Pulse EPR, 55Mn-ENDOR and ELDOR-detected NMR of the S2-state of the oxygen evolving complex in photosystem II.
Pulse EPR, 55Mn-ENDOR and ELDOR-detected NMR experiments were performed on the S2-state of the oxygen-evolving complex from spinach Photosystem II. The novel technique of random acquisition in ENDOR was used to suppress heating artefacts. Our data unambiguously shows that four Mn ions have significant hyperfine coupling constants. Numerical simulation of the 55Mn-ENDOR spectrum allowed the determination of the principal values of the hyperfine interaction tensors for all four Mn ions of the oxygen-evolving complex. The results of our 55Mn-ENDOR experiments are in good agreement with previously published data [Peloquin JM et al. (2000) J Am Chem Soc 122: 10926-10942]. For the first time ELDOR-detected NMR was applied to the S2-state and revealed a broad peak that can be simulated numerically with the same parameters that were used for the simulation of the 55Mn-ENDOR spectrum. This provides strong independent support for the assigned hyperfine parameters.


The V̇e/V̇co2 slope: a useful tool to evaluate the physiological status of children with congenital heart disease.
Cardiopulmonary exercise test (CPET) is becoming a key examination to assess physical capacity and disease severity in pediatric cardiology. The V̇e/V̇co2 slope has been increasingly used as a surrogate marker for morbidity and mortality in adult heart failure, pulmonary arterial hypertension, and for adult patients with congenital heart disease (CHD). Nevertheless, the use of the V̇e/V̇co2 slope in children remains limited in the absence of reference values and clearly identified clinical determinants. This study aimed to compare the V̇e/V̇co2 slope in a pediatric cohort with CHD to that of age- and gender-adjusted healthy controls. We also intended to identify the clinical and CPET variables associated with V̇e/V̇co2 slope in this population. This cross-sectional study was carried out between November 2010 and September 2015 in two tertiary care pediatric cardiology reference centers. A total of 700 children were enrolled (399 CHD and 301 healthy controls). The mean V̇e/V̇co2 slope was significantly higher in the CHD subjects than in healthy subjects (31.6 ± 4.8 vs. 29.3 ± 4.8; P < 0.001). The V̇e/V̇co2 slope was higher in children with significant pulmonary regurgitation, tricuspid regurgitation, right ventricular hypertension, and right ventricle outflow tract (RVOT) obstacle. In the CHD group, V̇e/V̇co2 slope increase was associated with body mass index, the presence of a RVOT obstacle, the number of cardiac catheter procedures, as well as low age, forced vital capacity, tidal volume, and [Formula: see text]. Increased V̇e/V̇co2 slope was predominantly in children with single ventricle and/or residual right heart abnormalities, suggesting that maldistribution of pulmonary blood flow during exercise is an important CHD-unique determinant of V̇e/V̇co2 slope.NEW & NOTEWORTHY Using V̇e/V̇co2 slope is useful for children with congenital heart disease. V̇e/V̇co2 slope is sensitive to pulmonary blood flow maldistribution during exercise, this concerns congenital heart disease with pulmonary regurgitation, tricuspid regurgitation, right ventricular hypertension, and right ventricle outflow tract obstacle. V̇e/V̇co2 slope is a good parameter to follow single ventricles and right heart residual lesions (tetralogy of Fallot; pulmonary atresia; truncus arteriosus…).


Anesthesia management in a patient with an abdominal aortic aneurysm undergoing liver transplantation: a case report.
We describe the anesthetic management in a 56-year old man with hepatocellular carcinoma and cirrhosis who underwent liver transplantation (LT). Pretransplantation workup showed a 3-cm wide by 10-cm long infrarenal abdominal aortic aneurysm (AAA) with chronic dissection. He subsequently underwent living donor LT. The total operative time was 12 hours. The systolic blood pressure was maintained at normal levels. Severe hypertension was not noted. Hypotension noted during the anhepatic phase was managed with increased volume infusion and small doses (0.1 mg) of intravenous phenylephrine. Metabolic acidosis and ionized hypocalcemia were corrected accordingly. Total blood loss was 460 mL. Blood or blood products were not given. The intravascular volume was replaced with 1400 mL of 5% albumin and 10,610 mL of crystalloid. Extubation was performed in the intensive care unit at 12 hours after the operation. The postoperative course was unremarkable. The patient is alive at 3 years after LT. Patients with AAA undergoing LT present a challenge to the anesthesiologist because among the risk factors for rupture, blood pressure is the only factor under his or her control during the operation. If blood loss can be kept to a minimum and hemodynamic stability achieved, a chronically small dissected AAA may not be a contraindication to LT.


The timing of haloperidol administration does not affect its prophylactic antiemetic efficacy.
Postoperative nausea and vomiting (PONV) occurs frequently after general anesthesia. We evaluated the timing of 2 mg iv doses of haloperidol on the efficacy of this drug as a prophylactic antiemetic for PONV. Ninety-four non-smoking female patients with a history of motion sickness and/or PONV (Apfel's simplified risk score=3; predicted incidence of PONV=60%) were eligible to participate in this randomized, double-blind study. Patients were divided into two groups. Group 1 patients received haloperidol 2 mg iv during induction of anesthesia. Group 2 patients received haloperidol 2 mg iv approximately 30 min before the end of surgery. The occurrence of nausea and vomiting, nausea scores, pain scores, sedation levels, and adverse effects (cardiac arrhythmias, and extrapyramidal effects) were recorded by a trained, blinded investigator. The percentages of patients who experienced PONV in the first 24 hr were similar for the two groups (Group 1=30%; Group 2=26%, P=0.645). The incidence of PONV was significantly smaller in both groups than that predicted (60%) according to the patients' underlying risks (Group 1, P=0.004; Group 2, P=0.001). Nausea scores, pain scores, sedation scores, emergence times, and time to first rescue treatment were similar in the two groups. No adverse effects attributable to the study medication were observed. The timing of administration of haloperidol 2 mg iv does not influence its antiemetic efficacy. The recovery profiles were also similar whether haloperidol was administered at the start, or at the end of surgery.


Catalytic asymmetric alkynylation of alpha-imino ester: a versatile approach to optically active unnatural alpha-amino acid derivatives.
The catalytic asymmetric introduction of alkynyl functionality to alpha-amino acid derivatives was realized by the direct addition of terminal alkynes to alpha-imino ester in the presence of chiral Cu(I) complex under mild reaction conditions. Owing to the rich chemistry to which alkyne can be subjected, the present system provides a remarkably versatile tool for the construction of optically active alpha-amino acid derivatives. Good yields and enantiomeric excess values were achieved with an array of terminal alkynes and challenging, biologically active, unnatural alpha-amino acid derivatives could be conveniently obtained.


Exclusion of the neuronal nicotinic acetylcholine receptor alpha7 subunit gene as a candidate for catatonic schizophrenia in a large family supporting the chromosome 15q13-22 locus.
The gene encoding the neuronal nicotinic acetylcholine receptor alpha7 subunit (CHRNA7) is located on chromosome 15q13.2. This region was suggested to be involved in the etiopathogenesis of: (a) schizophrenia combined with a neurophysiological deficit; (b) lithium-responsive bipolar disorder; and (c) familial catatonic schizophrenia (periodic catatonia). Therefore, members of a large family with periodic catatonia strongly supporting the chromosome 15q13-22 region were genotyped with polymorphic markers localized around the CHRNA7 locus. A recombination event distally of marker D15S144 leading to the exclusion of the CHRNA7 locus from this candidate region was detected in one branch of the pedigree. This result provides strong evidence that a gene located telomeric to CHRNA7 is causative for the pathogenesis of catatonic schizophrenia in this family.


The reproductive toxicology of male SD rats after PM2.5 exposure mediated by the stimulation of endoplasmic reticulum stress.
Evidence has shown that exposure to fine particulate matter (PM) contributed to poor semen quality in males. However, the reproductive toxicity and relevant molecular mechanisms of Particular Matter 2.5 (PM2.5) from different seasons are not well understood. In the present work, we intend to investigate the toxic effects of PM2.5 during summer and winter on reproductive cells and tissues and focus on endoplasmic reticulum stress (ERS) to illustrate the possible molecular mechanisms. Sprague Dawley (SD) rats were exposed to PM2.5 from the summer season (0.2, 0.6, and 1.5 mg/kg b. w.) and winter (0.3, 1.5, and 2.7 mg/kg b. w.) through intratracheal instillation. The exposure was performed once every 3 days and continued for 2 months. Sperm and reproductive organs (testis and epididymis) were collected from the animals to conduct toxicity evaluation and mechanism analysis. The data showed that sperm relative motility rates were remarkably decreased, while sperm malformation rates were significantly increased with exposure to the summer and winter PM2.5. In particular, the reproductive toxicity of winter PM2.5 in the highest dose group was significantly greater than that in the other PM2.5 exposure groups. The pathological results also showed that the rats in the winter PM2.5 group of 2.7 mg/kg b. w. had severe testicular tissue injury, as determined by haematoxylin and eosin (HE) staining. The apoptotic results obtained by terminal dUTP nick-end labelling (TUNEL) further suggested that summer and winter PM2.5 exposure promoted the testicular germ cell apoptosis. The reproductive toxicity of winter PM2.5 in the testis was stronger than that of summer PM2.5. In addition, the expressions of GRP78 and XBP-1, biomarkers of ERS, was enhanced under the conditions of PM2.5 exposure, and ERS-mediated apoptosis through the upregulation of CHOP and Caspase-12 in the epididymis and testis was activated. In conclusion, PM2.5 exposure induced reproductive toxicity in male SD rats by the stimulation of ERS.


Effects of oral quinidine on left ventricular performance in normal subjects and patients with congestive cardiomyopathy.
To evaluate the effects of oral quinidine therapy on left ventricular performance, 10 normal subjects and 8 patients with cardiomyopathy were studied with echocardiography at rest, after intravenous injection of atropine and during infusion with phenylephrine. The echocardiographic studies were performed during oral quinidine therapy and during placebo administration. In the normal subjects heart rate was significantly faster with quinidine than with placebo (74 +/- 8 (standard deviation) versus 68 +/- 9 beats/min, P less than 0.01), but there was no significant change in blood pressure or left ventricular size and performance. After administration of atropine, heart rate was identical with and without quinidine but the mean normalized velocity of left ventricular dimension shortening was significantly less with quinidine than with placebo (1.28 +/- 0.19 versus 1.44 +/- 0.21, P less than 0.01). During acute pressure loading with phenylephrine there was no difference in left ventricular size or performance during quinidine therapy. In the patients with cardiomyopathy, no significant differences in left ventricular function were detected with this protocol during quinidine therapy. It is concluded that oral quinidine therapy appears to have little adverse effect on left ventricular performance at rest or during acute pressure loading.


