[Establishment of biological assess for quality control of Fuzi based on determination of premature ventricular contractions in rats].
Aconiti Lateralis Radix (Fuzi) is a toxic traditional Chinese medicine with definite efficacy. In order to improve the quality control of its different prepared products and ensure the security in clinic, it is significant to establish a method of quality evaluation related to clinic adverse effects. Aiming at the important biological marker of early cardiac toxicity reaction, there was no method to detect it. In this manuscript, a novel approach for measuring the minimal toxic dose (MTD) of premature ventricular contractions (PVC) poisoning of rats was established. Then, the determination methodology and conditions were optimized to meet the needs of the quality and biological assessment, including animal sex, weight, stability of standards and test solutions. Using this method, the MTD value of different Fuzi products were determined, such as Heishunpian, Baifupian, Zhengfupian, Baofupian, and Paotianxiong. The results showed that the MTD of Fuzi was significantly decreased after detoxification processed (P<0.05) and the MTD of Heishunpian, Zhengfupian, Baofupian and Baifupian was as much as 15.76, 22.36, 19.65 and 20.97 times to that of unprocessed Shengfuzi. In addition, Paotianxiong could not induce PVC in rats, which indicated that Paotianxiong was nontoxic and safe.This method could appropriately reflects the cardiotoxity of Fuzi and its prepared samples. Together with the chemical composition analysis, the contents of diester alkaloids were explored including aconitine, mesaconitine and hypaconitine as well as monoester alkaloids in Fuzi and its prepared products were significantly associated with PVC. Furthermore, there may be some components undetermined facilitating arrhythmia to be worth exploring. This research provides an overall and comprehensive approach to diagnose early clinical cardiotoxity and control the quality of Fuzi, which could not only be a complementary solution for the chemical evaluation, but a new method to ensure its efficacy and security of clinical application.


Chimeric Ad5.F35 vector evades anti-adenovirus serotype 5 neutralization opposing GUCY2C-targeted antitumor immunity.
Adenovirus serotype 5 (Ad5) is a commonly used viral vector for transient delivery of transgenes, primarily for vaccination against pathogen and tumor antigens. However, endemic infections with Ad5 produce virus-specific neutralizing antibodies (NAbs) that limit transgene delivery and constrain target-directed immunity following exposure to Ad5-based vaccines. Indeed, clinical trials have revealed the limitations that virus-specific NAbs impose on the efficacy of Ad5-based vaccines. In that context, the emerging focus on immunological approaches targeting cancer self-antigens or neoepitopes underscores the unmet therapeutic need for more efficacious vaccine vectors. Here, we evaluated the ability of a chimeric adenoviral vector (Ad5.F35) derived from the capsid of Ad5 and fiber of the rare adenovirus serotype 35 (Ad35) to induce immune responses to the tumor-associated antigen guanylyl cyclase C (GUCY2C). In the absence of pre-existing immunity to Ad5, GUCY2C-specific T-cell responses and antitumor efficacy induced by Ad5.F35 were comparable to Ad5 in a mouse model of metastatic colorectal cancer. Furthermore, like Ad5, Ad5.F35 vector expressing GUCY2C was safe and produced no toxicity in tissues with, or without, GUCY2C expression. Importantly, this chimeric vector resisted neutralization in Ad5-immunized mice and by sera collected from patients with colorectal cancer naturally exposed to Ad5. These data suggest that Ad5.F35-based vaccines targeting GUCY2C, or other tumor or pathogen antigens, may produce clinically relevant immune responses in more (≥90%) patients compared with Ad5-based vaccines (~50%).


Thalassotalea mangrovi sp. nov., a bacterium isolated from marine mangrove sediment.
A novel Gram-stain-negative, strictly aerobic bacterium that has a rod-like shape with a single polar flagellum in the exponential phase of growth and a spherical or ovoid shape without a flagellum in the stationary phase was isolated from a mangrove wetland sediment sample collected at Beilun Estuary National Nature Reserve, Guangxi Province, PR China and designated strain ZS-4T. This strain grew optimally at pH 6.0-8.0, at a temperature of 37 °C and in the presence of 3-4 % (w/v) NaCl. Its polar lipid profile included phosphatidylethanolamine, phosphatidylglycerol, one unidentified aminophospholipid and two uncharacterized lipids. Ubiquinone 8 (Q-8) was the sole respiratory quinone and the cellular fatty acids were dominated by C17 : 1ω8c and C16 : 0. A phylogenetic analysis based on the 16S rRNA gene sequence showed that strain ZS-4T exhibited its highest similarities to the type strains Thalassotalea litorea HMF4135T (97.8 %) and Thalassotalea ponticola GJSW-36T (95.9 %). A whole genome-level comparison of strain ZS-4T with T. litorea MCCC 1K03283T revealed an average nucleotide identity value of 75.6 % and a calculated DNA-DNA hybridization value of 19.6 %. In addition, the genomic DNA G+C content of strain ZS-4T was 45.9 mol%. Thus, based on analyses of its morphology, physiology, fatty acid composition and 16S rRNA gene sequence, strain ZS-4T should be considered a novel species of the genus Thalassotalea, with the proposed name Thalassotaleamangrovi sp. nov. The type strain is ZS-4T (=KCTC 72399T=MCCC 1K03630T).


Ciglitazone, a PPARgamma agonist, ameliorates diabetic nephropathy in part through homocysteine clearance.
Diabetes and hyperhomocysteinemia (HHcy) are two independent risk factors for glomeruloslerosis and renal insufficiency. Although PPARgamma agonists such as ciglitazone (CZ) are known to modulate diabetic nephropathy, the role of CZ in diabetes-associated HHcy and renopathy is incompletely defined. We tested the hypothesis that induction of PPARgamma by CZ decreases tissue Hcy level; this provides a protective role against diabetic nephropathy. C57BL/6J mice were administered alloxan to create diabetes. Mice were grouped to 0, 1, 10, 12, and 16 wk of treatment; only 12- and 16-wk animals received CZ in drinking water after a 10-wk alloxan treatment. In diabetes, PPARgamma cDNA, mRNA, and protein expression were repressed, whereas an increase in plasma and glomerular Hcy levels was observed. CZ normalized PPARgamma mRNA and protein expression and glomerular level of Hcy, whereas plasma level of Hcy remained unchanged. GFR was dramatically increased at 1-wk diabetic induction, followed by hypofiltration at 10 wk, and was normalized by CZ treatment. This result corroborated with glomerular and preglomerular arteriole histology. A steady-state increase of RVR in diabetic mice became normal with CZ treatment. CZ ameliorated decrease bioavailability of NO in the diabetic animal. Glomerular MMP-2 and MMP-9 activities as well as TIMP-1 expression were increased robustly in diabetic mice and normalized with CZ treatment. Interestingly, TIMP-4 expression was opposite to that of TIMP-1 in diabetic and CZ-treated groups. These results suggested that diabetic nephropathy exacerbated glomerular tissue level of Hcy, and this caused further deterioration of glomerulus. CZ, however, protected diabetic nephropathy in part by activating PPARgamma and clearing glomerular tissue Hcy.


Are Strength Indicators and Skin Temperature Affected by the Type of Warm-Up in Paralympic Powerlifting Athletes?
(1) Background: the present study aimed to evaluate the effect of different types of warm-ups on the strength and skin temperature of Paralympic powerlifting athletes. (2) Methods: the participants were 15 male Paralympic powerlifting athletes. The effects of three different types of warm-up (without warm-up (WW), traditional warm-up (TW), or stretching warm-up (SW)) were analyzed on static and dynamic strength tests as well as in the skin temperature, which was monitored by thermal imaging. (3) Results: no differences in the dynamic and static indicators of the force were shown in relation to the different types of warm-ups. No significant differences were found in relation to peak torque (p = 0.055, F = 4.560, η2p = 0.246 medium effect), and one-repetition maximum (p = 0.139, F = 3.191, η2p = 0.186, medium effect) between the different types of warm-ups. In the thermographic analysis, there was a significant difference only in the pectoral muscle clavicular portion between the TW (33.04 ± 0.71 °C) and the WW (32.51 ± 0.74 °C) (p = 0.038). The TW method also presented slightly higher values than the SW and WW in the pectoral muscles sternal portion and the deltoid anterior portion, but with p-value > 0.05. (4) Conclusions: the types of warm-ups studied do not seem to interfere with the performance of Paralympic Powerlifting athletes. However, the thermal images showed that traditional warm-up best meets the objectives expected for this preparation phase.


[Vulnerability of management of waste from health services in João Pessoa, State of Paraíba (Brazil)].
The discussion in the academy on solid urban waste residues (UWR) is analyzed, with health service waste (HSW) being an integral part of UWR, not necessarily in terms of the amount generated, but due to the risk potential to collective and environmental health. The scope of the study was to verify the vulnerability of UWR handling in primary, secondary and tertiary healthcare units in João Pessoa in the state of Paraíba. This involved a quantitative, exploratory and descriptive investigation of the handling of UWR, using a checklist and analytical evaluation of the vulnerability of HSW handling for data collection. It was detected that 21.05% of the establishments failed to separate hazardous waste, 26.34% did not standardize plastic disposal bags, and 47.37% of the workers responsible for collection were not trained to handle HSW. It was concluded that the vulnerability of HSW handling in João Pessoa in the order of 48.02%, was significant, potentially contributing to the deterioration of environmental and collective health and leading to a public health problem.


