Development of novel in silico model for developmental toxicity assessment by using naïve Bayes classifier method.
Toxicological testing associated with developmental toxicity endpoints are very expensive, time consuming and labor intensive. Thus, developing alternative approaches for developmental toxicity testing is an important and urgent task in the drug development filed. In this investigation, the naïve Bayes classifier was applied to develop a novel prediction model for developmental toxicity. The established prediction model was evaluated by the internal 5-fold cross validation and external test set. The overall prediction results for the internal 5-fold cross validation of the training set and external test set were 96.6% and 82.8%, respectively. In addition, four simple descriptors and some representative substructures of developmental toxicants were identified. Thus, we hope the established in silico prediction model could be used as alternative method for toxicological assessment. And these obtained molecular information could afford a deeper understanding on the developmental toxicants, and provide guidance for medicinal chemists working in drug discovery and lead optimization.


Post-translational modifications on RNA-binding proteins: accelerators, brakes, or passengers in neurodegeneration?
RNA-binding proteins (RBPs) are critical players in RNA expression and metabolism, thus, the proper regulation of this class of proteins is critical for cellular health. Regulation of RBPs often occurs through post-translational modifications (PTMs), which allow the cell to quickly and efficiently respond to cellular and environmental stimuli. PTMs have recently emerged as important regulators of RBPs implicated in neurodegenerative disorders, in particular amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Here, we summarize how disease-associated PTMs influence the biophysical properties, molecular interactions, subcellular localization, and function of ALS/FTD-linked RBPs, such as FUS and TDP-43. We will discuss how PTMs are believed to play pathological, protective, or ambiguous roles in these neurodegenerative disorders.


Radiolabeled nano-peptides show specificity for an animal model of human PC3 prostate cancer cells.
Cancer has been investigated using various pre-targeting techniques or models focusing on radiobombesin analogues; however, both are not offered together. In this study, nano-bombesin labeling by a pre-targeting system was undertaken to develop an alternative approach for prostate tumor treatment. A two-step pre-targeting system utilizing a combination of streptavidin (SA), biotinylated morpholino (B-MORF), biotinylated BBN (B-BBN) with two different spacers (b-Ala and PEG), and a radiolabeled cMORF was evaluated in vitro and in vivo. Final conjugation conditions consisted of a 1:1:2 ratio of SA:B-MORF:B-BBN, followed by addition of 99mTc-cMORF to compensate for free MORF. In vitro binding experiments with prostate cancer cells (PC-3) revealed that total binding was time-dependent for the Ala spacer but not for the PEG spacer. The highest accumulation (5.06 ± 1.98 %) was achieved with 1 hour of incubation, decreasing as time progressed. Specific binding fell to 1.05 ± 0.35 %. The pre-targeting biodistribution in healthy Swiss mice was measured at different time points, with the best responses observed for 7-h and 15-h incubations. The effector, 99mTc-MAG3-cMORF, was administered 2 h later. Strong kidney excretion was always documented. The greatest tumor uptake was 2.58 ± 0.59 %ID/g at 7 h for B-bAla-BBN, with a region of interest (ROI) value of 3.9 % during imaging. The tumor/blood ratio was low due to the slow blood clearance; however, the tumor/muscle ratio was 5.95. The pre-targeting approach with a peptide was a viable concept. Further evaluation with modified sequences of MORF, including less cytosine, and additional test intervals could be worthwhile.


Studies in Chinese Tettigoniidae: Recent discoveries of Meconematinae katydids from Xizang,China (Tettigoniidae: Meconematinae).
In this work, we describe one new genus and 4 new species from Xizang, China: Allogrigoriora carinata gen sp. nov., Nigrimacula beybienkoi sp. nov., Sinoxizicus carinatus sp. nov. and Tamdaora curvicerca sp. nov. Besides the new taxa, 3 new combinations [Grigoriora cheni (Bey-Bienko, 1955) comb. nov.; Grigoriora kweichowensis (Tinkham, 1944) comb. nov.; Nefateratura bifurcata (Liu Bi, 1994) comb nov.] are also changed. All the specimens are deposited in Shanghai Entomological Museum, Chinese Academy of Sciences.


Induction of cytochrome P-450 in congenic C57BL/6J mice by isosafrole: lack of correlation with the Ah locus.
Isosafrole induction of cytochrome P-450 was compared in congenic strains of C57BL/6J mice, one of which expresses normal levels of the Ah receptor [B6(Ahb)], and another that does not contain a measurable receptor concentration [B6(Ahd)]. Using sucrose gradient analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) binding, an Ah receptor concentration of 69.1 +/- 3.8 fmol/mg protein was measured in the hepatic cytosol from B6(Ahb) mice, while no receptor could be detected in the cytosol from B6(Ahd) mice. Isosafrole treatment (75 mg/kg X 3 days) increased the total hepatic microsomal cytochrome P-450 content to the same extent in the two congenic strains. The level of microsomal monooxygenase induction in the isosafrole-treated B6(Ahd) mice was greater than that of B6(Ahb) mice for ethylmorphine N-demethylase and isosafrole metabolite-complex formation, the latter a measure of cytochrome P2-450. In the case of 7-ethoxycoumarin O-deethylase only the isosafrole-treated B6(Ahd) mice had elevated microsomal activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) also revealed a similar induction pattern for the two congenic strains, following isosafrole treatment. Thus, the isosafrole treated B6(Ahd) mice produced an equivalent or slightly larger induction of cytochrome P-450 than the B6(Ahb) mice, suggesting that there is no direct role for the Ah receptor in the regulation of these cytochrome P-450 monooxygenase activities by isosafrole.


Typology of Currently or Formerly Homeless Individuals Based on Their Use of Health and Social Services.
This study identified profiles among 455 currently or formerly homeless individuals in Quebec (Canada), based on health and social service use. Using latent class analysis, four profiles were identified that grouped individuals with: (1) few health problems, and using few case managers and family doctors, but with high frequency of psychiatric consultations, emergency department (ED) visits and hospitalizations; (2) chronic physical illnesses, having case managers and family doctors, but low frequency of ED visits and hospitalizations; (3) moderate health problems and little service use; and (4) multiple and complex health problems and high frequency of service use. These profiles suggest the following recommendations to more adequately meet patient needs: regarding Class 1, improved outreach services, more ED liaison nurses and peer navigation; Class 2: more family doctors and case managers; Class 3: higher family doctors; and Class 4: more assertive or intensive case management, harm reduction and permanent housing resources.


Functional analysis of two novel TBX5 variants present in individuals with Holt-Oram syndrome with different clinical manifestations.
Holt-Oram syndrome (HOS) is a rare disorder characterized by cardiac and upper-limb defects. Pathogenic variants in TBX5-a gene encoding a transcription factor important for heart and skeletal development-are the only known cause of HOS. Here, we present the identification and functional analysis of two novel TBX5 pathogenic variants found in two individuals with HOS presenting distinct phenotypes. The individual with the c.905delA variant has a severe cardiac phenotype but mild skeletal defects, unlike the individual with the c.246_249delGATG variant who has no cardiac problems but severe upper limbs malformations, including phocomelia. Both frameshift variants, c.246_249delGATG and c.905delA, generate mRNAs harbouring premature stop codons which, if not degraded by nonsense mediated decay, will lead to the production of shorter TBX5 proteins, p.Gln302Argfs*92 and p.Met83Phefs*6, respectively. Immunocytochemistry results suggest that both mutated proteins are produced and furthermore, like the wild-type protein, p.Gln302Argfs*92 mutant appears to be mainly localized in the nucleus, in contrast with p.Met83Phefs*6 mutant that displays a higher level of cytoplasmic localization. In addition, luciferase activity analysis revealed that none of the TBX5 mutants are capable of transactivating the NPPA promoter. In conclusion, our results provide evidence that both pathogenic variants cause a severe TBX5 loss-of-function, dramatically reducing its biological activity. The absence of cardiac problems in the individual with the p.Met83Phefs*6 variant supports the existence of other mechanisms/genes underlying the pathogenesis of HOS and/or the existence of an age-related delay in the development of a more serious cardiac phenotype. Further studies are required to understand the differential effects observed in the phenotypes of both individuals.


Analysis of the Leeds-Keio ligament for extensor mechanism repair: favourable mechanical and functional outcome.
Many techniques have been described for the challenging problem of extensor mechanism rupture, including the use of synthetic grafts. In 1994 Fujikawa et al. [J. Bone Surg. 1994:76B:200-203] reported excellent results using the Leeds-Keio polyester ligament. Patients were mobilised early, had few post operative complications and minimal extensor lag. This is the first independent assessment of this technique using both subjective and objective analysis of outcome. We reviewed 11 patients (12 knees) whose extensor mechanisms were repaired as described in the original paper. At a mean of 3 years after surgery there were no infections and no re-ruptures. Good functional recovery was observed with a mean Tegner activity score of 1.7, Lysholm score of 66 and Irrgang score of 58%. Indeed, 80% of working patients returned to their former employment. However, objective tests showed peak extensor torque recovered to only half normal strength. We conclude the results of this technique of repair compare favourably with other described techniques especially in complex cases.


Intestinal water and solute absorption studies: comparison of in situ perfusion with chronic isolated loops in rats.
The effects of lumenal glucose on jejunal water transport and the influence of glucose-induced water absorption on solute uptake from single-pass perfusions are compared in anesthetized rats in situ and isolated chronic loops in unanesthetized rats in vivo. While the magnitudes of solute membrane permeabilities are consistently higher in the chronic loop system, the effects on water transport and its promotion of jejunal solute uptake are comparable between the two experimental systems. The effect of glucose-induced water absorption on the enhanced/baseline jejunal uptake ratio of the hydrophilic drug, acetaminophen, is greater than that for the lipophilic drug, phenytoin, in both experimental systems. The fact that chronic loop effective solute permeabilities were equivalent to solute membrane permeabilities in situ is consistent with greater lumenal fluid mixing in vivo. In addition, in situ body temperature affects the uptake of phenytoin but not acetaminophen, water, or glucose. This suggests that active and paracellular solute transport is not compromised in situ, while membrane partitioning and diffusion of lipophilic species are more sensitive to experimental conditions.