What factors are associated with posttraumatic growth in head and neck cancer carers?
Researchers have recently called for more work to be conducted on positive outcomes and head and neck cancer. The purpose of this study was to investigate the factors associated with posttraumatic growth in head and neck cancer caregivers. 197 carers were surveyed. A descriptive cross-sectional survey design was used. In multivariate analysis the following factors were statistically significantly associated with increased posttraumatic growth: social support, longer time since diagnosis, increased worry about cancer and increased financial stress and strain stemming from caring. While HNC carers in the post-treatment phase of the illness trajectory can experience considerable psychological burdens, this study suggests that these burdens can lead some carers to experience growth and change and an expanded sense of themselves and their social worlds. Implications for nursing practice are discussed.


Sympathetic activation markedly reduces endothelium-dependent, flow-mediated vasodilation.
We sought to evaluate whether increased sympathetic outflow may interfere with flow-mediated dilation (FMD). Endothelial function, assessed as FMD, is frequently used as an intermediate end point in intervention studies. Many disease states with increased sympathetic tone are also characterized by endothelial dysfunction. Sixteen healthy volunteers underwent FMD studies with and without concomitant sympathetic stimulation. Intra-arterial nitroglycerin (NTG) infusion was used to assess endothelium-independent vasodilation. Pathophysiologically relevant sympathetic stimulation was achieved by baroreceptor unloading, using a lower body negative pressure box. In a subset of eight volunteers, this protocol was repeated during loco-regional alpha-adrenergic blockade by intra-arterial infusion of phentolamine (PE). Reactive hyperemic flow was assessed with strain-gauge phlethysmography. Overall, FMD responses (8.3 +/- 3.4%) were significantly attenuated by concomitant sympathetic stimulation (3.6 +/- 3.4%, p < 0.01). Loco-regional alpha-adrenergic blockade had no effect on baseline FMD responses (10.7 +/- 4.7%), whereas the attenuation by sympathetic stimulation was abolished completely during PE co-infusion (11.5 +/- 3.3%). During intra-arterial NTG infusions, arterial diameters relative to baseline were not significantly different between the four possible stages. Sympathetic stimulation, at a clinically relevant range, significantly impairs the FMD response by an alpha-adrenergic mechanism.


Impulsivity and Problem Drinking in College: Examining the Mediating Role of Sex-Related Alcohol Expectancies and Alcohol Use at Sex.
Dispositional impulsivity has been consistently implicated as a risk factor for problem drinking among college students and research suggests that this relationship may be explained in part by alcohol expectancies. A subset of alcohol expectancies, sex-related alcohol expectancies, is particularly linked to problem drinking among college students. The acquired preparedness model of risk postulates that people with dispositional impulsivity develop stronger sex-related alcohol expectancies, are subsequently more likely to drink at problematic levels in sexual situations, and thus, engage in more problem drinking. Using this model, the current study examined whether sex-related alcohol expectancies and alcohol use at sex mediated the relationship between impulsivity and problem drinking among college students. College students (N = 101) completed self-report measures of alcohol use, sex-related alcohol expectancies, and five dimensions of impulsivity: negative urgency, positive urgency, sensation seeking, lack of premeditation, and lack of perseverance. Two facets of impulsivity-sensation seeking and lack of premeditation-provided unique contributions to problem drinking. Sex-related alcohol expectancies significantly mediated the effects of lack of premeditation and sensation seeking on problem drinking. In support of the acquired preparedness model, the relationship between the impulsivity traits and problem drinking was serially mediated by sex-related alcohol expectancies and alcohol use at sex. Results suggest that sensation seeking and lack of premeditation continue to be areas of intervention for problem drinking among college students, and implicate sex-related alcohol expectancies as an area of intervention for alcohol use at sex and problem drinking.


IPPD-induced growth inhibition and its mechanism in zebrafish.
N-isopropyl-N-phenyl-1,4-phenylenediamine (IPPD) is used as a ubiquitous antioxidant worldwide, it is an additive in tire rubber easily discharged into the surrounding environment. At present, there is no study concerning the subacute toxicity of IPPD on fish. We used zebrafish embryos (2 h post-fertilization) exposed to IPPD for 5 days at concentrations of 0, 0.0012, 0.0120 and 0.1200 mg/L to investigate its toxic effects of embryonic development, disruption of growth hormone/insulin-like growth factor (GH/IGF) and hypothalamic-pituitary-thyroid (HPT) axis. The results showed that IPPD exposure decreased hatchability, weakened movement ability, reduced body length, and caused multiple types of deformities in zebrafish embryos. The expression of genes involved to GH/IGF and HPT axis were altered after exposure to IPPD in zebrafish larvae. Meanwhile, exposure to IPPD significantly decreased thyroxine (T4) and 3,5,3'-triiodothyronine (T3) contents in larvae, which indicated that HPT axis was in a disturbed state. Moreover, treatment of IPPD decreased the enzymatic activities of superoxide dismutase (SOD) and catalase (CAT) as well as levels of glutathione (GSH). While the contents of malondialdehyde (MDA) were elevated after exposure to IPPD. The present study thus demonstrated that IPPD induced oxidative stress, caused developmental toxicity and disrupted the GH/IGF and HPT axis of zebrafish, which could be responsible for developmental impairment and growth inhibition.


Urothelial Carcinoma: Update on Staging and Reporting, and Pathologic Changes Following Neoadjuvant Chemotherapies.
Staging and reporting of cancers of the urinary tract have undergone major changes in the past decade to meet the needs for improved patient management. Substantial progress has been made. There, however, remain issues that require further clarity, including the substaging of pT1 tumors, grading and reporting of tumors with grade heterogeneity, and following NAC. Multi-institutional collaborative studies with prospective data will further inform the accurate diagnosis, staging, and reporting of these tumors, and in conjunction with genomic data will ultimately contribute to precision and personalized patient management.


Multiple Beneficial Effects of High-dose BoNT-A Injection to Improve Wound Healing and Spasticity in a Patient With Hypoxic Encephalopathy Due to High-voltage Electrical Injury.
High-voltage electrical injuries remain a difficult challenge for physicians. The survivors often have complicated wounds over the trunk or extremities, and some of them may develop hypoxic encephalopathy. The emergence of spasticity following hypoxic encephalopathy may further interfere with the healing of wounds. The authors report the case of a 17-year-old male with strong spasticity of finger flexors graded 4 by the Modified Ashworth Scale (0-5) after electric shock. He also had a nonhealing wound on the flexor side of the left index finger after 6 weeks of standard wound care. The wound measured 0.3 cm × 0.3 cm × 0.2 cm in size. The authors hypothesized that wound healing was negatively affected by spasticity and expected the wound might heal gradually after reducing the strong spasticity of the index finger. The authors employed electrical stimulator for guidance and injected high-dose (50 units/muscle) botulinum toxin type A into the flexor digitorum superficialis and the flexor digitorum profundus of his left index finger. At 7 days following administration, focal spasticity of these muscles in the left index finger decreased from 4 to 1 on the Modified Ashworth Scale. At 21 days post administration, the wound healed completely. For patients with hypoxic encephalopathy due to high-voltage electrical injury, botulinum toxin type A injection may be an option of therapeutic approach for both reduction of spasticity and facilitation of wound healing.


Notch1 signaling promotes the maturation of CD4 and CD8 SP thymocytes.
Notch proteins regulate many developmental processes. Notch1 is highly expressed on thymocytes, but its role in regulating their development is not known. We show that activation of Notch1 signaling in CD4+CD8+ double positive thymocytes promotes the maturation of both CD4+ and CD8+ single positive thymocytes and that this occurs in the absence of interactions between the T cell receptor and MHC molecules expressed on thymic epithelial cells. We have also identified several genes that are transcriptionally regulated by Notch1 in T cells and show that they are upregulated during maturation into both single positive lineages. These observations suggest that Notch1 signaling plays a role in promoting maturation into both the CD4 and CD8 T cell lineages.


Obesity perceptions and documentation among primary care clinicians at a rural academic health center.
Obesity recognition in primary care is important to address the epidemic. We aimed to evaluate primary care clinician-reported documentation, management practices, beliefs and attitudes toward obesity compared to body mass index (BMI) calculation, obesity prevalence and actual documentation of obesity as an active problem in electronic health record in a rural academic center. Our target population for previously validated clinician survey was 56 primary care providers working at 3 sites. We used calendar year 2012 data for assessment of baseline system performance for metrics of documentation of BMI in primary care visits, and proportion of visits in patients with obesity with obesity as a problem. Standard statistical methods assessed the data. Survey response rate was 91%. Average age of respondents was 48.9 years and 62.7% were females. 72.5% clinicians reported having normal BMI. The majority of clinicians reported regularly documenting obesity as an active problem, and utilized motivational interviewing and basic good nutrition and healthy exercise. Clinicians identified lack of discipline and exercise time, access to unhealthy food and psychosocial issues as major barriers. Most denied disliking weight loss discussion or patients taking up too much time. In 21,945 clinic visits and 11,208 annual preventive care visits in calendar year 2012, BMI was calculated in 93% visits but obesity documentation as an active problem only 27% of patients meeting BMI criteria for obesity. Despite high clinician-reported documentation of obesity as an active problem, actual obesity documentation rates remained low in a rural academic medical center.


WNK4 is an Adipogenic Factor and Its Deletion Reduces Diet-Induced Obesity in Mice.
The with-no-lysine kinase (WNK) 4 gene is a causative gene in pseudohypoaldosteronism type II. Although WNKs are widely expressed in the body, neither their metabolic functions nor their extrarenal role is clear. In this study, we found that WNK4 was expressed in mouse adipose tissue and 3T3-L1 adipocytes. In mouse primary preadipocytes and in 3T3-L1 adipocytes, WNK4 was markedly induced in the early phase of adipocyte differentiation. WNK4 expression preceded the expression of key transcriptional factors PPARγ and C/EBPα. WNK4-siRNA-transfected 3T3-L1 cells and human mesenchymal stem cells showed reduced expression of PPARγ and C/EBPα and lipid accumulation. WNK4 protein affected the DNA-binding ability of C/EBPβ and thereby reduced PPARγ expression. In the WNK4-/- mice, PPARγ and C/EBPα expression were decreased in adipose tissues, and the mice exhibited partial resistance to high-fat diet-induced adiposity. These data suggest that WNK4 may be a proadipogenic factor, and offer insights into the relationship between WNKs and energy metabolism.