Synergistic effects of adjuvants, endotoxin, and fasting on induction of diabetes with multiple low doses of streptozocin in rats.
Three weekly intraperitoneal injections of complete Freund's adjuvant (CFA) and, 1 day later, low-dose streptozocin (STZ; 25 mg/kg i.p.) have been reported to cause immune destruction of beta-cells and a gradual onset of diabetes mellitus. In this study, male Lewis rats were injected intraperitoneally with CFA and 1 day later with low-dose STZ; these were repeated at weekly intervals for 3 wk. The incidence of diabetes mellitus (nonfasted plasma glucose greater than 200 mg/dl) in wk 1, 2, 3, and 4 was 50, 80, 93, and 100%, respectively. Rats receiving either CFA or STZ only did not develop diabetes. Injections of either the components of CFA (incomplete Freund's adjuvant and Mycobacterium butyricum), another granuloma-inducing organism (Listeria monocytogenes), or endotoxin before STZ induced diabetes, but the onset was slower and the diabetes was less severe than with CFA and STZ. Because intraperitoneal CFA injections caused peritoneal irritation, acute weight loss, and hypoglycemia on the day after injection, we examined whether fasting alone potentiated low-dose STZ. Fasting for 24 h before and 24 h after low-dose STZ caused diabetes that was similar in rapidity of onset and severity to that induced with CFA and STZ. Administration of CFA subcutaneously before STZ did not cause hypoglycemia or weight loss but did cause diabetes. Thus, the fasting effect of intraperitoneal CFA was not responsible for the induction of diabetes with CFA and STZ. These data indicate that immunologic adjuvants, endotoxin, and fasting all potentiate the diabetogenic action of low-dose STZ.(ABSTRACT TRUNCATED AT 250 WORDS)


[Neurohumoral changes induced by hemofiltration in congestive heart failure: physiopathological inferences].
Hemofiltration is a method suitable for rapid substraction of plasma water that generally allows reduction of circulating levels of norepinephrine. Using that non-pharmacological approach we investigated the mechanisms involved in the metabolism of the hormone as well as the hemodynamic correlates of a prompt and great fall of the sympathetic neurotransmitter in patients with chronic refractory congestive heart failure (CHF). In 23 patients with CHF, hemofiltration of 2983 +/- 1228 ml of plasma water (in 5 +/- 2 hours of treatment) increased urinary output by 606 +/- 415 ml in the day of the procedure as well as sodium excretion by 53 +/- 38 mEq/24 h; simultaneously, a mean fall in plasma norepinephrine concentration by 515 +/- 444 pg/ml was observed. These effects were prompt and persisted or even rose in the next 24 and 48 hours, not being related to changes in plasma renin activity, right atrial, wedge pulmonary artery and renal perfusion pressures and to the amount and duration of hemofiltration. Our data did not clarify the mechanism involved in the increase of the diuresis and for its coupling with the fall in plasma norepinephrine. Nevertheless, we found a strong and statistically significant correlation (r = 0.7; p less than 0.01) between percent changes from baseline values of norepinephrine and diuresis. It is therefore suggested that the same, still unknown, mechanism which increased urinary output also potentiated norepinephrine removal by the kidney: or that water reabsorption from extravascular spaces (triggered by hemofiltration and continued by increased diuresis) resulted in regression of organ congestion leading to an improved clearance of norepinephrine by different organs.(ABSTRACT TRUNCATED AT 250 WORDS)


Genetic analysis of the longevity of French sport horses in jumping competition.
The longevity of sport horses is an economic and ethical issue. The aim of this study was to analyze the duration of the competitive life of jumping horses in France to assess the potential for genetic evaluation and to propose rules of management for sport horses. Data included lifetimes spent in jumping competitions for the 209,296 horses born from 1968 onward with performances between 1972 and 2008; the data set contained 22% right-censored records. Longevity was measured in years. Discrete survival analysis included fixed effects of region of birth, month of birth, year of recording, age at first competition, interaction between sex and level of jumping performance as measured by the logarithm of earnings adjusted for sex, age, year, and random sire and maternal grand-sire effects. There were 16,668 sires and maternal grand-sires. All fixed effects were highly significant (P < 0.001). Management of the sports career had an important effect on longevity: against common belief, the younger the horse started competing, the longer it stayed in competition. For horses that started competing at an age of 6 yr, the risk of culling was 1.33-fold that of horses having started at 4 yr of age. The less success in competition, the greater was the chance for leaving competition, especially for horses without earnings. For a gelding without earnings, the risk of culling was 1.40-fold that of an average-performance gelding and 2.57-fold that of a top-rated gelding (performance at least 2 SD above the mean). Mares always had greater relative risk than geldings or stallions because they may be culled from competition to be used for breeding. The risk of culling for females was 1.45-fold that of a gelding with the same performance. The heritability of the length of competitive life was 0.10. Breeding values were predicted for sires, and 3,303 sires showed an accuracy greater than 0.60. Among these sires, 262 were used for breeding in 2008.


Nocturnal airflow obstruction, histamine, and the autonomic central nervous system in children with allergic asthma.
A study was carried out to investigate whether an imbalance in the autonomic nervous system or release of histamine, or both, is responsible for the nocturnal increase in airflow obstruction in asthmatic children. The study comprised 18 children with allergic asthma, nine with (group 1) and nine without (group 2) nocturnal airflow obstruction, and an age matched control group. All drugs were withheld for three days before and during the study. On day 4 each child was admitted to hospital and a series of measurements was made every four hours for 24 hours. These included measurements of the forced expiratory volume in one second (FEV1), heart rate and sinus arrhythmia gap from an electrocardiogram (an indirect measure of parasympathetic activity) and urine sampling for determination of catecholamine and N'-methylhistamine concentrations (measures of sympathetic activity and histamine release respectively). Urinary N'-methylhistamine excretion was significantly higher over the 24 hours in children in group 1 than in children in group 2, and overnight values were also significantly higher in children in group 1 than those in group 2. Mean (SEM) values (mumol/mol creatinine) were 154.6 (11.2) in group 1 and 110 (11.2) in group 2 for 2400-0400 hours samples and 139.2 (13.1) and 101.2 (10.6) 0400-0800 hours samples. There was no evidence of decreased sympathetic or increased parasympathetic activity in association with the nocturnal airflow obstruction; noradrenaline concentrations were increased in group 1. These observations indicate that nocturnal airflow obstruction is associated with increased release of histamine overnight.


Subcellular localization of CrmA: identification of a novel leucine-rich nuclear export signal conserved in anti-apoptotic serpins.
The cowpox virus-encoded anti-apoptotic protein cytokine response modifier A (CrmA) is a member of the serpin family that specifically inhibits the cellular proteins caspase 1, caspase 8 and granzyme B. In this study, we have used Flag- and yellow fluorescent protein (YFP)-tagged versions of CrmA to investigate the mechanisms that regulate its subcellular localization. We show that CrmA can actively enter and exit the nucleus and we demonstrate the role of the nuclear export receptor CRM1 in this shuttling process. CrmA contains a novel leucine-rich nuclear export signal (NES) that is functionally conserved in the anti-apoptotic cellular serpin PI-9. Besides this leucine-rich export signal, additional sequences mapping to a 103-amino-acid region flanking the NES contribute to the CRM1-dependent nuclear export of CrmA. Although YFP-tagged CrmA is primarily located in the cytoplasm, shifting its localization to be predominantly nuclear by fusion of a heterologous nuclear localization signal did not impair its ability to prevent Fas-induced apoptosis. We propose that nucleocytoplasmic shuttling would allow CrmA to efficiently target cellular pro-apoptotic proteins not only in the cytoplasm, but also in the nucleus, and thus to carry out its anti-apoptotic function in both compartments.


A 16-Year Cohort Analysis of Autism Spectrum Disorder-Associated Morbidity in a Pediatric Population.
Introduction: This chapter presents the analysis of physician-diagnosed International Classification of Diseases (ICD version 9) disorders and diseases associated with autism spectrum disorders (ASD) in a 16-year pediatric cohort. Materials and Methods: The sample (n = 47,180; 62% male) consisted of children in the Alberta Health Services Calgary Health Region catchment under the age of 3 years, who received any physician-assigned ICD 9 diagnosis before the age of three between April 1993 and December 31, 1994. There were 111 females and 609 males with ASD diagnosed at any time between 1993 and 2010. The results detail the 16-year odds ratio (OR) associations of ASD diagnosis within the major classes of international classification of diseases (ICD 9) stratified by age and sex in the cohort. Further, for those suffering from ASD and any other disorder or disease, the analysis presents by sex, age, and duration, the proportions of all index physician-assigned ICD diagnoses, arising significantly before and after the index ASD diagnosis. Results: The rate of treated ASD in the cohort was 1 in 65 and the 16-year population rate of ASD was 62 per 10,000. For males with an ASD over the 16 year period, the ORs were significantly greater than the value one for 15 of the 17 main ICD classes and for 10 of the main ICD classes for females. Different age strata presented a more specific account of the main ICD class OR profiles. More specifically, 28 ICD disorders significantly preceded and 95 ICD disorders significantly followed ASD for females. Thirty-eight ICD disorders significantly preceded and 234 ICD disorders significantly followed ASD for males. Conclusions: The results largely confirm past studies focusing on more constrained sets of ASD morbidity. The age-stratified ORs gauge the order of risk in time for the cohort. The proportions of specific ICD disorders arising before and after ASD may be useful in respect to informing basic ASD research and ASD clinical management. Limitations are discussed.


Evidence for a virus in non-A, non-B hepatitis transmitted via the fecal-oral route.
Typical acute hepatitis was reproduced in a human volunteer immune to hepatitis A virus (HAV) after oral administration of pooled stool extracts from presumed cases of epidemic non-A, non-B hepatitis. Markers of hepatitis B infection, anti-HAV IgM, and increase in total anti-HAV level were not detectable in the volunteer's sera during the course of infection. Spherical 27- to 30-nm virus-like particles were visualized by immune electron microscopy (IEM) in stool samples collected during preclinical and early postclinical phases. These particles banded in CsCl at a buoyant density of 1.35 g/cm3. They reacted in the IEM test with sera from individuals who had experienced two non-B hepatitis episodes but did not react with sera from routine anti-HAV IgM-positive hepatitis patients. Intravenous inoculation of cynomolgus monkeys with the virus-containing stool extract resulted in histopathologically and enzymatically confirmed hepatitis, excretion of virus-like particles, and antibody response to them.


[Import of dogs by public animal shelters and private animal protection societies].
Every year thousands of dogs from EU Member States and third countries are brought to Germany by private animal protectionists, animal protection societies and public animal shelters offering them for an average charge of 250,00 Euro each. In addition to violations of current legal regulations, there might also be real crime offence behind the matter. Moreover, people, especially children, are increasingly endangered by various infectious diseases imported to Germany and spreading throughout the country as well as by dogs suffering from bad deprivation damage. The fact that the new owners are often not told the truth about the dogs'foreign origin is another problem. They are often marked with a chip after being brought to Germany and only then they are provided with a national vaccination record or a German pet passport. Finally the question arises whether the "rescue" of dogs and finding a place for them in Germany is just "big business"--for animal protection societies, practitioners, dog shelters, dog psychologists and the media.


Antibody responses of naive cattle to two inactivated bovine viral diarrhoea virus vaccines, measured by indirect and blocking ELISAS and virus neutralisation.
Two groups of naive heifers were given primary courses of two inactivated bovine viral diarrhoea (BVD) virus vaccines licensed for use in the UK. Their humoral responses in serum and milk were assayed by means of an indirect ELISA detecting antibodies to structural viral glycoproteins, a blocking ELISA specific for antibodies to the non-structural protein NS2-3 and the virus neutralisation test (VNT). For each assay, the numbers of serum or milk samples testing positive at each sample point and the mean values were determined. In both vaccine groups, serum antibody responses were detected by the indirect ELISA and the VNT, with both the numbers of seropositive animals and mean values peaking five weeks after the second vaccination. In the 23 heifers vaccinated with Bovilis BVD, the mean NS2-3-specific ELISA values remained low throughout the trial, with no serum or milk samples testing positive. In the 24 heifers vaccinated with Bovidec, the mean NS2-3 responses peaked below the level of positivity five weeks after the second vaccination, before declining again; NS2-3-specific antibodies were detected in one serum sample and one milk sample from two heifers in this group. A pooled milk sample from each vaccine group tested negative by both ELISAS 12 weeks after the second vaccination.