Survival effect of pre-RT PET-CT on cervical cancer: Image-guided intensity-modulated radiation therapy era.
No large-scale, well-designed randomized study with a long-term follow-up has evaluated the survival effect of pretreatment 18-fluorodeoxyglucose positron emission tomography-computed tomography (18FDG-PET-CT) on patients with stage IB-IVA cervical cancer receiving image-guided intensity-modulated radiation therapy (IG-IMRT). This is the first head-to-head propensity score-matched, nationwide population-based cohort study evaluating this survival effect. The results revealed that pretreatment 18FDG-PET-CT might be associated with longer survival in patients with stage IB-IVA cervical cancer receiving radiotherapy or concurrent chemoradiotherapy, especially in the IG-IMRT era. No large-scale, well-designed randomized study with a long-term follow-up has evaluated the survival effect of pretreatment 18-fluorodeoxyglucose positron emission tomography-computed tomography (18FDG-PET-CT) on patients with stage IB-IVA cervical cancer receiving image-guided intensity-modulated radiation therapy (IG-IMRT). Therefore, in this propensity score-matched, population-based cohort study, we investigated these survival effects. We included 4167 patients with stage IB-IVA cervical cancer receiving radiotherapy (RT) or concurrent chemoradiotherapy (CCRT) through the IG-IMRT technique. The patients were categorized into two 1:2 propensity score-matched groups depending on whether they underwent pretreatment 18FDG-PET-CT, and their outcomes were compared. We included 2778 and 1389 patients with cervical cancer in the nonpretreatment and pretreatment PET-CT groups, respectively. Univariable and multivariable analyses revealed an association between pretreatment PET-CT and improved survival in the patients (in the adjusted model, the adjusted hazard ratio [aHR] was 0.88; 95% confidence interval [CI], 0.80-0.97: P = 0.010). Regardless of the cancer stage (early or advanced), pretreatment PET-CT was significantly superior to nonpretreatment PET-CT in terms of all-cause death (aHR, 0.78; 95% CI, 0.60-0.92; P = 0.013 and aHR, 0.90; 95% CI, 0.81-0.99; P = 0.039 for the early [IB-IIA] and advanced stages [IIB-IVA], respectively). Pretreatment 18FDG-PET-CT might be associated with longer survival in patients with stage IB-IVA cervical cancer receiving RT or CCRT, especially in the era of IG-IMRT.


Reactivity to endothelium-dependent and -independent vasoactive substances is maintained in coronary resistance vessels of the failing hamster heart.
Several studies suggest that regulation of coronary vasomotion is abnormal in heart failure, but controversies exist as to whether an altered endothelium-dependent dilation of the coronary vasculature contributes to these abnormalities. In the present study we evaluated the coronary reactivity to endothelium-dependent and -independent vasoactive substances in a model of chronic heart failure. Isolated hearts from > 200-day-old cardiomyopathic hamsters (UM-X7.1) and age-matched normal Syrian hamsters were mounted on a Langendorff apparatus and retrogradely perfused at constant pressure (100 mmHg). Heart rate, left ventricular developed pressure (LVP) and coronary flow (CF) were measured. A first group of normal and failing hearts were challenged with increasing doses of acetylcholine (Ach), 1-1000 nM, and sodium nitroprusside (SNP), 0.01-10 microM. A second group was challenged with Ach (100 nM) and SNP (1 microM) before and during infusion of the NO synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME). Finally, normal and failing hearts were challenged with Ach (100 nM) before and during infusion of indomethacin (10 microM) or tetraethylammonium (1 mM). Myocardial contractility and coronary flow were significantly impaired in failing hearts (LVP = 32 +/- 5 vs. 50 +/- 4 mmHg in normal hearts; CF = 4.7 +/- 0.6 vs. 6.7 +/- 0.6 ml/min in normal hearts). Coronary flow increases with increasing doses of Ach (EC50 = 84 +/- 15 nM for failing hearts and 100 +/- 3 nM for normal hearts, P = ns). At concentrations exceeding the EC50, failing hearts were more sensitive to the coronary dilator effects of Ach. The reduction in coronary flow elicited by L-NAME was similar in normal (-41 +/- 2%) and failing hearts (-40 +/- 3%); in both normal and failing hearts, the acetylcholine vasodilator response was not significantly affected by L-NAME. Indomethacin infusion resulted in a slight increase in coronary flow and significantly reduced the coronary dilator effects of acetylcholine. Tetraethylammonium had no significant effects on basal coronary perfusion of normal and failing hearts. However, in the latter group, acetylcholine vasodilator response was significantly attenuated. Results obtained in isolated failing hamster hearts allow us to conclude that (1) significant coronary dysfunctions are present in this model of chronic heart failure, (2) neither the NO-synthase nor the cyclo-oxygenase pathway contributes to these coronary dysfunctions, and (3) an adequate coronary vasodilator response appears to be present in this model of chronic heart failure.


Probing RNA structure in vivo.
RNA structure underpins many essential functions in biology. New chemical reagents and techniques for probing RNA structure in living cells have emerged in recent years. High-throughput, genome-wide techniques such as Structure-seq2 and DMS-MaPseq exploit nucleobase modification by dimethylsulfate (DMS) to obtain complete structuromes, and are applicable to multiple domains of life and conditions. New reagents such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), glyoxal, and nicotinoyl azide (NAz) greatly expand the capabilities of nucleobase probing in cells. Additionally, ribose-targeting reagents in selective 2'-hydroxyl acylation and primer extension (SHAPE) detect RNA flexibility in vivo. These techniques, coupled with crosslinking nucleobases in psoralen analysis of RNA interactions and structures (PARIS), provide new and diverse ways to elucidate RNA secondary and tertiary structure in vivo and genome-wide.


Management of mustard gas-induced limbal stem cell deficiency and keratitis.
To report the clinical findings and compare outcomes of different surgical techniques evolved over time in a large series of patients with delayed-onset mustard gas keratitis (MGK). Retrospective, comparative, interventional case series. Ninety Iranian male survivors (175 eyes) of Iraqi chemical warfare with chronic or delayed-onset MGK. The symptoms and clinical findings of patients are presented, and medical and surgical interventions to address dry eye, limbal ischemia and limbal stem cell deficiency (LSCD), and corneal involvements are explained. The results of limbal stem cell transplantation techniques (living-related conjunctival-limbal allograft [lrCLAL] versus keratolimbal allograft [KLAL]) as well as corneal transplantation techniques (penetrating keratoplasty [PK] versus lamellar keratoplasty [LK]) are compared in terms of clinical outcomes and graft survival rates. Ocular findings and appropriate surgical approach for LSCD and corneal involvements. A total of 175 eyes of 90 cases (all male) between 34 and 68 years of age were followed up for 101±30.3 months (range, 36-198 months). The most common ocular involvements were chronic blepharitis and dry eye. Conjunctival vascular abnormalities and limbal ischemia were observed in 27.4% and 29.7% of eyes, respectively. Limbal stem cell deficiency necessitating stem cell transplantation developed in 41.1% of eyes. The most common corneal sign was central and peripheral anterior stromal opacity (58.9%), followed by corneal stromal thinning (36.0%) and neovascularization (27.4%). Living-related conjunctival-limbal allograft was performed in 32 eyes, and KLAL was performed in 40 eyes. The rejection-free graft survival rate was 39.1% in the lrCLAL group and 80.7% in the KLAL group at month 40, with a mean length of 24.9 and 68.8 months, respectively (P = 0.02). Thirty eyes underwent PK and 51 underwent LK. Corneal graft failure was observed in 9 PK eyes and in 6 LK eyes. The rejection-free graft survival rate was 39.0% in the PK group and 90.3% in the LK group at month 28, with a mean length of 29.6 and 85.0 months, respectively (P<0.001). Chemical warfare victims who initially have mild symptoms ultimately may experience significant ocular involvements requiring surgical intervention. Limbal and corneal abnormalities can be managed best by KLAL and LK, respectively.


beta-Carotene promotes the development of NNK-induced small airway-derived lung adenocarcinoma.
beta-Carotene has shown cancer-preventive effects in preclinical studies while increasing lung cancer mortality in clinical trials. We have shown that beta-carotene stimulates cAMP signalling in vitro. Here, we have tested the hypothesis that beta-carotene promotes the development of pulmonary adenocarcinoma (PAC) in vivo via cAMP signalling. PAC was induced in hamsters with the carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), followed by beta-carotene for 1.5 years. Incidence, multiplicity and size of lung tumours were recorded, and phosphorylated CREB and ERK1/2 in tumour cells were determined by Western blots. Cyclic AMP in blood cells was analysed by immunoassays, retinoids in serum and lungs by HPLC. beta-Carotene increased lung tumour multiplicity, lung tumour size, blood cell cAMP, serum and lung levels of retinoids and induced p-CREB and p-ERK1/2 in lung tumours. Our data suggest that beta-carotene promotes the development of PAC via increased cAMP signalling.


Pituitary adenylate cyclase-activating peptide (PACAP) and its receptors in Mus musculus testis.
To enlighten the involvement of PACAP/receptors system in the control of mammal testis, we investigated the expression of PACAP and the localization of PACAP and its receptors PAC1, VPAC1, and VPAC2 in the testis of Mus musculus. By molecular and immunohistochemical investigations, we highlighted that PACAP and its receptors are widely represented in germ cells of Mus testis, particularly in spermatocytes I, spermatids, and spermatozoa, strongly suggesting their involvement in spermatogenesis process. Moreover, for the first time in the adult mouse testis we highlighted that PACAP is present within Leydig cells, as PACAP receptors, confirming its involvement in the control of steroidogenesis in mouse.


Construction and evaluation of a prognosis lncRNA model for hepatocellular carcinoma.
Current studies indicate that long non-coding RNA (lncRNA) is often abnormally expressed in hepatocellular carcinoma (HCC). We intend to generate a multi-lncRNA signal to improve the prognosis of HCC. By analyzing 12 pairs of HCC and adjacent normal mucosal tissues, 3900 differentially expressed lncrnas were identified as candidate biomarkers for the prognosis of HCC. Then, the 12-lncrna signature was constructed using the LASSO Cox regression method and verified in the TCGA training dataset. Finally, we established a novel 12-lncrna signature that was significantly associated with overall survival (OS) in the training data set. With the use of 12-lncrna markers, patients in the training cohort were divided into high-risk and low-risk groups with significant OV differences (P < .0001). Similar results were consistent in the TCGA verification dataset (P = .046). Multivariate Cox model was used to analyze and construct the risk scores of selected key lncRNA and AJCC stages. The results showed that, compared with AJCC stages, lncRNA-based risk scores were another important factor affecting the OS of patients. We found that risk scores based on lncRNA have a stronger prediction ability than the AJCC stage alone on 4-year OS. For 4-year survival rates, prediction combined with the lncRNA risk score and AJCC stage, model effectiveness (sensitivity and specificity) has reached to 0.750. To further explore the biological processes involved in prognostic lncRNA, all HCC samples in TCGA are divided into two groups according to the median lncRNA risk score, and analyzed the gene enrichment of high expression genes and low expression genes in KEGG data using goana in limma. The results suggest that the genes associated with tumor pathways, such as PI3K-Akt and ECM-receptor interaction, are highly expressed in the high risk group.