Feeding regimens after pyloromyotomy.
In a prospective randomized study three different feeding regimens after operation were compared in 74 babies with infantile hypertrophic pyloric stenosis: gradual regarding of feeds over 48 h (regimen 1), rapid regarding of feeds over 16 h (regimen 2), and initial starvation followed by full normal feeds at 24 h (regimen 3). No significant difference between the treatment groups was found either in episodes of vomiting after operation (regimen 1, 2.9 episodes in 21 patients; regimen 2, 3.6 episodes in 28 patients; regimen 3, 3.6 episodes in 25 patients) or in the mean duration of postoperative hospital stay (regimen 1, 59.3 h; regimen 2, 47.8 h; regimen 3, 56.7 h). We conclude that vomiting following pyloromyotomy is self-limiting and independent of the timetable or composition of the postoperative dietary regimen.


[Ablative and fractional lasers].
The use of pulsed or scanning Carbon Dioxide, and pulsed Erbium-YAG lasers allows the programmable and reproducible photocoagulation of thin layers of the epidermis and superficial dermis. Thermal damage depends on the type of laser and is greater with CO(2) lasers. The degree of neocollagenesis is proportional to the thermal damage and is better with CO(2) lasers. Their main indication is the correction of photoaged facial skin but they can also be used for corrective dermatology, e.g. for scars and genodermatosis. Results are highly satisfactory but the technique is invasive and the patient experiences a social hindrance of around two weeks. The fractional techniques such as Fraxel are used to treat non-adjacent microzones without ablation of the epidermis. Around 25 p. 100 of the affected region is treated per session without ablation of the epidermis. Each fraction is only mini-invasive and is performed under local anesthesia. Social hindrance is minimal. Nonetheless, the results are inferior to those obtained with ablative lasers, especially regarding deep wrinkles. The treatment is costly and four sessions are usually required to treat the whole affected area. Others regions of the face may also be treated. Encouraging results have been obtained with mélasma.


Exploring the use of mothers' own milk as oral care for mechanically ventilated very low-birth-weight preterm infants.
The purpose of this study was to explore the use of mothers' own milk (colostrums, transitional milk, and mature milk) as oral care in the ventilator-associated pneumonia (VAP)-prevention bundle of mechanically ventilated preterm infants weighing 1500 g or less. Mechanically ventilated preterm infants weighing 1500 g or less admitted to a regional level III NICU in the Gulf South between January 1, 2006, and December 31, 2009. Retrospective descriptive. Oral care with mothers' own milk was implemented as part of the VAP-prevention bundle in the neonatal intensive care unit in the fourth quarter of 2007. Using retrospective deidentified data retrieved from the electronic medical record, the primary and secondary outcome variables were collected among eligible infants (≤1500 g) admitted January 1, 2006, to December 31, 2007 (before implementation) and January 1, 2008, to December 31, 2009 (after implementation). Sample characteristics, including infant gestational age, birth weight, and gender, as well as maternal age, type of delivery, and incidence of maternal chorioamnionitis, were also collected. Data analysis included frequencies and distributions to summarize sample characteristics and variables of interest. Appropriate tests for differences were conducted on outcome variables between the before and after groups of the human milk oral care intervention. The feasibility outcome variable included nursing compliance with the oral care procedure. The safety outcome variable included record of any adverse events associated with the oral care procedure. The efficacy health outcomes included the rate of positive tracheal aspirates, positive blood cultures, the number of ventilator days, and length of stay. Infant age (26.1-26.6 weeks) and weight (840-863 g) were similar in the before (n = 70) and after (n = 68) sample subjects. There were no statistically significant differences in ventilator days, χ² (46, n = 115) = 46.22, P = .46, and length of stay, χ (75, n = 115) = 78.78, P = .36, between groups. Although the rate of positive tracheal aspirates and positive blood cultures reduced after implementation of oral care with mothers' own milk, these differences were not statistically significant (U(47) = 250, z = -7.1, P = .48; U(47) = 217.5, z = -1.44, P = .15). There were no statistically significant differences in the rates of positive tracheal aspirates and blood cultures after implementation of oral care with mothers' own milk. The findings of this study suggest that using mothers' own milk as part of the VAP-prevention bundle is a feasible and safe practice; however, further research is needed to determine the immunological benefits of this practice.


On the genetics and immunopathogenesis of COVID-19.
Most severe cases with COVID-19, especially those with pulmonary failure, are not a consequence of viral burden and/or failure of the 'adaptive' immune response to subdue the pathogen by utilizing an adequate 'adaptive' immune defense. Rather it is a consequence of immunopathology, resulting from imbalanced innate immune response, which may not be linked to pathogen burden at all. In fact, it might be described as an autoinflammatory disease. The Kawasaki-like disease seen in children with SARS-CoV-2 exposure might be another example of similar mechanism.


If the price is right: Cost-effectiveness of radiofrequency ablation versus thyroidectomy in the treatment of benign thyroid nodules.
Radiofrequency ablation is an emerging technology in the United States to treat benign thyroid nodules. The cost-effectiveness of radiofrequency ablation in comparison with traditional thyroidectomy is unknown. A patient-level state transition microsimulation decision model was constructed comparing radiofrequency ablation with lobectomy in the management of benign thyroid nodules. Our base case was a 45-year-old woman with a solitary 30-cm3 nodule. Estimates of health utilities, complications, and mortality were obtained from the literature, and costs were estimated using Medicare reimbursement data. The primary outcomes of interest included total cost, quality-adjusted life years, and incremental cost-effectiveness ratios. All model estimates were subjected to 1-way sensitivity analyses to identify factors that strongly influence cost-effectiveness. A probabilistic sensitivity analysis was run across 1 million simulations to gauge outcome confidence with a willingness-to-pay threshold set at $100,000/quality-adjusted life year. Radiofrequency ablation was assumed to cost $5,000, with an initial success rate of 78%. Patients with volume reduction ratio <50% underwent a second treatment of radiofrequency ablation. Radiofrequency ablation represented the dominant strategy, yielding 21.31 quality-adjusted life years for a total cost of $16,563 in comparison to lobectomy, which yielded 21.13 quality-adjusted life years for a total cost of $19,262. In a 1-way sensitivity analysis varying the cost of radiofrequency ablation across of range of values, the radiofrequency ablation strategy remained cost-effective until the cost of radiofrequency ablation exceeded $12,330 at willingness-to-pay $50,000 or $17,950 at willingness-to-pay $100,000. Radiofrequency ablation is a cost-effective strategy in the treatment of benign thyroid nodules but is most sensitive to the cost of radiofrequency ablation.


Conditional survival of patients with the four major histologic subgroups of lung cancer in Denmark.
The survival probability of patients with lung cancer is usually based on the extent of disease as assessed at the time of diagnosis. The discouraging 5-year survival is often reported (< 10%) without taking into account changes in the survival probability as time advances from diagnosis. Conditional survival estimates by sex, age, extent of disease, and histology were estimated for patients diagnosed with lung cancer in Denmark from 1943 to 1997. Survival probabilities were calculated by the Kaplan-Meier method, and cumulative survival estimates were used to derive conditional survival estimates. For every additional year survived, the probability of surviving the next 5 years increases from 33% (men) and 36% (women) after the first year, to 60% (men) and 67% (women) who have survived 5 years. The 5-year survival probability of patients younger than 49 years who had survived the first year was 33%, and increased to 81% after the fifth year. Corresponding estimates for 60- to 69-year-old patients were 23% and 52%. The conditional survival differed greatly among patients with localized and regional disease (29% and 10%, respectively) in the first year, but converged with time (52% and 47%, respectively) after 5 years. The conditional survival is similar in patients with squamous cell carcinoma, adenocarcinoma, and large-cell carcinoma, but is markedly lower in patients with small-cell carcinoma. For patients who have survived more than 1 year, the conditional survival probability provides a more accurate estimate of survival as compared with the conventional observed survival rates.


A comparison of the effects of pirenzepine and atropine on gastric acid secretion, salivary secretion and pupil diameter in the rat.
The ability of pirenzepine and atropine, given i.v., to inhibit gastric acid and salivary secretion and increase pupil diameter has been assessed in the rat. Pirenzepine had a similar potency against acid secretion, ED 50 0.71 (0.41 to 1.1) mg.kg-1, and salivary secretion, ED 50 (0.43 to .59) mg.kg-1, whilst its potency was less in the eye, ED 50 1.8 (1.6 to 2.1) mg.kg-1. Atropine however, was more potent in reducing salivary secretion, ED 50 0.012 (0.010 to 0.016) mg.kg-1 and increasing pupil diameter, ED 50 0.028 (0.025 to 0.031) mg.kg-1 than in inhibiting gastric acid secretion, ED 50 0.056 (0.037 to 0.083) mg.kg-1. Therefore, that quantity of pirenzepine which inhibits gastric acid secretion by 50% will have only a slight effect on the eye and will inhibit salivary secretion by a similar magnitude. In contrast, the amount of atropine required to inhibit acid secretion by 50% will significantly increase pupil diameter and abolish salivary secretion.


Cancer risk in patients with chronic urticaria: a population-based cohort study.
To investigate the relative risk of cancer among patients with chronic urticaria in the Taiwanese population. Retrospective population-based cohort study. The National Health Insurance Research Database of Taiwan from January 1, 1996, through December 31, 2008. A total of 12,720 patients with chronic urticaria, with long-term antihistamine use and no history of malignant tumors, autoimmune diseases, atopy, or allergic diseases. Relative cancer risk calculated by standardized incidence ratios. There were 704 cancers among chronic urticaria patients. An increased risk of cancer (standardized incidence ratio, 2.2; 95% CI, 2.0-2.3), especially hematologic malignant tumor (4.1; 3.1-5.4), was observed. The relative risk of cancer varied by age and was highest among those aged 20 to 39 years in comparison with the general population. Most cancer cases were detected within the first year of diagnosis. The risk of non-Hodgkin lymphoma was greatest (standardized incidence ratio, 4.4; 95% CI, 3.0-6.1) among the hematologic cancers. Patients with chronic urticaria are at increased risk of cancer, especially hematologic malignant tumors. Further studies are needed to delineate the associations.


Direct electrochemistry of cholesterol oxidase immobilized on gold nanoparticles-decorated multiwalled carbon nanotubes and cholesterol sensing.
An electrochemical cholesterol biosensor based on the direct electron transfer of cholesterol oxidase (ChOx) immobilized on gold nanoparticles-decorated multiwalled carbon nanotubes (GNPs-MWCNTs) was fabricated. GNPs-MWCNTs were prepared based on the reduction of HAuCl4 in the presence of carboxyl group functionalized MWCNTs. Transmission electron microscopy image shows that rounded gold nanoparticles with diameters of 6-10nm were decorated on carbon nanotube surfaces. ChOx was directly adsorbed on the nanocomposite modified glassy carbon electrode and protected by a Nafion film. Direct electrochemistry of ChOx on the electrode surface was obtained, proved by one pair of well-defined, quasi-reversible redox peaks in phosphate buffered saline. Under optimized conditions, the fabricated electrode displayed a linear response in the cholesterol concentration range from 0.0100 to 5.00 mmol L(-1) with a detection limit of 4.3 μmol L(-1) estimated at a signal-to-noise ratio of 3. The apparent Michaelis-Menten constant was measured to be 0.29 mmol L(-1), indicating that the immobilized ChOx on GNPs-MWCNTs matrix retained its native activity. The developed biosensor presented good selectivity, repeatability, reproducibility and stability. The concentration of free cholesterol in a human serum sample, detected by using the developed biosensor, is in good agreement with that determined by the well-established spectrophotometric method.