Binding of peptides that are specified by complementary RNAs.
An interesting pattern in the genetic code was reported previously [Blalock & Smith (1984) Biochem. Biophys. Res. Commun. 121, 203-207]. In the 5'-to-3' direction, codons for hydrophilic and hydrophobic amino acids are generally complemented by codons for hydrophobic and hydrophilic amino acids respectively. The average tendency of codons for 'unchanged' (slightly hydrophilic) amino acids was to be complemented by codons for 'unchanged' amino acids. We now show that the same pattern results when the complementary codon is read in the 3'-to-5' direction. This pattern is further shown to result in the interaction of peptides specified by complementary RNAs regardless of whether the amino acids are assigned in the 5'-to-3' or the 3'-to-5' direction. Here we demonstrate that peptides specified by complementary RNAs bind to each other with specificity and high affinity.


Degeneration in vivo of rat hippocampal neurons by wild-type Alzheimer amyloid precursor protein overexpressed by adenovirus-mediated gene transfer.
In an attempt to elucidate the pathological implications of intracellular accumulation of the amyloid precursor protein (APP) in postmitotic neurons in vivo, we transferred APP695 cDNA into rat hippocampal neurons by using a replication-defective adenovirus vector. We first improved the efficiency of adenovirus-mediated gene transfer into neurons in vivo by using hypertonic mannitol. When a beta-galactosidase-expressing recombinant adenovirus suspended in 1 M mannitol was injected into a dorsal hippocampal region, a number of neurons in remote areas were positively stained, presumably owing to increased retrograde transport of the virus. When an APP695-expressing adenovirus was injected into the same site, part of the infected neurons in the hippocampal formation underwent severe degeneration in a few days, whereas astrocytes near the injection site showed no apparent degeneration. These degenerating neurons accumulated different epitopes of APP, and beta/A4 protein (Abeta)-immunoreactive materials were undetected in the extracellular space. A small number of degenerating neurons showed nuclear DNA fragmentation. Electron microscopic examinations demonstrated that degenerating neurons had shrunken perikarya along with synaptic abnormalities. Microglial cells/macrophages were often found in close proximity to degenerating neurons, and in some cases they phagocytosed these neurons. These results suggest that intracellular accumulation of wild-type APP695 causes a specific type of neuronal degeneration in vivo in the absence of extracellular Abeta deposition.


Nuclear corepressor SMRT is a strong regulator of body weight independently of its ability to regulate thyroid hormone action.
Silencing Mediator of Retinoid and Thyroid Hormone Receptors (SMRT) and the nuclear receptor co-repressor1 (NCoR1) are paralogs and regulate nuclear receptor (NR) function through the recruitment of a multiprotein complex that includes histone deacetylase activity. Previous genetic strategies which deleted SMRT in a specific tissue or which altered the interaction between SMRT and NRs have suggested that it may regulate adiposity and insulin sensitivity. However, the full role of SMRT in adult mice has been difficult to establish because its complete deletion during embryogenesis is lethal. To elucidate the specific roles of SMRT in mouse target tissues especially in the context of thyroid hormone (TH) signaling, we used a tamoxifen-inducible post-natal disruption strategy. We found that global SMRT deletion causes dramatic obesity even though mice were fed a standard chow diet and exhibited normal food intake. This weight gain was associated with a decrease in energy expenditure. Interestingly, the deletion of SMRT had no effect on TH action in any tissue but did regulate retinoic acid receptor (RAR) function in the liver. We also demonstrate that the deletion of SMRT leads to profound hepatic steatosis in the setting of obesity. This is unlike NCoR1 deletion, which results in hepatic steatosis due to the upregulation of lipogenic gene expression. Taken together, our data demonstrate that SMRT plays a unique and CoR specific role in the regulation of body weight and has no role in TH action. This raises the possibility that additional role of CoRs besides NCoR1 and SMRT may exist to regulate TH action.


Effects of shared mental models in teams performing video-assisted thoracoscopic surgery lobectomy.
Important non-technical skills enable operating teams to establish shared mental models (SMMs). The importance of SMMs in regards to surgical performance and peri-operative outcomes remains to be investigated. The aim of this study was to explore whether shared mental models (SMMs) of team resources and the current situation, respectively, were predictive of technical skills, duration of surgery, and amount of intra-operative bleeding in video-assisted thoracoscopic surgery (VATS). A prospective multi-center observational study was conducted at four tertiary academic hospitals during VATS lobectomy procedures. Data included pre-operative and post-operative questionnaires answered by each of the six team members to measure the SMMs; thoracoscopic video recordings assessed using the previously validated VATS lobectomy Assessment Tool (VATSAT); surgery-related time stamps; and amount (volume) of intra-operative bleeding. Linear regression analyses were conducted to adjust for confounders. Fifty-eight lobectomy procedures were included. Median (interquartile range) VATSAT score was 33.3 (scale 8-40) duration of surgery 101 min (88-123), and amount of intra-operative bleeding 100 ml (20-150). The mean (± SD) of teams' SMMs of the current situation was 20 (± 5). They were not predictive of the surgeons' technical skills, but every one point increase in SMM score significantly predicted a 1 min 52 s decrease in duration of surgery and an 11% decrease in amount of bleeding. The SMMs of team resources were not predictive of any outcomes. VATS teams' superior SMMs of the current situation related to significantly shorter duration of surgery and decreased intra-operative bleeding, indicating an effect on team performance and patient care. NCT02999113 at http://www. gov .


Molecular identification of a peroxidase gene controlling body size in the entomopathogenic nematode Steinernema hermaphroditum.
The entomopathogenic nematode Steinernema hermaphroditum was recently rediscovered and is being developed as a genetically tractable experimental system for the study of previously unexplored biology, including parasitism of its insect hosts and mutualism with its bacterial endosymbiont Xenorhabdus griffiniae. Through whole-genome re-sequencing and genetic mapping we have for the first time molecularly identified the gene responsible for a mutationally defined phenotypic locus in an entomopathogenic nematode. In the process we observed an unexpected mutational spectrum following EMS mutagenesis in this species. We find that the ortholog of the essential C. elegans peroxidase gene skpo-2 controls body size and shape in S. hermaphroditum. We confirmed this identification by generating additional loss-of-function mutations in the gene using CRISPR-Cas9. We propose that the identification of skpo-2 will accelerate gene targeting in other Steinernema entomopathogenic nematodes used commercially in pest control, as skpo-2 is X-linked and males hemizygous for loss of its function can mate, making skpo-2 an easily recognized and maintained marker for use in co-CRISPR.


The first three weeks assessed by transvaginal color Doppler.
Transvaginal color Doppler has made possible to study ovarian and uterine perfusion in non-pregnant and pregnant patients, thus advancing the understanding of the early human development. RI of follicular blood flow starts decreasing prior to ovulation reaching its nadir at ovulation. It is considered that apart from hormonal factors the angiogenesis is also involved. The mature corpus luteum shows increased blood flow velocity in relation to preovulatory follicle. Comparing RI values of luteal blood flow of normal and ectopic pregnancy no difference was found. But in threatened, incomplete and missed abortions the resistance and pulsatility indices were significantly higher than in normal pregnancy. The follow up of the luteal flow might have a prognostic value in a group of patients with threatened abortion. In women with spontaneous cycles the day preceding the ovulation impedance to uterine flow velocity starts decreasing. Alterations in flow velocity patterns of the radial and spiral arteries in spontaneous ovulatory cycles are paralleling blood flow dynamics of the uterine arteries. In stimulated cycles RI increases the day before ovulation in both the uterine arteries and their branches. It seems that endometrial perfusion presents more accurate noninvasive assay of uterine receptivity than uterine artery perfusion alone. Endometrial receptivity is maximum during the time of peak luteal function during which implantation is most likely to occur. During the pregnancy impedance to blood flow decrease from the main uterine to the spiral arteries as well as with the advancing gestational age. The spiral arteries in pregnancy become the vessels with completely different haemodynamic characteristics in relation to other arteries of uteroplacental circulation. Color Doppler adds new information on perfusion and pathophysiological changes connected with the ectopic trophoblast implantation.


A new focus of Schistosoma haematobium/S. intercalatum hybrid in Cameroon.
Terminal-spined schistosome eggs were found in both stool and urine specimens of a high proportion of infected individuals in Kinding Njabi, a village located in a savannah/forest ecotone of Center Province, Cameroon. Infections with terminal-spined schistosomes are not common in that region and first appeared in Kinding Njabi in the mid-1980s. The village population, determined by house-to-house census in 1986, was 345. Of 302 stool samples examined by a thick smear technique, 58 (19%) were positive, and of 320 urine samples examined by sedimentation, 40 (12%) were positive. Twenty-six (37%) of a total of 71 infected persons passed eggs in both stool and urine. Egg size distribution and morphology were similar to previous reports for Schistosoma haematobium/S. intercalatum hybrids. Bulinus forskalii was the only snail found during extensive surveys in and around Kinding Njabi; infected snails were recovered from several temporary habitats. This focus is of particular interest because it represents the first report of transmission of hybrid schistosomes outside of the hybrid zone. Interviews with villagers and local health officials suggest that schistosomiasis was introduced by immigrants from Loum, a town approximately 100 km southwest of Kinding Njabi, where S. haematobium, S. intercalatum, and their hybrid are known to exist. Results from snail host specificity studies using the Kinding Njabi parasite showed that stool-derived miracidia were compatible only with B. forskalii, the exclusive host for S. intercalatum in Cameroon. Urine-derived miracidia were compatible only with B. truncatus, a S. haematobium host in Cameroon.(ABSTRACT TRUNCATED AT 250 WORDS)


Excitatory drive to spinal motoneurones is necessary for serotonin to modulate motoneurone excitability via 5-HT2 receptors in humans.
Serotonin modulates corticospinal excitability, motoneurone firing rates and contractile strength via 5-HT2 receptors. However, the effects of these receptors on cortical and motoneurone excitability during voluntary contractions have not been explored in humans. Therefore, the purpose of this study was to investigate how 5-HT2 antagonism affects corticospinal and motoneuronal excitability with and without descending drive to motoneurones. Twelve individuals (aged 24 ± 4 years) participated in a double-blind, placebo-controlled, crossover study, whereby the 5-HT2 antagonist cyproheptadine was administered. Transcranial magnetic stimulation (TMS) was delivered to the motor cortex to produce motor evoked potentials (MEPs), and electrical stimulation at the cervicomedullary junction was used to generate cervicomedullary motor evoked potentials (CMEPs) in the biceps brachii at rest and during a range of submaximal elbow flexions. Evoked potentials were also obtained after a conditioning TMS pulse to produce conditioned MEPs and CMEPs (100 ms inter-stimulus interval). 5-HT2 antagonism reduced maximal torque (p < 0.001), and compared to placebo, reduced unconditioned MEP amplitude at rest (p = 0.003), conditioned MEP amplitude at rest (p = 0.033) and conditioned MEP amplitude during contractions (p = 0.020). 5-HT2 antagonism also increased unconditioned CMEP amplitude during voluntary contractions (p = 0.041) but not at rest. Although 5-HT2 antagonism increased long-interval intracortical inhibition, net corticospinal excitability was unaffected during voluntary contractions. Given that spinal motoneurone excitability was only affected when descending drive to motoneurones was present, the current study indicates that excitatory drive is necessary for 5-HT2 receptors to regulate motoneurone excitability but not intracortical circuits.


