AV fistula failure (immaturity or stenosis) was 30%, with no significant difference between patients receiving rosuvastatin and those receiving the placebo (33.3% vs. 26.7%, p = .5731). Although not statistically significant, the administration of rosuvastatin might have increased the incidence of postoperative complications (2.99 vs. 2.39 event rate per 1000 patient-days; odds ratio, 1.33; p = .5986). Rosuvastatin showed no significant beneficial effects on the primary patency of AV fistulas in diabetic patients with stage 5 CKD, but might have been associated with the risk of drug-related complications.


Morphologic characterization of the anterior inferior cerebellar artery: a direct anatomic study.
An adequate knowledge of the anterior inferior cerebellar artery (AICA) is oriented to the morphological sciences, clinical management and surgical planning of the posterior fossa. We aimed to determine the morphology of AICA in a sample from Colombian population. We studied 92 AICA from fresh cadavers. For each specimen, the vertebral arteries were injected with 100 cc of semi-synthetic resin (a mixture of Palatal E210® BASF 80 cc and Styrene 20 cc) dyed with mineral red. The biometrics and morphological variables of AICA were registered. AICA originated at 9.9 ± 3.2 mm from the vertebrobasilar junction. In 12 samples (8.1%), we observed a common trunk between AICA and posterior inferior cerebellar artery, which presented a caliber of 1.56 ± 0.23 mm and a length of 11.3 Â ± 3. 53 mm. In 80 (51.3%) specimens, AICA was originated from the proximal segment of basilar artery, while in 76 (48.7%) of them emerged from the medium segment. The AICA bifurcation distance from its origin was less than 20 mm in 20.5% of cases; between 20 and 40 mm in 62.3%. In its trajectory, AICA passed ventral to the facial nerve in 85 samples (53.2%), dorsal to the facial nerve in 68 samples (43.6%) and between the roots in 5 samples (3.2%). The origin of the AICA from the proximal segment of the basilar artery is confirmed in this study, which disagrees with reports that point out its origin in the middle segment.


A child with resistance to thyroid hormone without thyroid hormone receptor gene mutation: a 20-year follow-up.
We report here the 20-year follow-up study of a male subject diagnosed at 15 months of age as a sporadic case of pituitary resistance to thyroid hormone on the combination of clinical hyperthyroidism, elevated serum thyroid hormone (TH) levels and inappropriate thyrotropin (TSH). On D-thyroxine (D-T(4)) therapy from 30 months of age to 12.5 years, hyperactivity and hyperthyroid signs and symptoms as well as growth abnormalities improved, serum L-thyroxine (L-T(4)) enantiomer normalized, and basal and stimulated TSH decreased significantly without complete suppression. After 8 years off D-T(4), at 20 years of age, clinical status was normal despite persisting high TH levels and inappropriate TSH. Evolution of serum markers of TH action and echocardiography measurements followed up from 15 months to 20 years of age either in basal condition or on triiodothyronine (T(3)), as well as the sequential determination of bone mineral density suggest differences in the tissue responses to T(3): normal in bone with a high remodelling rate, heterogeneity for various hepatic markers, and decreased at heart level. No mutations were found in the coding sequence of TRbeta1, TRbeta2, TRalpha1, RXRgamma, SMRT, NCoR1, and NCoA1. In this patient the putative long-term effects of the persisting high bone resorption are unknown.


Association Between Diabetes and the Risk of Kidney Cancer: Systematic Review and Meta-Analysis.
Diabetes is a risk factor for several types of cancer, but the specific relationship between diabetes and kidney cancer is not well understood. We conducted a search strategy in scientific databases for case-control and cohort studies on this topic. We analyzed 17 studies and found that diabetes was significantly associated with the risk of developing kidney cancer and that this risk was slightly stronger for women and for people living in Asia. These findings were not influenced by obesity, cigarette smoking, or hypertension. We conclude that diabetes is an independent risk factor for the development of kidney cancer.


