Economic comparison of systemic antimicrobial therapies for metritis in dairy cows.
Metritis is a prevalent disease with effects on production, reproduction, and survival, thereby affecting dairy farm profitability. A component of the cost of metritis is antimicrobial therapy. Some antimicrobials result in milk withhold that adds to the cost of disease. The objectives were (1) to determine cost of metritis for 2 antimicrobial treatments using a herd budget that includes costs associated with incidence of concurrent diseases, milk production and reproduction losses, and removal from the herd and (2) to apply sensitivity analysis to determine the cost of different scenarios. Cows with metritis from a previous study assigned randomly to receive ampicillin (AMP, n = 259), an antimicrobial that requires milk withhold, or ceftiofur (CEFT, n = 269), an antimicrobial with no milk withhold, were used for the economic analysis. A group of cows with no metritis (NOMET, n = 268), matched by parity and calving day, served as the baseline for comparison. The incidence of other diseases in the first 60 d postpartum, culling and death, reproductive performance, discarded milk, milk yield, total milk sold per cow, and residual cow value were used as responses. The economic analysis considered the costs associated with therapy, reproductive management, discarded milk, estimated DM consumed, income from saleable milk, and the residual cow value at 300 d postpartum or earlier if the cow was removed from the herd. Sensitivity analyses were performed considering 3 scenarios for milk and feed prices. The incidence of diseases other than metritis (NOMET, 30.4%; AMP, 45.4%; CEFT, 34.0%) and days in the hospital (NOMET, 2.7 d; AMP, 8.6 d; CEFT, 3.5 d) were greater for cows treated with AMP than CEFT. Treatment did not affect the risk of leaving the herd (NOMET, 15.5%; AMP, 15.0%; CEFT, 19.1%). The 21-d pregnancy rate was lower for cows with metritis but did not differ between AMP and CEFT (NOMET, 24.9%; AMP, 18.9%; CEFT, 17.0%). Milk yield was greater for cows without metritis than those with metritis and greater for AMP than CEFT (NOMET, 33.7 kg/d; AMP, 32.5 kg/d; CEFT, 31.2 kg/d). Cost of metritis did not differ with choice of therapy, but it increased as milk price increased. When both milk and feed prices were the average values considered ($0.44/kg and $0.26/kg, respectively), the costs of a case of metritis for AMP and CEFT were, respectively, $344 and $410 when milk was discarded and $267 and $406 when milk was fed to calves. Cost of therapy for AMP and CEFT represented 16.6 and 24.6% of the total cost of metritis when milk was discarded and 21.5 and 24.8% of the total cost of metritis when milk was fed to calves. The largest component of cost of metritis for both therapies was the reduced income from milk minus feed cost, ranging from 40.0 to 56.7%. Collectively, metritis is an expensive disease, and choice of antimicrobial therapy did not influence survival, reproduction, or cost of the disease.


Socioeconomic inequalities in risk of congenital anomaly.
To investigate socioeconomic inequalities in the risk of congenital anomalies, focusing on risk of specific anomaly subgroups. A total of 858 cases of congenital anomaly and 1764 non-malformed control births were collected between 1986 and 1993 from four UK congenital malformation registers, for the purposes of a European multicentre case control study on congenital anomaly risk near hazardous waste landfill sites. As a measure of socioeconomic status, cases and controls were given a value for the area level Carstairs deprivation index, by linking the postcode of residence at birth to census enumeration districts (areas of approximately 150 households). Risk of non-chromosomal anomalies increased with increasing socioeconomic deprivation. The risk in the most deprived quintile of the deprivation index was 40% higher than in the most affluent quintile. Some malformation subgroups also showed increasing risk with increasing deprivation: all cardiac defects, malformations of the cardiac septa, malformations of the digestive system, and multiple malformations. No evidence for socioeconomic variation was found for other non-chromosomal malformation groups, including neural tube defects and oral clefts. A decreasing risk with increasing deprivation found for all chromosomal malformations and Down's syndrome in unadjusted analyses, occurred mainly as a result of differences in the maternal age distribution between social classes. Our data, although based on limited numbers of cases and geographical coverage, suggest that more deprived populations have a higher risk of congenital anomalies of non-chromosomal origin and some specific anomalies. Larger studies are needed to confirm these findings and to explore their aetiological implications.


Is a randomized trial of antioxidants in the primary prevention of Alzheimer disease warranted?
Alzheimer disease is a common condition that severely affects both persons with the illness and their families. Prevention of Alzheimer disease is an urgent priority, but study of potentially modifiable risk factors for the illness is at an early stage. Laboratory studies suggest that oxidative mechanisms may be involved in the pathogenesis of Alzheimer disease and raise the possibility that antioxidant nutrients could be used in disease prevention. Observational studies suggest that antioxidant nutrients may have protective effects against a number of other common chronic diseases, including cardiovascular disease and cancer. Because the protective effects, if any, of antioxidant nutrients are likely to be small to moderate in magnitude, large-scale randomized trials of primary prevention will likely be necessary to resolve this issue, and a major question is how quickly to progress to expensive and time consuming trials needed to provide more definitive evidence. This is a difficult question, but the severity of the disease, the current absence of preventive strategies and the likelihood that observational studies alone will not provide clear resolution of the issue all suggest that it may be prudent to strongly consider such trials in the near future.


Platelet and monocyte variables in homocystinuria due to cystathionine-beta-synthase deficiency.
To gain insight into the mechanisms responsible for enhanced thromboxane (TX) A2 biosynthesis in homozygous homocystinuria due to cystathionine-beta-synthase deficiency (CBSD), we measured a series of platelet and monocyte variables in 9 homozygous and 8 obligate heterozygous CBSD patients and evaluated their relationships to thromboxane formation, as reflected by urinary excretion of its major metabolite, 11-dehydro-TXB2 (TXM). Consistent with our previous data, homozygous CBSD patients showed abnormally high TXM excretion (1175 +/- 236 pg/mg creatinine vs. 284 +/- 39 in control subjects; p < 0.001). Significantly higher TXM excretion was also found in obligate heterozygotes (755 +/- 450 pg/mg creatinine; p < 0.05 vs. control subjects). All platelet and monocyte variables fell within the normal range in CBSD patients and none showed a correlation with TXM excretion (p always > 0.05). Our results argue against abnormalities of platelet and monocyte function being responsible for the abnormally high in vivo TXA2 biosynthesis in homocystinuria due to CBSD.


Health Maintenance and Vaccination of Patients With Inflammatory Bowel Disease: Practice and Perception of Responsibility of Gastroenterologists vs Primary Care Providers.
Background: Although it is well established that patients with inflammatory bowel disease (IBD) are at increased risk of complicating diseases and vaccination-preventable infections, whether gastroenterologists (GIs) or primary care providers (PCPs) assume responsibility for these patients' health maintenance is not clear. Methods: We anonymously surveyed a convenience sample of 94 PCPs and 61 GIs at Saint Louis University School of Medicine in St. Louis, MO, about their practice and perception of the health maintenance and vaccination of patients with IBD. Results: Response rates were 82% and 93% for GIs and PCPs, respectively. GIs were as likely as PCPs to screen for smoking (88% vs 89%) and were significantly less likely to screen for depression/anxiety (24% vs 54%) or to provide pertussis (14% vs 44%) or diphtheria (20% vs 48%) vaccines. GIs were significantly more likely than PCPs to assess for colonoscopy need (94% vs 80%); to screen for nonmelanoma skin cancer (62% vs 14%), melanoma (56% vs 7%), osteoporosis (72% vs 51%), or tuberculosis (94% vs 44%); to prescribe calcium/vitamin D (74% vs 53%); to perform nutritional assessment (78% vs 33%); or to provide hepatitis A (60% vs 39%) or hepatitis B (86% vs 56%) vaccines. GIs were as likely as PCPs (64% vs 75%) to perceive that PCPs should order vaccinations and significantly more likely to perceive that GIs should track vaccinations (58% vs 16%) and other health maintenance issues (90% vs 49%). We found positive associations between performing the various health maintenance and vaccination tasks and the perception of responsibility. Conclusion: Several health maintenance aspects are inadequately addressed by GIs and PCPs, in part because of conflicting perceptions of responsibility. Clear guidelines and better GI/PCP communication are required to ensure effective health maintenance for patients with IBD.


Inhibitory effects of pH5 enzyme from non-lactating bovine mammary gland on various stages of protein synthesis in the rat liver amino acid-incorporating system.
1. pH5 enzyme from non-lactating bovine mammary gland was found to contain potent inhibitors of protein synthesis in the rat liver cell-free system. These inhibitors affect (a) formation of aminoacyl-tRNA where tRNA represents transfer RNA, (b) transfer of labelled amino acids from rat liver amino[(14)C]acyl-tRNA to protein in rat liver polyribosomes, and (c) incorporation of (14)C-labelled amino acids into peptide by rat liver polyribosomes supplemented with rat liver pH5 enzyme. 2. Increasing amounts of pH5 enzyme from bovine mammary gland progressively inhibited the incorporation of labelled amino acids into protein by a complete incorporating system from rat liver. Approx. 80% inhibition was observed at a concentration of 2mg. of protein of pH5 enzyme from bovine mammary gland. The inhibitory effect of the bovine pH5 enzyme fraction could not be overcome by the addition of increasing amounts of rat liver pH5 enzyme. 3. Fractionation of bovine pH5 enzyme with ammonium sulphate into four fractions showed that all the fractions inhibited the incorporation of (14)C-labelled amino acids in the rat liver system, but to varying extents. The highest inhibition observed (90%) was exhibited by the 60%-saturated-ammonium sulphate fraction. 4. Heat treatment of bovine pH5 enzyme at various temperatures caused only a partial loss of its inhibitory effect on labelled amino acid incorporation by the rat liver system. Treatment at 105 degrees for 5min. resulted in the bovine pH5 enzyme fraction losing 30% of its inhibitory activity. 5. pH5 enzyme from bovine mammary gland strongly inhibited the charging of rat liver tRNA in the presence of its own pH5 enzymes. 6. The transfer of labelled amino acids from rat liver amino[(14)C]acyl-tRNA to protein in a system containing rat liver polyribosomes and pH5 enzyme was almost completely inhibited by bovine pH5 enzyme at a concentration of 2mg. of protein of the enzyme fraction. 7. One of the inhibitors of various stages of protein synthesis in rat liver present in bovine pH5 enzyme was identified as an active ribonuclease, and the second inhibitor present was shown to be tRNA.