Stress-induced dendritic remodeling in the prefrontal cortex is circuit specific.
Chronic stress exposure has been reported to induce dendritic remodeling in several brain regions, but it is not known whether individual neural circuits show distinct patterns of remodeling. The current study tested the hypothesis that the projections from the infralimbic (IL) area of the medial prefrontal cortex (mPFC) to the basolateral nucleus of the amygdala (BLA), a pathway relevant to stress-related mental illnesses like depression and post-traumatic stress disorder, would have a unique pattern of remodeling in response to chronic stress. The retrograde tracer FastBlue was injected into male rats' BLA or entorhinal cortex (EC) 1 week prior to 10 days of immobilization stress. After cessation of stress, FastBlue-labeled and unlabeled IL pyaramidal neurons were loaded with fluorescent dye Lucifer Yellow to visualize dendritic arborization and spine density. As has been previously reported, randomly selected (non-FastBlue-labeled) neurons showed stress-induced dendritic retraction in apical dendrites, an effect also seen in EC-projecting neurons. In contrast, BLA-projecting neurons showed no remodeling with stress, suggesting that this pathway may be particularly resilient against the effects of stress. No neurons showed stress-related changes in spine density, contrasting with reports that more dorsal areas of the mPFC show stress-induced decreases in spine density. Such region- and circuit-specificity in response to stress could contribute to the development of stress-related mental illnesses.


Use of a platelet-rich plasma-collagen scaffold as a bioenhanced repair treatment for management of partial cruciate ligament rupture in dogs.
Dogs are commonly affected with cruciate ligament rupture (CR) and associated osteoarthritis (OA), and frequently develop a second contralateral CR. Platelet rich plasma (PRP) is a component of whole blood that contains numerous growth factors, which in combination with a collagen scaffold may act to promote bioenhanced primary repair of ligament. This study tested the hypothesis that treatment of partial stable CR stifles with an intra-articular collagen scaffold and PRP would decrease the disease progression, synovitis and risk of complete CR over a 12-month study period. We conducted a prospective cohort study of 29 client-owned dogs with an unstable stifle due to complete CR and stable contralateral stifle with partial CR. All dogs were treated with tibial plateau leveling osteotomy (TPLO) on the unstable stifle and a single intra-articular application of PRP-collagen in the stable partial CR stifle. Dogs were evaluated at the time of diagnosis, and at 10-weeks and 12-months after treatment. We evaluated correlation between both development of complete CR and time to complete CR with diagnostic tests including bilateral stifle radiographs, 3.0 Tesla magnetic resonance (MR) imaging, and bilateral stifle arthroscopy. Additionally, histologic evaluation of synovial biopsies, C-reactive protein (CRP) concentrations in serum and synovial fluid, and synovial total nucleated cell count, were determined. Results indicated that a single application of PRP-collagen in partial CR stifles of client owned dogs is not an effective disease-modifying therapy for the prevention of progression to complete CR. Radiographic effusion, arthroscopic evaluation of cranial cruciate ligament (CrCL) damage, and MR assessment of ligament fiber tearing in partial CR stifles correlated with progression to complete CR over the 12-month follow-up period. We determined that the best predictive model for development of complete CR in PRP-collagen treated partial CR stifles included variables from multiple diagnostic modalities.


Pneumonia cluster in a boarding school--implications for influenza control.
Streptococcus pneumoniae is a common cause of community acquired pneumonia (CAP). Influenza infection increases susceptibility to S. pneumoniae infection in adults but this link is less well described in children. We report on an outbreak of CAP affecting 25 previously well adolescents in a New South Wales boarding school. S. pneumoniae 1 was confirmed in two cases. During this period, the school also experienced an influenza outbreak with an influenza-like illness attack rate peaking at 27% in Year 8 students. A planned school closure may have contributed to controlling the outbreak. Boarding schools are vulnerable to outbreaks of respiratory illness and strategies for limiting this risk are required.


Composite tissue allografting (CTA).
Composite tissue allografting (CTA) is a relatively new term that appears with increased frequency in the medical and surgical literature. The recent successful allografts, or homotransplantations, of hands has sparked a resurgence of research, both experimental and clinical. Plastic surgeons have long been interested--particularly in the field of skin grafts for extensive burns and wounds--but have been replaced by the organ transplant teams and immunogeneticists. The skin, one of the most strongly antigenic organs, has required such high levels of immunosuppressive drugs to date, that elective transplants of skin and other composite tissues have received little attention.


Integrin β7-mediated regulation of multiple myeloma cell adhesion, migration, and invasion.
Integrin-β7 (ITGB7) mRNA is detected in multiple myeloma (MM) cells and its presence is correlated with MAF gene activation. Although the involvement of several integrin family members in MM-stoma cell interaction is well documented, the specific biologic functions regulated by integrin-β7 in MM are largely unknown. Clinically, we have correlated integrin-β7 expression in MM with poor survival outcomes post autologous stem cell transplantation and postsalvage therapy with bortezomib. Functionally, we have found that shRNA-mediated silencing of ITGB7 reduces MM-cell adhesion to extra-cellular matrix elements (fibronectin, E-cadherin) and reverses cell-adhesion-mediated drug resistance (CAM-DR) sensitizing them to bortezomib and melphalan. In addition, ITGB7 silencing abrogated MM-cell transwell migration in response to SDF1α gradients, reduced vessel density in xenografted tumors, and altered MM cells in vivo homing into the BM. Mechanistically, ITGB7 knockdown inhibited focal adhesion kinase (FAK) and Src phosphorylation, Rac1 activation, and SUMOylation, reduced VEGF production in MM-BM stem cell cocultures and attenuated p65-NF-κB activity. Our findings support a role for integrin-β7 in MM-cell adhesion, migration, and BM homing, and pave the way for a novel therapeutic approach targeting this molecule.


Comparison of two intraosseous devices in adult patients in the emergency setting: a pilot study.
To compare two intraosseous (IO) insertion devices in terms of safety and ease of use in patients who need urgent vascular access in the emergency setting following failed attempts for intravenous lines. This prospective, randomized clinical study compared two different IO access devices in adults (≥16 years of age or weighing>40 kg) admitted to our emergency department with difficult peripheral venous access and in need of urgent intervention. The findings were compared in terms of the rates of successful insertion of spring-loaded impact-driven adult BIG and the battery-driven EZ-IO devices on the first attempt, difficulty of use, time taken to complete the insertion, and complication rates. A total of 52 patients were enrolled and randomized for the study. IO access was performed in 26 patients using the BIG device and in 26 patients using the EZ-IO device. The rates of successful insertion of BIG and EZ-IO devices on the first attempt were 92.3 and 84.6%, respectively (P=0.668). The time taken to complete the procedure was significantly greater in the EZ-IO group (5.2±2.2 s) compared with the BIG group (2.8±1.2 s) (P<0.001). Difficulty of use was scored using the visual analogue scale, which yielded the mean visual analogue scale for EZ-IO and BIG (25.4±12.6 and 8.6±6.4 mm, respectively) (P<0.001). No mechanical problems or technique-related complications were encountered in any of the groups. Both EZ-IO and BIG are shown to be reliable and safe methods for insertion of intravascular access in emergency conditions.


Role of SmcHD1 in establishment of epigenetic states required for the maintenance of the X-inactivated state in mice.
X inactivation in mammals is regulated by epigenetic modifications. Functional deficiency of SmcHD1 has been shown to cause de-repression of X-inactivated genes in post-implantation female mouse embryos, suggesting a role of SmcHD1 in the maintenance of X inactivation. Here, we show that de-repression of X-inactivated genes accompanied a local reduction in the enrichment of H3K27me3 in mouse embryonic fibroblasts deficient for SmcHD1. Furthermore, many of these genes overlapped with those having a significantly lower enrichment of H3K27me3 at the blastocyst stage in wild type. Intriguingly, however, depletion of SmcHD1 did not compromise the X-inactivated state in immortalized female mouse embryonic fibroblasts, in which X inactivation had been established and maintained. Taking all these findings together, we suggest that SmcHD1 facilitates the incorporation of H3K27me3 and perhaps other epigenetic modifications at gene loci that are silenced even with the lower enrichment of H3K27me3 at the early stage of X inactivation. The epigenetic state at these loci would, however, remain as it is at the blastocyst stage in the absence of SmcHD1 after implantation, which would eventually compromise the maintenance of the X-inactivated state at later stages.