Effects of Limbic stimulation and the microinontophoresis of aminergic drugs on the single unit activity of hypothalamic neurosecretory cells (author's transl).
Attempts were made to investigate the chemical nature of hypothalamic afferents from the extrahypothalamic structures which participate in the control of hypothalamic neurosecretory cells. Experimental procedures were discribed in the preceding report, except that of the electrical stimulation and the microinontophoresis. Some part of data obtained in this study were referred in the preceding report for improvement of statistic reliability. For electrical stimulation, side-by-side bipolar electrodes were placed in the medial preoptic area (MPO), medial amygdala (mAMYG) and dorsal hippocampus (dHPC) from the parietal surface of the brain and fastened to the cranium by dental resin. Microintophoresis of norepinephrine (NE), dopamine (DA) and acetylcholine (Ach) was carried out by using five-barrelled glass pipette on the antidromically activated units in the medical basal hypothalamus by the median eminence stimulation. Extracellular potentials were recorded by the central barrel of each electrode. One of the outer barrels was filled with physiological saline and used for control purpose. (1) Eelctrical stimulation of the MPO with train pulses of 0.2 msec duration and 300 muA intensity in 100 HZ for 5 sec, induced facilitation and inhibition in 26.4% and 9.4% of 106 tests, respectively. By the mAMYG stimulation with 500 muA, facilitation was seen in 18.8% and inhibition in 10.4%, of 96 tests. The HPC stimulation induced facilitation in 10.0% and inhibition in 23.3% of 90 tests. (2) Facilitatory effect of the MPO was most frequently seen in the ovariectomized and estrogen primed rats (58.8% of the tests). In contrast, the inhibitory effect of the dHPC was striking in the diestrus-I (40.0%). (3) Microiontophoresis of NE induced facilitation in 60.0% and inhibition in 10.6% of 66 identified cells. DA induced facilitation in 45%, and inhibition in 11.7% of 60 tests. Ach induced facilitation in 41.4%, and inhibition in 13.8% of 58 tests. (4) By NE infusion, facilitation was most commonly seen in the proestrus (70.6%). Inhibition was most readily elicited by Ach infusion on the diestrus-I (30%). Facilitative effect of DA was eminent in the diestrus-I (81.8%). (5) Successive administration of NE and DA on identical units revealed that in 56.0%, the units were responsive to only one agent and the other was effectless. 25.5% of the units responded to both of NE and DA. (6) The conceidence of effects, either facilitation or inhibition, between that of electrical stimulation of the microinontophoresis, was calculated on 46 units. The facilitatory effect of the MPO was most commonly mimicked by NE infusion (81.8%), and the inhibitory effect of the dHPC by Ach infusion (46.7%). NE also mimicked facilitatory effect of mAMYG stimulation (73.3%). (7) Intraventricular infusion of 20 mug of NE induced significant increase of serum LH (p less than 0.05 to saline) when measured by radioimmunoassay, whereas DA resulted in a decrease of LH (p less than 0.01).


Successful treatment of COVID-19 using extracorporeal membrane oxygenation, a case report.
Since the end of 2019, COVID-19 has been prevalent in Wuhan, China, and has been rapidly spreading to mainland China. Currently, more than 80,000 people have been infected, of which over 10,000 were severely ill and had characteristics of dyspnea and hypoxemia about one week after onset. Severe patients had rapidly progressed to acute respiratory distress syndrome (ARDS), causing multiple organ failures and even death, with a mortality rate of nearly 4.3%. The treatment of severe COVID-19 patients has been rarely reported. This study reported a successful example of a severe COVID-19 patient with extracorporeal membrane oxygenation (ECMO) technology in our hospital. This experience revealed that the early application of ECMO can dramatically promote the recovery of severe COVID-19 patients.


Methods for analysing individual changes in sick-leave diagnoses over time.
Several methodological challenges arise when attempting to analyse individual data on changes of sick-leave diagnoses over several years. Sick-leave spells for a person can recur, have different sick-leave diagnoses, and both these aspects are dependent of previous episodes, the numbers of repeated periods vary across subjects, and standard statistical methods are not valid for variables on nominal scales, e.g. sick-leave diagnoses. Our aim was to ascertain whether the number and pattern of changes in sick-leave diagnoses are associated with future disability pension (DP) and to test methods for analysis of repeated measurements on nominal data. Data from a 12-year prospective cohort study of the 8000 sick-leave periods of the 213 persons aged 25-34 who, in 1985, had a new sick-leave spell 28 days with back diagnoses were used. We used entropies, uncertainty coefficients adjusted for repeated measurements, and transition matrices to examine the changes in sick-leave diagnoses that occurred during follow up. In the 12 years 22% were granted DP and they had changed sick-leave diagnosis less frequently and more often had new sick-leave periods with musculoskeletal diagnoses than the others. The variation in diagnoses and the degree of dependence between consecutive diagnoses were associated with DP. Many tools in statistics are based on linear methods that require numerical variables, but such methods are not valid for repeated measurements on discrete variables on nominal scales, as for sick-leave diagnosis. In such cases, it can be beneficial to use tools that are applied in statistical information theory.