Rituximab induction immunotherapy for first-line low-tumor-burden follicular lymphoma: survival analyses with 7-year follow-up.
The purpose of this study was to report long-term results of rituximab induction monotherapy in patients with low-tumor-burden follicular lymphoma (LTBFL). Of 49 first-line LTBFL patients who received weekly doses of rituximab (375 mg/m(2)), 46 have been followed with a long-term analysis of clinical and molecular responses. Best clinical response (at any staging within a year following treatment) was 80%, 24 (52%) patients had complete or unconfirmed complete response, 13 (28%) had partial response and 9 (20%) had stable or progressive disease. Of 31 patients having a positive bcl2-JH rearrangement, 15 (48%) became negative following treatment. After 83.9 months of follow-up (95% confidence interval 6.4-92.8 months), the median progression-free survival is 23.5 months and overall survival (OS) is 91.7%. Five patients died (one progression, one myelodysplasia, one diffuse large B-cell lymphoma and two solid tumors). Seven patients (15%) are progression-free including five who are bcl2 informative. No unexpected long-term adverse event has been observed. A significant proportion of patients remain progression-free 7 years after a single 4-dose rituximab treatment in first-line LTBFL. The 7-year overall survivalOS is very high in this selected population of patients.


Clinicopathologic Characteristics of Centrilobular Injury in Pediatric Liver Transplantation.
Centrilobular injury (CLI) is defined as the presence of perivenular mononuclear inflammation, hepatocyte dropout, and extravasated erythrocytes. In pediatric liver allografts, CLI has been associated with advanced fibrosis and chronic rejection (CR). We sought to better characterize the clinicopathologic features of CLI in the setting of T cell-mediated rejection (TCMR) and its association with complement component 4d (C4d) deposition. A total of 206 posttransplant pediatric patients (491 allograft liver biopsies) were available from 2000 to 2018, of which 63 patients (102 biopsies) showed evidence of TCMR and were included in the study. Of the patients, 35 (55.6%) had CLI on their initial episode of TCMR; those patients with CLI were significantly associated with the type of immunosuppression treatment (P = 0.03), severity of TCMR (P < 0.001), higher gamma-glutamyltransferase (P = 0.01), and advanced fibrosis (P = 0.03). There was a trend to shorter time interval from transplantation to presentation of CLI compared with those without CLI (P = 0.06). No difference was observed in graft or overall survival in the patients with CLI. In 20 patients with CLI, additional biopsies were available; in 45% of these patients, CLI was a persistent/recurrent finding. C4d deposition was noted in 12% of all biopsies (6 patients) with CLI. No significant correlation was noted in C4d deposition and CLI, CR, or graft/overall survival. In conclusion, CLI, although not significantly associated with worse graft survival, was significantly associated with severe TCMR and degree of fibrosis, which highlights the importance of active clinical management and follow-up for these patients.


Axillar ultrasound of the glenohumeral joint.
A new ultrasonographic method for detection of intraarticular effusion in the glenohumeral joint was developed. The distance between the humerus and the joint capsule is measured at the axilla with the humerus in 90 degrees abduction. Effusion was detected by ultrasonography in 10 of 11 joints in which effusion was confirmed by intraarticular puncture. Intraarticular injection of physiological NaC1 solution could be seen as a change in the scan. The mean ultrasonographic distance between the head of humerus and the joint capsule was 2.4 mm (SD 0.5 mm) in 80 shoulder joints of 40 healthy adults. Thus, intrasynovial effusion can be suspected if the distance is 3.5 mm (mean + 2 SD) or more, or the difference between both sides is 1 mm or more.


Autophagy-mediated insulin receptor down-regulation contributes to endoplasmic reticulum stress-induced insulin resistance.
Endoplasmic reticulum (ER) stress is associated with obesity-induced insulin resistance, yet the underlying mechanisms remain to be fully elucidated. Here we show that ER stress-induced insulin receptor (IR) down-regulation may play a critical role in obesity-induced insulin resistance. The expression levels of IR are negatively associated with the ER stress marker C/EBP homologous protein (CHOP) in insulin target tissues of db/db mice and mice fed a high-fat diet. Significant IR down-regulation was also observed in fat tissue of obese human subjects and in 3T3-L1 adipocytes treated with ER stress inducers. ER stress had little effect on IR tyrosine phosphorylation per se but greatly reduced IR downstream signaling. The ER stress-induced reduction in IR cellular levels was greatly alleviated by the autophagy inhibitor 3-methyladenine but not by the proteasome inhibitor N-benzoyloxycarbonyl (Z)-Leu-Leu-leucinal (MG132). Inhibition of autophagy prevented IR degradation but did not rescue IR downstream signaling, consistent with an adaptive role of autophagy in response to ER stress-induced insulin resistance. Finally, chemical chaperone treatment protects cells from ER stress-induced IR degradation in vitro and obesity-induced down-regulation of IR and insulin action in vivo. Our results uncover a new mechanism underlying obesity-induced insulin resistance and shed light on potential targets for the prevention and treatment of obesity-induced insulin resistance and type 2 diabetes.


Oxidation of dihydrotestosterone by human cytochromes P450 19A1 and 3A4.
Dihydrotestosterone is a more potent androgen than testosterone and plays an important role in endocrine function. We demonstrated that, like testosterone, dihydrotestosterone can be oxidized by human cytochrome P450 (P450) 19A1, the steroid aromatase. The products identified include the 19-hydroxy- and 19-oxo derivatives and the resulting Δ(1,10)-, Δ(5,10)-, and Δ(9,10)-dehydro 19-norsteroid products (loss of 19-methyl group). The overall catalytic efficiency of oxidation was ~10-fold higher than reported for 3α-reduction by 3α-hydroxysteroid dehydrogenase, the major enzyme known to deactivate dihydrotestosterone. These and other studies demonstrate the flexibility of P450 19A1 in removing the 1- and 2-hydrogens from 19-norsteroids, the 2-hydrogen from estrone, and (in this case) the 1-, 5β-, and 9β-hydrogens of dihydrotestosterone. Incubation of dihydrotestosterone with human liver microsomes and NADPH yielded the 18- and 19-hydroxy products plus the Δ(1,10)-dehydro 19-nor product identified in the P450 19A1 reaction. The 18- and 19-hydroxylation reactions were attributed to P450 3A4, and 18- and 19-hydroxydihydrotestosterone were identified in human plasma and urine samples. The change in the pucker of the A ring caused by reduction of the Δ(4,5) bond is remarkable in shifting the course of hydroxylation from the 6β-, 2β-, 1β-, and 15β-methylene carbons (testosterone) to the axial methyl groups (18, 19) in dihydrotestosterone and demonstrates the sensitivity of P450 3A4, even with its large active site, to small changes in substrate structure.


Tissue expression patterns identify mouse cilia genes.
In mammals, cilia are critical for development, sensation, cell signaling, sperm motility, and fluid movement. Defects in cilia are causes of several congenital syndromes, providing additional reasons to identify cilia-related genes. We hypothesized that mRNAs selectively abundant in tissues rich in highly ciliated cells encode cilia proteins. Selective abundance in olfactory epithelium, testes, vomeronasal organ, trachea, and lung proved to be an expression pattern uniquely effective in identifying documented cilia-related genes. Known and suspected cilia-related genes were statistically overrepresented among the 99 genes identified, but the majority encoded proteins of unknown function, thereby predicting new cilia-related proteins. Evidence of expression in a highly ciliated cell, the olfactory sensory neuron, exists for 73 of the genes. In situ hybridization for 17 mRNAs confirmed expression of all 17 in olfactory sensory neurons. Most were also detected in vomeronasal sensory neurons and in neighboring tissues rich in ciliated cells such as respiratory epithelium. Immunoreactivity for one of the proteins identified, Spa17, colocalized with acetylated tubulin in the cilia layer of the olfactory epithelium. In contrast, the ciliary rootlet protein, Crocc, was located in discrete structures whose position was consistent with the dendritic knobs of the olfactory sensory neurons. A compilation of >2,000 mouse genes predicted to encode cilia-related proteins revealed a strong correlation (R = 0.99) between the number of studies predicting a gene's involvement in cilia and documented evidence of such involvement, a fact that simplifies the selection of genes for further study of the physiology of cilia.


Practical Entanglement Estimation for Spin-System Quantum Simulators.
We present practical methods to measure entanglement for quantum simulators that can be realized with trapped ions, cold atoms, and superconducting qubits. Focusing on long- and short-range Ising-type Hamiltonians, we introduce schemes that are applicable under realistic experimental conditions including mixedness due to, e.g., noise or temperature. In particular, we identify a single observable whose expectation value serves as a lower bound to entanglement and that may be obtained by a simple quantum circuit. As such circuits are not (yet) available for every platform, we investigate the performance of routinely measured observables as quantitative entanglement witnesses. Possible applications include experimental studies of entanglement scaling in critical systems and the reliable benchmarking of quantum simulators.


Metabolic Syndrome Patients Have Lower Levels of Adropin When Compared With Healthy Overweight/Obese and Lean Subjects.
Metabolic syndrome (MetS), a cluster of cardiometabolic risk factors, is a challenging public health issue. The aim of current study was to test the hypothesis that concentrations of plasma adropin and leptin differ between patients with MetS and comparable age- and sex-matched control groups. This case-control study involved 153 subjects (51 per group). The study group included obese subjects with MetS and the two control groups included weight-matched subjects without MetS ("healthy": obese) and normal weight subjects without MetS. Body composition parameters were measured using bioelectrical impedance analysis. Plasma levels of adropin, leptin, and their ratio were measured. Leptin was significantly different between obese patients with/without MetS groups and normal weight subjects. Patients with MetS had higher levels of leptin (14 ± 12.4) compared with those without MetS (11.2 ± 9.3 vs. 7 ± 7.1 obese and normal weight without MetS, respectively; p = .002). Compared with healthy obese and normal weight subjects, MetS subjects had lower levels of plasma adropin ( p < .001) and a lower plasma adropin to leptin ratio ( p < .001), which remained significant when adjusted for body fat mass by analysis of covariance ( p < .001). This study demonstrates low levels of adropin are correlated with MetS and hence identify it as a potentially protective agent against MetS development. Variation in adropin levels may partly explain the "healthy obese" phenomenon.


Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape.
Delivery of short interfering RNAs (siRNAs) remains a key challenge in the development of RNA interference (RNAi) therapeutics. A better understanding of the mechanisms of siRNA cellular uptake, intracellular transport and endosomal release could critically contribute to the improvement of delivery methods. Here we monitored the uptake of lipid nanoparticles (LNPs) loaded with traceable siRNAs in different cell types in vitro and in mouse liver by quantitative fluorescence imaging and electron microscopy. We found that LNPs enter cells by both constitutive and inducible pathways in a cell type-specific manner using clathrin-mediated endocytosis as well as macropinocytosis. By directly detecting colloidal-gold particles conjugated to siRNAs, we estimated that escape of siRNAs from endosomes into the cytosol occurs at low efficiency (1-2%) and only during a limited window of time when the LNPs reside in a specific compartment sharing early and late endosomal characteristics. Our results provide insights into LNP-mediated siRNA delivery that can guide development of the next generation of delivery systems for RNAi therapeutics.


Endobronchial inflammatory pseudotumor: a case report.
Inflammatory pseudotumor of the lung is a rare form of benign neoplasia and is generally characterized by a solitary pulmonary nodule. The endobronchial presentation is uncommon. Conservative surgery remains the treatment of choice, and surgeons should always strive to achieve tumor-free margins due to the possibility of local recidivism. This article reports the case of a 36-year-old male patient with recurrent attacks of wheezing and cough. The patient underwent successful bronchoplasty for the resection of an endobronchial inflammatory pseudotumor.


Intrauterine transfusion for fetal anemia due to red blood cell alloimmunization: 14 years experience in Leuven.
The purpose of this study is to report on the pregnancy and neonatal outcome of intrauterine transfusion (IUT) for red blood cell (RBC-)alloimmunization. Retrospective cohort study of all IUT for RBC-alloimmunization in the University Hospital of Leuven, between January 2000 and January 2014. The influence of hydrops, gestational age and technique of transfusion on procedure related adverse events were examined. 135 IUTs were performed in 56 fetuses. In none of the cases fetal or neonatal death occurred. Mild adverse events were noted in 10% of IUTs, whereas severe adverse events occurred in 1.5%. Hydrops and transfusion in a free loop were associated with an increased risk of adverse events whereas gestational age (GA) at transfusion after 34 weeks was not. Median GA at birth was 35.6 weeks and 9% was born before 34 weeks. Besides phototherapy 65.4% required additional neonatal treatment for alloimmune anemia. Non-hematologic complications occurred in 23.6% and were mainly related to preterm birth. In experienced hands, IUT for RBC-alloimmunization is a safe procedure in this era. Patients should be referred to specialist centers prior to the development of hydrops. IUT in a free loop of cord and unnecessary preterm birth are best avoided.


The effect of RAAS inhibitors on acute hypoxemic respiratory failure and in-hospital mortality in the hypertensive Covid-19 patients.
We have aimed to investigate the relationship between use of angiotensin-converting-enzyme inhibitor (ACEI) or angiotensin-receptor-blocker (ARB) drugs and acute hypoxemic respiratory failure (AHRF) and in-hospital mortality in hypertensive Covid-19 patients. Consecutive 1345 patients diagnosed with Covid-19 between April and October 2020 who met inclusion criteria were divided into two groups based on presence and absence of AHRF and mortality. The groups were compared regarding epidemiological, clinical, radiological, laboratory findings and treatments methods. The patient groups ACEI, ARB and other antihypertensive drugs (non-ACEI/ARB) were compared regarding same parameters. Median age was 68 (60-76) years in the patient group including 805 (59.9.1%) females. Of the patients, 475 (35.3%), 644 (47.9%) and 226 (16.8%) were using ACEIs, ARBs and non-ACEI/ARB, respectively. AHRF and in-hospital mortality developed in 1053 (78.3%) and 290 (21.6%) patients, respectively. Age, gender, coronary artery disease, diabetes mellitus (DM), neutrophil, lymphocyte, creatinine, D-dimer, C-reactive protein (CRP), ACEI, beta blocker and aspartate transaminase (AST) found statistically significant in the univariable logistic regression performed to identify independent predictors of mortality were included multivariable logistic regression model. Age (OR: 1.066, 95%CI: 1.049-1.083; p < .001), DM (OR: 1.682, 95%CI: 1.238-2.286; p = .001), neutrophil (OR: 1.041, 95%CI: 1.007-1.077; p = .019), creatinine (OR: 1.178, 95%CI: 1.048-1.325; p = .006), CRP (OR: 1.008, 95%CI: 1.006-1.010; p < .001), ACEI (OR: 0.718, 95%CI: 0.521-0.988; p = .042), AST (OR: 1.005, 95%CI: 1.001-1.010; p = .010) were found associated with in-hospital mortality. In our study, it was not detected clinically significant difference between three groups with regard to their relation with in-hospital mortality.


Silver nanoparticle-doped polyvinyl alcohol coating as a medium for surface-enhanced Raman scattering analysis.
A simple polymer substrate for inducing Surface-Enhanced Raman Scattering (SERS) has been investigated. This SERS substrate consists of a solid support, such as a glass slide covered with polyvinyl alcohol (PVA) impregnated with fine silver nanoparticles. The preparation simply involves mixing aqueous PVA polymer with solid AgNO3 to produce a solution that can be easily spin coated on the glass substrate and dried to obtain a hard translucent coating. Aqueous solution of FeSO4.7H2O was used to reduce Ag+ ions to silver nanoparticles. The effects of various experimental conditions of sample preparation were investigated in order to improve the Raman enhancement efficiency of the substrate. The overall substrate performance was evaluated with the use of biologically important compounds: benzoic acid, p-amino benzoic acid, pyridine and dopamine. The spectral features of these compounds closely matched with those reported in literature. The use of the polymer matrix made the SERS substrate resistant to scratching, therefore, improving it to be more suitable for field applications. The hydrophilic nature of the polymer provides additional advantages for probing biological samples. The shelf-life of the dried, unreduced substrates is at least one month.


Kakkonto Inhibits Cytokine Production Induced by Rhinovirus Infection in Primary Cultures of Human Nasal Epithelial Cells.
Rhinovirus (RV) is a primary etiologic agent of common cold that can subsequently acutely exacerbate bronchial asthma or chronic obstructive pulmonary disease. Kakkonto (Ge-gen-tang in Chinese), one of the most frequently prescribed traditional Japanese (Kampo) medicines, is used for treating common cold, shoulder stiffness, or inflammatory diseases of the upper body. Previous experimental studies have indicated that kakkonto exerts antiviral and anti-inflammatory effects on the influenza virus and the human respiratory syncytial virus. However, there is a lack of reports investigating the efficacy of kakkonto in RV infection. Hence, the aim of the current study was to investigate the effects of kakkonto on RV infection of human nasal epithelial (HNE) cells. HNE cells obtained via endoscopic sinus surgery were cultured and infected with RV14, with or without kakkonto treatment. The supernatants from the cells were collected, and the RV14 titer and cytokine levels were assessed. Reverse transcription-polymerase chain reaction was performed to determine the amount of viral RNA, while the level of nuclear factor kappa B (NF-κB) subunits in the nucleus was assessed by enzyme-linked immunosorbent assay. Although kakkonto treatment did not reduce RV14 titer or RNA levels, indicating that it did not inhibit RV14 proliferation, it was found to reduce the production of specific pro-inflammatory cytokines, including interleukin (IL)-8, tumor necrosis factor (TNF)-α, and monocyte chemotactic protein-1 (MCP-1). Unlike that observed with the kakkonto extract, none of the crude drugs contained in kakkonto reduced IL-8 level. Furthermore, though kakkonto treatment significantly reduced p50 levels, it did not impact the p65 subunit of NF-κB. These results indicated that kakkonto can inhibit inflammation caused by RV infection and may exert an immunomodulatory effect on HNE cells. This is the first report to elucidate the effects of kakkonto extract on RV infection in primary cultures of HNE cells, providing evidence that kakkonto may act as an effective therapy for RV infection and subsequent airway inflammation.


SIGNing a symbiotic treaty with gut microbiota.
Beneficial microbes hold great promise for the treatment of a wide range of immune and inflammatory disorders. In this issue of The EMBO Journal, Lightfoot and colleagues report how the food-grade bacterium Lactobacillus acidophilus helps the immune system to limit experimental colitis in mice through interaction between SIGNR3 and surface layer protein A (SlpA) in L. acidophilus. These results pave the way for future development of novel therapies for inflammatory diseases, including inflammatory bowel disease.


Ovarian morphology and the concentration of steroids during the oestrus cycle of sheep actively immunized against androstenedione.
Ewes were immunized against androstenedione-11 alpha-hemisuccinyl-bovine serum albumin (androstenedione-11-BSA) or BSA alone (controls). The ovaries were examined macroscopically and ovarian venous blood was collected at laparotomy by in-situ cannulation of the ovarian vein on Day 10 of the oestrous cycle, or by needle puncture of the ovarian vein 36 h after prostaglandin treatment. Follicular fluid was also collected from follicles greater than 5 mm diameter in some ewes. Ovaries and adrenal glands were collected and examined histologically. Oestrone, oestradiol-17 beta, androstenedione, testosterone and progesterone were measured by radioimmunoassays. Immunization against androstenedione-11-BSA was followed by an increase in the number of surface follicles greater than 3 mm in diameter and in ovulation rate. These changes were accompanied by an increased production of androgens (especially androstenedione) and of oestrogens during the preovulatory period. There were no observed changes in the adrenal cortex or pituitary glands, but the ovaries were heavier in immunized ewes and luteinized follicles were present in the ovaries of 1 of 4 immunized ewes. These results show that immunization against androstenedione-11-BSA leads to increased ovulation rate and suggest two possible mechanisms: (1) interference with the negative feedback effects of oestradiol-17 beta and/or (2) a reduction in the incidence of follicular atresia.


Diffusion-weighted image and MR spectroscopic analysis of a case of MELAS with repeated attacks.
We report the clinical and MR manifestations of an 18 year-old girl with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome. Recurrent status epilepticus caused reversible cytotoxic edema on diffusion-weighted images (DWI). Initial and one month follow-up MR spectroscopy, after seizure control, showed some discrepancies in the ratio of metabolites. N-acetylaspartate (NAA) partially recovered (NAA/creatine (Cr) ratio: 1.27-->1.84). This was because of a normalization of decreased NAA due to cellular dysfunction as a result of status epilepticus. A low ratio of NAA/Cr due to abnormal mitochondria remained in the decreased state. Reversible NAA/Cr ratios in the acute lesion suggested that NAA reflects the neuronal function as well as the level of neuronal structural damage. The altered NAA/Cr ratio better correlated with the abnormal signal intensity area of T2-weighted images (T2WI) and DWI than the lactate (Lac)/Cr ratio. With conservative treatment with anti-epileptics not accompanied by coenzyme Q or sodium dichloroacetate, lactate persistently increased (Lac/Cr ratio: 1.01-->1.21) because of the continued production of lactate in cells with respiratory deficiency, which is the main pathology of MELAS.