Anti-cytokine targeted therapies for ANCA-associated vasculitis.
Anti-neutrophilic cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) are a group of rare auto-inflammatory diseases that affects mainly small vessels. AAV includes: granulomatosis with polyangiitis (GPA), microscopic polyangiitis (MPA) and eosinophilic granulomatosis with polyangiitis (EGPA). Anti-cytokine targeted therapy uses biological agents capable of specifically targeting and neutralising cytokine mediators of the inflammatory response. To assess the benefits and harms of anti-cytokine targeted therapy for adults with AAV. We searched the Cochrane Central Register of Controlled Trials (2019, Issue 7), MEDLINE and Embase up to 16 August 2019. We also examined reference lists of articles, clinical trial registries, websites of regulatory agencies and contacted manufacturers. Randomised controlled trials (RCTs) or controlled clinical trials of targeted anti-cytokine therapy in adults (18 years or older) with AAV compared with placebo, standard therapy or another modality and anti-cytokine therapy of different type or dose. We used standard methodological procedures expected by Cochrane. We included four RCTs with a total of 440 participants (mean age 48 to 56 years). We analysed the studies in three groups: 1) mepolizumab (300 mg; three separate injections every four weeks for 52 weeks) versus placebo in participants with relapsing or refractory EGPA; 2) belimumab (10 mg/kg on days 0, 14, 28 and every 28 days thereafter until 12 months after the last participant was randomised) or etanercept (25 mg twice a week) with standard therapy (median 25 months) versus placebo with standard therapy (median 19 months) in participants with GPA/MPA; and 3) infliximab (3 mg/kg on days 1 and 14, before the response assessment on day 42) versus rituximab (0.375g/m2 on days 1, 8, 15 and 22) in participants with refractory GPA for up to 12 months. None of the studies were assessed as low risk of bias in all domains: one study did not report randomisation or blinding methods clearly. Three studies were at high risk and one study was at unclear risk of bias for selective outcome reporting. One trial with 136 participants with relapsing or refractory EGPA compared mepolizumab with placebo during 52 weeks of follow-up and observed one death in the mepolizumab group (1/68, 1.5%) and none in the placebo group (0/68, 0%) (Peto odds ratio (OR) 7.39, 95% confidence interval (CI) 0.15 to 372.38; low-certainty evidence). Low-certainty evidence suggests that more participants in the mepolizumab group had ≥ 24 weeks of accrued remission over 52 weeks compared to placebo (27.9% versus 2.9%; risk ratio (RR) 9.5, 95% CI 2.30 to 39.21), and durable remission within the first 24 weeks sustained until week 52 (19.1% mepolizumab versus 1.5% placebo; RR 13.0, 95% CI 1.75 to 96.63; number needed to treat for an additional beneficial outcome (NNTB) 6, 95% Cl 4 to 13). Mepolizumab probably decreases risk of relapse (55.8% versus 82.4%; RR 0.68, 95% CI 0.53 to 0.86; NNTB 4, 95% CI 3 to 9; moderate-certainty evidence). There was low-certainty evidence regarding similar frequency of adverse events (AEs): total AEs (96.9% versus 94.1%; RR 1.03, 95% CI 0.96 to 1.11), serious AEs (17.7% versus 26.5%; RR 0.67, 95% CI 0.35 to 1.28) and withdrawals due to AEs (2.9% versus 1.5%; RR 2.00, 95% CI 0.19 to 21.54). Disease flares were not measured. Based on two trials with different follow-up periods (mean of 27 months for etanercept study; up to four years for belimumab study) including people with GPA (n = 263) and a small group of participants with MPA (n = 22) analysed together, we found low-certainty evidence suggesting that adding an active drug (etanercept or belimumab) to standard therapy does not increase or reduce mortality (3.4% versus 1.4%; Peto OR 2.45, 95% CI 0.55 to 10.97). Etanercept may have little or no effect on remission (92.3% versus 89.5%; RR 0.97, 95% CI 0.89 to 1.07), durable remission (70% versus 75.3%; RR 0.93, 95% CI 0.77 to 1.11; low-certainty evidence) and disease flares (56% versus 57.1%; RR 0.98, 95% CI 0.76 to 1.27; moderate-certainty evidence). Low-certainty evidence suggests that belimumab does not increase or reduce major relapse (1.9% versus 0%; RR 2.94, 95% CI 0.12 to 70.67) or any AE (92.5% versus 82.7%; RR 1.12, 95% CI 0.97 to 1.29). Low-certainty evidence suggests a similar frequency of serious or severe AEs (47.6% versus 47.6%; RR 1.00, 95% CI 0.80 to 1.27), but more frequent withdrawals due to AEs in the active drug group (11.2%) compared to the placebo group (4.2%), RR 2.66, 95% CI 1.07 to 6.59). One trial involving 17 participants with refractory GPA compared infliximab versus rituximab added to steroids and cytotoxic agents for 12 months. One participant died in each group (Peto OR 0.88, 95% CI, 0.05 to 15.51; 11% versus 12.5%). We have very low-certainty evidence for remission (22% versus 50%, RR 0.44, 95% Cl 0.11 to 1.81) and durable remission (11% versus 50%, RR 0.22, 95% CI 0.03 to 1.60), any severe AE (22.3% versus 12.5%; RR 1.78, 95% CI 0.2 to 16.1) and withdrawals due to AEs (0% versus 0%; RR 2.70, 95% CI 0.13 to 58.24). Disease flare/relapse and the frequency of any AE were not reported. We found four studies but concerns about risk of bias and small sample sizes preclude firm conclusions. We found moderate-certainty evidence that in patients with relapsing or refractory EGPA, mepolizumab compared to placebo probably decreases disease relapse and low-certainty evidence that mepolizumab may increase the probability of accruing at least 24 weeks of disease remission. There were similar frequencies of total and serious AEs in both groups, but the study was too small to reliably assess these outcomes. Mepolizumab may result in little to no difference in mortality. However, there were very few events. In participants with GPA (and a small subgroup of participants with MPA), etanercept or belimumab may increase the probability of withdrawal due to AEs and may have little to no impact on serious AEs. Etanercept may have little or no impact on durable remission and probably does not reduce disease flare.


Clinical physiology of dopa dyskinesia.
Levodopa-induced dyskinesias are clinically heterogeneous, both in appearance and timing with respect to dose. Electromyogram observations indicate that levodopa-induced dyskinesias are comprised of irregular bursts of either synchronous or asynchronous neuronal firing in antagonist muscles. Studies of the blink reflex and spontaneous blinking have provided useful neurophysiologic information on brainstem function that is sensitive to changes in brain dopamine concentrations. The blink rate is reduced in Parkinson's disease (PD) and increased with dopamine treatment. The blink rate in patients with levodopa-induced dyskinesias, however, has been shown to be faster than that in optimally treated PD patients and normal individuals. These results suggest that dyskinesias are associated with a relative hyperdopaminergic state. However, there appears to be no correlation of dopaminergic benefit to the parkinsonian symptoms, indicating perhaps that there are several dopaminergic systems, including one responsible for motor function and one for dyskinesia. Alternatively, it may be that the pattern of neural firing influences dyskinesias, while the average firing rate may be responsible for motor benefits.


Assembly of Metal-Phenolic Networks onto Microbubbles for One-Step Generation of Functional Microcapsules.
The one-step assembly of metal-phenolic networks (MPNs) onto particle templates can enable the facile, rapid, and robust construction of hollow microcapsules. However, the required template removal step may affect the refilling of functional species in the hollow interior space or the in situ encapsulation of guest molecules during the formation of the shells. Herein, a simple strategy for the one-step generation of functional MPNs microcapsules is proposed. This method uses bovine serum albumin microbubbles (BSA MBs) as soft templates and carriers, enabling the efficient pre-encapsulation of guest species by leveraging the coordination assembly of tannic acid (TA) and FeIII ions. The addition of TA and FeIII induces a change in the protein conformation of BSA MBs and produces semipermeable capsule shells, which allow gas to escape from the MBs without template removal. The MBs-templated strategy can produce highly biocompatible capsules with controllable structure and size, and it is applicable to produce other MPNs systems like BSA-TA-CuII and BSA-TA-NiII . Finally, those MBs-templated MPNs capsules can be further functionalized or modified for the loading of magnetic nanoparticles and the pre-encapsulation of model molecules through covalence or physical adsorption, exhibiting great promise in biomedical applications.


Nanovalve activation by surface-attached photoacids.
Proton transfer caused by excitation of a photoacid attached to the surface of a mesoporous silica nanoparticle activates a nanovalve and causes release of trapped molecules. The protonation of an aniline-based stalk releases a noncovalently bound cyclodextrin molecule that blocked a pore. The results show that pH-responsive molecular delivery systems can be externally controlled using light.


Critical elements of pediatric liver cancer surgery.
The appropriate management of pediatric liver malignancies, primarily hepatoblastoma and hepatocellular carcinoma, requires an in depth understanding of contemporary preoperative risk stratification, experience with advanced hepatobiliary surgery, and a good relationship with one's local or regional liver transplant center. While chemotherapy regimens have become more effective, operative indications more well-defined, and overall survival improved, the complexity of liver surgery in small children provides ample opportunity for protocol violation, inadequate resection, and iatrogenic morbidity. These guidelines represent the distillation of contemporary literature and expert opinion as a means to provide a framework for preoperative planning and intraoperative decision-making for the pediatric surgeon.


A flow interruption device for measurement of airway resistance.
Unlike conventional methods, the interrupter method for measuring airway resistance is non-invasive and requires minimal patient co-operation. It can therefore be applied in critically ill patients, acute asthmatics, neonates, pre-school children, geriatric patients and unconscious patients. The method is based on transient interruption of airflow at the mouth for a brief period during which alveolar pressure equilibrates with mouth pressure. Measurement of mouth pressure is used to estimate alveolar pressure prior to interruption and the ratio of this to flow prior to interruption gives airway resistance. Using the interrupter method we have developed a portable device for measuring airway resistance which is simple to use and gives a direct instantaneous reading. Measurements of airway resistance obtained using the new device were compared with those obtained using conventional body plethysmograph methods in 43 adult patients. A close correlation was seen (r = 0.86). The two methods appear equally sensitive in detecting changes in airway resistance following bronchodilator therapy. The device has been used successfully in pre-school children unable to co-operate with conventional methods.


Synthetic, structural and biochemical studies of polynuclear platinum(II) complexes with heterocyclic ligands.
"Non-classical" di- and trinuclear Pt(II) complexes with polydentate nitrogen ligands; ionic [(PtCl(2))(2)(tptz)(2)(mu-PtClNCPh)]Cl (1) [tptz =2,4,6-tris(2-pyridyl)-1,3,5-triazine], [(PtCl(2))(2)(bptz)(2)(mu-Pt)]Cl(2) (2) [bptz = 3,6-bis(2-pyridyl)-1,2,4,5-tetrazine] and neutral [(PtCl(2))(2)(tptz)(2)(mu-PtCl(2))](H(2)O)(4) (3), [(PtCl(2))(2)(mu-tppz)](CHCl(3)) (4) [tppz = 2,3,5,6-tetra(2-pyridyl)pyrazine] complexes, have been prepared and structurally characterized. The neutral tptz and tppz complexes present three and two separate PtCl(2) moieties, respectively, in a cis position, presumably acting in a bifunctional mode towards DNA; the cationic tptz and bptz complexes contain monofunctional and bifunctional bridging Pt(II) moieties, respectively, (other Pt(II) moieties in the complexes are bifunctional). All complexes were tested for their biological activity. Both tptz complexes, neutral and ionic, show a potent cytotoxic activity and reduced cell viability in a concentration-dependent manner that was evaluated in a panel of different cancer cell lines: human HT29 colon-rectal carcinoma, HepG2 hepatoma, MDA-MB-231 breast cancer and MG63 osteosarcoma cells; their activity was higher than cisplatin, IC50 values have been calculated for the active compounds and flow cytometric analysis for the tptz complexes performed. Therefore, these new platinum drugs warrant further investigation into their antitumor activity against different types of tumors.