Occurrence of endocrine-disrupting chemicals (EDCs) in river water and sediment of the Mahakam River.
The study was performed to examine the occurrence of endocrine disrupting chemicals (EDCs), including four steroid estrogens, one plasticizer, and three preservatives in the Mahakam River, Indonesia. The physicochemical analysis of river water and sediment quality parameters were determined as well as the concentration of EDCs. The range of values for pH, total dissolved solids (TDS), dissolved oxygen (DO), biochemical oxygen demand (BOD), total suspended solids (TSS), nitrate, ammonium, phosphate, and oil/grease in river water and sediment were higher than recommended limits prescribed by the World Health Organization's Guidelines for Drinking-water Quality (GDWQ). Bisphenol A (BPA) was the most widely found EDC with the highest concentration level at 652 ng/L (mean 134 ng/L) in the river water and ranged from ND (not detected) to 952 ng/L (mean 275 ng/L) in the sediment. Correlation analysis to investigate the relationship between the EDCs' concentrations in water and sediment also revealed a significant correlation (R2 = 0.93) between the EDCs' concentrations. High concentrations of EDCs are found in urban and residential areas because these compounds are commonly found in both human and animal bodies, resulting in the disposal of EDCs into canals and rivers in urban and suburban areas, as well as livestock manure and waste that is generated from intensive livestock farming around the suburban area.


A comparative study of matrix metalloproteinase and aggrecanase mediated release of latent cytokines at arthritic joints.
Latent cytokines are engineered by fusing the latency associated peptide (LAP) derived from transforming growth factor-β (TGF-β) with the therapeutic cytokine, in this case interferon-β (IFN-β), via an inflammation-specific matrix metalloproteinase (MMP) cleavage site. To demonstrate latency and specific delivery in vivo and to compare therapeutic efficacy of aggrecanase-mediated release of latent IFN-β in arthritic joints to the original MMP-specific release. Recombinant fusion proteins with MMP, aggrecanase or devoid of cleavage site were expressed in CHO cells, purified and characterised in vitro by Western blotting and anti-viral protection assays. Therapeutic efficacy and half-life were assessed in vivo using the mouse collagen-induced arthritis model (CIA) of rheumatoid arthritis and a model of acute paw inflammation, respectively. Transgenic mice with an IFN-regulated luciferase gene were used to assess latency in vivo and targeted delivery to sites of disease. Efficient localised delivery of IFN-β to inflamed paws, with low levels of systemic delivery, was demonstrated in transgenic mice using latent IFN-β. Engineering of latent IFN-β with an aggrecanase-sensitive cleavage site resulted in efficient cleavage by ADAMTS-4, ADAMTS-5 and synovial fluid from arthritic patients, with an extended half-life similar to the MMP-specific molecule and greater therapeutic efficacy in the CIA model. Latent cytokines require cleavage in vivo for therapeutic efficacy, and they are delivered in a dose dependent fashion only to arthritic joints. The aggrecanase-specific cleavage site is a viable alternative to the MMP cleavage site for the targeting of latent cytokines to arthritic joints.


Europium(III) tetrakis(β-diketonate) complex as an ionic liquid: a calorimetric and spectroscopic study.
An intrinsic photoluminescent ionic liquid based on europium(III) tetrakis(β-diketonate) complex with a tetraalkylphosphonium as counterion was synthesized. Calorimetric measurements showed a melting point at 63 °C, which allows the ionic liquid classification. When cooling the material from the liquid state, metastable supercooled ionic liquid is obtained, as seen from NMR spectroscopy as well. Eu(III) photoluminescence is clearly observed while the absorption spectra of the ligand is dominant, showing the antenna effect. This was confirmed with submicrosecond time scale luminescence spectroscopy, where a rise of Eu(III) emission is observed with the correspondent decay of the ligand excited state. Temperature effects in the photoluminescence are also shown, being prominent above the melting point where the intensity decreases with Arrhenius behavior. Eu(III) luminescence decays also show features characteristic of energy migration between homologue Eu(III) species. Solvent effects were also studied by NMR and Luminescence spectroscopies, highlighting that the nucleophilicity of organic solvents such as n-alcohols leads to a coordination with Eu(III), which ultimately compromises the stability of the complex.