Prevention of ankle sprains.
Ankles are sprained when supported on an unstable foundation, while too rididly fixed to the playing surface, or when forced into unnatural positions by extrinsic muscle tightness. The unstable foundation may be the shoe itself, a chuck-hole, or another player's foot. Undue fixation may be by 1-inch mud cleats, baseball spikes, or a modern wrestling mat. When these circumstances occur, heel cord tightness may alter the ankle's response. Thus, prevention of ankle sprains may be by modification of any of these factors.


Sedimentation velocity analysis of highly heterogeneous systems.
This article discusses several improvements to the van Holde-Weischet (vHW) method [Biopolymers 17 (1978) 1387] that address its capability to deal with sedimentation coefficient distributions spanning a large range of s values. The method presented here allows the inclusion of scans early and late in the experiment that ordinarily would need to be excluded from the analysis due to ultracentrifuge cell end effects. Scans late in the experiment are compromised by the loss of a defined plateau region and by back-diffusion from the bottom of the cell. Early scans involve partial boundaries that have not fully cleared the meniscus. In addition, a major refinement of the algorithm for determining the boundary fractions is introduced, taking into account different degrees of radial dilution for different species in the system. The method retains its desirable model-independent properties (the analysis of sedimentation data does not require prior knowledge of a user-imposed model or range of sedimentation coefficients) and reports diffusion-corrected s value distributions, which can be presented either in a histogram format or the traditional integral distribution format. Data analyzed with the traditional vHW method are compared with those of the improved method to demonstrate the benefit from the added information in the analysis.


Psychotic symptoms in psychiatric inpatients with dementia of the Alzheimer and vascular types.
Demented patients often manifest psychotic symptoms including delusion, hallucination and misidentification. This report attempts to describe the frequencies and characteristics of such symptoms in psychiatric inpatients with dementia, and to identify the characteristics of dementia with and without psychotic symptoms. The study included 78 demented patients who were consecutively admitted to the psychiatric ward. Their psychotic symptoms were obtained from semistructured interview and the assessment using BEHAVE AD. Delusion and hallucination were rated as "present" if the descriptions were consistent with the definition of Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R). Misidentification was defined individually. There were 54 patients with DAT and 24 patients with VD. The frequency of psychotic symptoms in DAT and VD was 71.8%. The frequency of delusion in DAT and VD was 59.0%, misidentification, 33.3% and hallucination, 25.6%. Demented patients with psychotic symptoms were older (p = 0.037) than those without psychotic symptoms. Psychotic symptoms are common in psychiatric inpatients with DAT and VD, the most common being delusional. Delusion of theft is the most common one. Demented patients with psychotic symptoms were older than those without.


Testicular tumour in non-palpable undescended testis: A rare presentation and laparoscopic management in a 4-year-old.
Testicular tumour in an intra-abdominal undescended testis is a rare finding. We report a case of mature teratoma in an intra-abdominal testis of a 4-year-old boy. He presented with nonpalpable testis on the right side. Abdominal ultrasound was normal. Diagnostic laparoscopy revealed a large tumour arising from an intra-abdominal right testis, and histologic analysis identified a mature teratoma. The tumour was completely removed laparoscopically, and there was no recurrence at follow-up 1 year later.


A Mendelian randomization study on the causal association of circulating cytokines with colorectal cancer.
Circulating cytokines have been associated with colorectal cancer (CRC). However, their causal correlation remains undetermined. This investigation uses genetic data to evaluate the mechanism that links circulating cytokines and CRC via Mendelian Randomization (MR). A two-sample MR evaluation was carried out to investigate the mechanism associating circulating cytokines and CRC in individuals of European ancestry. The Genome-wide association studies statistics, which are publically accessible, were used. Eligible instrumental SNPs that were significantly related to the circulating cytokines were selected. Multiple MR analysis approaches were carried out, including Simple Mode, inverse variance weighted (IVW), MR-Egger, Weighted Mode, Weighted Median, and MR pleiotropy residual sum and outlier (MR-PRESSO) methods. The evidence supporting the association of genetically predicted circulating levels with the increased risk of CRC was revealed; these included vascular endothelial growth factor (OR = 1.352, 95% CI: 1.019-1.315, P = 0.024), interleukin-12p70 (OR = 1.273, 95% CI: 1.133-1.430, P = 4.68×10-5), interleukin-13 (OR = 1.149, 95% CI: 1.012-1.299, P = 0.028), interleukin-10 (OR = 1.230, 95% CI: 1.013-1.493, P = 0.037), and interleukin-7 (OR = 1.191, 95% CI: 1.023-1.386 P = 0.024). Additionally, MR analysis negative causal association between macrophage colony stimulating factor and CRC (OR = 0.854, 95% CI: 0.764-0.955, P = 0.005). The data from Simple Mode, Weighted Median, MR-Egger, and Weighted Mode analyses were consistent with the IVW estimates. Furthermore, the sensitivity analysis indicated that the presence of no horizontal pleiotropy to bias the causal estimates. This investigation identified a causal association between circulating cytokines levels risk of CRC and may provide a deeper understanding of the pathogenesis of CRC, as well as offer promising leads for the development of novel therapeutic targets for CRC.


[Methodical approaches to evaluation of the safety of veterinary preparation application].
Evaluation of toxicologic hazards associated with the use of veterinary preparations against ecto- and endoparasites of animals and poultry with the purpose of prophylaxis and treatment, evaluation of industrial safety and determination of the upper limits of their residue in live-stock products and the environment, underlies protection of population's and operators' health from the effects of the drugs. The article shows the difference between the acute toxicity parameters of cypermethrin isomers and demonstrates that they enter the organism chiefly through the skin.


Follow-up in train-the-trainer continuing medical education events.
The purpose of train-the-trainer (TTT) programs within the context of continuing medical education (CME) is to help facilitators acquire and/or enhance their skills at leading CME sessions. The provision of follow-up is one feature of successful CME workshops over which CME providers have some control. Follow-up is defined as any encounter between participants and workshop leaders, following an initial workshop or other development session, and is designed to enhance, maintain, reinforce, transfer, extend, or support the learning from the original workshop. In this article, we elaborate on the use of audio teleconferences to provide follow-up for a TTT workshop in Saskatchewan, a largely rural province in western Canada. The teleconferences began 6 weeks after the workshop and were held at approximately 6-week intervals, with five conference calls in total. Each lasted about 45 minutes. Participants were interviewed to determine their view of the value of the teleconferences. Participants reported learning from the teleconferences and feeling more prepared to conduct CME sessions due to their participation in the teleconferences. Participants missed teleconferences only for extenuating circumstances (e.g., emergency deliveries). We have found that audio teleconferences allow for and encourage professional discussion that is crucial to changing practices. They are an effective way to incorporate follow-up to TTT workshops when participants travel great distances to attend.


Caspase-2 impacts lung tumorigenesis and chemotherapy response in vivo.
Caspase-2 is an atypical caspase that regulates apoptosis, cell cycle arrest and genome maintenance, although the mechanisms are not well understood. Caspase-2 has also been implicated in chemotherapy response in lung cancer, but this function has not been addressed in vivo. Here we show that Caspase-2 functions as a tumor suppressor in Kras-driven lung cancer in vivo. Loss of Caspase-2 leads to enhanced tumor proliferation and progression. Despite being more histologically advanced, Caspase-2-deficient tumors are sensitive to chemotherapy and exhibit a significant reduction in tumor volume following repeated treatment. However, Caspase-2-deficient tumors rapidly rebound from chemotherapy with enhanced proliferation, ultimately hindering long-term therapeutic benefit. In response to DNA damage, Caspase-2 cleaves and inhibits Mdm2 and thereby promotes the stability of the tumor-suppressor p53. Caspase-2 expression levels are significantly reduced in human lung tumors with wild-type p53, in agreement with the model whereby Caspase-2 functions through Mdm2/p53 regulation. Consistently, p53 target genes including p21, cyclin G1 and Msh2 are reduced in Caspase-2-deficient tumors. Finally, we show that phosphorylation of p53-induced protein with a death domain 1 leads to Caspase-2-mediated cleavage of Mdm2, directly impacting p53 levels, activity and chemotherapy response. Together, these studies elucidate a Caspase-2-p53 signaling network that impacts lung tumorigenesis and chemotherapy response in vivo.


[Mechanistic issues and prevention strategies targeting occupational carcinogenesis].
Carcinogenesis can be visualized either as a multistep process (initiation, promotion, progression, invasion, and metastasis) or as a continuum of mutagenic and mitogenic events, with the contribution of epigenetic mechanisms. The exponential growth of the neoplastic mass explains the importance of secondary prevention (early diagnosis) and of tertiary prevention. Primary prevention, which was successful in controlling occupational cancers, aims at minimizing exposures to carcinogens in healthy subjects and at favoring the intake of chemopreventive agents with dietary and pharmacological agents. Besides chemical carcinogens, often in the form of complex mixtures, the workplace may involve exposures to physical agents, such as sunlight and artificial illumination systems delivering UV radiation, or to biological agents, such as chronic viral infections (HBV, HCV, and HIV) associated with cancers. A controversial issue is the occurrence of threshold doses for carcinogens in the workplace and the environment.


The combination of ISCOMATRIX adjuvant and TLR agonists induces regression of established solid tumors in vivo.
The development of therapeutic vaccines for treatment of established cancer has proven challenging. Cancer vaccines not only need to induce a robust tumor Ag-specific immune response but also need to overcome the tolerogenic and immunosuppressive microenvironments that exist within many solid cancers. ISCOMATRIX adjuvant (ISCOMATRIX) is able to induce both tumor Ag-specific cellular and Ab responses to protect mice against tumor challenge, but this is insufficient to result in regression of established solid tumors. In the current study, we have used B16-OVA melanoma, Panc-OVA pancreatic, and TRAMP-C1 prostate cancer mouse tumor models to test therapeutic efficacy of ISCOMATRIX vaccines combined with other immune modulators. The coadministration of an ISCOMATRIX vaccine with the TLR3 agonist, polyinosinic-polycytidylic acid, and TLR9 agonist, CpG, reduced tumor growth in all tumor models and the presence of ISCOMATRIX in the formulation was critical for the therapeutic efficacy of the vaccine. This vaccine combination induced a robust and multifunctional CD8(+) T cell response. Therapeutic protection required IFN-γ and CD8(+) T cells, whereas NK and CD4(+) T cells were found to be redundant. ISCOMATRIX vaccines combined with TLR3 and TLR9 agonists represent a promising cancer immunotherapy strategy.