Personalized at-home neurofeedback compared to long-acting methylphenidate in children with ADHD: NEWROFEED, a European randomized noninferiority trial.
Neurofeedback is considered a promising intervention for the treatment of attention-deficit hyperactivity disorder (ADHD). NEWROFEED is a prospective, multicentre, randomized (3:2), reference drug-controlled trial in children with ADHD aged between 7 and 13 years. The main objective of NEWROFEED was to demonstrate the noninferiority of personalized at-home neurofeedback (NF) training versus methylphenidate in the treatment of children with ADHD. The NF group (n = 111) underwent eight visits and two treatment phases of 16 to 20 at-home sessions with down-training of the theta/beta ratio (TBR) for children with high TBR and enhancing the sensorimotor rhythm (SMR) for the others. The control group (n = 67) received optimally titrated long-acting methylphenidate. The primary endpoint was the change between baseline and endpoint in the Clinician ADHD-RS-IV total score in the per-protocol population (90 NF/59 controls). US National Institute of Health, ClinicalTrials.gov #NCT02778360. Our study failed to demonstrate noninferiority of NF versus methylphenidate (mean between-group difference 8.09 90% CI [8.09; 10.56]). However, both treatment groups showed significant pre-post improvements in core ADHD symptoms and in a broader range of problems. Reduction in the Clinician ADHD-RS-IV total score between baseline and final visit (D90) was 26.7% (SMD = 0.89) in the NF and 46.9% (SMD = 2.03) in the control group. NF effects increased whereas those of methylphenidate were stable between intermediate and final visit. Based on clinicians' reports, the effects of at-home NF were inferior to those of methylphenidate as a stand-alone treatment.


Validating the Performance of the Modified Early Obstetric Warning System Multivariable Model to Predict Maternal Intensive Care Unit Admission.
To evaluate the performance of the Modified Early Obstetric Warning System (MEOWS) to predict maternal ICU admission in an obstetric population. Case-control study. Two maternity units in Vancouver, Canada, one with ICU facilities, between January 1, 2000, and December 31, 2011. Pregnant or recently delivered (≤6 weeks) women admitted to the hospital for >24 hours. Three control patients were randomly selected per case and matched for year of admission. Retrospective, observational, case-control validation study investigating the physiologic predictors of admission in the 24-hour period preceding either ICU admission >24 hours (cases) or following admission (control patients). Model performance was assessed based on sensitivity, specificity, and predictive values. Forty-six women were admitted to the ICU for >24 hours (0.51/1000 deliveries); the study included 138 randomly selected control patients. There were no maternal deaths in the cohort. MEOWS had high sensitivity (0.96) but low specificity (0.54) for ICU admission >24 hours, whereas ≥1 one red trigger maintained sensitivity (0.96) and improved specificity (0.73). Altering MEOWS trigger parameters may improve the accuracy of MEOWS in predicting ICU admission. Formal modelling of a MEOWS scoring system is required to support evidence-based care.


Vaccination with a genotype 1 modified live vaccine against porcine reproductive and respiratory syndrome virus significantly reduces viremia, viral shedding and transmission of the virus in a quasi-natural experimental model.
The present study assessed the efficacy of vaccination against genotype 1 porcine reproductive and respiratory syndrome virus (PRRSV) in terms of reduction of the transmission. Ninety-eight 3-week-old piglets were divided in two groups: V (n=40) and NV (n=58) that were housed separately. V animals were vaccinated with a commercial genotype 1 PRRSV vaccine while NV were kept as controls. On day 35 post-vaccination, 14 NV pigs were separated and inoculated intranasally with 2 ml of a heterologous genotype 1 PRRSV isolate ("seeder" pigs, SP). The other V and NV animals were distributed in groups of 5 pigs each. Two days later, one SP was introduced into each pen to expose V and NV to PRRSV. Sentinel pigs were allocated in adjacent pens. Follow-up was of 21 days. All NV (30/30) became viremic after contact with SP while only 53% of V pigs were detected so (21/40, p<0.05). Vaccination shortened viremia (12.2±4 versus 3.7±3.4 days in NV and V pigs, respectively, p<0.01). The 50% survival time for becoming infected (Kaplan-Meier) for V was 21 days (CI95%=14.1-27.9) compared to 7 days (CI95%=5.2-8.7) for NV animals (p<0.01). These differences were reflected in the R value as well: 2.78 (CI95%=2.13-3.43) for NV and 0.53 (CI95%=0.19-0.76) for V pigs (p<0.05). All sentinel pigs (10/10) in pens adjacent to NV+SP pens got infected compared to 1/4 sentinel pigs allocated contiguous to a V+SP pen. These data show that vaccination of piglets significantly decrease parameters related to PRRSV transmission.


Using the consolidated framework for implementation research to identify recruitment barriers and targeted strategies for a shared decision-making randomized clinical trial in pediatric sickle cell disease.
Recruitment is often a barrier in clinical trials that include minoritized populations, such as individuals with sickle cell disease. In the United States, the majority of people with sickle cell disease identify as Black or African American. In sickle cell disease, 57% of the United States trials that ended early did so due to low enrollment. Thus, there is a need for interventions that improve trial enrollment in this population. After lower-than-expected recruitment during the first 6 months of the Engaging Parents of Children with Sickle Cell Anemia and their Providers in Shared-Decision-Making for Hydroxyurea trial, a multi-site study for young children with sickle cell disease, we collected data to understand barriers and used the Consolidated Framework for Implementation Research to categorize them and guide the development of targeted strategies. Study staff used screening logs and coordinator and principal investigator calls to identify recruitment barriers that were then mapped onto Consolidated Framework for Implementation Research constructs. Targeted strategies were implemented during Months 7-13. Recruitment and enrollment data were summarized before (Months 1-6) and during the implementation period (Months 7-13). During the first 13 months, 60 caregivers (M = 30.65 years; SD = 6.35) enrolled in the trial. Most caregivers primarily self-identified as female (n = 54, 95%) and African American or Black (n = 51, 90%). Recruitment barriers mapped onto three Consolidated Framework for Implementation Research constructs: (1) Process barriers (i.e. no identified "site champion" and poor recruitment planning at several sites); (2) Inner setting barriers (i.e. limited communication, low relative study priority at several sites); and (3) Outer setting barriers (i.e. poor patient attendance at clinic appointments). Targeted strategies to improve recruitment included (1) principal investigator site visits and retraining on recruitment procedures to address process barriers; (2) increased frequency of communication through all coordinator, site principal investigator, and individual site calls to address inner setting barriers; and (3) development and implementation of no-show procedures for clinic appointments to address outer setting barriers. After implementation of the recruitment strategies, the number of caregivers identified for pre-screening increased from 54 to 164, and enrollment more than tripled from 14 to 46 caregiver participants. Consolidated Framework for Implementation Research constructs guided the development of targeted strategies that increased enrollment. This reflective process reframes recruitment challenges as the responsibility of the research team rather than characterizing minoritized populations as "difficult" or "hard to reach." Future trials including patients with sickle cell disease and minoritized populations may benefit from this approach.


Antibody profiling of bloodstains.
The detection and measurement of antibodies in bloodstains represents a departure from the classical blood grouping used routinely in forensic serology. The antibodies in question are those associated with atopic allergy and infections by micro-organisms and other parasites. The production of these antibodies is not primarily under genetic control but reflects an individual's reaction to his environment. This paper describes the explicit information which may be obtained from an antibody profile, the high discriminating power and other advantages of the method.


Changes in C-reactive protein associated with surgical treatment of mandibular fractures.
C-reactive protein (CRP) levels were determined in 80 patients (67 male, 13 female), each with a single mandibular fracture in either the symphysis, body, or angle region. All patients underwent osteosynthesis (63 with rigid plate fixation following the AO/ASIF principle, 17 with miniplates). C-reactive protein levels were measured on admission and daily during hospitalization. Preoperative CRP levels (mean, 28.5 mg/L) varied according to the time that had elapsed since injury. After surgery, there was always an increase in CRP level. The level reached its maximum (mean, 73.2 mg/L) on the second day after operation. Postoperative CRP levels were significantly influenced by the time between trauma and surgery. Significant differences were also observed with different types of fixation. Fixation with rigid plates was associated with smaller increases in CRP level than was fixation with miniplates. The location of the fracture, associated condylar fractures, the use of maxillomandibular fixation, the presence or absence of a tooth at the fracture site, and the surgical approach did not affect the CRP levels. The significance of CRP measurement for interpretation of postoperative situations is discussed and examples of cases in which there was infection are described.


Medication use and adherence among elderly Medicare beneficiaries with diabetes enrolled in Part D and retiree health plans.
Employer-based retiree drug benefits have long been viewed as the gold standard of drug coverage for elderly Medicare beneficiaries. The question for policy makers is whether beneficiaries enrolled in Part D plans exhibit drug utilization patterns comparable with those seen in retiree plans. To compare utilization patterns for antidiabetic agents, renin-angiotensin-aldosterone system inhibitors, and antihyperlipidemics by elderly Medicare beneficiaries with diabetes enrolled in Medicare prescription drug plans (PDPs) and retiree health plans (RHPs). A random 5% sample (N=45,613) of elderly diabetic patients with continuous 2006 PDP enrollment was selected from Medicare files and compared with a similar sample of elderly RHP enrollees from MarketScan (N=211,919) on any use, duration of therapy, and medication possession ratio for each drug class. Adjusted comparisons were made on samples (N=16,859 each) using propensity score matching. Drug utilization and adherence rates were high in both groups. In propensity score adjusted comparisons, prevalence rates for PDP enrollees were within 2.2% of the level of RHP enrollees for antidiabetic agents and renin-angiotensin-aldosterone system inhibitors, but differed sharply for antihyperlipidemics (61% vs. 69%; P<0.0001). There were no clinically meaningful differences between PDP and RHP enrollees in duration of therapy or medication possession ratio for any drug class. When otherwise similar Medicare beneficiaries with diabetes have drug coverage, the source of benefits has little effect on use and adherence with most (but not all) drugs recommended in diabetes guidelines.


Preparation and properties of a neurotoxin purified from the venom of black widow spider (Latrodectus mactans tredecimguttatus).
A neurotoxin of the venom of the spider Latrodectus mactans tredecimguttatus, has been prepared in a homogeneous form and examined by a variety of techniques. The protein has a molecular weight of 130 000 and its toxicity in mice is about 49 000 LD50 mg pure protein/g body weight. The toxin releases norepinephrine from synaptosomes prepared from rat brain and shows most of the toxic effects of the crude venom preparation.