[Influencing factors on dampproof effect of microcapsules and study on mechanism].
The influence of spray drying technology on the damp proof effect of microcapsules and the mechanism were studied. The microcapsules prepared with different spray drying parameters have been put in certain surroundings for 12 hours, then the hygroscopic curves were gotten; the mechanism was studied from the following aspects: solvent residue, film's shrink and particle size. The damp proof effect enhanced with the increase of inlet air temperature and the decrease of flow rate and air pressure. The properties of the wall, the solvent residue and particle size can influence the damp proof effect of the microcapsules. The physical properties of microcapsules are different because of the different spray drying parameters, which lead to different damp proof effect of microcapsules.


Plasma-Lyte 148 and Plasma-Lyte 148 + 5% glucose compatibility with commonly used critical care drugs.
Plasma-Lyte is a balanced, crystalloid intravenous fluid which has been shown to avoid the hyperchloremic metabolic acidosis associated with 0.9% sodium chloride. Data on physical, pH and chemical compatibility with other medicines are essential. The compatibility of adrenaline, dobutamine, dopamine, furosemide, midazolam, morphine and milrinone with Plasma-Lyte 148 (PLA) and Plasma-Lyte 148 with 5% glucose (PLA-G) was investigated. Control solutions were 0.9% sodium chloride and 5% glucose. Chemical stability was defined as < 5% concentration change with high-performance liquid chromatography (HPLC). Physical compatibility was assessed by checking for colour changes and precipitate formation. The pH of the admixtures was considered acceptable if between 5 and 9 at all time points. Six repeats were carried out for HPLC, 2 for physical compatibility checks and pH measurements, with all admixtures being tested at 0, 2 and 24 h after mixing. All combinations were found to be chemically stable at 0, 2 and 24 h apart from furosemide with PLA-G at 24 h and midazolam with PLA or PLA-G at both 2 and 24 h. Only midazolam was physically incompatible when mixed with both Plasma-Lyte solutions. The pH remained stable in all admixtures, although not all pH values recorded were within the range of 5-9. All drugs excluding furosemide and midazolam were shown to be chemically, physically and pH stable at the tested concentrations when diluted with PLA and PLA-G.


The relationship between cocaine self-administration and actigraphy-based measures of sleep in adult rhesus monkeys.
Clinical trials show that chronic cocaine users suffer from sleep disturbances and preclinical research has shown that acute sleep deprivation increases the rate of cocaine self-administration in rats. This study examined the effect of cocaine self-administration on behavioral indices of sleep and alternatively the effect of sleep disruption on cocaine-maintained responding by rhesus monkeys. Seven adult rhesus monkeys, fitted with Actical® activity monitors, were trained to respond under a concurrent choice paradigm with food (three 1.0-g pellets) and cocaine (0.003-0.3 mg/kg) or saline presentation. For each monkey, the lowest preferred dose of cocaine (>80% cocaine choice) was determined. Activity data were analyzed during lights out (2000-0600) to determine sleep efficiency, sleep latency, and total activity counts. Subsequently, the monkeys' sleep was disrupted (every hour during lights-out period) the night prior to food-cocaine choice sessions. Self-administration of the preferred dose of cocaine resulted in a significant decrease in sleep efficiency, with a significant increase in total lights-out activity. Sleep disruption significantly altered behavioral indices of sleep, similar to those seen following cocaine self-administration. However, sleep disruption did not affect cocaine self-administration under concurrent choice conditions. Based on these findings, cocaine self-administration does appear to disrupt behavioral indices of sleep, although it remains to be determined if treatments that improve sleep measures can affect future cocaine taking.


Derivation of monoclonal antibodies to human somatomedin C/insulin-like growth factor I.
Somatomedin C, also called insulin-like growth factor I (Sm-C/IGF-I), is a highly conserved polypeptide required for the proliferation of many cell types. Since several attempts in our laboratory to recover monoclonal antibody-secreting hybrids to this peptide by the direct fusion of hyperimmunized splenocytes with myeloma cells had been unsuccessful, we modified our approach by coculturing hyperimmunized BALB/c splenocytes and a small amount of the antigen for 5 days prior to fusion with the P3X63Ag.8.653 myeloma cell line. Of 88 microcultures at risk, specific antibody was detected in 24. Two clones were expanded in ascites fluid and characterized as to isotype, affinity, and specificity. Both were IgG1,kappa and bound human Sm-C/IGF-I with affinity constants of 1.09 and 1.01 X 10(10) liter/mol, respectively. Both clones were quite specific for Sm-C/IGF-I with inconsequential binding to insulin-like growth factor II, multiplication-stimulating activity, any of the chymotryptic fragments of Sm-C/IGF-I, insulin preparations, hGH, hTSH, mEGF, or mouse albumin. In vitro boosting after primary in vivo immunization appears to provide monoclones of an IgG isotype in contrast to primary in vitro immunization, which reportedly favors an IgM isotype. The antibodies produced in this study have proved to be extraordinarily useful in defining the physiologic role of Sm-I/IGF-I with immunoneutralization techniques and in the purification of human Sm-C/IGF-I by affinity chromatography.


A yellowness index for use in dentistry.
To develop an equation that predicts the perceptual yellowness of teeth. A large set of new psychophysical yellowness data were generated from an experiment where 500 participants each ranked a set of 58 shade guide samples. Two existing equations (WIO and b*) and one new equation (YIO) were evaluated by comparing their values for the 58 shade guide tabs with the psychophysical data. Coefficient of determination (r2), '% wrong decisions', and STRESS were used as measures of performance. The YIO equation was optimized using these data to maximize the r2 value. A validation set of psychophysical data was prepared in an experiment where 40 participants each ranked 5 sets of 9 samples that were viewed on an emissive display. The candidate equations were evaluated using these data and the r2, %WD, and STRESS metrics. All three metrics YIO, WIO and b* were strongly correlated with perceptual yellowness. YIO and WIO both showed stronger correlation than b*. A new yellowness equation YIO has been developed to correlate with tooth yellowness. It is suggested that tooth yellowness and whiteness are highly related concepts.


Exploring the possibility of parents' broad internalizing phenotype acting through passive gene-environment correlations on daughters' disordered eating.
Twin studies demonstrate significant environmental influences and a lack of genetic effects on disordered eating before puberty in girls. However, genetic factors could act indirectly through passive gene-environment correlations (rGE; correlations between parents' genes and an environment shaped by those genes) that inflate environmental (but not genetic) estimates. The only study to explore passive rGE did not find significant effects, but the full range of parental phenotypes (e.g., internalizing symptoms) that could impact daughters' disordered eating was not examined. We addressed this gap by exploring whether parents' internalizing symptoms (e.g., anxiety, depressive symptoms) contribute to daughters' eating pathology through passive rGE. Participants were female twin pairs (aged 8-14 years; M = 10.44) in pre-early puberty and their biological parents (n = 279 families) from the Michigan State University Twin Registry. Nuclear twin family models explored passive rGE for parents' internalizing traits/symptoms and twins' overall eating disorder symptoms. No evidence for passive rGE was found. Instead, environmental factors that create similarities between co-twins (but not with their parents) and unique environmental factors were important. In pre-early puberty, genetic factors do not influence daughters' disordered eating, even indirectly through passive rGE. Future research should explore sibling-specific and unique environmental factors during this critical developmental period.


Molecular analysis of a recurrent glioblastoma treated with bevacizumab.
We treated a case of recurrent glioblastoma (GBM) with bevacizumab and assessed its effect biologically. A 55-year-old man with a left frontal lobe GBM was experiencing recurrence 7 months postoperation. We administered bevacizumab concomitant with temozolomide (TMZ). Follow-up magnetic resonance imaging (MRI) showed dramatic but temporal tumor reduction; however, the patient died of re-recurrent disease 6 months after beginning bevacizumab. We obtained an autopsy and analyzed the detailed molecular change. In the autopsy specimen, the quantity of microvessels was significantly reduced. Vascular endothelial growth factor receptor (VEGFR) 1 and VEGFR2 were downregulated, most likely due to a negative feedback mechanism by blocking of VEGF signaling. Matrix metalloproteinase (MMP)-2 and membrane-type 1 MMP were upregulated, resulting in the higher activation of MMP-2 in the autopsy specimen. MIB-1 staining index and phosphorylation levels of p44/42-mitogen-activated protein kinase did not change, whereas phosphorylated protein kinase B (Akt) was decreased in the autopsy specimen, suggesting compensation and/or amplification of other proliferative signaling pathways such as suppression of apoptosis signaling. Consequently, bevacizumab might inhibit the VEGF autocrine loop, which then causes a change in molecular expression related not only to enhancement of tumor invasion but also maintenance of tumor proliferation.


Current topics on sample preservation. A report on the progress of the ESA Topical Team. Preservation of fixed and non-fixed samples during space experimentation.
The existence of preservation problems is one of the most important consequences of Space Biological Research. The Topical Team is critically analyzing the currently performed procedures and is establishing the bases for a recommendation on new procedures, capable of overcoming the present constraints.


Designing Image Analysis Pipelines in Light Microscopy: A Rational Approach.
With the progress of microscopy techniques and the rapidly growing amounts of acquired imaging data, there is an increased need for automated image processing and analysis solutions in biological studies. Each new application requires the design of a specific image analysis pipeline, by assembling a series of image processing operations. Many commercial or free bioimage analysis software are now available and several textbooks and reviews have presented the mathematical and computational fundamentals of image processing and analysis. Tens, if not hundreds, of algorithms and methods have been developed and integrated into image analysis software, resulting in a combinatorial explosion of possible image processing sequences. This paper presents a general guideline methodology to rationally address the design of image processing and analysis pipelines. The originality of the proposed approach is to follow an iterative, backwards procedure from the target objectives of analysis. The proposed goal-oriented strategy should help biologists to better apprehend image analysis in the context of their research and should allow them to efficiently interact with image processing specialists.