[The response of free alpha-subunit of glycoprotein hormones to LH-RH administration (author's transl)].
The responses of immunoreactive free alpha-subunit of glycoprotein hormones to LH-RH administration were studied in normal men and women, and in patients with hypergonadotropic hypogonadism, hypogonadotropic hypogonadism, trophoblastic disease and isolated ectopic alpha-subunit producing tumor. In patients with hypergonadotropic hypergonadism, basal levels of serum alpha-subunit were elevated and the responses to LH-RH were also excessive compared to those of normal men and women. Conversely, in hypogonadotropic hypogonadism, basal levels of alpha-subunit were significantly low and its response to LH-RH was barely detectable. The response of alpha-subunit to constant intravenous infusion of LH-RH (1 microgram/kg/h) was studied in 4 normal men. Both LH and alpha-subunit revealed biphasic patterns of elevation. Its releasing pattern suggests the possibility that two pools of gonadotropin are involved in the production and secretion of alpha-subunit. In patients with trophoblastic disease secreting low levels of hCG (18 mIU/ml), the responses of alpha-subunit as well as pituitary gonadotropin to LH-RH were normal. However, in cases of high concentrations of hCG (1000 mIU/ml), the responses of alpha-subunit and gonadotropin were suppressed. After the administration of LH-RH to a patient with an isolated ectopic alpha-subunit producing tumor, the serum concentration of pituitary gonadotropin increased within the normal range, although that of alpha-subunit did not show a significant change. These results suggest that the production of alpha-subunit by tumors may be autonomous in contrast with a regulatory production in the pituitary.


Multiple and sensitive SERS detection of cancer-related exosomes based on gold-silver bimetallic nanotrepangs.
Exosomes are endogenous vesicles of cells, and can be used as important biomarkers for cancers. In this work, we developed a sensitive and reliable SERS sensor for simultaneous detection of multiple cancer-related exosomes. The SERS detection probes were made of bimetallic SERS-active nanotags, gold-silver-silver core-shell-shell nanotrepangs (GSSNTs), which were composed of bumpy surface nanorod (gold nanotrepang, GNT) cores and bilayer silver shells, and decorated with linker DNAs, which were complementary to the aptamer targeting exosomes. Three kinds of SERS detection probes were designed via the adoption of different Raman reporter molecules and linker DNAs. The capture probes were prepared by modifying specific aptamers of the target exosomes on magnetic beads (MBs). In the absence of target exosomes, SERS detection probes were coupled with MBs via specific DNA hybridization for use as aptamer-based SERS sensors. In the presence of target exosomes, the aptamer specifically recognized and captured the exosomes, and GSSNTs were subsequently released into the supernatant. Therefore, attenuated SERS signals were detected on the MBs, indicating the presence of target exosomes. The proposed aptamer-based SERS sensor is expected to be a facile and sensitive method for the multiplex detection of cancer biomarkers and has potential future applications in clinical diagnosis.


Life-cycling and uncovering cation-trapping evidence of a monolithic inorganic electrochromic device: glass/ITO/WO3/LiTaO3/NiO/ITO.
The visualization of the microstructure change and of the depth of lithium transport inside a monolithic ElectroChromic Device (ECD) is realized using an innovative combined approach of Focused Ion Beam (FIB), Secondary Ion Mass Spectrometry (SIMS) and Glow Discharge Optical Emission Spectroscopy (GDOES). The electrochemical and optical properties of the all-thin-film inorganic ECD glass/ITO/WO3/LiTaO3/NiO/ITO, deposited by magnetron sputtering, are measured by cycling voltammetry and in situ transmittance analysis up to 11 270 cycles. A significant degradation corresponding to a decrease in the capacity of 71% after 2500 cycles and of 94% after 11 270 cycles is reported. The depth resolved microstructure evolution within the device, investigated by cross-sectional cutting with FIB, points out a progressive densification of the NiO layer upon cycling. The existence of irreversible Li ion trapping in NiO is illustrated through the comparison of the compositional distribution of the device after various cycles 0, 100, 1000, 5000 and 11 270. SIMS and GDOES depth profiles confirm an increase in the trapped Li content in NiO as the number of cycles increases. Therefore, the combination of lithium trapping and apparent morphological densification evolution in NiO is believed to account for the degradation of the ECD properties upon long term cycling of the ECD.


IGF-1 and PDGF-bb suppress IL-1β-induced cartilage degradation through down-regulation of NF-κB signaling: involvement of Src/PI-3K/AKT pathway.
Interleukin-1β (IL-1β) is a pro-inflammatory cytokine that plays a key role in the pathogenesis of osteoarthritis (OA). Growth factors (GFs) capable of antagonizing the catabolic actions of cytokines may have therapeutic potential in the treatment of OA. Herein, we investigated the potential synergistic effects of insulin-like growth factor (IGF-1) and platelet-derived growth factor (PDGF-bb) on different mechanisms participating in IL-1β-induced activation of nuclear transcription factor-κB (NF-κB) and apoptosis in chondrocytes. Primary chondrocytes were treated with IL-1β to induce dedifferentiation and co-treated with either IGF-1 or/and PDGF-bb and evaluated by immunoblotting and electron microscopy. Pretreatment of chondrocytes with IGF-1 or/and PDGF-bb suppressed IL-1β-induced NF-κB activation via inhibition of IκB-α kinase. Inhibition of IκB-α kinase by GFs led to the suppression of IκB-α phosphorylation and degradation, p65 nuclear translocation and NF-κB-regulated gene products involved in inflammation and cartilage degradation (COX-2, MMPs) and apoptosis (caspase-3). GFs or BMS-345541 (specific inhibitor of the IKK) reversed the IL-1β-induced down-regulation of collagen type II, cartilage specific proteoglycans, β1-integrin, Shc, activated MAPKinase, Sox-9 and up-regulation of active caspase-3. Furthermore, the inhibitory effects of IGF-1 or/and PDGF-bb on IL-1β-induced NF-κB activation were sensitive to inhibitors of Src (PP1), PI-3K (wortmannin) and Akt (SH-5), suggesting that the pathway consisting of non-receptor tyrosine kinase (Src), phosphatidylinositol 3-kinase and protein kinase B must be involved in IL-1β signaling. The results presented suggest that IGF-1 and PDGF-bb are potent inhibitors of IL-1β-mediated activation of NF-κB and apoptosis in chondrocytes, may be mediated in part through suppression of Src/PI-3K/AKT pathway, which may contribute to their anti-inflammatory effects.


New library construction method for single-cell genomes.
A central challenge in sequencing single-cell genomes is the accurate determination of point mutations, phasing of these mutations, and identifying copy number variations with few assumptions. Ideally, this is accomplished under as low sequencing coverage as possible. Here we report our attempt to meet these goals with a novel library construction and library amplification methodology. In our approach, single-cell genomic DNA is first fragmented with saturated transposition to make a primary library that uniformly covers the whole genome by short fragments. The library is then amplified by a carefully optimized PCR protocol in a uniform and synchronized fashion for next-generation sequencing. Each step of the protocol can be quantitatively characterized. Our shallow sequencing data show that the library is tightly distributed and is useful for the determination of copy number variations.


Biosynthesis of drug metabolites and quantitation using NMR spectroscopy for use in pharmacologic and drug metabolism studies.
The contribution of drug metabolites to the pharmacologic and toxicologic activity of a drug can be important; however, for a variety of reasons metabolites can frequently be difficult to synthesize. To meet the need of having samples of drug metabolites for further study, we have developed biosynthetic methods coupled with quantitative NMR spectroscopy (qNMR) to generate solutions of metabolites of known structure and concentration. These quantitative samples can be used in a variety of ways when a synthetic sample is unavailable, including pharmacologic assays, standards for in vitro work to help establish clearance pathways, and/or as analytical standards for bioanalytical work to ascertain exposure, among others. We illustrate five examples of metabolite biosynthesis and qNMR. The types of metabolites include one glucuronide and four oxidative products. Concentrations of the isolated metabolite stock solutions ranged from 0.048 to 8.3 mM, with volumes from approximately 0.04 to 0.150 ml in hexadeutarated dimethylsulfoxide. These specific quantified isolates were used as standards in the drug discovery setting as substrates in pharmacology assays, for bioanalytical assays to establish exposure, and in variety of routine absorption, distribution, metabolism, and excretion assays, such as protein binding and determining blood-to-plasma ratios. The methods used to generate these materials are described in detail with the objective that these methods can be generally used for metabolite biosynthesis and isolation.


The effects of norepinephrine infusion on the circulating lymphocyte counts of post-open heart surgery patients.
In this retrospective study employing chart reviews, 75 open heart surgery patients (OHSPs) were divided into 3 groups of 25 patients. Group 1 received no intravenous (i.v.) norepinephrine (NE) after surgery. Group 2 and group 3 received a minimum of 0.028 mcg/kg/min of i.v. NE for 6-24 h and greater than 24 h, respectively. In the 3 groups, preoperative lymphocyte counts were compared to counts obtained on postoperative days 1 and 2. The results showed lower lymphocyte counts on postoperative day 2 in group 3 subjects, who received NE for 24 h or more, compared to subjects of the other groups who received no NE or 6-24 h of NE (p < 0.05). There was also evidence that preoperative use of beta-blocking agents significantly affected the change in lymphocyte counts from day 1 to day 2 in both groups receiving NE. Furthermore, postoperative infections were more prevalent in group 3 than the other 2 groups (p < 0.05). The lower lymphocyte counts and higher infection rate, however, may be linked to lower postoperative blood pressure and increased number of intensive care unit days in group 3. Further investigation is warranted to elucidate the effects of i.v. NE administration on the lymphocyte counts of OHSPs and to reduce infections in those receiving NE.


Postprandial hyperinsulinaemic hypoglycaemia and type 1 diabetes mellitus.
A patient with severe postprandial hyperinsulinaemic hypoglycaemia (PPHH) for 4 years developed type 1 diabetes mellitus. She had no insulin or insulin receptor antibodies but was positive for islet cell and glutamic acid decarboxylase (GAD) antibodies. PPHH prior to the onset of type 1 diabetes mellitus has not been previously described and may be a prodrome of type 1 diabetes mellitus.