Long-term renal function in patients undergoing surgical reconstruction for tubercular cicatrized bladder.
We evaluated the long-term renal function in patients after surgical reconstruction for tuberculous contracted bladder (TBC) and determined factors associated with decreased renal function (RF) during follow up. We reviewed the records of 61 patients who underwent augmentation cystoplasty (AC) or orthotopic neobladder (ONB) for TBC between June 1994 and August 2019 in our institute. The estimated glomerular filtration rate (eGFR) was calculated preoperatively at initial presentation, before augmentation and at various intervals during follow up. Renal function decrease was defined as a defined as new-onset stage-3A Chronic kidney disease(CKD) or upstaging of pre-operative CKD stage 3A in follow-up. Multivariable analysis was done to evaluate the association of clinicopathological features and postoperative complications with decreased renal function. We analyzed 39 patients who had a minimum follow-up of 1-year post reconstruction. At a median follow-up of 52 months (IQR 31-103 months), 16/39 patients developed RF decrease. In univariate analyses, initial eGFR, and associated ureteric stricture in contralateral renal unit were significantly associated with new-onset renal insufficiency (p < 0.001 each). On multivariable analysis, only initial presenting eGFR (p < 0.001) was an independent predictor of new-onset renal insufficiency. ROC cut-off levels for eGFR at presentation predicting the primary end point of RF decrease was 45 ml/min. Decreased renal function is noted in most patients during long term follow-up after surgical reconstruction for TBC. After controlling for preoperative and postoperative risk factors, patients with initial presenting GFR < 45 ml/min are at greater risk of a decline in renal function following reconstruction.


Specific activation of the V5 brain area by auditory motion processing: an fMRI study.
Previous neuroimaging studies devoted to auditory motion processing have shown the involvement of a cerebral network encompassing the temporoparietal and premotor areas. Most of these studies were based on a comparison between moving stimuli and static stimuli placed at a single location. However, moving stimuli vary in spatial location, and therefore motion detection can include both spatial localisation and motion processing. In this study, we used fMRI to compare neural processing of moving sounds and static sounds in various spatial locations in blindfolded sighted subjects. The task consisted of simultaneously determining both the nature of a sound stimulus (pure tone or complex sound) and the presence or absence of its movement. When movement was present, subjects had to identify its direction. This comparison of how moving and static stimuli are processed showed the involvement of the parietal lobules, the dorsal and ventral premotor cortex and the planum temporale during auditory motion processing. It also showed the specific recruitment of V5, the visual motion area. These results suggest that the previously proposed network of auditory motion processing is distinct from the network of auditory localisation. In addition, they suggest that the occipital cortex can process non-visual stimuli and that V5 is not restricted to visual processing.


Identification and Potential Value of Candidate Genes in Patients With Non-obstructive Azoospermia.
To explore the candidate genes involved in the pathogenesis of non-obstructive azoospermia (NOA) using bioinformatics analysis and experimental verification. The gene expression profiles (GSE9210) were downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were identified. We performed function enrichment analyses, constructed protein-protein interaction (PPI) network and identified hub genes. Further, the miRNA-hub genes regulatory network was constructed. Finally, the expression level of CEP55 was verified using RT-qPCR and Western blot, and its diagnostic value was analyzed by the receiver operating characteristic (ROC) curve. 626 DEGs were identified, including 11 upregulated and 615 downregulated genes. Function enrichment analyses showed that these DEGs were significantly enriched in spermatogenesis, fertilization, meiotic cell cycle, flagellated sperm motility, sperm capacitation, spermatid nucleus differentiation and male meiotic nuclear differentiation. The top 10 hub genes were identified including CCNB2, BUB1, TOP2A, BIRC5, CENPF, PBK, NCAPG, DLGAP5, NUF2 and CEP55. In the miRNAs prediction, the hsa-miRNA-449a, hsa-miRNA-34c-5p and hsa-miRNA-34b-5p may be implicated in NOA. In the validation stage, the expression level of CEP55 was significantly decreased in patients with NOA compared to patients with OA. ROC analysis showed that CEP55 had a good diagnostic value for NOA and the combination of CEP55, FSH and mean testicular volume enhanced the prediction performance. This study identified key genes associated with NOA and their biological functions. Furthermore, CEP55 might play an important role in the pathogenesis of NOA, which will provide novel insights into the targeting therapy of NOA.


Liposome-mediated gene transfer to lung isografts.
Our objective were to determine the feasibility, efficacy, and safety of in vivo and ex vivo liposome-mediated gene transfer to lung isografts. Fischer rats were divided into three main groups: (1) Nontransplant setting: Liposome-chloramphenicol acetyl transferase cDNA was intravenously injected, and lungs were harvested at different time points: 2, 6, 12, and 24 hours; 2, 5, 8, and 21 days (n = 3). Chloramphenicol acetyl transferase activity was determined in lungs, hearts, livers, and kidneys. The distribution and type of transfected cells were evaluated by in situ hybridization. Lung toxicity was assessed by arterial oxygen tension, histology, and tumor necrosis factor-alpha levels. (2) In vivo graft transfection: Left lungs were transplanted 6 hours, 4 hours, and 15 minutes after intravenous injection and were assessed for chloramphenicol acetyl transferase activity and arterial oxygen tension on postoperative day 2. (3) Ex vivo graft transfection: Grafts were infused ex vivo with either 660 micrograms (n = 3) or 330 micrograms (n = 3) of DNA complexed to liposomes and stored at 10 degrees C for 4 hours. Chloramphenicol acetyl transferase activity was assessed 44 hours after transplantation. Transgene expression was detected in endothelial cells, macrophages, and interstitial cells. Chloramphenicol acetyl transferase activity was present as early as 2 hours, increased significantly between 6 hours and 8 days, and then decreased to minimal levels by 21 days. Chloramphenicol acetyl transferase activity was greatest in donor lungs and hearts and minimal in livers and kidneys. Arterial oxygen tension was normal in treated animals. Inflammation was minimal, and tumor necrosis factor-alpha levels increased only sevenfold in treated animals. In vivo and ex vivo liposome-mediated gene transfer to lung isografts allows significant transgene expression with minimal effects on graft function.


Aortoesophageal fistula secondary to placement of an esophageal stent: emergent treatment with cyanoacrylate and endovascular stent graft.
We report on N-butyl 2-cyanoacrylate embolization and subsequent endovascular stent graft placement for the treatment of an aortoesophageal fistula secondary to placement of an esophageal stent. A 53-year-old man with lung cancer was admitted with massive hematemesis due to the formation of an aortoesophageal fistula 20 days after esophageal stent placement. Injection of N-butyl 2-cyanoacrylate into the aortoesophageal fistula was performed as emergent treatment for this hemodynamically unstable condition, and an endovascular stent graft was subsequently placed via the right femoral artery. The patient was well without hematemesis until he died of pneumonia 45 days later. Cyanoacrylate embolization and subsequent endovascular stent graft placement for the treatment of massive hemorrhage caused by an aortoesophageal fistula is a prompt, effective method and can be an alternative to surgical repair.


Disease monitoring programs of rare genetic diseases: transparent data sharing between academic and commercial stakeholders.
It has recently been suggested that registries for rare neuromuscular diseases should be formed and governed exclusively by physicians and patients in an effort to limit conflicts of interest. Enacting such an approach would not only be challenging logistically and financially, but it would also exclude the involvement of sponsors, who are an integral component of drug development within the current compliance framework. Therefore, as an alternative to traditional registries, we propose the use of a better collaborative model for post-marketing follow-up that includes all stakeholders. We developed the concept of Disease Monitoring Programs (DMPs), which are designed to monitor disease manifestations over a 10-year period whether on a sponsored drug or not, and ensure consistent collection, ownership sharing and governance of data.


Effects of L-carnitine administration on VO2max and the aerobic-anaerobic threshold in normoxia and acute hypoxia.
Seven healthy young male adults were subjected to a total of 56 tests to ascertain the effects of L-carnitine (L-C) and a placebo (P) on ventilation, O2 intake (VO2), CO2 output, heart rate, blood pressure and serum lactic acid, non-esterified fatty acid, glycerol and glucose during strenuous and aerobic/anaerobic threshold-level treadmill exercise. The tests were made in conditions of normoxia (O2 = 20.9%) and hypoxia (O2 = 13.0%, equivalent to 3,500 m above sea level). The only clear difference was in the respiratory quotient (RQ = 0.883, SD 0.025 vs 0.904, SD 0.035) after L-C and P administration respectively (P less than 0.01), under normal oxygenation and 0.861, SD 0.052 following L-C vs 0.926, SD 0.040 after P (P less than 0.01) in acute hypoxia at VO2 levels around the anaerobic threshold. The lower RQ values of the L-C-treated subjects during hypoxia indicate a lower rate of carbohydrate transformation.


Towards markerless surgical tool and hand pose estimation.
: Tracking of tools and surgical activity is becoming more and more important in the context of computer assisted surgery. In this work, we present a data generation framework, dataset and baseline methods to facilitate further research in the direction of markerless hand and instrument pose estimation in realistic surgical scenarios. : We developed a rendering pipeline to create inexpensive and realistic synthetic data for model pretraining. Subsequently, we propose a pipeline to capture and label real data with hand and object pose ground truth in an experimental setup to gather high-quality real data. We furthermore present three state-of-the-art RGB-based pose estimation baselines. : We evaluate three baseline models on the proposed datasets. The best performing baseline achieves an average tool 3D vertex error of 16.7 mm on synthetic data as well as 13.8 mm on real data which is comparable to the state-of-the art in RGB-based hand/object pose estimation. : To the best of our knowledge, we propose the first synthetic and real data generation pipelines to generate hand and object pose labels for open surgery. We present three baseline models for RGB based object and object/hand pose estimation based on RGB frames. Our realistic synthetic data generation pipeline may contribute to overcome the data bottleneck in the surgical domain and can easily be transferred to other medical applications.