[Outpatient external dacryocystorhinostomy under regional anesthesia and sedation].
To assess the rate of early complications of outpatient external dacryocystorhinostomy (DCR) and patient satisfaction with the anesthetic technique. This prospective study enrolled 58 patients undergoing external DCR. We analyzed demographic variables, ASA physical status, level of sedation achieved, postoperative pain, systemic complications, intraoperative bleeding, duration of surgery, time until discharge home, and patient and surgeon satisfaction with the anesthetic technique. The mean (SD) level of satisfaction was 4.85 (0.80) points on the Iowa Satisfaction With Anesthesia Scale (ISAS). A positive association was found between postoperative pain and a lower ISAS score. There was also a positive association between use of rescue analgesia in the early postoperative period and a lower ISAS score. Mean blood loss per procedure was 178.9 (108.2) mL. The rate of minor systemic complications was 15.5%. The surgeon's rating of conditions in the surgical field was excellent or good in 89.6% of the cases. External DCR can be performed on an outpatient basis within a reasonable safety margin and with a low early postoperative complication rate. Patient satisfaction with anesthesia was high. Provision of preoperative information about the meaning of sedation, postoperative analgesia, and surgical bleeding are aspects to improve in this practice setting.


Associations of Current, Childhood, and Adolescent Milk Intake with Serum Insulin-like Growth Factor (IGF)-1 and IGF Binding Protein 3 Concentrations in Adulthood.
Previous studies have shown that insulin-like growth factor (IGF)-1 levels are positively and IGF binding protein (IGFBP)-3 levels negatively associated with risk of certain cancers. Also, dietary factors may influence the IGF system. We aimed to analyze the associations of current, childhood and adolescent milk intake with IGF-1 levels, IGFBP-3 levels and IGF-1:IGFBP-3 molar ratio in adulthood. Multivariable linear regression analyses by sex and race/ethnicity were performed using cross-sectional data from the Third National Health and Nutrition Examination Survey. A total of 5,805 participants were included in the analyses. Adult IGF-1 levels and IGF-1:IGFBP-3 molar ratio had significant inverse associations (P-trend = 0.02) with adolescent milk intake in non-Hispanic white men, but not in men of other race/ethnicities or in women. There were no associations between current or childhood milk intake and IGF-1 levels or IGF-1:IGFBP-3 molar ratio in adulthood. Current milk intake and childhood milk intake had significant positive associations (P-trend = 0.02) with adult IGFBP-3 levels in non-Hispanic white and non-Hispanic black women, respectively, but no associations were observed in Mexican American women or in men. Overall, there were long-term and short-term associations between milk intake and IGF-1 and IGFBP-3 levels, but the associations varied by race/ethnicity and sex.


Oxytocin increases generosity in humans.
Human beings routinely help strangers at costs to themselves. Sometimes the help offered is generous-offering more than the other expects. The proximate mechanisms supporting generosity are not well-understood, but several lines of research suggest a role for empathy. In this study, participants were infused with 40 IU oxytocin (OT) or placebo and engaged in a blinded, one-shot decision on how to split a sum of money with a stranger that could be rejected. Those on OT were 80% more generous than those given a placebo. OT had no effect on a unilateral monetary transfer task dissociating generosity from altruism. OT and altruism together predicted almost half the interpersonal variation in generosity. Notably, OT had twofold larger impact on generosity compared to altruism. This indicates that generosity is associated with both altruism as well as an emotional identification with another person.


A benchmarking of workflows for detecting differential splicing and differential expression at isoform level in human RNA-seq studies.
Over the last few years, RNA-seq has been used to study alterations in alternative splicing related to several diseases. Bioinformatics workflows used to perform these studies can be divided into two groups, those finding changes in the absolute isoform expression and those studying differential splicing. Many computational methods for transcriptomics analysis have been developed, evaluated and compared; however, there are not enough reports of systematic and objective assessment of processing pipelines as a whole. Moreover, comparative studies have been performed considering separately the changes in absolute or relative isoform expression levels. Consequently, no consensus exists about the best practices and appropriate workflows to analyse alternative and differential splicing. To assist the adequate pipeline choice, we present here a benchmarking of nine commonly used workflows to detect differential isoform expression and splicing. We evaluated the workflows performance over different experimental scenarios where changes in absolute and relative isoform expression occurred simultaneously. In addition, the effect of the number of isoforms per gene, and the magnitude of the expression change over pipeline performances were also evaluated. Our results suggest that workflow performance is influenced by the number of replicates per condition and the conditions heterogeneity. In general, workflows based on DESeq2, DEXSeq, Limma and NOISeq performed well over a wide range of transcriptomics experiments. In particular, we suggest the use of workflows based on Limma when high precision is required, and DESeq2 and DEXseq pipelines to prioritize sensitivity. When several replicates per condition are available, NOISeq and Limma pipelines are indicated.