[Physicians as leaders. Evaluation of a leadership training program for physicians (Medled)].
In September 1990, 40 doctors started a two year management training programme consisting of six one week courses organised by the Norwegian Medical Association. The programme was evaluated, and this article presents the results of two of the methods, used for evaluation: the first an interview with 12 participants, and the second a questionnaire answered by participants, colleagues at the same department and colleagues at other hospitals (a control group) before and after the course. Compared with colleagues from the same department and from the control group the participants had changed their attitude towards important questions of management. They had become more concerned about the importance of motivating the staff, that the department should give priority to the hospital's objectives in preference to own interests, and that the ultimate responsibility for the department should reside in one person and this person should be a doctor with a relevant specialty. The results of the questionnaire survey are confirmed by the results of the interviews. In addition the results of the interviews suggest that, during the management training programme, the participants had changed from an individual-oriented to a more group-oriented style of management.


Conservative transcription in three steps visualized in a double-stranded RNA virus.
Endogenous RNA transcription characterizes double-stranded RNA (dsRNA) viruses in the Reoviridae, a family that is exemplified by its simple, single-shelled member cytoplasmic polyhedrosis virus (CPV). Because of the lack of in situ structures of the intermediate stages of RNA-dependent RNA polymerase (RdRp) during transcription, it is poorly understood how RdRp detects environmental cues and internal transcriptional states to initiate and coordinate repeated cycles of transcript production inside the capsid. Here, we captured five high-resolution (2.8-3.5 Å) RdRp-RNA in situ structures-representing quiescent, initiation, early elongation, elongation and abortive states-under seven experimental conditions of CPV. We observed the 'Y'-form initial RNA fork in the initiation state and the complete transcription bubble in the elongation state. These structures reveal that de novo RNA transcription involves three major conformational changes during state transitions. Our results support an ouroboros model for endogenous conservative transcription in dsRNA viruses.


Serum alkaline phosphatase and inorganic phosphorus values in spinal cord injury patients with heterotopic ossification.
The blood chemistry was studied in 140 spinal cord injury (SCI) patients (acute injury ward), including 18 patients who developed heterotopic ossification (HO). Comparisons between the HO and non-HO groups were made to determine if the alkaline phosphatase (AP), inorganic phosphorus (P), or calcium (Ca) levels were of diagnostic value. The results showed that AP, P, and Ca by themselves were of little help in the diagnosis of HO. However, the combination of elevated AP and P was significant, especially if both were consistently elevated. There were no significant differences between the HO and non-HO groups concerning completeness or level of spinal injury.


The acidic glycosaminoglycans in gasric mucosa of athyroid rats: The effects of L-tri-iododthyronine.
Hyaluronic acid is increased about 12%, whereas dermatan sulfate is decreased about 34%, in gastric mucosa of athyroid rats when compared with controls. Administration of 2 doses of L-tri-iodothyronine reversed the effects of thyroidectomy.


Stereotypic behavioral response of rat fetuses to acute hypoxia is altered by maternal alcohol consumption.
Transient compression of the umbilical cord has been implicated as a potential cause of unexplained brain damage and/or fetal death. We induced acute fetal hypoxia by compressing the umbilical cord of the fetal rat with a microvascular clamp. Direct observation of fetal behavior immediately after cord compression identified a three-phase behavioral response to acute hypoxia, consisting of an initial suppression in motor activity, a brief period of hyperactivity, and a secondary suppression of movement. All three phases occurred in less than 2 minutes from the onset of hypoxia. The stereotypic hypoxic response of rat fetuses may be adaptive as a means of removing mechanical sources of umbilical cord compression. Among fetuses exposed to alcohol in utero, the response to hypoxia was diminished in intensity and duration. Because of their altered hypoxic response, alcohol-exposed fetuses may therefore be at greater risk from transient cord compression in utero.


Influence of sub-lethal crude oil concentration on growth, water relations and photosynthetic capacity of maize (Zea mays L.) plants.
Maize tolerance potential to oil pollution was assessed by growing Zea mays in soil contaminated with varying levels of crude oil (0, 2.5 and 5.0 % v/w basis). Crude oil contamination reduced soil microflora which may be beneficial to plant growth. It was observed that oil pollution caused a remarkable decrease in biomass, leaf water potential, turgor potential, photosynthetic pigments, quantum yield of photosystem II (PSII) (Fv/Fm), net CO2 assimilation rate, leaf nitrogen and total free amino acids. Gas exchange characteristics suggested that reduction in photosynthetic rate was mainly due to metabolic limitations. Fast chlorophyll a kinetic analysis suggested that crude oil damaged PSII donor and acceptor sides and downregulated electron transport as well as PSI end electron acceptors thereby resulting in lower PSII efficiency in converting harvested light energy into biochemical energy. However, maize plants tried to acclimate to moderate level of oil pollution by increasing root diameter and root length relative to its shoot biomass, to uptake more water and mineral nutrients.


The use of balloons for uterine cervical ripening is associated with an increased risk of umbilical cord prolapse: population based questionnaire survey in Japan.
To clarify whether the use of balloons for cervical ripening is associated with the incidence of umbilical cord prolapse. A postal questionnaire survey was distributed in Japan. Cases of umbilical cord prolapse occurring during labor in association with the use of balloons for cervical ripening between 2007 and 2011 in Japan were analyzed. Answers from 942 institutions were obtained. The subjects included 369 patients with fore-lying or prolapse of the umbilical cord among a total of 2,037,460 deliveries. Among the singleton vertex cases, fore-lying or prolapse of the umbilical cord during labor were observed in 88 (0.005%) of 1,891,189 deliveries not associated with the use of balloons for cervical ripening and in 93 (0.064%) of 146,271 deliveries associated with the use of balloons for cervical ripening (Odds ratio 13.67, 95% confidence interval 10.21, 18.30). All types of balloons were significantly associated with the occurrence of fore-lying or prolapse of the umbilical cord. A total of 39% of cases of umbilical cord prolapse occurred during manual or spontaneous balloon removal, while 53% of cases occurred after a while not directly associated with balloon removal. The risk of umbilical cord prolapse was significantly increased during the use of balloons for cervical ripening, especially in cases involving the use of disk-type and ball-type balloons filled with large amounts of water.


Management of tuberculosis in pregnancy and the newborn.
Tuberculosis in pregnant women and their offspring is a serious and sometimes life-threatening infection. A high index of suspicion is a prerequisite for proper diagnosis. Early detection is essential since treatment is successful in these patients.


Gastrokine 1 functions as a tumor suppressor by inhibition of epithelial-mesenchymal transition in gastric cancers.
Gastrokine 1 (GKN1) plays an important role in the gastric mucosal defense mechanism and also acts as a functional gastric tumor suppressor. The specific aim of this study was to determine the molecular mechanisms underlying GKN1 tumor suppressor activity in the progression of gastric cancers. We examined the effect of GKN1 on epithelial-mesenchymal transition (EMT) and cell migration in GKN1-transfected and recombinant GKN1-treated AGS gastric cancer cells using in vitro wound healing, microchemotaxis, and invasion assays. In GKN1-transfected AGS cells, we observed inhibition of cell migration and invasion in wound healing, transwell and Matrigel assay. Also, GKN1-transfected and recombinant GKN1-treated AGS cells showed decreased levels of ROS and expression of phosphatidylinositol 3-kinase (PI3K)/Akt pathway proteins, concomitant with re-expression of E-cadherin and decreased expression of cytoplasmic and nuclear expression of β-catenin, slug, snail, fibronectin, and vimentin. These data suggest that the GKN1 gene may play an important role in the progression of sporadic gastric cancers via inhibition of EMT and cancer cell migration.


Significance of the 'maturation' of metastases from germ cell tumours after intensive chemotherapy.
A comparison between primary and metastatic germ cell tumours from 38 male patients showed that 19 of 24 metastases with residual differentiated teratoma after adequate therapy came from tumours with teratoma as a component of the primary. The correlation between the presence of teratoma in the primary and the metastases is statistically significant (P less than 0.01) and supports the view that the so called 'maturation' of germ cell tumours is due to selective destruction of anaplastic components in tumours which have already shown an inherent capacity for differentiation. Elevation of the serum concentrations of HCG and AFP on presentation with disseminated disease was significantly related to the presence of morphologically identifiable trophoblast and yolk sac elements respectively in the primary tumours (P less than 0.001). Histological identification and specific mention of teratomatous, trophoblastic and yolk sac elements in reporting germ cell tumours is therefore useful since their presence in the primary correlates with the morphology in the metastases.


Cytochemical and biochemical characterization of neurosecretory material in the brain of an annelid, Ophryotrocha puerilis (Polychaeta).
In the brain of the protandric annelid Ophryotrocha, prominent axon terminals accumulate neurosecretory material in female-phase individuals. This material is not present in females after they have had social contact for 2 days and in males. After treatment with Formalin or glyoxylic acid, the axon terminals in isolated females are brightly fluorescent. The injection of reserpine into the coelomic cavity of such females prevents the development of fluorescence in the nerve endings. In an HPLC study of brains containing axon terminals filled with neurosecretory material, catecholamines were found to be present in large quantities. In specimens with empty axon terminals, the amount of catecholamines was significantly less. Thus the secretory material appears to contain catecholamines. The results presented suggest dopamine to be the major component of the neurosecretory material.


Phase I study of cisplatin and nanoparticle albumin-bound-paclitaxel combined with concurrent radiotherapy in locally advanced esophageal squamous cell carcinoma.
This phase I study aimed to assess the safety, dose-limiting toxicity (DLT), maximum tolerated dose (MTD) and preliminary effect of nanoparticle albumin-bound (nab)-paclitaxel in combination with concurrent chemoradiotherapy in patients with locally advanced esophageal squamous cell carcinoma (ESCC). Patients with locally advanced ESCC who were ineligible or refused surgery were enrolled. Nab-paclitaxel (60 mg/m2 , 75 mg/m2 , and 90 mg/m2 ) and cisplatin (25 mg/m2 ) were administered intravenously weekly on days 1, 8, 15, 22, and 29 on the basis of the 3 + 3 dose escalation method. The total dose of radiation was 50-64 Gy. The primary endpoint was the safety of chemotherapy. The study enrolled 12 patients across three dose levels. No treatment-related deaths occurred. One patient in the 60 mg/m2 dose level occurred dose-limiting Grade 3 febrile neutropenia. No DLT was found in the 90 mg/m2 dose level thus the MTD was not reached. The phase II study's recommended dose was 75 mg/m2 based on the available preclinical and clinical data including pharmacokinetics, pharmacodynamics, efficacy, and toxicity. The frequent hematologic toxicities were leukocytopenia (Grade 1-2 of 66.7% and Grade 3-4 of 33.3%), neutropenia (Grade 1-2 of 91.7% and Grade 3-4 of 8.3%). Nonhematologic toxicities were mild and manageable. Overall response rate (ORR) of all patients achieved 100%. Weekly schedule of cisplatin and nab-paclitaxel in combination with concurrent radiotherapy showed manageable toxicities and promising antitumor activity in patients with locally advanced ESCC. The recommended dose of nab-paclitaxel for further studies is 75 mg/m2 .