Tumor necrosis factor-alpha and interferon-gamma induce inflammasome-mediated corneal endothelial cell death.
Chronic corneal endothelial cell (CEC) loss results in corneal edema and vision loss in conditions such as pseudophakic bullous keratopathy (PBK), Fuchs' dystrophy, and corneal graft failure. Low CEC density has been associated with an elevation of intraocular pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α and interferon (INF)-γ. These cytokines are capable of triggering pyroptosis, a programmed cell death mechanism mediated by the inflammasome, prompting the activation of the pro-inflammatory cytokine interleukin (IL)-1β, the perpetuation of inflammation, and subsequent damage of corneal endothelial tissue. Therefore, the purpose of this study was to determine the deleterious contribution of the inflammasome and pyroptosis to CEC loss. CECs from human donor corneas were treated ex vivo with TNF-α and IFN-γ for 48 h. Levels of caspase-1 and IL-1β were then assayed by ELISA, and the expression of caspase-1 and gasdermin-D (GSDM-D) were confirmed by immunofluorescence. Endothelial cell damage was analyzed by a lactate dehydrogenase (LDH) release assay, and oxidative stress was determined by measuring the levels of reactive oxygen species (ROS) in the culture media. Inflammasome activation and oxidative stress were elevated in CECs following exposure to TNF-α and IFN-γ, which resulted in cell death by pyroptosis as determined by LDH release which was inhibited by the caspase-1 inhibitor Ac-YVAD-cmk. CEC death is induced by the pro-inflammatory cytokines TNF-α and IFN-γ, which contribute to inflammasome activation. Moreover, the inflammasome is a promising therapeutic target for the treatment of chronic CEC loss.


Natterin an aerolysin-like fish toxin drives IL-1β-dependent neutrophilic inflammation mediated by caspase-1 and caspase-11 activated by the inflammasome sensor NLRP6.
Natterin is an aerolysin-like pore-forming toxin responsible for the toxic effects of the venom of the medically significant fish Thalassophryne nattereri. Using a combination of pharmacologic and genetic loss-of-function approaches we conduct a systematic investigation of the regulatory mechanisms that control Natterin-induced neutrophilic inflammation in the peritonitis model. Our data confirmed the capacity of Natterin to induce a strong and sustained neutrophilic inflammation leading to systemic inflammatory lung infiltration and revealed overlapping regulatory paths in its control. We found that Natterin induced the extracellular release of mature IL-1β and the sustained production of IL-33 by bronchial epithelial cells. We confirmed the dependence of both ST2/IL-33 and IL-17A/IL-17RA signaling on the local and systemic neutrophils migration, as well as the crucial role of IL-1α, caspase-1 and caspase-11 for neutrophilic inflammation. The inflammation triggered by Natterin was a gasdermin-D-dependent inflammasome process, despite the cells did not die by pyroptosis. Finally, neutrophilic inflammation was mediated by non-canonical NLRP6 and NLRC4 adaptors through ASC interaction, independent of NLRP3. Our data highlight that the inflammatory process dependent on non-canonical inflammasome activation can be a target for pharmacological intervention in accidents by T. nattereri, which does not have adequate specific therapy.


The effect of breed and age on the gloss of chicken eggshells.
The monitoring of eggshell quality is important mainly in terms of production economy. Eggshell appearance is one of the most characteristics, influencing the purchasing behavior of consumers. Besides numerous eggshell appearance quality (color, shape, etc.), gloss is an important trait to reflect the eggshell appearance. In this study, 2 experiments were conducted to investigate the effect of breed and age on the gloss of eggshells. In experiment 1, we compared the eggshell gloss of 7 chicken breeds. In experiment 2, 105 Wanan (WA) chickens were raised, and 1 egg was collected from each individual at 26, 32, 40, and 50 wks of age. Eggshell gloss, color (L*, a*, b*), cuticle coverage (ΔE*ab), and thickness were measured. The results of experiment 1 showed that the average gloss values were highly variable among different breeds, and the highest was found in WA (gloss unit [GU] = 8.12), almost 2.5 folds as many as the lowest in Rhode Island Red (GU = 3.23). Also, the eggshell gloss of the local chicken breeds was significantly higher than the highly selected lines of egg-type chicken breeds (P < 0.001). In experiment 2, the results showed that gloss ranged from 9.08 GU to 12.12 GU with a variation of 28.38 to 39.71%. It fluctuated with the increasing age of hens and had the peak value at 26 wk. But, the correlation analysis between eggshell gloss and other eggshell quality traits were very low (-0.07 to 0.25). This study laid a foundation for improving the uniformity and intensity of eggshell gloss for breeders.


Adolescent archaics or adult moderns? Le Moustier 1 as a model for estimating the age at death of fragmentary supraorbital fossils in the modern human origins debate.
This study documents and examines selected implications of the adolescent supraorbital anatomy of the Le Moustier 1 Neandertal. Le Moustier's supraorbital morphology conforms to that expected of an adolescent Neandertal but indicates that significant development of the adult Neandertal torus occurs late in ontogeny. As the best preserved adolescent from the Late Pleistocene, Le Moustier 1's anatomy is used to help distinguish adolescent from adult anatomy in two cases of fragmentary supraorbital fossils, the Vindija late Neandertals and KRM 16425 from Klasies River Mouth (South Africa). It has been suggested that the modern-like aspects of the Vindija and Klasies supraorbital fossils are a function of developmental age rather than evolution. Although Le Moustier 1's anatomy does indicate that two of the Vindija fossils are adolescent; these two fossils have already been excluded from studies that demonstrate transitional aspects of the Vindija adult supraorbitals. Results of an analysis of KRM 16425 in light of Le Moustier 1 are more ambiguous. KRM 16425 is clearly not a Neandertal, but its morphology suggests that it may be an adolescent form of such late archaic Africans like Florisbad or Ngaloba. Both the Vindija and Klasies River Mouth cases highlight the need to be wary of confusing adolescent anatomy with modernity.


Preparation of protein loaded poly(D,L-lactide-co-glycolide) microparticles for the antigen delivery to dendritic cells using a static micromixer.
The cellular immune response against tumors, viruses, or intracellular bacteria requires adequate antigen delivery to professional phagocytes, their processing and the presentation of antigenic peptides to T-cells. Biodegradable microparticles to enhance antigen phagocytosis and the response of cytotoxic lymphocytes have been proposed. The aim of the present study was to formulate poly(lactide-co-glycolide) (PLGA) microparticles using a w/o/w solvent evaporation procedure in order to obtain suitable vehicles for vaccination. Bovine serum albumin bearing fluorescein isothiocyanate (FITC-BSA) was used as a model antigen. For microparticle preparation a static micromixer was employed. Microparticles of 2-3 microm can be produced with good reproducibility by applying high flow rates at the micromixer. Microparticles with a smooth surface and only one pore were observed using scanning electron microscopy (SEM). Confocal laser scanning microscopy (CLSM) allowed localisation of the FITC-BSA near the surface of the microparticle. Microencapsulation of FITC-BSA did not altered the polymer characteristics, as determined by measuring the glass transition temperature. Additionally we could determine residual methylene chloride, employed as solvent in microparticle preparation, to be less than 1/1000 of the USP and Ph. Eur. limit. The microparticles described herein were able to deliver the model antigen to human dendritic cells (DC).


Evaluation of the sensitivity and specificity of diagnostic criteria for sepsis in dogs.
To evaluate the ability of various individual criteria and grouped criteria to diagnose sepsis in dogs. Prospective acquisition of clinical data. Client-owned dogs; 30 septic and 320 nonseptic. Rectal temperature, heart rate, respiratory rate, white blood cell (WBC) count with percent bands, platelet count, and serum glucose concentration were obtained on day 0. True sepsis was determined on days 0 to 3 according to the following criteria: (1) histological, microbiological, and/or gross confirmation of infection, and (2) systemic illness caused by infection. Data were analyzed by the Mann-Whitney U test and multiple logistic regression. Sensitivity and specificity were calculated. The mean temperature, heart rate, WBC count, and percent bands were greater, whereas the mean platelet count was less in septic compared with nonseptic dogs. There was no difference in respiratory rate or glucose concentration. WBC/bands were the best individual criterion for the diagnosis of sepsis (sensitivity 87%; specificity 69%). The sensitivity and specificity of the grouped criteria (> or = two of four; temperature, heart rate, respiratory rate, WBC) varied according to ranges of normal used. Multiple logistic regression resulted in little improvement in the sensitivity/specificity of these diagnostic criteria for the diagnosis of sepsis. These criteria are useful for the diagnosis of sepsis when limits are used that result in a high sensitivity (e.g., 97%). The high sensitivity was associated with a low false-negative and a high false-positive rate; sepsis was overdiagnosed with these grouped criteria. These criteria may be used for a sensitive, but nonspecific, diagnosis of sepsis in dogs.


Safety of D-dimer testing as a stand-alone test for the exclusion of deep vein thrombosis as compared with other strategies.
Essentials The aim of deep vein thrombosis (DVT) diagnostic work-up is to maximize both safety and efficiency. We explored whether D-dimer is safe and efficient as a stand-alone test to exclude DVT. Our findings suggest it is a safe, efficient and simplified diagnostic strategy. The safety of age-adjusted D-dimer as a stand-alone test requires further investigation. SUMMARY: Background Several strategies for safely excluding deep vein thrombosis (DVT) while limiting the number of imaging tests have been explored. Objectives To determine whether D-dimer testing could safely and efficiently exclude DVT as a stand-alone test, and evaluate its performance as compared with strategies that incorporate the Wells score and age-adjusted D-dimer. Patients/Methods We included consecutive outpatients referred with suspected DVT to the Emergency Department at Østfold Hospital, Norway. STA-Liatest D-Di PLUS D-dimer was analyzed for all patients. Patients with a D-dimer level of ≥ 0.5 μg mL-1 were referred for compression ultrasonography (CUS). In patients with a D-dimer level of < 0.5 μg mL-1 , no further testing was performed and anticoagulation was withheld. Patients were followed for 3 months for venous thromboembolism (VTE). Results Of the 913 included patients, 298 (33%) had a negative D-dimer result. One hundred and seventy-three patients (18.9%) were diagnosed with DVT at baseline. One of 298 patients had DVT despite having a negative D-dimer result, resulting in a failure rate of 0.3% (95% confidence interval [CI] 0.1-1.9%). Adding the modified Wells score would have yielded a failure rate of 0.0% (95% CI 0.0-1.8%) while necessitating 87 more CUS examinations. Age-adjusted D-dimer as a stand-alone test would have necessitated 80 fewer CUS examinations than fixed D-dimer as a stand-alone test, at the cost of a failure rate of 1.6% (95% CI 0.7-3.4%). Conclusions This outcome study shows that a negative high-sensitivity D-dimer result safely excludes DVT in an outpatient population, and necessitates fewer CUS than if used in combination with Wells score. The safety of stand-alone age-adjusted D-dimer needs further assessment in prospective outcome studies.