3D full-field strain in bone-implant and bone-tooth constructs and their morphological influential factors.
The biomechanics of bone-tooth and bone-implant interfaces affects the outcomes of several dental treatments, such as implant placement, because bone, tooth and periodontal ligament are living tissues that adapt to the changes in mechanical stimulations. In this work, mechanical testing coupled with micro-CT was performed on human cadaveric mandibular bone-tooth and bone-implant constructs. Using digital volume correlation, the 3D full-field strain in bone under implant loading and tooth loading was measured. Concurrently, bone morphology and bone-implant and bone-tooth contact were also measured through the analysis of micro-CT images. The results show that strain in bone increased when a tooth was replaced by a dental implant. Strain concentration was observed in peri-implant bone, as well as in the buccal bone plate, which is also the clinically-observed bone resorption area after implant placement. Decreasing implant stability measurements (resonance frequency analysis and torque test) indicated increased peri-implant strain, but their relationships may not be linear. Peri-implant bone strain linearly increased with decreasing bone-implant contact (BIC) ratio. It also linearly decreased with increasing bone-tooth/bone-implant contact ratio. The high strain in the buccal bone plate linearly increased with decreasing buccal bone plate thickness. The results of this study revealed 3D full-field strain in bone-tooth and bone-implant constructs, as well as their several morphological influential factors.


Ultrastructural studies of beef heart mitochondria. 3. The inequality of gross morphological change and oxidative phosphorylation.
The relationships between membranes and intramembrane compartments of isolated heart mitochondria are inadequately defined to express the induced morphological changes associated with the structural organization. The inner membrane and matrix are the major structural entities which undergo transformation upon alteration of metabolism or incubation conditions. To better express these morphological changes within a mitochondrion, two inner membranes plus enclosed matrix are defined as an inmerix (plural inmerices). Three general morphological forms of mitochondria can be distinguished by the size and shape of inmerices. These are distended, condensed, and coalesced inmerixal configurations. Hypotonic conditions and P(i) in isotonic sucrose generate distended configurations. This P(i) distention is apparently dependent on utilization of energy. It does not occur under anaerobic conditions. Oxidizable substrates generate condensed configurations. ADP and dADP generate coalesced configurations and stop formation of condensed and distended inmerixal configurations in the absence of inhibitors. ADP coalescence is apparently not dependent on an energy input. It occurs under aerobic and anaerobic conditions, and in isotonic and hypotonic media. Atractyloside completely inhibits the effects of ADP on inmerixal membranes whereas oligomycin does not. Distention by P(i) is unaffected by the two inhibitors. Distended inmerices, without added P(i) (12 mM and 62 mM sucrose), are coalesced by ADP. These studies indicate that coalescence of inmerixal membranes probably reflects the consequences of specific stoichiometric binding or translocation of adenine nucleotides.


Passive transfer of modest titers of potent and broadly neutralizing anti-HIV monoclonal antibodies block SHIV infection in macaques.
It is widely appreciated that effective human vaccines directed against viral pathogens elicit neutralizing antibodies (NAbs). The passive transfer of anti-HIV-1 NAbs conferring sterilizing immunity to macaques has been used to determine the plasma neutralization titers, which must be present at the time of exposure, to prevent acquisition of SIV/HIV chimeric virus (SHIV) infections. We administered five recently isolated potent and broadly acting anti-HIV neutralizing monoclonal antibodies (mAbs) to rhesus macaques and challenged them intrarectally 24 h later with either of two different R5-tropic SHIVs. By combining the results obtained from 60 challenged animals, we determined that the protective neutralization titer in plasma preventing virus infection in 50% of the exposed monkeys was relatively modest (∼1:100) and potentially achievable by vaccination.


Mapping the research utilization field in nursing.
The recent increase in interest in the field of research utilization, often embedded in the notions of evidence-based practice, presents a rich opportunity to advance the field in nursing. While an extensive literature on the subject exists in nursing, close examination reveals that much of it is opinion and anecdotal literature, and that sustained and programmatic theory building and testing in this field has been sporadic at best. This article maps the field of research utilization, proposing that we focus on major areas of inquiry: scientific, historical, and philosophical foundations, synthesis, determinants, policy, interventions to increase research utilization, and outcomes. In so doing, alternative ways of viewing and conceptualizing this field are possible. In conducting the kinds of studies and supporting the kinds of programs identified in this map, nursing, in collaboration with appropriate partners, can significantly advance the field of research dissemination and utilization studies and practice at many levels in the health system.


A Rare Mutation in LMNB2 Associated with Lipodystrophy Drives Premature Cell Senescence.
Many proteins are causative for inherited partial lipodystrophies, including lamins, the essential constituents of the nuclear envelope scaffold called the lamina. By performing high throughput sequencing on a panel of genes involved in lipodystrophies, we identified a heterozygous mutation in LMNB2 gene (c.700C > T p.(Arg234Trp)) in a female patient presenting early onset type II diabetes, hypertriglyceridemia, and android fat distribution. This mutation is rare in the general population (frequency 0.013% in GnomAD) and was predicted pathogenic by a set of pathogenicity prediction software. Patient-derived fibroblasts showed nuclear shape abnormalities and premature senescence features, which are two typical cellular phenotypes associated with laminopathies. Moreover, we observed an atypical aggregation of lamin B2 in nucleoplasm, which co-distributes with emerin and lamin A/C, along with an abnormal distribution of lamin A/C at the nuclear envelope. Finally, reducing lamin B2 expression level by siRNA targeted toward LMNB2 transcripts resulted in decreased nuclear anomalies and senescence-associated beta-galactosidase, suggesting a role of the mutated protein in the occurrence of the observed cellular phenotype. Altogether, these results suggest that mutations in lamin B2 could produce premature senescence and partial lipodystrophy features as observed with certain mutants of lamin A/C.


[Neuron non-electrical functions].
Classical histological preparations of the metasympathetic nervous system treated with silver salts were compared with isolated neuron cultures to study non-electrical functions of the neurons. A hypothesis of a tissue trophic effect of the nervous tissue's receptors, is advanced. Proteolytic enzymes were shown to be a growth factor for surrounding tissues and to play a role of trophic agents. A permanent restructurisation seems to occur in formed plexuses of adult healthy animals.


Impact of Mutation Rate and Selection at Linked Sites on DNA Variation across the Genomes of Humans and Other Homininae.
DNA diversity varies across the genome of many species. Variation in diversity across a genome might arise from regional variation in the mutation rate, variation in the intensity and mode of natural selection, and regional variation in the recombination rate. We show that both noncoding and nonsynonymous diversity are positively correlated to a measure of the mutation rate and the recombination rate and negatively correlated to the density of conserved sequences in 50 kb windows across the genomes of humans and nonhuman homininae. Interestingly, we find that although noncoding diversity is equally affected by these three genomic variables, nonsynonymous diversity is mostly dominated by the density of conserved sequences. The positive correlation between diversity and our measure of the mutation rate seems to be largely a direct consequence of regions with higher mutation rates having more diversity. However, the positive correlation with recombination rate and the negative correlation with the density of conserved sequences suggest that selection at linked sites also affect levels of diversity. This is supported by the observation that the ratio of the number of nonsynonymous to noncoding polymorphisms is negatively correlated to a measure of the effective population size across the genome. We show these patterns persist even when we restrict our analysis to GC-conservative mutations, demonstrating that the patterns are not driven by GC biased gene conversion. In conclusion, our comparative analyses describe how recombination rate, gene density, and mutation rate interact to produce the patterns of DNA diversity that we observe along the hominine genomes.


Tolerability of combined treatment with verapamil and beta-blockers in angina resistant to monotherapy.
We have used a combination of a beta-blocker and verapamil to treat 42 consecutive patients with angina resistant to either agent alone. Patients with heart failure, heart block or uncontrolled hypertension were excluded. The mean duration of follow-up was 6.5 months. Thirty-six patients (81%) reported an improvement and the number of angina attacks was reduced from 17/week to 5/week. Side effects necessitated withdrawal of one or both drugs in 6 patients, 2 of whom developed bradyarrhythmias not solely related to drug treatment. The most common complication was mild left ventricular failure (6) treated by reducing or stopping the beta-blocker. The data suggest that the combination of verapamil and a beta-blocker may be used in a relatively unselected group of patients with difficult angina. However, as dosage adjustment and close observation may be necessary to minimise side effects, the use of this combination should be limited to hospital practice.


Enjoyment and Intensity of Physical Activity in Immersive Virtual Reality Performed on Innovative Training Devices in Compliance with Recommendations for Health.
The aim of the study is to assess the enjoyment and intensity of physical exercise while practicing physical activity (PA) in immersive virtual reality (IVR) using innovative training devices (omni-directional Omni treadmill and Icaros Pro flight simulator). The study also contains the results of subjective research on the usefulness of such a form of PA in the opinion of users. In total, 61 adults (10 women and 51 men) took part in the study. To assess the enjoyment level (EL) Interest/Enjoyment subscale of Intrinsic Motivation Inventory (IMI) was used. Exercise intensity was assessed during 10-min sessions of active video games (AVGs) in IVR based on heart rate (HR). The average enjoyment level during physical exercise in IVR on the tested training devices was high (Omni 5.74 points, Icaros 5.60 points on a 1-7 Likert scale) and differed significantly in favor of PA on Omni. In the opinion of the majority of participants, AVGs in IVR on the tested devices constitute a sufficiently useful form of PA to meet the needs of leisure time activities, and they can even replace some forms of physical effort performed in a classic way. The intensity of PA during games on training devices was at the level recommended for health benefits for 80.55% (Omni) and 50.77% (Icaros Pro) of its duration. Based on the conducted research, it can be assumed that AVGs in IVR using a multi-directional treadmill and a flight simulator can be an effective tool for increasing participation in health-oriented PA.


Transient receptor potential ankyrin 1 channels in the bladder mediate low temperature elicited bladder overactivity in rats.
This study aimed to investigate whether pathways involving transient receptor potential ankyrin 1 (TRPA1) channels in the urinary bladder mediate the bladder overactivity elicited by exposure to a low temperature in rats. At postnatal week 10, female Sprague-Dawley (SD) rats were intraperitoneally injected with the TRPA1 channel antagonist, HC030031, at room temperature (RT) and subsequently exposed to low temperature (LT). Bladder specimens treated with HC030031 were evaluated for contractions through cumulative addition of the TRPA1 channel agonist trans-cinnamaldehyde. Two days before cystometric investigation, small interfering RNA (siRNA) targeting TRPA1 was transfected into urinary bladders. Then, cystometric investigations were performed on rats subjected to TRPA1 siRNA transfection at both RT and LT. Expression of TRPA1 channels in the urinary bladder was assessed through immunohistochemistry and real-time reverse transcription-polymerase chain reaction. At RT, micturition patterns were unaffected by HC030031 treatment. However, upon exposure to LT, rats treated with HC030031 exhibited a reduction of LT-elicited bladder overactivity, as evidenced by inhibited decreases in voiding interval, micturition volume, and bladder capacity. Additionally, HC030031 inhibited trans-cinnamaldehyde-induced contractions. Immunohistochemical analysis showed the presence of TRPA1 channels in the urinary bladder. Notably, rats with TRPA1 siRNA-transfected bladders could partially inhibit bladder overactivity during LT exposure. These findings indicate that pathways involving TRPA1 channels expressed in the urinary bladder could mediate the LT-elicited bladder overactivity.


