Hypothermia combined with neuroprotective adjuvants shortens the duration of hospitalization in infants with hypoxic ischemic encephalopathy: Meta-analysis.
Objective: Therapeutic hypothermia (TH) is the current standard of care for neonatal hypoxic-ischemic encephalopathy (HIE), yet morbidity and mortality remain significant. Adjuvant neuroprotective agents have been suggested to augment hypothermic-mediated neuroprotection. This analysis aims to identify the classes of drugs that have been used in combination with hypothermia in the treatment of neonatal HIE and determine whether combination therapy is more efficacious than TH alone. Methods: A systematic search of PubMed, Embase and Medline from conception through December 2022 was conducted. Randomized- and quasi-randomized controlled trials, observational studies and retrospective studies evaluating HIE infants treated with combination therapy versus TH alone were selected. Primary reviewers extracted information on mortality, neurodevelopmental impairment and length of hospitalization for meta-analyses. Effect sizes were pooled using a random-effects model and measured as odds ratio (OR) or mean difference (MD) where applicable, and 95% confidence intervals (CI) were calculated. Risk of bias was assessed using the tool from the Cochrane Handbook for Systematic Reviews of Interventions. Results: The search strategy collected 519 studies, 16 of which met analysis inclusion criteria. HIE infants totaled 1,288 infants from included studies, 646 infants received some form of combination therapy, while 642 received TH alone. GABA receptor agonists, NMDA receptor antagonists, neurogenic and angiogenic agents, stem cells, glucocorticoids and antioxidants were identified as candidate adjuvants to TH that have been evaluated in clinical settings compared to TH alone. Length of hospitalization was significantly reduced in infants treated with combination therapy (MD -4.81, 95% CI [-8.42. to -1.19], p = .009) compared to those treated with TH alone. Risk of mortality and neurodevelopmental impairment did not differ between combination therapy and TH alone groups. Conclusion: Compared to the current standard of care, administration of neuroprotective adjuvants with TH reduced the duration of hospitalization but did not impact the risk of mortality or neurodevelopmental impairment in HIE infants. Meta-analysis was limited by a moderate risk of bias among included studies and small sample sizes. This analysis highlights the need for preclinical trials to conduct drug development studies in hypothermic settings to identify relevant molecular targets that may offer additive or synergistic neuroprotection to TH, and the need for larger powered clinical trials to determine the dose and timing of administration at which maximal clinical benefits are observed for adjuvant neuroprotectants.


Orexin neurons inhibit sleep to promote arousal.
Humans and animals lacking orexin neurons exhibit daytime sleepiness, sleep attacks, and state instability. While the circuit basis by which orexin neurons contribute to consolidated wakefulness remains unclear, existing models posit that orexin neurons provide their wake-stabilizing influence by exerting excitatory tone on other brain arousal nodes. Here we show using in vivo optogenetics, in vitro optogenetic-based circuit mapping, and single-cell transcriptomics that orexin neurons also contribute to arousal maintenance through indirect inhibition of sleep-promoting neurons of the ventrolateral preoptic nucleus. Activation of this subcortical circuit rapidly drives wakefulness from sleep by differentially modulating the activity of ventrolateral preoptic neurons. We further identify and characterize a feedforward circuit through which orexin (and co-released glutamate) acts to indirectly target and inhibit sleep-promoting ventrolateral preoptic neurons to produce arousal. This revealed circuitry provides an alternate framework for understanding how orexin neurons contribute to the maintenance of consolidated wakefulness and stabilize behavioral state.


Protein metabolism and related body function: mechanistic approaches and health consequences.
The development and maintenance of body composition and functions require an adequate protein intake with a continuous supply of amino acids (AA) to tissues. Body pool and AA cellular concentrations are tightly controlled and maintained through AA supply (dietary intake, recycled from proteolysis and de novo synthesis), AA disposal (protein synthesis and other AA-derived molecules) and AA losses (deamination and oxidation). Different molecular regulatory pathways are involved in the control of AA sufficiency including the mechanistic target of rapamycin complex 1, the general control non-derepressible 2/activating transcription factor 4 system or the fibroblast growth factor 21. There is a tight control of protein intake, and human subjects and animals appear capable of detecting and adapting food and protein intake and metabolism in face of foods or diets with different protein contents. A severely protein deficient diet induces lean body mass losses and ingestion of sufficient dietary energy and protein is a prerequisite for body protein synthesis and maintenance of muscle, bone and other lean tissues and functions. Maintaining adequate protein intake with age may help preserve muscle mass and strength but there is an ongoing debate as to the optimal protein intake in older adults. The protein synthesis response to protein intake can also be enhanced by prior completion of resistance exercise but this effect could be somewhat reduced in older compared to young individuals and gain in muscle mass and function due to exercise require regular training over an extended period.