Serum hsa-circ-0025244 as a biomarker in Chinese occupational mercury-exposed population and mediate apoptosis through JNK/p38 MAPK signaling pathway.
circRNAs have been recognized as biomarkers of numerous diseases. We would like to explore the expression pattern and molecular mechanisms of circRNAs in the Chinese occupational mercury-exposed population. The workers from a thermometer manufacturing plant and lamp factory in Jiangsu province of China were recruited in 2016. Blood samples were collected from the subjects with chronic mercury poisoning group, mercury absorption group, and the healthy controls. The differentially expressed circRNAs (DECRs) between the three groups were screened from serum samples using a circRNA microarray. The significant DECRs were validated by qRT-PCR, and their respective diagnostic values for mercury poisoning and mercury absorption were analyzed by receiver operating characteristic (ROC) curves. For in vitro experiments, 293T cells were treated with different doses of HgCl2 to determine the half-lethal concentration. The cells were transfected with the siRNA construct or expression plasmid of circRNA. The expression levels of JNK, p38, and caspase family proteins were analyzed by Western blotting. hsa_circ_0025244 was up-regulated in the mercury poisoning and absorption groups compared to the control group (P < 0.05), and positively correlated with the urine mercury levels (P < 0.05). The area under the ROC curve (AUC) of hsa_circ_0025244 for diagnosing occupational mercury poisoning was 0.748, indicating moderate accuracy (P < 0.001). Moreover, the diagnostic accuracy of occupational mercury absorption was high (P < 0.001) with an AUC of 0.918. Knockdown of hsa_circ_0025244 in 293T cells significantly reduced the expression levels of JNK/p38, and caspase family proteins compared to that in the control cells (P < 0.01), and its overexpression led to opposite effects (P < 0.05). hsa_circ_0025244 is a potential biomarker for mercury exposure and mediates mercury-induced apoptosis in 293T cells by activating the JNK/p38 MAPK signaling pathway.


A reservoir of foraging decision variables in the mouse brain.
In any given situation, the environment can be parsed in different ways to yield decision variables (DVs) defining strategies useful for different tasks. It is generally presumed that the brain only computes a single DV defining the current behavioral strategy. Here to test this assumption, we recorded neural ensembles in the frontal cortex of mice performing a foraging task admitting multiple DVs. Methods developed to uncover the currently employed DV revealed the use of multiple strategies and occasional switches in strategy within sessions. Optogenetic manipulations showed that the secondary motor cortex (M2) is needed for mice to use the different DVs in the task. Surprisingly, we found that regardless of which DV best explained the current behavior, M2 activity concurrently encoded a full basis set of computations defining a reservoir of DVs appropriate for alternative tasks. This form of neural multiplexing may confer considerable advantages for learning and adaptive behavior.


Effectiveness of Vegetated Drainage Ditches for Domestic Sewage Effluent Mitigation.
Plant species have an important role in eco-ditches; however, the Michaelis-Menten kinetic parameters of nutrient uptake, growth rate and purification efficiency of ditch plants and their influences on domestic sewage treatment efficiency are still unclear. Growth rates of all nine species, but especially Lemna gibba, Cladophora and Myriophyllum verticillatum were best in undiluted domestic sewage as opposed to a mixture of domestic sewage. Performance of species to accumulate nutrients was not only species-specific, but was also affected by both sewage treatments. Removal efficiency of nutrients was dependent on both plant species and treatment. Uptake kinetic parameters were significantly affected by both nutrient form and plant species. The maximum uptake rate (Vmax) of NH4-N was higher than NO3-N. Similarly, Km values for NH4-N were greater than NO3-N. These results could be used to identify plants for sewage treatment efficiency and enhance water quality in eco-ditch treatment systems.


Screening for Down syndrome with the femur length/biparietal diameter ratio: a new twist of the data.
The purpose of this study was to determine the value of discordant morphometric measurements as identifiers of Down syndrome by evaluating the relationship of biparietal diameter, femur length, biparietal diameter/femur length ratio, and cephalic index between a group of fetuses with trisomy 21 and a control population. Biometric measurements from 48 fetuses with trisomy were reviewed and compared with 107 normal fetuses of similar gestational age. Data were analyzed in 2-week gestational age intervals to determine the effect of gestational age on ultrasonographic detection of Down syndrome. Outcome measures were subject to least-squares linear regression and the t test for analysis. A positive relationship between abnormal morphometric measurements and fetuses with Down syndrome was detected but only during specific weeks of pregnancy. Although it appears that biometric measurements may be useful for Down syndrome, further study is needed before its widespread introduction into clinical practice.


Per and poly-fluoroalkyl substances (PFAS) as a contaminant of emerging concern in surface water: A transboundary review of their occurrences and toxicity effects.
Per and poly-fluoroalkyl substances (PFAS) have been recognized as contaminants of emerging concerns by the United States Environmental Protection Agency (US EPA) due to their environmental impact. Several advisory guidelines were proposed worldwide aimed at limiting their occurrences in the aquatic environments, especially for perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA). This review paper aims to provide a holistic review in the emerging area of PFAS research by summarizing the spatiotemporal variations in PFAS concentrations in surface water systems globally, highlighting the possible trends of occurrences of PFAS, and presenting potential human health impacts as a result of PFAS exposure through surface water matrices. From the data analysis in this study, occurrences of PFOA and PFOS in many surface water matrices were observed to be several folds higher than the US EPA health advisory level of 70 ng/L for lifetime exposure from drinking water. Direct discharge and atmospheric deposition were identified as primary sources of PFAS in surface water and cryosphere, respectively. While global efforts focused on limiting usages of long-chain PFAS such as PFOS and PFOA, the practices of using short-chain PFAS such as perfluorobutanoic acid (PFBA) and perfluorobutane sulfonic acid (PFBS) and PFAS alternatives increased substantially. These compounds are also potentially associated with adverse impacts on human health, animals and biota.


Regulation of prothoracicotropic hormone release by juvenile hormone in the penultimate and last instar larvae of Mamestra brassicae.
The penultimate instar larvae of Mamestra brassicae have one ecdysteroid peak 2 days after ecdysis which induces larval ecdysis. Allatectomy deleted this peak and caused a larger peak 7 days later just before precocious pupation, in a pattern similar to the ecdysteroid titer of the last instar. Application of juvenile hormone (JH) immediately after allatectomy restored the normal larval pattern and larval ecdysis. The brain of the normal Day 1 or 2 penultimate instar larva had higher prothoracicotropic hormone (PTTH) activity than that of the allatectomized larva. Therefore, JH in the penultimate instar activates the brain to secrete and/or produce PTTH. Allatectomy of last instar larvae prolonged the period from gut purge to pupation by causing a delay of the ecdysteroid peak necessary for pupation. Application of JH restored the normal timing of ecdysteroid release. Thus, JH is important for the occurrence of the ecdysteroid surge on Day 7, since its absence delayed this surge by a day.


Insulin resistance and the metabolic syndrome in obese French children.
To estimate the frequency of the metabolic syndrome (MS) and of the insulin resistance syndrome (IRS) in overweight or obese French children and to determine the risk factors. A total of 308 overweight and obese children [166 girls, 142 boys, aged 7-17 years; median body mass index (BMI) 4.7 standard deviation (SD) (Q1-Q3: 3.9-5.8) adjusted for age and sex] were included. The frequency of the MS was assessed with the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III) criteria and the frequency of the IRS with World Health Organization (WHO) criteria. The overall frequency of MS and IRS was 15.9% and 42.5%, respectively. The most common component, after abdominal obesity (95.8%) and IR (71.8%), was elevated systolic blood pressure (28.6%). The frequency of glucose tolerance disorders was low (3.6%). The frequency of MS was independently influenced by homeostatic model assessment (HOMA) (P = 0.06) and waist-to-hip ratio (P = 0.09), whereas the frequency of IRS was influenced by adiposity (degree of obesity: P = 0.02; waist-to-hip ratio: P = 0.05), puberty (P = 0.05) and mother's BMI (P = 0.01). Ethnicity had no effect on either MS or IRS. Metabolic complications and IR are frequent in overweight and obese children whereas the frequency of glucose tolerance disorders is very low. IRS is more prevalent than MS, indicating a major role of IR, which could precede the other metabolic complications in obese children. IRS is a relevant marker for the risk of type 2 diabetes (T2D) and cardiovascular complications in obese European children.


The effect of increased hip flexion using stirrups on lower-extremity venous flow: a prospective observational study.
Patient positioning during surgeries for colorectal cancer may represent an unrecognized risk factor for deep venous thrombosis. Twelve healthy control patients were positioned supine with knee flexion at 90°. Duplex ultrasound examined common femoral vein (CFV) and proximal femoral vein diameter, peak systolic velocity, and volume flow with hip flexion at 0°, 30°, 60°, and 90°. Data were analyzed using the paired t test. In the CFV, hip flexion to 90° was associated with a significant increase in mean volume flow when compared with hip flexion at 0° (.59 vs .36 L/min; P = .05) and 30° (.59 vs .35 L/min; P = .038). In both the CFV and proximal femoral vein, increased hip flexion was associated with significantly reduced vessel diameter and increased peak systolic velocity. Intraoperative positioning of the lower extremities represents a modifiable risk factor for deep venous thrombosis. When stirrups are used, hip flexion of 90° maximizes venous drainage from the legs.


The role of lipoproteins in the degradation of platelet-activating factor.
Measuring both platelet-stimulating activity and liberation of acetate, the capacity of serum and individual lipoproteins to degrade the platelet-activating factor (PAF) was studied. The highest degrading effect relative to the protein content was found in very low density lipoproteins (VLDL) and in low density lipoproteins (LDL). The effect is about 10- and 100-fold higher than that of high density lipoproteins (HDL) and serum, respectively. In lipoprotein deficient serum (LPDS) less than 5% of serum activity is detectable. Considerable individual variations are observed measuring the degradation of PAF under standard conditions in plasma from 37 healthy volunteers. Moreover, this activity is shown to correlate strongly with the plasma concentration of LDL. On the other hand, a significant negative correlation was found between the PAF-degrading capacity and the plasma concentration of HDL. In contrast, the PAF-degradation is unrelated to the concentration of plasma triglycerides. The results point to a possible role of plasma lipoproteins in regulating the degradation of PAF released into the circulation.


New mechanism of transforming growth factor-beta signaling in hepatoma: Dramatic up-regulation of tumor initiating cells and epidermal growth factor receptor expression.
Transforming growth factor-beta (TGF-beta) has dual activity in tumor cells. We studied the effect of TGF-beta on tumor-initiating cells (TICs), which are similar in self-renewal and differentiation features to normal adult stem cells. We used side population (SP) cells that exclude DNA binding dye Hoechst 33342 to obtain TICs, studied the differences in the kinetics of the SP cell response to TGF-beta treatment between hepatic tumor cell lines, and performed gene analysis. SP cells from all cell lines have higher proliferative ability compared to non-SP cells and they are drug resistant. TGF-beta treatment increased the percentage of SP cells (%SP) and the survival rate; chemotherapeutic drug resistance developed only in K-251 SP cells. Gene analysis showed that TGF-beta up-regulated epidermal growth factor receptor (EGFR) only in K-251 cells. There were no EGFR mutations in K-251, which had been reported in lung cancer. Knockdown of Smad4 using the small-interfering RNA technique in K-251 cells inhibited EGFR overexpression and significantly decreased the %SP. In contrast, the JNK inhibitor had little effect on EGFR expression or the %SP. TGF-beta treatment of K-251 cells causes tumor progression and the anti-cancer drug resistant phenotype by increasing SP.