Singing for your supper: sentinel calling by kleptoparasites can mitigate the cost to victims.
Parasitism generally imposes costs on victims, yet many victims appear to tolerate their parasites. We suggest that in some cases this may be because parasites provide victims with mitigating benefits, paradoxically giving rise to selection for advertisement rather than concealment by parasites. We investigate this possibility using the interaction between an avian kleptoparasite, the fork-tailed drongo (Dicrurus adsimilis), and one of its victims, the pied babbler (Turdoides bicolor). Combining field observations and a playback experiment, we demonstrate that a conspicuous vocal signal broadcast by drongos perched waiting to steal food from foraging babblers allows the latter to improve their own foraging efficiency, although not to the same extent as that experienced in response to conspecific sentinel calling. We argue that "sentinel" calling by drongos may originally have arisen as a means of manipulating babblers: because babblers find more food items and venture into the open more in response to these vocalizations, drongos are presented with more kleptoparasitism opportunities. However, the resulting benefit to babblers could be sufficient to reduce selection for the evolution of defenses against drongos, and the current situation may represent a rare example of an interspecific relationship in transition from a parasitism to a mutualism.


Correlation-based evaluation of visual performance to reduce the statistical error of visual acuity.
Ophthalmologists evaluate visual acuity tests by the number of correctly recognized optotypes (usually letters) in the different lines of an eye chart. This probability-based scoring results in significant statistical error that can only be decreased by the time-consuming analysis of a larger number of optotypes. In this paper, we present a new, more precise correlation-based scoring method that takes the degree of misidentification into consideration too, rather than the mere fact of it. According to our experimental results, this new method decreases the uncertainty error by 28% if using the same number of optotypes at a given letter size or requires half the optotype number to produce the same error as that of probability-based scoring.


Unmasking a cognitive vulnerability to depression: how lapses in mental control reveal depressive thinking.
This research tested the idea that a cognitive vulnerability to depression can be concealed by thought suppression and revealed when cognitive demands undermine mental control. Depressive, at-risk, and nondepressive participants unscrambled sentences that could from either positive or depressive statements. Half of the participants also received a cognitive load. The results indicated that without a load, at-risk participants showed little evidence of depressive thinking, producing a similar rate of positive statements as did nondepressive individuals and a lower percentage of negative statements than did depressive participants. However, the cognitive load caused an increase in at-risk participants' production of negative statements, revealing a previously undetected tendency toward negative thinking that made them resemble depressive participants. As predicted, this effect was especially pronounced among individuals who routinely engaged in thought suppression.


Prognostic factors of renal cell carcinoma with extension into inferior vena cava.
To evaluate the prognosis of our series of patients with renal cell carcinoma (RCC) and tumor thrombus involving inferior vena cava (IVC) treated with nephrectomy and thrombectomy. In 46 patients with unilateral RCC extending into IVC who underwent nephrectomy and thrombectomy (T3b in 38 patients, T3c in 6, T4 in 2, N+ in 15, M1 in 21), overall and cancer-specific survival rates were estimated, and the univariable and multivariable analysis were carried out to determine the prognostic factors among age, gender, performance status, fever, inflammatory laboratory parameters, nodal and distant metastasis, tumor thrombus level, pathological parameters and postoperative interferon-alpha administration. The median age was 66.5 (range 35-79) years. The median follow-up was 18.0 (mean 36.7 +/- 38.7) months. The overall and cancer-specific 5-year survival rates were 32.9% and 40.0%, respectively. The univariate analysis revealed that fever (hazard ratio: HR 4.03), C-reactive protein (HR 4.89), grade of tumor cell (HR 3.83), and lymph node metastasis (HR 5.99) were independent prognostic factors of cause-specific survival in all patients. The multivariate analysis demonstrated that lymph node metastasis (HR 4.13) was the only independent prognostic factor of cause-specific survival. The extension level or postoperative interferon-alpha administration did not influence the prognosis of patients with tumor thrombus involving IVC. Aggressive surgery should be considered first in RCC patients with any levels of tumor thrombus. However, patients with both IVC involvement and nodal metastasis showed significantly poor prognosis, and development of novel intensive multidisciplinary therapies will be needed.


Visualization and quantification of anastomotic perfusion in colorectal surgery using near-infrared fluorescence.
Anastomotic leakage (AL) is one of the most troublesome complications in colorectal surgery. Recently, near-infrared fluorescence (NIRF) imaging has been used intraoperatively to detect sentinel lymph nodes and visualize the blood supply at the region of interest (ROI). The aim of this study was to evaluate the role of visualization and quantification of bowel perfusion around the anastomosis using NIRF system in predicting AL. A prospective study was conducted on patients who had laparoscopic surgery for colorectal cancer at our institution. Perfusion of the anastomosis was evaluated with NIRF imaging after intravenous injection of indocyanine green (ICG). The time course of fluorescence intensity was recorded by an imaging analyzer We measured the time from ICG injection to the beginning of fluorescence (T0), maximum intensity (Imax), time to reach Imax (Tmax), time to reach Imax 50% ([Formula: see text]) and slope (S) after the anastomosis. Tumor locations were as follows; cecum: 2, ascending colon: 2, transverse colon: 7, descending colon: 1, sigmoid colon: 2, rectosigmoid colon: 3 and rectum: 6 (one case with synchronous cancer). All operations were performed laparoscopically. Four patients were diagnosed with or suspected to have AL (2 patients with grade B anastomotic leakage after low anterior resection, 1 patient with minor leakage in transverse colon resection and 1 patient needing re-anastomosis intraoperatively in transverse colon resection). T0 was significantly longer in the AL group than in patients without AL (64.3 ± 27.6 and 18.2 ± 6.6 s, p = 2.2 × 10-3). Perfusion of the anastomosis could be successfully visualized and quantified using NIRF imaging with ICG. T0 might be a useful parameter for prediction of AL.


Physical exam and occult post-traumatic vascular lesions: implications for the evaluation and management of arterial injuries in modern warfare in the endovascular era.
Based on complexity of extremity wounds sustained in recent combat, arteriography had been used routinely in evaluations for delayed or occult arterial injuries. This report aims to quantitatively analyze the sensitivity and specificity of physical exam (PE) in predicting the presence of these injuries. United States service members sustaining extremity trauma in the Global War on Terrorism were evacuated to our medical center and evaluated by a senior vascular surgeon. Those with an abnormal PE, at risk based on wounding patterns, or previously treated for vascular injury underwent arteriography. Data from each patient were prospectively entered into a Vascular Injury registry. Comprehensive information about the injuries, interventions, arteriogram results, and any periprocedural complications were analyzed. Twenty-five endovascular or open surgical interventions were performed in forty-six of 99 patients that had lesions on arteriography. Seventy-three patients had a normal PE, of which 36 had lesions that prompted 6 subsequent interventions. Twenty-two of 26 patients with an abnormal PE had lesions that prompted 19 interventions. For PE, sensitivity was 38%, specificity was 90%, and positive predictive value (PPV) and negative predictive value (NPV) were 85% and 51%, respectively. In proximity injuries, PPV improved to 100%, but was only 15% sensitive with a NPV of 60%. In conclusion normal PE did not reliably predict post-traumatic arterial lesions in these military extremity injuries. These lesions are amenable to endovascular therapies, and should be considered in cases of complex trauma involving high amounts of energy, penetrating mechanisms, or wounding patterns in proximity to named vessels.


Meta-analysis indicates that the European GWAS-identified risk SNP rs1344706 within ZNF804A is not associated with schizophrenia in Han Chinese population.
Recent genetic association studies have implicated several candidate susceptibility variants for schizophrenia among general populations. Rs1344706, an intronic SNP within ZNF804A, was identified as one of the most compelling candidate risk SNPs for schizophrenia in Europeans through genome-wide association studies (GWASs) and replications as well as large-scale meta-analyses. However, in Han Chinese, the results for rs1344706 are inconsistent, and whether rs1344706 is an authentic risk SNP for schizophrenia in Han Chinese is inconclusive. Here, we conducted a systematic meta-analysis of rs1344706 with schizophrenia in Chinese population by combining all available case-control samples (N = 12), including a total of 8,982 cases and 12,342 controls. The results of our meta-analysis were not able to confirm an association of rs1344706 A-allele with schizophrenia (p = 0.10, odds ratio = 1.06, 95% confidence interval = 0.99-1.13). Such absence of association was further confirmed by the non-superiority test (p = 0.0003), suggesting that rs1344706 is not a risk SNP for schizophrenia in Han Chinese. Detailed examinations of individual samples revealed potential sampling bias in previous replication studies in Han Chinese. The absence of rs1344706 association in Han Chinese suggest a potential genetic heterogeneity in the susceptibility of schizophrenia on this locus and also demonstrate the difficulties in replicating genome-wide association findings of schizophrenia across different ethnic populations.


Motorcycle right-of-way accidents--a literature review.
The most typical automobile-motorcycle collision take places when an automobile manoeuvres into the path of an approaching motorcycle by violating the motorcycle's right of way (ROW). The present paper provides a comprehensive review of past research that examined motorcycle ROW accidents. Articles and publications were selected for relevance and research strength through a comprehensive search of major databases such as Transportation Research Information Services (TRIS), Compendex, and Medline. Two major causes of such a crash scenario are the lack of motorcycle conspicuity and motorist's speed/distance judgment error, respectively. A substantial number of studies have manipulated physical characteristics of motorcycles and motorcyclists to enhance conspicuity, along with research addressing motorists' gap-acceptance behaviours and arrival time judgments when confronting motorcycles. Although various conspicuity aids have proven effective, some researchers reported that motorcyclist's/motorcycle's brightness per se may be less important as a determinant of conspicuity than brightness contrast between the motorcyclists and the surroundings. Larger vehicles tended to be judged to arrive sooner than motorcycles. Such a speed/distance judgment error is likely attributable to some psychological effects such that larger automobiles appear more threatening than motorcycles. Older motorists particularly have difficulties in accurately estimating the distance and the speed of an approaching motorcycle. Research examining the effects of conspicuity measures on motorists' speed/distance judgments when confronting motorcycles has been rather inconclusive. Past research offers valuable insight into the underlying motorcycle ROW crash mechanisms. However, with ageing society and a rapid change in traffic composition (e.g., more larger motorcycles) in recent years, prior research findings should be updated. The present study finally provides recommendations for future research on motorcycle ROW accidents.