Actively transcribed rRNA genes in S. cerevisiae are organized in a specialized chromatin associated with the high-mobility group protein Hmo1 and are largely devoid of histone molecules.
Synthesis of ribosomal RNAs (rRNAs) is the major transcriptional event in proliferating cells. In eukaryotes, ribosomal DNA (rDNA) is transcribed by RNA polymerase I from a multicopy locus coexisting in at least two different chromatin states. This heterogeneity of rDNA chromatin has been an obstacle to defining its molecular composition. We developed an approach to analyze differential protein association with each of the two rDNA chromatin states in vivo in the yeast Saccharomyces cerevisiae. We demonstrate that actively transcribed rRNA genes are largely devoid of histone molecules, but instead associate with the high-mobility group protein Hmo1.


Twenty-four-month outcomes of inflammatory choroidal neovascularisation treated with intravitreal anti-vascular endothelial growth factors: a comparison between two treatment regimens.
There is still no established treatment regimen for eyes with inflammatory choroidal neovascularisation (iCNV) treated with intravitreal anti-vascular endothelial growth factor (VEGF) injections. This study compared the 24-month outcomes of two treatment regimens of anti-VEGF injections in eyes with iCNV. Eyes with iCNV treated with anti-VEGF injections were divided into two groups: eyes treated with a loading phase of 3 monthly injections and then re-treated as needed (LOADING group) and eyes treated as needed from the beginning (PRN group). Visual acuity (VA), number of injections and iCNV recurrences at 24 months were compared between the groups. Eighty-two eyes were included, 42 in the LOADING and 40 in the PRN group. Baseline VA (mean(SD)) was 57.3 (15.8) letters in the LOADING vs 60.7 (15.6) letters in the PRN group (p=0.32). The VA (mean (95% CI)) increased at 3 months (+14.8 (10.6 to 18.9) and +11.2 (6.4 to 16) letters in the LOADING and PRN group, respectively) and remained significantly higher than baseline over the entire follow-up in both groups (all p<0.001). At 24 months, there was no difference in VA between the LOADING and PRN group (72.3 (14.0) vs 74.7 (11.3) letters, p=0.36) but the LOADING group received significantly more injections (median (Q1-Q3)) than the PRN (4.5 (3-7) vs 2.5 (2-3.2), p<0.0001). The iCNV recurrences were similar in both groups. iCNV responded well to anti-VEGF with significant and sustained VA improvement. The loading phase did not confer any advantage in terms of outcomes. PRN regimen from the beginning was as effective as more intensive treatment.


Effect of high-pressure D2 and H2 annealing on LFN properties in FD-SOI pTFET.
Tunneling field-effect transistors (TFETs) are a promising candidate for the next generation of low-power devices, but their performance is very sensitive to traps near the tunneling junction. This study investigated the effects of high-pressure deuterium (D2) annealing and hydrogen (H2) annealing on the electrical performance and low-frequency noise (LFN) of a fully depleted silicon-on-insulator p-type TFET. Without high-pressure annealing, the typical noise power spectral density exhibited two Lorentzian spectra that were affected by fast and slow trap sites. With high-pressure annealing, the interface trap density related to fast trap sites was reduced. The passivation of traps near the tunneling junction indicates that high-pressure H2 and D2 annealing improves the electrical performance and LFN properties, and it may become a significant and necessary step for realizing integrated TFET technology in the future.


Smilax china root extract as a novel Glucose- 6-phosphate dehydrogenase inhibitor for the treatment of hepatocellular carcinoma.
A novel therapeutic strategy for cancer treatment is to target altered tumor metabolism. Glucose- 6-phosphate dehydrogenase (G6PD) has been recently discovered to be implicated in apoptosis and angiogenesis, making it an excellent target in cancer treatment. The current study aimed to screen the plant extracts library to find potent hits against G6PD through enzymatic assay. Protein expression was induced by IPTG and purified using Ni-NTA columns after transformation of the pET-24a-HmG6PD plasmid into E. coli BL21-DE3 strain. An enzymatic assay was established by using purified rG6PD protein, for the screening of G6PD inhibitors. Out of 46 plant extracts screened, the sixteen plant extracts have shown inhibitory activity against the G6PD enzyme. At doses from 1 to 4 µg/ml, this extract demonstrated concentration-dependent inhibition of G6PD with an IC50 value of I.397 µg/ml. Moreover, the anticancer activity evaluation against HepG2 cells determined Smilax china as a potent inhibitor of cancer cells (IC50 value of 16.017 μg/ml). The acute and subacute toxicities were not observed in mice with various concentrations (50, 100, 200 and 2000 mg/kg). Furthermore, to identify the compounds from Smilax china as G6PD inhibitors, a literature-based phytochemical investigation of Smilax china was conducted, and sixty compounds were docked against the NADP+ and G6P binding sites of G6PD. The results of this study showed that three compounds were Scirpusin A, Smilachinin and Daucosterol with MolDock Score of -156.832, -148.215, and -145.733 respectively, against NADP+ binding site of G6PD. Conclusively, Smilax china root extract could be a safer drug candidate for the treatment of hepatocellular carcinoma.


Comparison of fentanyl and morphine in laser surgery for retinopathy of prematurity.
To compare complication rates of the analgesics fentanyl and morphine in preterm infants undergoing laser therapy for retinopathy of prematurity (ROP). In this observational study, the medical records of consecutive preterm neonates undergoing laser treatment of ROP from June 2007 through September 2010 were retrospectively reviewed. Because a fentanyl-based infusion protocol was initiated in November 2009, there was approximately the same number of treatment sessions with morphine and with fentanyl. In both groups, midazolam was used additionally on a case-by-case basis. Analgesia type, complications, and vital signs were documented at 5-minute intervals for all surgeries. The primary outcome was change in ventilation status. Secondary complications included change in temperature and incidence of apneic, bradycardic, and desaturation events. A total of 35 patients were included, with 17 in the morphine group (mean gestational age, 24.8 weeks; mean birth weight, 661 g) and 18 in the fentanyl group (mean gestational age, 24.4 weeks; mean birth weight, 681 g). Overall worsening of ventilation status was noted in 29% of patients in the morphine group and 6% of patients in the fentanyl group (P = 0.08; 95% confidence interval, -2% to 48%). Temperature instability (outside of 36.5° to 37.4°C range) was noted in 6% of patients in the morphine group and no patients in the fentanyl group. Apneic events were 3.2 times more common and bradycardic events 1.5 times more common in the morphine group. We found no difference in safety parameters for fentanyl infusion or morphine for analgesia in preterm infants undergoing ROP laser therapy in the neonatal intensive care unit setting. Although estimates of complication rates suggest that fentanyl may be safer, further study is needed to confirm this premise.


Mechanisms of oxidant-mediated cell injury. The glycolytic and mitochondrial pathways of ADP phosphorylation are major intracellular targets inactivated by hydrogen peroxide.
Inhibition of ADP phosphorylation by both glycolysis and mitochondria in P388D1 cells exposed to H2O2 is described. Net glucose uptake and lactate production were inhibited by oxidant exposure (ED50 = 50-100 microM). Glycolysis was specifically inactivated at the glyceraldehyde-3-phosphate dehydrogenase step by three independent mechanisms: (a) direct inactivation of the intracellular enzyme (ED50 approximately equal to 100 microM); (b) reduction of the intracellular concentration and redox potential of its nicotinamide cofactors; and (c) a cytosolic pH shift further from the enzyme optima. Consistent with inhibition of glycolysis at the glyceraldehyde-3-phosphate dehydrogenase step, a rise in the intracellular concentration of glyceraldehyde 3-phosphate, dihydroxyacetone phosphate, and fructose 1,6-bisphosphate was observed. The calculated combined inhibition of glyceraldehyde-3-phosphate dehydrogenase activity could be reasonably correlated with the depression in glycolytic flux rate with the appropriate modeling. The steady-state contribution by mitochondria to the total intracellular ATP pool was indirectly determined by the use of various metabolic inhibitors and was found to rapidly decline following exposure to 300-800 microM H2O2. The inhibition of ADP phosphorylation appeared to be related more to the direct inhibition of the ATPase-synthase complex rather than to the diminished capacity of the respiratory chain for coupled electron transport. Both the estimated rates of ADP phosphorylation by glycolysis and mitochondria and the estimated rate of ATP hydrolysis by ongoing metabolism were utilized to model the approximate decline in intracellular ATP expected at 15-min exposure to various H2O2 concentrations. Theoretical calculations and the measured intracellular ATP status were in good agreement. Oxidant exposure for 15 min resulted in dose-dependent killing of the cells (ED50 = 500 microM), indicating a close correlation between H2O2-mediated loss of intracellular ATP and cell viability. The possible contribution of impaired energy homeostasis during oxidant-mediated injury to the process of cell dysfunction and death is discussed.


Relative motion of selected carpal bones: a kinematic analysis of the normal wrist.
The relative motion of selected carpal bones and the radius was studied using five cadaver specimens labeled with metal markers to precisely quantitate their motions. Data was obtained by means of a combination of orthoradiography, sonic digitization, and computer analysis. We conclude that the wrist functions as two carpal rows with the distal row bones relatively tightly bound to one another and the proximal row bones less so but still moving together. Therefore, we theorize that the proximal row functions as a variable geometry intercalated segment between the distal row and the radius-triangular fibrocartilage.


Cardiac arrest induced by muscarinic or adenosine receptors agonists is reversed by DPCPX through double mechanism.
In the right atrium (RA), adenosine and acetylcholine inhibit the pacemaker function of the sinoatrial node and induce cardiac arrest. Pre-incubation of receptor antagonists is known to inhibit the cardiac arrest induced by these agonists; however, the effect of antagonist administration after established cardiac arrest has not been described. Therefore, we assessed whether specific receptor antagonists could revert cardiac arrest induced by adenosine and muscarinic receptors activation. RA isolated from adults Wistar rats were mounted in an organ bath containing Krebs solution. Cardiac arrest was induced by adenosine or ATP (1mM), the A1 adenosine receptor agonist CPA (0.1-1µM), and muscarinic receptor agonists, carbachol (0.3-1µM) and acetylcholine (1mM). After establishing the cardiac arrest, the A1 adenosine receptor antagonist DPCPX (0.3-30µM), the muscarinic receptor antagonist atropine (10nM to 100µM) or the phosphodiesterase inhibitor IBMX (10-300µM) were incubated in order to check for the return of spontaneous contractions. DPCPX reversed the cardiac arrest induced by adenosine, ATP and CPA. In addition, atropine reversed the cardiac arrest induced by carbachol. Unexpectedly, DPCPX also reversed the cardiac arrest induced by carbachol. Similarly to DPCPX, the phosphodiesterase inhibitor IBMX reversed the cardiac arrest induced by adenosine, CPA and carbachol. The antagonism of adenosine and acetylcholine receptors activation, as well as phosphodiesterase inhibition, are able to revert cardiac arrest. DPCPX restore spontaneous contractions via the selective antagonism of A1 adenosine receptor and through a secondary mechanism likely related to phosphodiesterase inhibition.