Excitatory amino acid induced oligodendrocyte cell death in vitro: receptor-dependent and -independent mechanisms.
Oligodendroglia play an important role in axonal conduction in the CNS and are sensitive to oxidative toxicity induced by glutamate in the absence of ionotropic glutamate receptors. In this study, oligodendrocyte signalling cascades were examined, in response to glutamate-induced oxidative injury and to excitotoxicity. Rat cortical oligodendrocytes, differentiated in culture, were highly vulnerable to glutamate-induced cell death. Competitive inhibition of cystine uptake and increased oxidative stress appeared responsible for this death, and caused an accumulation of intracellular peroxides as well as chromatin fragmentation and condensation. Glutamate receptor subtype agonists (quisqualate, ibotenate) known to inhibit cystine uptake were cytotoxic, but not NMDA itself; moreover, glutamate receptor antagonists were not protective. Oligodendrocytes were also vulnerable to overactivation of glutamate receptors, as kainic acid and AMPA proved to be toxic. AMPA toxicity required the presence of cyclothiazide, suggesting rapid desensitization of AMPA receptors. Glutamate-induced oxidative stress and kainate/AMPA receptor stimulation activated the mitogen-activated protein kinase (MAP kinase) pathway, as well as the transcription factor ELK. However, MAP kinase kinase inhibitors only protected against injury from glutamate-induced oxidative stress. Oligodendrocytes were sensitive to oxygen-glucose deprivation injury as well, in a MAP kinase dependent fashion. Glutamate toxicity may conceivably be operative in neuropathological conditions that disrupt neuronal/oligodendrocyte interactions in axons, e.g. multiple sclerosis and ischaemia-reperfusion injury.


Genetic analysis of mutations at the Glued locus and interacting loci in Drosophila melanogaster.
A genetic analysis of the dominant mutation Glued that perturbs the development of the normal axonal architecture of the fly's visual system was undertaken. Ten new alleles at this locus were identified and characterized. Two complementation groups that were identified failed to complement the original allele, suggesting that it is a double mutant or that it resides at a complex locus. Several of the new alleles display visual-system abnormalities similar to those of the original mutation. Seven of the eight members of one complementation group are embryonic/early larval lethals, like the original mutation. The other allele in this group is temperature sensitive. Homozygous mutant adults exhibit a temperature-sensitive female sterile phenotype. Unsuccessful attempts to recover genetic mosaics carrying clones of cells homozygous for some of these mutations revealed that they are either essential for the viability of individual cells or that they affect some other fundamental cellular function, such as mitosis or the ability to participate in tissue level organization, which prevents them from being recovered in adult mosaics. This also indicates that these mutations do not specifically affect neural cells. A number of X-ray- and EMS-induced partial and complete phenotypic "revertants" of the original allele have also been isolated as material for comparative analysis of visual system development. All "revertants" that alter the abnormal eye phenotype towards the wild type have similar impact on the organization of the optic lobe.


Effect of enhanced detailing and mass media on community use of oral rehydration salts and zinc during a scale-up program in Gujarat and Uttar Pradesh.
The Clinton Health Access Initiative implemented a program from 2012-2016 to increase use of oral rehydration salts (ORS) and zinc to treat diarrhea in children under five in three states in India: Gujarat, Madhya Pradesh, and Uttar Pradesh. The program interventions included detailing and development of a rural supply chain to reach private rural health care providers, training of Accredited Social Health Activists (ASHAs), technical support to the state governments, and a mass media campaign targeted at caregivers. In Gujarat and Uttar Pradesh, some of the program activities, such as detailing and ASHA trainings, were targeted to high-burden focal districts, thus providing an opportunity to study their effect compared to statewide activities that covered all districts, such as the mass media campaign. Our study aimed to estimate the effect of activities on ORS and zinc use. Household surveys were conducted at two points during the program and in both focal and non-focal districts. We used a difference-in-difference quasi-experimental approach to estimate the effect of the enhanced activities in focal districts and mass media campaign on the odds of a child being treated with ORS and zinc. Focal district interventions were associated with a significant increase in the odds of a diarrhea episode receiving ORS in Gujarat and Uttar Pradesh. Living in focal districts increased the odds of receiving ORS in Gujarat and Uttar Pradesh by factors of 3.42 (95% CI = 1.39-8.33) and 2.29 (95% CI = 1.19-4.39), respectively. Focal district interventions were also associated with 15.02 (95% CI = 2.97-75.19) greater odds of receiving both ORS and zinc in Gujarat. In Uttar Pradesh, where the mass media campaign was focused, exposure to the campaign further modified the odds of receiving ORS and combined ORS and zinc by 1.38 (95% CI = 1.04-1.84) and 1.57 (95% CI = 1.01-2.46), respectively. Comprehensive public and private provider interventions combined with mass media are effective strategies for increasing ORS and zinc use.


Metagenomic analyses of bacterial endophytes associated with the phyllosphere of a Bt maize cultivar and its isogenic parental line from South Africa.
Genetic modification of maize with Bacillus thuringiensis (Bt) cry proteins may predispose shifts in the bacterial endophytes' community associated with maize shoots. In this study, the diversity of bacterial endophytes associated with a Bt maize genotype (Mon810) and its isogenic non-transgenic parental line were investigated at pre-flowering (50 days) and post-flowering (90 days) developmental stages. PCR-DGGE and high throughput sequencing on the Illumina MiSeq sequencer were used to characterize bacterial 16S rRNA gene diversity in leaves, stems, seeds and tassels. PCR-DGGE profile revealed similarity as well as differences between bacterial communities of shoots in both cultivars and at both developmental stages. A total of 1771 operational taxonomic units (OTUs) were obtained from the MiSeq and assigned into 14 phyla, 27 classes, 58 orders, 116 families and 247 genera. Differences in alpha and beta diversity measures of OTUs between the phyllospheres of both genotypes were not significant (P > .05) at all developmental stages. In all cultivars, OTU diversity reduced with plant development. OTUs belonging to the phyla Proteobacteria were dominant in all maize phyllospheres. The class Gammaproteobacteria was dominant in Bt maize while, Alphaproteobacteria and Actinobacteria were dominant in non-Bt maize phyllospheres. Differences in the abundance of some genera, including Acidovorax, Burkerholderia, Brachybacterium, Enterobacter and Rhodococcus, whose species are known beneficial endophytes were observed between cultivars. Hierarchical cluster analysis further suggests that the bacterial endophyte communities of both maize genotypes associate differently (are dissimilar). Overall, the results suggest that bacterial endophytes community differed more across developmental stages than between maize genotypes.


The Amagugu Intervention: A Conceptual Framework for Increasing HIV Disclosure and Parent-Led Communication about Health among HIV-Infected Parents with HIV-Uninfected Primary School-Aged Children.
Advances in access to HIV prevention and treatment have reduced vertical transmission of HIV, with most children born to HIV-infected parents being HIV-uninfected themselves. A major challenge that HIV-infected parents face is disclosure of their HIV status to their predominantly HIV-uninfected children. Their children enter middle childhood and early adolescence facing many challenges associated with parental illness and hospitalization, often exacerbated by stigma and a lack of access to health education and support. Increasingly, evidence suggests that primary school-aged children have the developmental capacity to grasp concepts of health and illness, including HIV, and that in the absence of parent-led communication and education about these issues, HIV-exposed children may be at increased risk of psychological and social problems. The Amagugu intervention is a six-session home-based intervention, delivered by lay counselors, which aims to increase parenting capacity to disclose their HIV status and offer health education to their primary school-aged children. The intervention includes information and activities on disclosure, health care engagement, and custody planning. An uncontrolled pre-post-evaluation study with 281 families showed that the intervention was feasible, acceptable, and effective in increasing maternal disclosure. The aim of this paper is to describe the conceptual model of the Amagugu intervention, as developed post-evaluation, showing the proposed pathways of risk that Amagugu aims to disrupt through its intervention targets, mechanisms, and activities; and to present a summary of results from the large-scale evaluation study of Amagugu to demonstrate the acceptability and feasibility of the intervention model. This relatively low-intensity home-based intervention led to: increased HIV disclosure to children, improvements in mental health for mother and child, and improved health care engagement and custody planning for the child. The intervention model demonstrates the potential for disclosure interventions to include pre-adolescent HIV education and prevention for primary school-aged children.


A human laboratory study of the effects of quetiapine on subjective intoxication and alcohol craving.
The available treatments for alcoholism are only modestly effective, and patients vary widely in their treatment response. Quetiapine, an atypical antipsychotic medication with antagonist activity at D1 and D2, 5-HT(1A) and 5-HT(2A), H(1), and α1 and α2 receptors was shown to promote abstinence, reduce drinking days, and reduce heavy drinking days in a 12-week double-blind placebo-controlled trial. Although quetiapine represents one of the promising pharmacotherapies for the treatment of alcoholism, its mechanisms of action are poorly understood. The objective of this study is to elucidate the biobehavioral mechanisms of action of quetiapine for alcoholism, by examining its effects on subjective intoxication and craving. A total of 20 non-treatment-seeking alcohol-dependent individuals were randomized to one of the following conditions in a double-blind, placebo-controlled design: (1) quetiapine (400 mg/day); or (2) matched placebo. Participants were on the target medication dose (or matched placebo) for 4 weeks during which they completed weekly assessments of drinking, sleep, mood, and anxiety. Participants completed two counterbalanced intravenous placebo-alcohol administration sessions as well as cue-reactivity assessments. Analyses revealed a significant effect of quetiapine in reducing craving during the alcohol administration, the alcohol cue-exposure, and the weekly reports of alcohol craving. Quetiapine was also found to reduce subjective intoxication and alcohol-induced sedation during the alcohol administration paradigm. This study contributes critical new information about mechanisms of response to quetiapine for alcoholism, which, in turn, can inform larger-scale studies and ultimately, clinical practice.


Insertions of a novel class of transposable elements with a strong target site preference at the r locus of maize.
The r locus of maize regulates anthocyanin synthesis in various tissues of maize through the production of helix-loop-helix DNA binding proteins capable of inducing expression of structural genes in the anthocyanin biosynthetic pathway. The complex r variant, R-r: standard (R.r), undergoes frequent mutation through a variety of mechanisms including displaced synapsis and crossing over, and intrachromosomal recombination. Here we report a new mechanism for mutation at the R-r complex: insertion of a novel family of transposable elements. Because the elements were first identified in the R-p gene of the R-r complex, they have been named P instability Factor (PIF). Two different PIF elements were cloned and found to have identical sequences at their termini but divergent internal sequences. In addition, the PIF elements showed a marked specificity of insertion sites. Six out of seven PIF-containing derivatives examined had an element inserted at an identical location. Two different members of the PIF element family were identified at this position. The seventh PIF-containing derivative examined had the element inserted at a distinct position within r. Even at this location, however, the element inserted into a conserved target sequence. The timing of PIF excision is unusual. Germinal excision rates can range up to several percent of progeny. Yet somatic sectors are rare, even in lines exhibiting high germinal reversion rates.