Calcium inhibition of cardiac adenylyl cyclase. Evidence for two distinct sites of inhibition.
Increasing the free calcium concentration from 10(-8) M to 10(-4) M inhibited cardiac sarcolemmal adenylyl cyclase activated by the addition of 5 X 10(-4) M forskolin or 1 X 10(-4) M GTP or Gpp(NH)p. The calcium inhibition curve in the presence of all three activators was shallow and best fit by a two site model of high affinity (less than 1.0 microM) and low affinity (greater than 0.1 mM). Gpp(NH)p appeared to decrease the sensitivity of adenylyl cyclase to inhibition by calcium at the high affinity site. Similar inhibition constants were obtained with each of the activators. Calmodulin content of native freeze-thaw vesicles was 76.2 +/- 14.2 ng/mg. Treatment of the vesicles with 1 mM EGTA to remove calmodulin significantly reduced calmodulin content to 19.7 +/- 1.35 ng/mg. This treatment had no significant effect on the calcium inhibition profile. Increasing free calcium to 3 X 10(-6) M was shown to have no effect on the EC50 estimated for either Gpp(NH)p or forskolin but did slightly increase the EC50 estimated for Mg2+ in the presence of maximal concentrations of either activator. Nevertheless, maximally stimulating concentrations of Mg2+ were unable to overcome calcium inhibition. Pretreatment of sarcolemmal membranes with pertussis toxin was shown to have no significant effect on calcium inhibition of adenylyl cyclase. The results suggest that the overall inhibitory action of calcium was most likely calmodulin independent and involved a direct interaction with the catalytic subunit at two distinct sites of high and low affinity. At the low affinity site calcium most likely competes with Mg2+ for an allosteric divalent cation binding site.(ABSTRACT TRUNCATED AT 250 WORDS)


Sensitivity reaction to the cinnamonaldehyde component of toothpaste.
Sixteen patients developed a variety of oral lesions following a change in the toothpaste they used. Mucosal biopsy demonstrated features consistent with application of a topical medicament and patch testing, towards the constituents of the toothpastes, indicated the flavouring agent cinnamonaldehyde as being the likely responsible agent. Avoidance of the implicated toothpastes resulted in a considerable improvement in clinical signs and symptoms, whereas rechallenge in ten patients resulted in recurrence of symptoms in eight patients. It is concluded that sensitivity to the cinnamonaldehyde constituent of certain toothpastes, although uncommon, should be considered as a possible aetiological factor in patients presenting with non-specific oral changes.


Effects of morphine administration on catecholamine levels in rat brain; specific reduction of epinephrine concentration in hypothalamus.
The effects of morphine administration on concentrations of epinephrine, norepinephrine and dopamine were examined in the rat brain. Morphine injection reduced the epinephrine level only in the hypothalamus, while the norepinephrine level was reduced in the hypothalamus, medulla, and locus coeruleus. The dopamine concentration was elevated in all regions examined. These changes were blocked by administration of naloxone. Repeated injection of morphine for 14 days did not affect any catecholamine level. In naloxone-induced withdrawal, epinephrine was most markedly depleted in hypothalamus. These observations suggest that the epinephrine level in hypothalamus is affected by morphine acting on opioid receptors.


Effects of on-campus and off-campus smartphone overdependence prevention programs among university students.
To evaluate effects of self-determination theory-based on-campus and off-campus prevention programs on smartphone overdependence among university students. This was a pre-posttest quasi-experimental study with a nonequivalent control group. Seventy-eight students were recruited as participants. They were allowed to choose either an experiment group or a control group (CG). On-campus smartphone overdependence prevention program was provided to participants in experimental group 1 (EG1), while on-campus program combined with off-campus prevention camp was provided to those in experimental group 2 (EG2). Instruments used in this study included a smartphone overdependence self-diagnosis scale, a basic psychological needs scale, and a self-regulation ability scale. Data collection was performed at baseline, immediately after intervention, at one month and three months after intervention. Data were analyzed using mixed analysis of covariance (ANCOVA). Focus group interview (FGI) was performed for qualitative evaluation. After the intervention, smartphone overdependence and basic psychological needs exhibited significant interactions between group and time. Smartphone overdependence scores decreased in EG1 and EG2, but increased in CG (F = 4.56, p = .001). Basic psychological needs improved in EG1 and EG2, but deteriorated in CG (F = 5.04, p = .009). FGIs reveaaled that participants strived to control their smartphone usage through individual efforts and by interacting with new friends in college even after completing the program. In this study, on-campus only program and combined intervention of on- and off-campus programs were both effective in maintaining and managing smartphone use. However, participants perceived that the off-campus program provided an opportunity to apply the theory learned on-campus to the real world.


Anhedonia and suicidal ideation in young people with early psychosis: Further findings from the 2-year follow-up of the ReARMS program.
Hedonic deficits have been extensively studied in schizophrenia, but little is known about their association with suicidal ideation in early psychosis. The aim of this research was to examine the relationship between anhedonia and suicidal thoughts across a 2-year follow-up period in people with First Episode Psychosis (FEP) and at Ultra High Risk (UHR) of psychosis. Ninty-six UHR and 146 FEP, aged 13-35 years, completed the Comprehensive Assessment of At-Risk Mental States (CAARMS) and the Beck Depression Inventory-II (BDI-II). The BDI-II "Anhedonia" subscale score to assess anhedonia and the CAARMS "Depression" item 7.2 subscore to measure depression were used across the 2 years of follow-up. Hierarchical regression analyses were performed. No difference in anhedonia scores between FEP and UHR individuals was found. In the FEP group, a significant enduring association between anhedonia and suicidal ideation was found at baseline and across the follow-up, independent of clinical depression. In the UHR subgroup, the enduring relationship between anhedonia and suicidal thoughts were not completely independent from depression severity. Anhedonia is relevant in predicting suicidal ideation in early psychosis. Specific pharmacological and/or psychosocial interventions on anhedonia within specialized EIP program could reduce suicide risk overtime.


Real-life evaluation of a COVID-19 rapid antigen detection test in hospitalized children.
Rapid antigen detection (RAD) tests for the detection of SARS-CoV-2 are simpler, faster, and less expensive than the reverse-transcription polymerase chain reaction (RT-PCR) that is currently considered the gold standard for the diagnosis of coronavirus disease 2019 (COVID-19). The objective of this study was to determine the performance of the PANBIO COVID-19 Ag RAD (Abbott) test, a lateral flow immunoassay that detects the nucleocapsid protein, using as a reference RT-PCR method the Cobas®8800 System (Roche Diagnostics). This prospective study was conducted in a tertiary Children's Hospital and included individuals aged ≤16 years with COVID-19-related symptoms or epidemiological criteria for COVID-19. Two nasopharyngeal samples were collected to perform the PANBIO RAD test and RT-PCR. Of 744 children included, 51 (6.86%) had a positive RT-PCR result. The RAD test detected 42 of 51 PCR-positive children while there were no false-positive results. The overall sensitivity and specificity were 82.35% (95% CI, 71.9%-92.8%) and 100%, respectively. Sensitivity was >95% in symptomatic children. The assay performed poorly in asymptomatically infected children. In agreement with previous studies in adults, the PANBIO RAD test can be useful in screening for COVID-19 in children admitted with symptoms suggestive of the disease, especially in the first days of the illness.


Interdisciplinary care of pediatric oncology patients: A survey of clinicians in Central America and the Caribbean.
Quality cancer care depends on interdisciplinary communication. This study explored the communication practices of interdisciplinary clinicians, the types of healthcare services for which they engage in interdisciplinary collaboration, and the association between interdisciplinary care and perceived quality of care, as well as job satisfaction. We conducted a survey of interdisciplinary clinicians from cancer centers in Guatemala, Honduras, Panama, El Salvador, and Haiti. The survey included 68 items including previously validated tools and novel questions. Total 174 interdisciplinary clinicians completed the survey: nurses (n = 60), medical subspecialists (n = 35), oncologists (n = 22), psychosocial providers (n = 20), surgeons (n = 12), pathologists (n = 9), radiologists (n = 9), and radiation oncologists (n = 5). Oncologists reported daily communication with nurses (95%) and other oncologists (91%). While 90% of nurses reported daily communication with other nurses, only 66% reported daily communication with oncologists, and more than 50% of nurses reported never talking to pathologists, radiologists, radiation oncologists, or surgeons. Most clinicians described interdisciplinary establishment of cancer treatment goals and prognosis (84%), patient preferences (81%), and determination of first treatment modality (80%). Clinicians who described more interdisciplinary collaboration had higher job satisfaction (p = .04) and perceived a higher level of overall quality of care (p = .004). Clinicians in these limited resource settings describe strong interdisciplinary collaboration contributing to higher job satisfaction and perceived quality of care. However, nurses in these settings reported more limited interdisciplinary communication and care. Additional studies are necessary to further define clinical roles on interdisciplinary care teams and their associations with patient outcomes.


Osteopontin: a potential biomarker for heart failure and reverse remodeling after left ventricular assist device support.
Left ventricular assist device (LVAD) support in end-stage heart failure (HF) leads to recovery of the patient's condition, size reduction of cardiomyocytes, and also volume reduction and change in the composition of the extracellular matrix (ECM). Myocardial expression of ECM osteopontin (OPN) protein increases with the severity of HF. We analyzed whether OPN messenger RNA expression in heart tissue and/or OPN protein in plasma are associated with reverse remodeling during LVAD support. Plasma and heart tissue specimens of 22 end-stage HF patients before and after LVAD implantation and subsequent heart transplantation (HTx) were used to determine the concentrations of OPN protein (EIA) and OPN messenger RNA (mRNA) by quantitative polymerase chain reaction. Immunohistochemistry (IHC) and in situ hybridization (ISH) were performed to locate OPN protein and mRNA. The high OPN protein levels in plasma of HF patients did not differ significantly before and after LVAD support in ischemic heart disease (IHD) (pre-LVAD 167 ± 32 ng/ml; post-LVAD 165 ± 28 ng/ml) and in dilated cardiomyopathy (DCM) (pre-LVAD 99 ± 12 ng/ml; post-LVAD (142 ± 6 ng/ml). The OPN plasma levels after HTx decreased to control levels (IHD, 48 ± 6; DCM, 40 ± 5; control, 31 ± 3 ng/ml). In contrast, expression of OPN mRNA in heart biopsy specimens decreased significantly after LVAD support (the relative quantity decreased > 90% in IHD and 50% in DCM). ISH and IHC revealed that OPN was present in cardiomyocytes and in the ECM. Levels of OPN mRNA in the myocardium of HF patients showed a significant decrease after LVAD support but OPN protein expression did not. LVAD support only induced a decrease of OPN plasma levels in individual patients, whereas OPN plasma levels reduced significantly in all patients after HTx.