Risk factors for contact lens-related microbial keratitis in Singapore.
To investigate independent risk factors for contact lens-related microbial keratitis in Singapore and estimate their impact on disease load. Cases were contact lens wearers presenting to Singapore National Eye Centre with microbial keratitis between 2008 and 2010. Community contact lens wearers were recruited as controls. All wearers completed a previously validated questionnaire describing contact lens wear history, hygiene and compliance habits, and demographics. Risk factors significant in univariate analysis (P<0.2) were evaluated in a multivariate model. In all, 58 cases of microbial keratitis and 152 contemporaneous controls were identified. When controlling for other variables, Chinese had a 7 × lower risk compared with other races (95% CI: 2.3-21.3, P=0.001). Those aged between 25 and 44 years were at 3 × increased risk compared with younger wearers (95% CI: 1.1-9.6, P=0.04). Occasional overnight contact lens wear (less often than one night per week) was associated with a 4 × higher risk (95% CI: 1.2-15.4, P=0.03) compared with daily use. Not washing hands before handling was associated with a 13 × increased risk (95% CI: 1.9-84.8, P=0.008). Use of multipurpose solution A carried a 16 × higher risk compared with hydrogen peroxide (95% CI: 1.5-174.0, P=0.02). The combined PAR% for modifiable risk factors (occasional overnight wear, not washing of hands, and MPS A) was 82%. Consistent with previous findings, independent risk factors for contact lens-related microbial keratitis include poor hand hygiene, occasional overnight wear, and type of lens care solution. Prolonged overnight or extended contact lens use was infrequent in this population.


The yin and yang of neurotrophin action.
Neurotrophins have diverse functions in the CNS. Initially synthesized as precursors (proneurotrophins), they are cleaved to produce mature proteins, which promote neuronal survival and enhance synaptic plasticity by activating Trk receptor tyrosine kinases. Recent studies indicate that proneurotrophins serve as signalling molecules by interacting with the p75 neurotrophin receptor (p75NTR). Interestingly, proneurotrophins often have biological effects that oppose those of mature neurotrophins. Therefore, the proteolytic cleavage of proneurotrophins represents a mechanism that controls the direction of action of neurotrophins. New insights into the 'yin and yang' of neurotrophin activity have profound implications for our understanding of the role of neurotrophins in a wide range of cellular processes.


When to collect resting-state data: The influence of odor on post-task resting-state connectivity.
The human brain networks at rest represent spontaneous activity that is highly correlated between different brain regions. Previous studies have shown that these resting-state networks are flexible and dynamic, and they can be affected by performance of different types of tasks. Moreover, it has been suggested that the re-activation of a task-related brain network during rest promotes learning and improves the expertise on that task. However, it is still unclear whether the presence of different sensory information in the on-task state affects functional connectivity in subsequent resting-state fMRI even though the perception of the sensory information did not induce significant behavioral effects. To clarify this issue, we compared pre- and post-task resting-state fMRI of two groups of participants performing the same task either with an odor context (ODOR group) or without an odor context (AIR group). Seed-based functional connectivity analyses were performed with orbitofrontal cortex, piriform cortex and working-memory core network as seeds. The results showed that an odor context presented during an encoding task induced significant changes in the functional connectivity only within the olfactory network of the post-task resting-state compared to the same post-task situation without previous odor context. No significant difference in functional connectivity were found for the working-memory core network. This evidence emphasizes how the sensory context, in which a task is performed, is relevant for understanding the observed changes of functional connectivity during rest.


Effects of dexamethasone on expression and maintenance of cartilage in serum-containing cultures of calvaria cells.
The effects of dexamethasone on the ability of cells enzymatically isolated from 21-day fetal rat calvaria to produce cartilage in vitro has been investigated. Primary cultures of single-cell suspensions of rat calvaria were grown for up to 28 days in vitro in alpha-minimal essential medium containing 15% fetal bovine serum, 50 micrograms/ml ascorbic acid, 10 mM Na beta-glycerophosphate and dexamethasone at concentrations of 1 microM to 1 nM. Two types of nodules were present in dexamethasone-containing cultures. One has been characterized previously as bone (Bellows et al. 1986). The second morphologically resembled hyaline cartilage, possessed a strong Alcian blue-positive matrix and contained type-II, but not type-I, collagen. Both bone and cartilaginous nodules were spatially distinct and developed in isolation from each other. Cartilaginous nodules were found in the highest number at a dexamethasone concentration of 100 nM. Time-course experiments revealed that while the number of bone nodules increased continuously at least to day 28, the number of cartilaginous nodules remained constant after cultures had reached confluency. When cells were isolated separately from frontal and parietal bones and sutural regions, the greatest number of cartilaginous nodules developed from parietal bones. Since 21-day fetal rat calvaria contains 2 distinct patches of cartilage at the periphery of the parietal bones, it seems likely that this cartilaginous tissue is the origin of the cartilage cells. The results demonstrate that cultures of rat calvaria cells contain chondrocytes and possibly chondroprogenitor cells that are distinct from osteoprogenitors.(ABSTRACT TRUNCATED AT 250 WORDS)


[Synthesis of the capsular polysaccharide of Streptococcus pneumoniae type 14. 1. Synthesis of a derivative of the tetrasaccharide repetitive link].
A branched tetrasaccharide derivative of the repeating unit of the capsular polysaccharide of Streptococcus pneumoniae type 14 has been synthesised by condensation of per-O-benzoylated 1,2-O-(1-cyano)ethylidene derivative of lactose with 6-O-tritylated methyl lactosaminide.


Incidental Findings and Normal Anatomical Variants on Brain MRI in Children for Primary Headaches.
When MRI scans of the brain are obtained for evaluation of primary headaches in children, incidental findings and anatomical variants are commonly present. After a review of the prevalence, 11 types are presented.


Lingua-epiglottis position predicts glossopharyngeal obstruction in patients with obstructive sleep apnea hypopnea syndrome.
The objective of the study was to investigate the relationship between lingua-epiglottis position and glossopharyngeal obstruction in patients with obstructive sleep apnea hypopnea syndrome (OSAHS). One hundred and four patients with OSAHS diagnosed by polysomnography (PSG) were enrolled. Lingua-epiglottis position was visualized using endoscopy and classified into three types. Spiral CT imaging of the upper respiratory tract was performed to measure the cross-sectional area and inner diameter of the glossopharyngeal airway. The PSG was repeated after nasopharyngeal tube insertion (NPT-PSG). The NPT-PSG results, CT-measured data and incidence of stenosis were compared among the different lingua-epiglottis position groups. Obstructive sleep apnea hypopnea syndrome patients with different lingua-epiglottis positions had similar demographics. As lingua-epiglottis position type varied from type I to type III, cross-sectional area and inner diameter of the glossopharyngeal area decreased, glossopharyngeal airway stenosis rate increased, and apnea hypopnea index measured by NPT-PSG increased. The lowest oxygen saturation decreased. Lingua-epiglottis position was significantly related to glossopharyngeal obstruction. Lingua-epiglottis position should be used in clinical practice for the preliminary assessment of glossopharyngeal obstruction.


Policies and services in residential substance abuse programs: comparisons with psychiatric programs.
This article describes the Policy and Service Characteristics Inventory (PASCI), which assesses the policies and services of hospital and community-based substance abuse and psychiatric residential programs for adults. Developed using data from 96 representative programs, the PASCI has nine dimensions that have good internal consistency, are relatively independent, and are quite stable. In comparison to psychiatric programs (N = 43), substance abuse programs (N = 53) had more restrictive admissions policies and were less tolerant of problematic resident behavior; provided less individual choice and privacy, but more formal structures for resident control and policy clarity; and offered less daily living assistance. Compared to community-based substance abuse programs, hospital-based programs had less restrictive admissions policies, provided residents with less choice and control but more policy clarity, and offered more health and treatment and social-recreational services. The PASCI dimensions were related to substance abuse programs' organizational features such as size and staffing, and to program-level outcomes such as the proportion of patients who successfully complete the program. Potential applications of the PASCI and the full inventory of which it is a part are discussed.


Safety and efficacy of fingolimod in clinical practice: The experience of an academic center in the Middle East.
Few published studies addressed real-world clinical experience with fingolimod especially in the Middle East region. To review our clinical experience with fingolimod at a specialized academic MS center in Lebanon. All patients treated with fingolimod at the MS Center between October 2011 and January 2015 were retrospectively identified. A total of 122 patients were included. The first dose observation was uneventful in 98.8% of patients. Annualized relapse rate decreased from 1.16 pre-treatment to 0.29 post-treatment representing a relative risk reduction of 75% (p<0.0001). The proportion of patients with no new T2 or enhancing lesions was 66.3%. Seventy-six (62.3%) patients experienced adverse events with lymphopenia, increase liver enzymes, urinary tract infections and fatigue being the most common. Our cohort confirms the effectiveness and safety of fingolimod in a real world setting.


Effect of application of dairy manure, effluent and inorganic fertilizer on nitrogen leaching in clayey fluvo-aquic soil: A lysimeter study.
Dairy farm manure and effluent are applied to cropland in China to provide a source of plant nutrients, but there are concerns over its effect on nitrogen (N) leaching loss and groundwater quality. To investigate the effects of land application of dairy manure and effluent on potential N leaching loss, two lysimeter trials were set up in clayey fluvo-aquic soil in a winter wheat-summer maize rotation cropping system on the North China Plain. The solid dairy manure trial included control without N fertilization (CK), inorganic N fertilizer (SNPK), and fresh (RAW) and composted (COM) dairy manure. The liquid dairy effluent trial consisted of control without N fertilization (CF), inorganic N fertilizer (ENPK), and fresh (FDE) and stored (SDE) dairy effluent. The N application rate was 225kgNha-1 for inorganic N fertilizer, dairy manure, and effluent treatments in both seasons. Annual N leaching loss (ANLL) was highest in SNPK (53.02 and 16.21kgNha-1 in 2013/2014 and 2014/2015, respectively), which were 1.65- and 2.04-fold that of COM, and 1.59- and 1.26-fold that of RAW. In the effluent trial (2014/2015), ANLL for ENPK and SDE (16.22 and 16.86kgNha-1, respectively) were significantly higher than CF and FDE (6.3 and 13.21kgNha-1, respectively). NO3- contributed the most (34-92%) to total N leaching loss among all treatments, followed by dissolved organic N (14-57%). COM showed the lowest N leaching loss due to a reduction in NO3- loss. Yield-scaled N leaching in COM (0.35kgNMg-1 silage) was significantly (P<0.05) lower than that in the other fertilization treatments. Therefore, the use of composted dairy manure should be increased and that of inorganic fertilizer decreased to reduce N leaching loss while ensuring high crop yield in the North China Plain.