Asymptomatic shedding of herpes simplex virus from the cervix and lesion site during pregnancy. Correlation of antepartum shedding with shedding at delivery.
Asymptomatic shedding of herpes simplex virus (HSV) was detected in 0.83% of 955 cultures obtained from pregnant women with culture-proved recurrent genital HSV infections during pregnancy; seven (2.3%) of 299 women had asymptomatic shedding. In addition, one (2.3%) of 42 pregnant women with recurrent genital infection in the past but no attacks during pregnancy shed HSV when asymptomatic. Shedding occurred more frequently from the usual lesion site than from the cervix. The virus was not isolated at delivery from women with asymptomatic antepartum shedding. When an active lesion was present, concomitant shedding of HSV from the cervix occurred in seven (3.6%) of 193 pregnant women with vulvar lesions and in one (2.1%) of 47 women with lesions remote from the vulva. Ninety-two percent of the latter lesions were caused by type 2 HSV. In women who have had recurrent genital HSV infections in the past, asymptomatic shedding occurs in those with active attacks during pregnancy as well as in those asymptomatic throughout pregnancy; however, asymptomatic shedding during the antepartum period does not predict asymptomatic shedding at delivery.


Synaptotagmin VII restricts fusion pore expansion during lysosomal exocytosis.
Synaptotagmin is considered a calcium-dependent trigger for regulated exocytosis. We examined the role of synaptotagmin VII (Syt VII) in the calcium-dependent exocytosis of individual lysosomes in wild-type (WT) and Syt VII knockout (KO) mouse embryonic fibroblasts (MEFs) using total internal reflection fluorescence microscopy. In WT MEFs, most lysosomes only partially released their contents, their membrane proteins did not diffuse into the plasma membrane, and inner diameters of their fusion pores were smaller than 30 nm. In Syt VII KO MEFs, not only was lysosomal exocytosis triggered by calcium, but all of these restrictions on fusion were also removed. These observations indicate that Syt VII does not function as the calcium-dependent trigger for lysosomal exocytosis. Instead, it restricts the kinetics and extent of calcium-dependent lysosomal fusion.


Developmental and structural studies of an intracellular lipid binding protein expressed in the ileal epithelium.
Enterocytes located in the pig distal small intestine (ileum) contain a cytosolic protein that is homologous to two proteins that are also synthesized in these cells: intestinal and "liver" fatty acid-binding proteins (I- and L-FABPc, respectively). To begin to investigate the functional interrelationships of these three proteins, we compared their patterns of tissue-specific expression and developmental regulation in the mouse. Blot hybridization analyses of RNA prepared from 12 adult tissues revealed that this mRNA was confined to the small intestine. Unlike I- and L-FABPc mRNA, which are most abundant in the proximal jejunum, this mRNA is most abundant in the ileum. While I- and L-FABPc gene transcription commence in late fetal life coincident with initial cytodifferentiation of the mouse gut epithelium, the ileal gene is activated later, at the suckling/weaning transition (postnatal day 12). The ileal location and developmental pattern of expression suggested that this protein may play a role in the intracellular transport of bile salts in the ileal epithelium. To test this hypothesis, we expressed the porcine ileal peptide (PIP) in Escherichia coli, purified it to apparent homogeneity, and analyzed its binding properties for bile acids and fatty acids using 13C NMR spectroscopy. Like I-FABPc, PIP binds palmitate and oleate with a 1:1 molar stoichiometry. However, unlike I-FABPc PIP binds chenodeoxycholate. In addition, the presence of chenodeoxycholate blocks fatty acid binding to PIP, but not to I-FABPc. E. coli-derived PIP was subsequently crystallized with and without chenodeoxycholic acid. All crystals are orthorhombic in the P2(1)2(1)2(1) space group. The unit cell dimensions are a = 36.15 A, b = 50.13 A, and c = 67.18 A.


A Crude Extract Preparation and Optimization from a Genomically Engineered Escherichia coli for the Cell-Free Protein Synthesis System: Practical Laboratory Guideline.
With the advancement of synthetic biology, the cell-free protein synthesis (CFPS) system has been receiving the spotlight as a versatile toolkit for engineering natural and unnatural biological systems. The CFPS system reassembles the materials necessary for transcription and translation and recreates the in vitro protein synthesis environment by escaping a physical living boundary. The cell extract plays an essential role in this in vitro format. Here, we propose a practical protocol and method for Escherichia coli-derived cell extract preparation and optimization, which can be easily applied to both commercially available and genomically engineered E. coli strains. The protocol includes: (1) The preparation step for cell growth and harvest, (2) the thorough step-by-step procedures for E. coli cell extract preparation including the cell wash and lysis, centrifugation, runoff reaction, and dialysis, (3) the preparation for the CFPS reaction components and, (4) the quantification of cell extract and cell-free synthesized protein. We anticipate that the protocol in this research will provide a simple preparation and optimization procedure of a highly active E. coli cell extract.


Aneurysm of sinus of Valsalva dissecting into interventricular septum: a late complication of aortic valve replacement.
A 43-year-old man who had a Carbomedics prosthetic aortic valve replacement in 1997 was admitted to our hospital with complaints of shortness of breath and dyspnea on exertion in 2000. The patient was hospitalized due to atrioventricular (AV) complete block and a permanent pacemaker was implanted. At that time echocardiography indicated an aneurysm at the left sinus of Valsalva. In 2003, the patient was re-admitted to our clinic with complaints of shortness of breath and fatigue. Echocardiography showed a sinus of Valsalva aneurysm dissecting into interventricular septum. Operation confirmed dissection of the interventricular septum and communication between this cavity and the aneurysm of the left sinus of Valsalva. The postoperative course was uneventful and the patient was discharged in a satisfactory condition. This is the first reported case of aneurysm of the sinus of Valsalva dissecting into interventricular septum late and complicating aortic valve replacement.


Small molecule semiconductors for high-efficiency organic photovoltaics.
Organic photovoltaic cells (OPVs) are a promising cost-effective alternative to silicon-based solar cells, and possess light-weight, low-cost, and flexibility advantages. Significant progress has been achieved in the development of novel photovoltaic materials and device structures in the last decade. Nowadays small molecular semiconductors for OPVs have attracted considerable attention, due to their advantages over their polymer counterparts, including well-defined molecular structure, definite molecular weight, and high purity without batch to batch variations. The highest power conversion efficiencies of OPVs based on small molecular donor/fullerene acceptors or polymeric donor/fullerene acceptors are up to 6.7% and 8.3%, respectively, and meanwhile nonfullerene acceptors have also exhibited some promising results. In this review we summarize the developments in small molecular donors, acceptors (fullerene derivatives and nonfullerene molecules), and donor-acceptor dyad systems for high-performance multilayer, bulk heterojunction, and single-component OPVs. We focus on correlations of molecular chemical structures with properties, such as absorption, energy levels, charge mobilities, and photovoltaic performances. This structure-property relationship analysis may guide rational structural design and evaluation of photovoltaic materials (253 references).


Cytotoxicity and anti-Leishmania amazonensis activity of Citrus sinensis leaf extracts.
Leishmania amazonensis is the main agent of diffuse cutaneous leishmaniasis, a disease characterized by lesional polymorphism and the commitment of skin surface. Previous reports demonstrated that the Citrus genus possess antimicrobial activity. This study evaluated the anti-L. amazonensis activity of Citrus sinensis (L.) Osbeck (Rutaceae) extracts. Citrus sinensis dried leaves were subjected to maceration with hexane (CH), ethyl acetate (CEA), dichloromethane/ethanol (CD/Et - 1:1) or ethanol/water (CEt/W - 7:3). Leishmania amazonensis promastigotes were treated with C. sinensis extracts (1-525 μg/mL) for 120 h at 27 °C. Ultrastructure alterations of treated parasites were evaluated by transmission electron microscopy. Cytotoxicity of the extracts was assessed on RAW 264.7 and J774.G8 macrophages after 48-h treatment at 37 °C using the tetrazolium assay. In addition, Leishmania-infected macrophages were treated with CH and CD/Et (10-80 μg/mL). CH, CD/Et and CEA displayed antileishmanial activity with 50% inhibitory activity (IC50) of 25.91 ± 4.87, 54.23 ± 3.78 and 62.74 ± 5.04 μg/mL, respectively. Parasites treated with CD/Et (131.2 μg/mL) presented severe alterations including mitochondrial swelling, lipid body formation and intense cytoplasmic vacuolization. CH and CD/Et demonstrated cytotoxic effects similar to that of amphotericin B in the anti-amastigote assays (SI of 2.16, 1.98 and 1.35, respectively). Triterpene amyrins were the main substances in CH and CD/Et extracts. In addition, 80 μg/mL of CD/Et reduced the number of intracellular amastigotes and the percentage of infected macrophages in 63% and 36%, respectively. The results presented here highlight C. sinensis as a promising source of antileishmanial agents.


Risk factors and prognostic variables for survival of foals with radiographic evidence of pulmonary disease.
The medical records of 163 neonatal foals that had thoracic radiographs taken within 48 hours of admission to a referral hospital were reviewed. The objectives of this study were (1) to identify risk factors for the development of thoracic radiographic changes and (2) to identify prognostic indicators for survival in foals with radiographic evidence of pulmonary disease. Failure of transfer of passive immunity (IgG concentration < or = 400 mg/dL) was the only risk factor for radiographic evidence of respiratory disease identified by multivariate analysis. Hypoxemic patients (PaO2 < or = 60 mm Hg) were 4.9 times more likely to reveal radiographic abnormalities in a subset of foals for which arterial blood gas results were available. Foals with a serum creatinine concentration > 1.7 mg/dL upon presentation, dyspnea, and a history of dystocia were significantly more likely to die based on the multivariate statistical outcome analysis. An anion gap > or = 20 mEq/dL was strongly associated with nonsurvival in a subset of foals with arterial blood gas results. These hematologic and biochemical variables can be readily obtained during the initial evaluation of sick foals. The presence of a high anion gap appeared to have the greatest clinical impact and may be a useful prognostic indicator in foals with radiographic evidence of respiratory disease. In contrast, the majority of physical examination variables, including evaluation of tachypnea, abnormal respiratory sounds, fever, weakness, and milk reflux from the nares, which are usually obtained during the general respiratory evaluation of foals, were unrelated to outcome.