Thermal Profiles: Novel phenotypic measurements of animal growth and metabolic efficiency.
The costs of production for high density protein and the impacts food production have on the environment are becoming increasingly important issues in animal agriculture. The objective of the present study was to investigate the use of novel thermal profiles including a Thermal Efficiency Index (TEI) on the ability to identify efficient animals in a fraction of the time and at a significantly lower cost of conventional feed station and performance technology. Three hundred and fourty four high performance Duroc sires from a genetic nucleus herd were used in the study. The animals were monitored for feed consumption and growth performance using conventional feed station technology for a 72 day period. Animals were monitored in these stations between approximately 50 kg and 130 kg live body weight. An infrared thermal scan was performed on the animals at the end of the performance test by collecting automated dorsal thermal images and using these biometrics to measure both bio-surveillance values and a thermal phenotypic profile including the TEI (mean dorsal temperature /body weight 0.75). The thermal profile values were significantly correlated (r = 0.40, P < 0.0001) with a current industry best practice for performance in Residual Intake and Gain (RIG). The data from the current study suggest these rapid, real time, cost effective values for TEI constitute a useful precision farming tool for the animal industries to reduce the cost of production and green house gas (GHG) impact for high density protein production.


Transperitoneal migration of multiple ova in a didelphic uterus: a case report.
Transperitioneal migration of the ova takes place frequently. This condition is often diagnosed during ectopic pregnancies. There are no case reports of such an occurrence in a multiple pregnancy with a didelphic uterus.


Unambiguous and accurate measurement of scalar coupling constants through a selective refocusing NMR experiment.
Scalar coupling plays an important role in the analysis of molecular structure and dynamics. A great number of nuclear magnetic resonance (NMR) selective refocusing experiments, such as 2D G-SERF and PSYCHEDELIC, were developed to extract scalar coupling constants involving a selected proton from overlapped spectra. However, intense axial peaks occur in this type of experiments, leading to possible ambiguity in the assignment of spectral peaks and subsequent accurate measurement of 1H-1H scalar coupling constants. Here, a method based on selective coherence transfer and PSYCHEDELIC module is designed to acquire absorption-mode selective refocusing spectrum while suppressing intense axial peaks. Therefore, unambiguous and accurate measurement of scalar coupling constants involving the selectively excited proton can be achieved. The performances of the proposed method are demonstrated on several samples.


Simplified high yield TAILS terminomics using a new HPG-ALD 800K-2000 polymer with precipitation.
Identification of the N terminus of proteins in complex samples has been accelerated in recent years by the increased power of mass spectrometers, bioinformatic tools and biochemical methods. We developed Terminal Amine Isotopic Labeling of Substrates (TAILS), a powerful method for the isolation, identification and quantification of N terminal peptides comprising the N terminome from proteome in vitro analyses, cultured cells and tissues. TAILS depletes internal tryptic peptides so enriching for the natural and cleaved neo N-terminal peptides present in complex proteome samples. To do so N-terminal peptides are blocked at the protein level by isotopically labeled amine reactive reagents, then digested with trypsin. TAILS relies on a water-soluble 100-kDa highly-branched polyglycerol polymer (HPG-ALD) developed in our laboratories to bind and deplete the sample of tryptic peptides. Polymer-bound internal tryptic peptides are separated from the blocked N-terminal peptides by spin filters. Here we describe a revised TAILS protocol that uses a newly developed 800-kDa HPG-ALD polymer that allows precipitation of the polymer post-depletion. Precipitation has multiple advantages over spin filters including maximal N-terminal peptide recovery and faster high-throughput N-terminome identification and quantification in any sample type.


Synthesis and in vitro antitubercular activity of some 1-[(4-sub)phenyl]-3-(4-{1-[(pyridine-4-carbonyl)hydrazono]ethyl}phenyl)thiourea.
Various isonicotinyl hydrazones were prepared by reacting isonicotinyl hydrazide [INH] with 1-(4-acetylphenyl)-3-[(4-sub)phenyl]thiourea and were tested for their antimycobacterial activity in vitro against Mycobacterium tuberculosis H37Rv and INH-resistant M. tuberculosis using the BACTEC 460 radiometric system. Among the synthesized compounds, 1-(4-fluorophenyl)-3-(4-{1-[(pyridine-4-carbonyl)-hydrazono]ethyl}phenyl)thiourea (4d) was found to be the most potent compound with a minimum inhibitory concentration of 0.49 microM against M. tuberculosis H37Rv and INH-resistant M. tuberculosis. When compared to INH, 4d was found to be 3 and 185 times more active against M. tuberculosis H37Rv and INH-resistant M. tuberculosis, respectively, with a selectivity index of >300.


ncFO: A Comprehensive Resource of Curated and Predicted ncRNAs Associated with Ferroptosis.
Ferroptosis is a form of regulated cell death driven by the accumulation of lipid hydroperoxides. Regulation of ferroptosis might be beneficial to cancer treatment. Non-coding RNAs (ncRNAs) are a class of RNA transcripts that generally cannot encode proteins and have been demonstrated to play critical roles in regulating ferroptosis. Herein, we developed ncFO, the ncRNA-ferroptosis association database, to document the manually curated and predicted ncRNAs that are associated with ferroptosis. Collectively, ncFO contains 90 experimentally verified entries, including 46 microRNAs (miRNAs), 21 long non-coding RNAs (lncRNAs), and 17 circular RNAs (circRNAs). In addition, ncFO also incorporates two online prediction tools based on the regulation and co-expression of ncRNA and ferroptosis genes. Using default parameters, we obtained 3260 predicted entries, including 598 miRNAs and 178 lncRNAs, by regulation, as well as 2,592,661 predicted entries, including 967 miRNAs and 9632 lncRNAs, by ncRNA-ferroptosis gene co-expression in more than 8000 samples across 20 cancer types. The detailed information of each entry includes ncRNA name, disease, species, tissue, target, regulation, publication time, and PubMed identifier. ncFO also provides survival analysis and differential expression analysis for ncRNAs. In summary, ncFO offers a user-friendly platform to search and predict ferroptosis-associated ncRNAs, which might facilitate research on ferroptosis and discover potential targets for cancer treatment. ncFO can be accessed at http://www.jianglab.cn/ncFO/.


Time-lag of urinary and salivary cortisol response after a psychological stressor in bonobos (Pan paniscus).
Cortisol is often measured as a marker for stress. Therefore, a profound validation of the time-lag between the stressor and the increase and peak in cortisol levels is needed. No study measured both the urinary and salivary cortisol time-lag after a psychological stressor. In this study, we used a frequent sampling study design to (1) describe the urinary and salivary cortisol pattern during a control day; and (2) characterize the induced excretion pattern of urinary and salivary cortisol after a psychological stressor in six zoo-housed bonobos. Liquid chromatography-tandem mass spectrometry was used to analyze 71 urine and 162 saliva samples collected on a control and a test day. We found that the time-lag between the stressor and the maximal cortisol concentration was similar in urine and saliva (160 min after the stressor). However, salivary cortisol after the stressor did show a faster and steeper increase than urinary cortisol. We also show inter-individual variation in the baseline and stress levels of cortisol, which should be considered in future cortisol studies. Our research highlights the importance of validation studies to confirm relevant sampling windows for cortisol sampling in order to obtain biologically meaningful results.


A prevalent mutation for galactosemia among black Americans.
To define the mutation causing galactosemia in patients of black American origin who have no galactose-1-phosphate uridyltransferase (GALT) activity in erythrocytes but good clinical outcome. We discovered a mutation caused by a C-->T transition at base-pair 1158 of the GALT gene that results in a serine-to-leucine substitution at codon 135 (S135L). We developed a method with which to screen populations for its prevalence. We compared galactose-1-phosphate uridyltransferase among erythrocytes, leukocytes, and transformed lymphoblasts, as well as total body oxidation of D-(13C)-galactose to 13CO2 among three genotypes for GALT (S135L/S135L, Q188R/Q188R, and Normal/Normal). We found a 48% prevalence of the S135L mutation among 17 black American patients with classic galactosemia and a 1% prevalence in a population of 50 black Americans without galactosemia. The S135L mutation was not found in 84 white patients with G/G galactosemia nor in 87 white control subjects without galactosemia. We found normal whole body oxidation of D-(13C)-galactose by the patient homozygous for S135L and various degrees of enzyme impairment among different tissues. The S135L mutation in the GALT gene is a prevalent cause of galactosemia among black patients. Because GALT activity varies in different tissues of patients homozygous for S135L, they may have a better clinical outcome than patients who are homozygous for Q188R when both are treated from infancy.


Generation of a human induced pluripotent stem cell line from a Bietti crystalline corneoretinal dystrophy patient with CYP4V2 mutations.
Bietti crystalline corneoretinal dystrophy (BCD) is an autosomal recessively inherited progressive retinal disease. Here, we describe the generation and characterization of a human induced pluripotent stem cell (hiPSC) line of BCD patient with CYP4V2 mutations. The reprogramming of this iPSC line was performed from skin fibroblast by using the Sendai-virus based approach. The established hiPSC line retained the disease-associated mutations and showed normal karyotype, pluripotency and differentiation capacity.


Non-invasive measurement of intraocular pressure by near-infrared spectroscopy.
To assess a new non-contact method for non-invasive measurement of intraocular pressure (IOP) using near-infrared spectroscopy. Experimental study. Eleven porcine eyes were cannulated into the anterior chamber by a needle connected with a water column. Measuring the near-infrared spectrum in diffuse reflectance out of the anterior chamber by near-infrared spectroscopy, IOP was determined and compared with the height of the water column which was varied between 0 cm and 100 cm. In total, 363 near-infrared spectrometric measurement values were obtained. Correlating the near-infrared spectrometric measurements with the height of the water column showed a relationship with a correlation coefficient of r(2) = .858 and a steepness of the regression line of .96. The results of the present pilot study suggest that near-infrared spectrometric tonometry may present a new technology for non-invasive and non-contact IOP measurement.