Acute neurotrophic keratitis with trigeminal trophic syndrome after craniotomy.
We report the case of a child who, shortly after undergoing suboccipital craniotomy for resection of a medullary cavernoma, developed corneal and conjunctival epithelial breakdown of the right eye with ipsilateral facial hypoesthesia as well as erosions and crusting of the eyelids, nostril, and lips on the right side. This combination of findings likely results from acute injury to the ipsilateral trigeminal ganglion, leading to acute neurotrophic keratitis and trigeminal trophic syndrome.


A model of prediction system for adverse cardiovascular reactions by calcineurin inhibitors among patients with renal transplants using gene-based single-nucleotide polymorphisms.
The application of pharmacogenomic information to diagnostic assays is expected to improve the prediction of drug efficacy and toxicity, leading to appropriate therapeutic regimens for individual patients. Cardiovascular events are common and severe adverse drug reactions (ADRs) among transplant patients treated with calcineurin inhibitors (CNIs). We conducted case-control association studies using 50,947 gene-based single-nucleotide polymorphisms (SNPs) to identify genetic variations that might be associated with cardiovascular risk factors in 72 renal transplant recipients with CNI therapy. The overall incidence of cardiovascular events was 13.9% (10/72) among patients receiving cyclosporine or tacrolimus; arrhythmias in six patients (8.3%), ischemic heart diseases in two patients (2.8%), and heart failure in two patients (2.8%). On the basis of results of the genome-wide association studies, we attempted to establish a scoring system to predict individual risks for cardiovascular toxicity of cyclosporine and tacrolimus. Estimation of the predictive performance was carried out by the use of internal leave-one-out cross-validation test. When we combined arrhythmia, ischemic heart disease and heart failure cases as subjects with a cardiotoxicity phenotype, nine of ten ADR patients and 50 of 62 non-ADR patients were correctly classified into the respective categories using the top eight SNPs. In addition, the proportion of individuals in the control population (n=246) with scores over the cut-off (11.0%) was close to the cardiovascular ADR frequency (8.3%) among renal transplant patients in the previous clinical study. Our results open the possibility that prediction of CNI-induced cardiovascular complications can lead to better prognosis and quality of life among kidney-transplant patients, and to improved immunosuppressive regimens.


Arsenic trioxide suppressed mantle cell lymphoma by downregulation of cyclin D1.
Mantle cell lymphoma (MCL) is aggressive with poor prognosis. Due to t(11;14)(q13;q32), cyclin D1 is overexpressed. The in vitro activities of arsenic trioxide (As2O3) in MCL were investigated. In MCL lines Jeko-1 and Granta-519, As2O3 induced dose-dependent and time-dependent increases in apoptosis accompanied by cyclin D1 suppression. Downregulation of cyclin D1 resulted in decreased retinoblastoma protein phosphorylation, which led to repressed G1 progression to S/G2 phases. As2O3 did not affect cyclin D1 gene transcription. Instead, As2O3 activated glycogen synthase kinase-3beta (by tyrosine-216 phosphorylation) and IkappaB kinase alpha/beta (by serine-176/180 phosphorylation), both of which phosphorylated cyclin D1 at threonine-286, leading to its poly-ubiquitination and degradation in the proteasome. These observations were recapitulated partly in primary MCL samples obtained from patients refractory to conventional treatment. Our findings suggested that As2O3 might be clinically useful in MCL.


A peptidases-resistant glycosylated analogue of substance P-(5-11). Specificity towards substance P receptors.
Glycosylated analogues of the C-terminal heptapeptide of substance P either free or blocked on the N-terminal glutamine were synthesized in order to develop a metabolically stable peptide that would have an increased specificity for one type of receptor. Of the analogue described, (N-alpha-Boc-beta-D-Glc-p (1----5) Gln) -Gln-Phe-Phe-Gly-Leu-Met-NH2 is highly resistant to degradation on exposure to rat hypothalamic slices. This glycosylated peptide is about one third as potent as substance P in eliciting contractions of the guinea-pig ileum and is almost devoided of affinity for the 125I-Bolton Hunter-SP specific binding sites on rat brain synaptosomes.


A Simple In Vitro Assay to Measure the Activity of Geranylgeranyl Diphosphate Synthase and Other Short-Chain Prenyltransferases.
Most carotenoids are C40 metabolites produced from C20 geranylgeranyl diphosphate (GGPP). The enzymes that produce this precursor, GGPP synthases (GGPPS), are members of the short-chain prenyltransferase (SC-PT) family. SC-PTs are enzymes that catalyze the sequential head-to-tail addition of one or more C5 molecules of isopentenyl diphosphate (IPP) to dimethylallyl diphosphate (DMAPP) with the concomitant release of pyrophosphate (PPi). SC-PTs produce linear isoprenyl diphosphates of up to C20 (GGPP) that serve as precursors for many groups of isoprenoids with a wide range of essential biological functions in Eucarya, Bacteria, and Archaea. Enzymatic analysis of SC-PT activity normally requires complex, laborious and expensive methods such as radioactivity-based assays or liquid chromatography-mass spectrometry (LC-MS). Here we describe a fast and inexpensive spectrophotometric protocol for determining the kinetic parameters of SC-PTs in purified enzyme preparations, using an adapted assay for PPi quantification. We developed the method using the Arabidopsis thaliana GGPPS11 enzyme, which produces geranylgeranyl diphosphate for the synthesis of carotenoids in the chloroplast.


Effect of cytochalasin D on the growth, encystation, and multinucleation of Entamoeba invadens.
The effect of cytochalasin D, a specific inhibitor of microfilaments, on the growth, encystation, and multinucleation of Entamoeba invadens was examined. Cytochalasin D blocked the growth of axenic E. invadens strain IP-1 in a dose-dependent manner, which suggests that the drug is effective against this species of Entamoeba as well as against E. histolytica strain HM1: IMSS as previously demonstrated. Encystation of E. invadens as induced in vitro was also inhibited by cytochalasin D. This is the first evidence of the participation of microfilaments in the encystation process. Concentrations of cytochalasin D effective for the inhibition of encystation were lower than those effective for the inhibition of growth. Trophozoites grown with cytochalasin D became multinucleate; more than three nuclei per cell were observed in 71% of trophozoites grown in the presence of the drug as opposed to only 5% of those grown in the absence of the drug. Also, trophozoites grown with cytochalasin D produced multinucleate cysts following their transfer to encystation medium. Encystation with cytochalasin D was more strongly inhibited among trophozoites grown in the presence of the drug than among those grown in the absence of the drug. Also, encystation without cytochalasin D was less frequently observed among trophozoites grown in the presence of the drug than among those grown in the absence of the drug. Thus, the multinucleation of trophozoites induced by cytochalasin D had an inhibitory effect on their encystation.


Murine teratocarcinoma-derived neuronal cultures.
This chapter describes the culture and propagation of murine embryonic stem cells, F9 and P19 and strategies for differentiation of these stem cells into neurons. Protocols focus on maintenance and propagation of these cells and routine procedures employed for differentiation into neuronal cells. Additional protocols are also described for obtaining enriched populations of mature neurons from P19 cells and differentiation of F9 cells into serotonergic or catecholaminergic neurons.The protocols described herein can be employed for dissection of the pathways such as gliogenesis and neurogenesis that are involved in differentiation of pluripotent stem cells such as F9 and P19 into glial cells or terminally differentiated neurons.


Toxoflavin contamination in rice samples from rice processing complexes in South Korea.
Toxoflavin contamination was investigated in broken rice produced as a by-product of domestic rice processing complexes (RPCs) in 2011 in South Korea. Of the 68 RPCs investigated, toxoflavin contamination was confirmed in 12 from three provinces: Gangwon, Gyeonggi, and Gyeongsang. Isolation of toxoflavin-producing bacteria independent of toxoflavin contamination was also performed in this study. We obtained 25 toxoflavin-producing bacterial isolates from rice samples; these samples were collected from the same 12 RPCs mentioned above. All 25 toxoflavin-producing bacteria were identified as Burkholderia glumae by 16S rRNA gene sequencing. Toxoflavin-producing ability differed slightly among the 25 isolates, but they all inhibited rice seed germination and induced seed rot. This is the first report of toxoflavin contamination and the toxin-producing bacterium B. glumae in broken rice produced during the rice milling process. Because toxoflavin has stable physical properties even above a boiling temperature of 100°C, it can pose a problem even if rice is cooked or processed. These results will serve as baseline data aiding comprehensive management of toxoflavin contamination during the post-harvest storage and processing of rice.


Interaction between inflammation-related gene polymorphisms and cigarette smoking on the risk of myocardial infarction in the Physician's Health Study.
Inflammation is known to be a major component of atherosclerosis, and cigarette smoking is known to induce a systemic inflammatory response. We therefore, investigated possible gene-environment interactions between various inflammation-related gene polymorphisms and cigarette smoking on the risk of myocardial infarction (MI) in the Physician's Health Study (PHS), a cohort of initially healthy middle-aged men. We used a nested case-control design consisting of 522 MI cases and 2,089 controls derived from PHS. Eleven inflammatory polymorphisms were studied using logistic regression analysis: eotaxin (ala23thr), intercellular adhesion molecule 1 (gly241arg), interleukin-4 (582C>T), interleukin-4 receptor (ile75val, gln576arg), interleukin-6 (-174G>C), interleukin-10 (-571C>A), P-selectin (val640leu, thr756pro, ser330asn), and vascular cell adhesion molecule 1 (-1594T>C). Interactions of smoking with all the three modes of inheritance (additive, dominant, recessive) were tested. Statistically significant (P<0.05) interaction terms were found for interleukin-4 receptor (ile75val), with odds ratios of 0.52 (95%CI:0.29-0.95) for Ile-Val and 0.34 (95%CI:0.14-0.83) for Val-Val, compared to the wildtype Ile-Ile; for interleukin-6 (-174G>C) with odds ratios of 2.16 (1.14-4.09) for GC and 0.81 (0.31-2.12) for CC, compared to the wildtype GG; and for P-selectin (ser330asn) with odds ratios of 0.48 (0.24-0.95) for Ser-Asn and 1.08 (0.29-3.93) for Asn-Asn, compared to the wildtype Ser-Ser, with these effects occurring only among the smokers. These data raise the possibility of interaction between the smoking status and certain inflammatory polymorphisms on the risk of MI in men. However, these results should be interpreted with caution due to the potential for false positive results that can arise from analyses with multiple comparisons.