Electrochemical consideration on the optimum pH of bilirubin oxidase.
Steady-state current-potential curves were obtained for the direct electron transfer (DET) of bilirubin oxidase (BOD) at a highly oriented pyrolytic graphite electrode, and the theoretical analysis based on nonlinear regression enabled us to determine the formal redox potential (E degrees') of BOD in a wide pH range of 2.0 to 8.5. Cyclic voltammetric measurements were also performed for substrates, including p-phenylenediamine (PPD), o-aminophenol (OAP), and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and their E degrees ' values or the anodic peak potentials (for OAP) were determined at various pH values. The difference in the redox potentials between BOD and substrates (DeltaE degrees') showed a maximum at pH 6.5 to 8.0, pH 6.5 to 8.0, and pH 3.5 to 4.5 for PPD, OAP, and ABTS, respectively. These pH ranges should be thermodynamically most favorable for the electron transfer between BOD and the respective substrates. In practice, the pH ranges showing a maximum DeltaE degrees' corresponded well with the optimum pH values for the O(2) reduction activity of BOD: pH 6.5 to 7.5, pH 8.0 to 8.5, and pH 4.0 for PPD, OAP, and ABTS, respectively. Thus, it was suggested that DeltaE degrees ' should be one of the primary factors determining the activity of BOD with the substrates.


Coordinator-based systems for secondary prevention in fragility fracture patients.
The underlying causes of incident fractures--bone fragility and the tendency to fall--remain under-diagnosed and under-treated. This care gap in secondary prevention must be addressed to minimise both the debilitating consequences of subsequent fractures for patients and the associated economic burden to healthcare systems. Clinical systems aimed at ensuring appropriate management of patients following fracture have been developed around the world. A systematic review of the literature showed that 65% of systems reported include a dedicated coordinator who acts as the link between the orthopaedic team, the osteoporosis and falls services, the patient and the primary care physician. Coordinator-based systems facilitate bone mineral density testing, osteoporosis education and care in patients following a fragility fracture and have been shown to be cost-saving. Other success factors included a fracture registry and a database to monitor the care provided to the fracture patient. Implementation of such a system requires an audit of existing arrangements, creation of a network of healthcare professionals with clearly defined roles and the identification of a 'medical champion' to lead the project. A business case is needed to acquire the necessary funding. Incremental, achievable targets should be identified. Clinical pathways should be supported by evidence-based recommendations from national or regional guidelines. Endorsement of the proposed model within national healthcare policies and advocacy programmes can achieve alignment of the objectives of policy makers, professionals and patients. Successful transformation of care relies upon consensus amongst all participants in the multi-disciplinary team that cares for fragility fracture patients.


An Approach to the Evaluation of Incidentally Identified Bone Lesions Encountered on Imaging Studies.
This article discusses the strengths and weaknesses of the various anatomic and molecular imaging techniques in the evaluation of unexpected bone lesions. An approach to the imaging evaluation of chondroid, osteoblastic, and osteolytic lesions as well as focal marrow abnormalities is reviewed.


Optimal conduit for diabetic patients: propensity analysis of radial and right internal thoracic arteries.
Multiple arterial grafts, in addition to the left internal thoracic artery, improve long-term survival after coronary artery bypass grafting (CABG); yet, the use of this procedure remains low for both the right internal thoracic artery (RITA) and the radial artery (RA). To identify the optimal arterial conduit to deploy for revascularization of diabetic patients, we compared the outcomes for RA and RITA grafts to the circumflex coronary. From January 1, 1995, to December 31, 2011, 908 consecutive diabetic patients underwent first-time, isolated CABG (99% on-pump), 659 with the RA and 502 with the RITA, respectively, in two affiliated hospitals. Data were prospectively collected, and late mortality was determined from the Social Security Death Index. Propensity matching, based on preoperative and operative variables, identified 202 matched pairs from each group. Long-term survival was similar for matched patients. Mortality, myocardial infarction, reoperation for bleeding, stroke, sepsis, and renal failure were not significantly different between groups. However, deep sternal wound infection (p<0.035) and respiratory failure (p<0.048) favored the RA group, in which the total major adverse events were significantly fewer (p=0.002). In diabetic patients undergoing multivessel revascularization with either RA or RITA grafts to the circumflex coronary, long-term survival is similar. However, RA patients experienced significantly fewer respiratory or sternal wound adverse events. The RA is the preferred conduit to extend to more diabetic patients the recognized survival benefit of a multiple arterial graft strategy.


Employing RNA viruses to fight cancer: novel insights into oncolytic virotherapy.
Within recent decades, viruses that specifically target tumor cells have emerged as novel therapeutic agents against cancer. These viruses do not only act via their cell-lytic properties, but also harbor immunostimulatory features to re-direct the tumor microenvironment and stimulate tumor-directed immune responses. Furthermore, oncolytic viruses are considered to be superior to classical cancer therapies due to higher selectivity towards tumor cell destruction and, consequently, less collateral damage of non-transformed healthy tissue. In particular, the field of oncolytic RNA viruses is rapidly developing since these agents possess alternative tumor-targeting strategies compared to established oncolytic DNA viruses. Thus, oncolytic RNA viruses have broadened the field of virotherapy facilitating new strategies to fight cancer. In addition to several naturally occurring oncolytic viruses, genetically modified RNA viruses that are armed to express foreign factors such as immunostimulatory molecules have been successfully tested in early clinical trials showing promising efficacy. This review aims to provide an overview of the most promising RNA viruses in clinical development, to summarize the current knowledge of clinical trials using these viral agents, and to discuss the main issues as well as future perspectives of clinical approaches using oncolytic RNA viruses.


Lovastatin induces fibroblast apoptosis in vitro and in vivo. A possible therapy for fibroproliferative disorders.
Diseases associated with pathological fibroproliferation represent a major cause of morbidity and mortality. Despite the importance of this class of disorders, current therapy is of limited value, and no therapy is available to reduce the fibroblast population size within existing fibrotic lesions. In this regard, constitutive expression of growth-promoting genes can sensitize cells to undergo apoptosis. Studies in our laboratory have demonstrated that lovastatin potently induces apoptosis in fibroblasts constitutively expressing Myc, and that lung fibroblasts isolated from fibrotic lesions constitutively express growth-promoting genes. In this study, we sought to determine if nontransformed lung fibroblasts would manifest susceptibility to lovastatin-induced apoptosis similar to that observed in fibroblasts ectopically expressing Myc. Here we show that clinically achievable concentrations of lovastatin induce apoptosis in normal and fibrotic lung fibroblasts in vitro, as evidenced by acridine orange staining, terminal transferase nick end translation (TUNEL), and DNA laddering. Apoptosis of human lung fibroblasts was dose- and time-dependent, and blocked by exogenous mevalonic acid. Furthermore, apoptosis was associated with decreased levels of mature Ras, a molecule directly implicated in fibroblast rescue from apoptosis. The ability of lovastatin to induce fibroblast apoptosis in vivo was examined using a guinea pig wound chamber model. Lovastatin (5 microM, 8 d) reduced granulation tissue formation in the wound chambers by 64.7%, with associated ultrastructural evidence of fibroblast apoptosis. These findings support further study of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors as potential therapy for patients with fibroproliferative disorders.


Blocking of ERK1 and ERK2 sensitizes human mesothelioma cells to doxorubicin.
Malignant mesotheliomas (MM) have a poor prognosis, largely because of their chemoresistance to anti-cancer drugs such as doxorubicin (Dox). Here we show using human MM lines that Dox activates extracellular signal-regulated kinases (ERK1 and 2), causally linked to increased expression of ABC transporter genes, decreased accumulation of Dox, and enhanced MM growth. Using the MEK1/2 inhibitor, U0126 and stably transfected shERK1 and shERK2 MM cell lines, we show that inhibition of both ERK1 and 2 sensitizes MM cells to Dox. U0126 significantly modulated endogenous expression of several important drug resistance (BCL2, ABCB1, ABCC3), prosurvival (BCL2), DNA repair (BRCA1, BRCA2), hormone receptor (AR, ESR2, PPARγ) and drug metabolism (CYP3A4) genes newly identified in MM cells. In comparison to shControl lines, MM cell lines stably transfected with shERK1 or shERK2 exhibited significant increases in intracellular accumulation of Dox and decreases in cell viability. Affymetrix microarray analysis on stable shERK1 and shERK2 MM lines showed more than 2-fold inhibition (p ≤ 0.05) of expression of ATP binding cassette genes (ABCG1, ABCA5, ABCA2, MDR/TAP, ABCA1, ABCA8, ABCC2) in comparison to shControl lines. Moreover, injection of human MM lines into SCID mice showed that stable shERK1 or shERK2 lines had significantly slower tumor growth rates in comparison to shControl lines after Dox treatment. These studies suggest that blocking ERK1 and 2, which play critical roles in multi-drug resistance and survival, may be beneficial in combination with chemotherapeutic drugs in the treatment of MMs and other tumors.


PTP1B is a negative regulator of interleukin 4-induced STAT6 signaling.
Protein tyrosine phosphatase 1B (PTP1B) is a ubiquitously expressed enzyme shown to negatively regulate multiple tyrosine phosphorylation-dependent signaling pathways. PTP1B can modulate cytokine signaling pathways by dephosphorylating JAK2, TYK2, and STAT5a/b. Herein, we report that phosphorylated STAT6 may serve as a cytoplasmic substrate for PTP1B. Overexpression of PTP1B led to STAT6 dephosphorylation and the suppression of STAT6 transcriptional activity, whereas PTP1B knockdown or deficiency augmented IL-4-induced STAT6 signaling. Pretreatment of these cells with the PTK inhibitor staurosporine led to sustained STAT6 phosphorylation consistent with STAT6 serving as a direct substrate of PTP1B. Furthermore, PTP1B-D181A "substrate-trapping" mutants formed stable complexes with phosphorylated STAT6 in a cellular context and endogenous PTP1B and STAT6 interacted in an interleukin 4 (IL-4)-inducible manner. We delineate a new negative regulatory loop of IL-4-JAK-STAT6 signaling. We demonstrate that IL-4 induces PTP1B mRNA expression in a phosphatidylinositol 3-kinase-dependent manner and enhances PTP1B protein stability to suppress IL-4-induced STAT6 signaling. Finally, we show that PTP1B expression may be preferentially elevated in activated B cell-like diffuse large B-cell lymphomas. These observations identify a novel regulatory loop for the regulation of IL-4-induced STAT6 signaling that may have important implications in both neoplastic and inflammatory processes.


Estrogen metabolism and the malignant potential of human papillomavirus immortalized keratinocytes.
Increased 16alpha-hydroxylation of estradiol has been shown to be associated with heightened cancer risk in estrogen responsive tissue. Certain types of human papillomavirus (HPV) are cofactors for cancer in the cervix, an estrogen sensitive tissue. We have demonstrated that estradiol and 16alpha-hydroxyestrone increased the number of cells positive for proliferating cell nuclear antigen in HPV immortalized keratinocytes, the in vitro correlate of the premalignant keratinocyte. These estrogens caused the abnormal proliferation and anchorage independent growth, which correlates with malignant conversion. Indole-3-carbinol, a phytochemical in cruciferous vegetables known to preferentially induce 2-hydroxylation with minimal effect on 16alpha-hydroxylation, markedly blocked the ability of estradiol to increase anchorage independent growth. The results indicate that 16alpha-hydroxyestrone increases the malignant phenotype of HPV immortalized keratinocytes. However, indole-3-carbinol will block this response.


Factors affecting interspecific differences in genetic divergence among populations of Anolis lizards in Cuba.
Geographical patterns and degrees of genetic divergence among populations differ between species, reflecting relative potentials for speciation or cladogenesis and differing capacities for environmental adaptation. Identification of factors that contribute to genetic divergence among populations is important to the understanding of why some species exhibit greater interpopulation genetic divergence. In this study, we calculated the mean pairwise genetic distances among populations as species' average genetic divergence by a phylogeny using nuclear and mitochondrial genes of 303 individuals from 33 Cuban Anolis species and estimated species ages by another phylogeny using nuclear and mitochondrial genes of 51 Cuban and 47 non-Cuban Anolis species. We identified factors that influence species' differences in genetic divergence among 26 species of Anolis lizards from Cuba. Species ages, environmental heterogeneity within species ranges, and ecomorph types were considered as factors affecting average genetic divergences among populations. The phylogenies presented in this study provide the most comprehensive sampling of Cuban Anolis species to date. The phylogeny showed more conservative evolution of Anolis ecomorphs within Cuba and identified twig anoles as a monophyletic group. Subsequent Phylogenetic Generalized Least Squares (PGLS) analyses showed that species age was positively correlated with species' average genetic divergence among populations. Although previous studies have focused on factors affecting genetic divergence within species, the present study showed for the first time that species differences in genetic divergence could be largely affected by species age.


Neurocognitive development in 5- to 16-year-old North American children: a cross-sectional study.
Executive functions are thought to be the latest functions to mature. However, this view has not been tested by assessing simultaneously memory, perception of emotions, visuospatial perception, and visuoconstructional skills. NEPSY II norm data from 1000 5- to 16-year-old U.S. children were obtained. Fifteen NEPSY II subtests with no floor or ceiling effects in any age group and no major changes in task type were selected. The 16-year level was attained at age 12 to 13 in all subtests with two exceptions: social perception (age 10 to 11) and narrative memory (age 14). Trend analyses showed that development was rapid in the age range 5 to 9 years followed by a deceleration in the rate of development. Peak performances were reached at 14 to 16 years but later in some subtests representing executive functions, verbal memory, and visuospatial performance. Thus, the study specified developmental time tables of neurocognitive functions. It demonstrated that not only executive functions but also verbal memory and visuospatial performance continue to develop beyond age 16.


Interleukin-29 binds to melanoma cells inducing Jak-STAT signal transduction and apoptosis.
Interleukin-29 (IL-29) is a member of the type III IFN family that has been shown to have antiviral activity and to inhibit cell growth. Melanoma cell lines were tested for expression of the IL-29 receptor (IL-29R) and their response to IL-29. Expression of IL-28R1 and IL-10R2, components of IL-29R, was evaluated using reverse transcription-PCR. A combination of immunoblot analysis and flow cytometry was used to evaluate IL-29-induced signal transduction. U133 Plus 2.0 Arrays and real-time PCR were used to evaluate gene expression. Apoptosis was measured using Annexin V/propridium iodide staining. In situ PCR for IL-29R was done on paraffin-embedded melanoma tumors. Both IL-28R1 and IL-10R2 were expressed on the A375, 1106 MEL, Hs294T, 18105 MEL, MEL 39, SK MEL 5, and F01 cell lines. Incubation of melanoma cell lines with IL-29 (10-1,000 ng/mL) led to phosphorylation of signal transducer and activator of transcription 1 (STAT1) and STAT2. Microarray analysis and quantitative reverse transcription-PCR showed a marked increase in transcripts of IFN-regulated genes after treatment with IL-29. In the F01 cell line, bortezomib-induced and temozolomide-induced apoptosis was synergistically enhanced following the addition of IL-29. In situ PCR revealed that IL-10R2 and IL-28R1 were present in six of eight primary human melanoma tumors but not in benign nevi specimens. In conclusion, IL-29 receptors are expressed on the surface of human melanoma cell lines and patient samples, and treatment of these cell lines with IL-29 leads to signaling via the Jak-STAT pathway, the transcription of a unique set of genes, and apoptosis.


MicroRNA in central nervous system trauma and degenerative disorders.
MicroRNAs (miRNAs) are a novel class of small noncoding RNAs that negatively regulate gene expression at the posttranscriptional level by binding to the 3'-untranslated region of target mRNAs leading to their translational inhibition or sometimes degradation. MiRNAs are predicted to control the activity of at least 20-30% of human protein-coding genes. Recent studies have demonstrated that miRNAs are highly expressed in the central nervous system (CNS) including the brain and spinal cord. Although we are currently in the initial stages of understanding how this novel class of gene regulators is involved in neurological biological functions, a growing body of exciting evidence suggests that miRNAs are important regulators of diverse biological processes such as cell differentiation, growth, proliferation, and apoptosis. Moreover, miRNAs are key modulators of both CNS development and plasticity. Some miRNAs have been implicated in several neurological disorders such as traumatic CNS injuries and neurodegenerative diseases. Recently, several studies suggested the possibility of miRNA involvement in neurodegeneration. Identifying the roles of miRNAs and their target genes and signaling pathways in neurological disorders will be critical for future research. miRNAs may represent a new layer of regulators for neurobiology and a novel class of therapeutic targets for neurological diseases.


[Diagnosis and treatment for Paget's breast cancer: state of the art].
Breast cancer is the most common cancer in women. According to WHO experts in the world each year there are revealed from 800,000 up to 1 million new cases of breast cancer. In the structure of malignant tumors morbidity among female population, in Russia in 2012 breast cancer ranked the first place (20.7%) and remained the leading malignant pathology in women. Paget's breast cancer is a rare form of breast cancer that occurs in the mouth of the excretory ducts of the nipple, which characterized by lesion of the nipple and large ducts, often with the formation of tumor in the breast. This rare abnormality occurs in 0.5-5% of all cases of breast cancer.


Next-generation sequence analysis of cancer xenograft models.
Next-generation sequencing (NGS) studies in cancer are limited by the amount, quality and purity of tissue samples. In this situation, primary xenografts have proven useful preclinical models. However, the presence of mouse-derived stromal cells represents a technical challenge to their use in NGS studies. We examined this problem in an established primary xenograft model of small cell lung cancer (SCLC), a malignancy often diagnosed from small biopsy or needle aspirate samples. Using an in silico strategy that assign reads according to species-of-origin, we prospectively compared NGS data from primary xenograft models with matched cell lines and with published datasets. We show here that low-coverage whole-genome analysis demonstrated remarkable concordance between published genome data and internal controls, despite the presence of mouse genomic DNA. Exome capture sequencing revealed that this enrichment procedure was highly species-specific, with less than 4% of reads aligning to the mouse genome. Human-specific expression profiling with RNA-Seq replicated array-based gene expression experiments, whereas mouse-specific transcript profiles correlated with published datasets from human cancer stroma. We conclude that primary xenografts represent a useful platform for complex NGS analysis in cancer research for tumours with limited sample resources, or those with prominent stromal cell populations.


Binding of two bis-bipyridine minor groove binders to a DNA template in the presence of Cu2+ ions.
Some diseases are associated with abnormally extended regions of triplet repeats. These repeating regions are an attractive target for both diagnostic and therapeutic goals. In an attempt to approach to this goal, we have focused on establishment of an allosteric binding mechanism, in which the binding of the ligand promotes the next ligand binding. In the previous study, we already reported that the ligand having the bipyridine unit for binding with Cu(2+) and the Hoechst33258 for binding to A(3)T(3) site displayed Cu(2+)- mediated assembly on the DNA with two A(3)T(3) sites. In this study, we synthesized the new ligands containing two bipyridine units attached to Hoechst33258 by different length linkers. It was expected that the bipyridine-Cu(2+) complexation would enhance assembly of a number of the ligand on the DNA sequence with repeating regions. UV spectroscopy has been used to demonstrate the binding of these ligands to a DNA template mediated by the complexation of Cu(2+) ions.


[Importance of a second opinion in breast surgical pathology and therapeutic implications].
to evaluate the agreement between histopathologic diagnoses of breast lesions made by general pathologists and by a specialist in breast pathology. a cohort retrospective study comparing histopathologic diagnoses of 329 cases of breast lesions received in consultation for a second opinion was carried out. The material received for consultation included slides (152 cases), paraffin blocks (59 cases) or slides and blocks (118 cases). Cases were reviewed and the original diagnoses and diagnoses from a specialist in breast pathology were compared. The main diagnoses, nuclear grade of ductal carcinoma in situ, and the histopathologic grade of invasive mammary carcinomas were evaluated. The kappa index and percentual concordance were used in the statistical analyses. a moderate agreement was observed between the original histopathologic diagnoses and the second opinion (kappa index=0.48; percentual concordance=59.9%). The diagnosis of malignancy was confirmed in 185/225 cases (82.2%) and diagnosis of benign lesions was confirmed in 89/104 cases (85.6%). The highest agreement was observed in the diagnosis of invasive mammary carcinomas (81%) and the highest disagreement was observed among diagnoses of ductal carcinoma in situ with microinvasion (74%), lobular carcinoma in situ (70%), and atypical epithelial hyperplasias (61%). There was a moderate agreement in the nuclear grade of ductal carcinoma in situ (kappa index=0.52; percentual concordance=68.8%), and good concordance in the histologic grade of invasive carcinomas (kappa index=0.61; percentual concordance=74.3). the results show higher concordance rate in the diagnosis of invasive carcinomas and lower concordance in the diagnosis of ductal carcinoma in situ with microinvasion and premalignant breast lesions, especially lobular neoplasia in situ, and atypical epithelial hyperplasias.