Functional genomics uncovers three glucosyltransferases involved in the synthesis of the major sweet glucosides of Stevia rebaudiana.
Stevia rebaudiana leaves accumulate a mixture of at least eight different steviol glycosides. The pattern of glycosylation heavily influences the taste perception of these intensely sweet compounds. The majority of the glycosides are formed by four glucosylation reactions that start with steviol and end with rebaudioside A. The steps involve the addition of glucose to the C-13 hydroxyl of steviol, the transfer of glucose to the C-2' and C-3' of the 13-O-glucose and the addition of glucose to the hydroxyl of the C-4 carboxyl group. We used our collection of ESTs, an UDP-glucosyltransferase (UGT)-specific electronic probe and key word searches to identify candidate genes resident in our collection. Fifty-four expressed sequence tags (ESTs) belonging to 17 clusters were found using this procedure. We isolated full length cDNAs for 12 of the UGTs, cloned them into an expression vector, and produced recombinant enzymes in Escherichia coli. An in vitro glucosyltransferase activity enzyme assay was conducted using quercetin, kaempferol, steviol, steviolmonoside, steviolbioside, and stevioside as sugar acceptors, and (14)C-UDP-glucose as the donor. Thin layer chromatography was used to separate the products and three of the recombinant enzymes produced labelled products that co-migrated with known standards. HPLC and LC-ES/MS were then used to further define those reaction products. We determined that steviol UGTs behave in a regioselective manner and propose a modified pathway for the synthesis of rebaudioside A from steviol.


Synchronous colorectal liver metastases: the "surgeon as oncologist" perspective.
Colon cancer presenting with synchronous metastatic disease to the liver creates challenging clinical scenarios. Traditionally, patients with resectable synchronous colorectal liver metastases (CRLM) were treated initially by resection of the primary tumor followed by an arbitrary but often prolonged duration of postoperative chemotherapy. In patients without progression of disease while receiving this postoperative chemotherapy, surgery for CRLM was then offered. In recent years this algorithm has been questioned, and in this review we describe the rationale for this classic approach as well as data that suggests that this model should be reconsidered. Evolving definitions of resectability of CRLM as well as technical advances in liver surgery have increased the number of patients eligible for potentially curative resections. There still exists controversies regarding the role of chemotherapy, its timing, the feasibility of simultaneous CRLM and primary tumor resection, and how to treat the asymptomatic primary in the context of CRLM. While it is a valuable treatment in unresectable CRLM, the role of prehepatectomy chemotherapy in resectable CRLM should be reconsidered as its survival benefits have not been well established and there are reports of chemotherapy related morbidity following hepatic resection. Although simultaneous primary tumor and CRLM resection has been traditionally viewed as prohibitively morbid, recent data suggests that it may be safe in carefully selected patients with limited volume of metastatic disease. The traditional paradigm of treating synchronous CRLM (SCRLM) should be reconsidered. Randomized controlled trials are necessary to help clarify the role and timing of chemotherapy in the treatment of SCRLM.


Is MtnE, the fifth Drosophila metallothionein, functionally distinct from the other members of this polymorphic protein family?
Metallothioneins (MTs) are a super-family of small, Cys-rich, non-homologous proteins that bind metal ions through the formation of metal-thiolate bonds. Although universally ubiquitous, they exhibit distinct metal-binding preferences, either for divalent (Zn-thioneins) or monovalent (Cu-thioneins) metal ions. Drosophila constitutes a bizarre exception, since it is currently the only case of metazoans synthesizing only Cu-thioneins, which are similar to the paradigmatic yeast Cup1 protein. Until recently, the Drosophila MT system was assumed to be composed of 4 isoforms (MtnA, MtnB, MtnC and MtnD), all of them responsive to heavy metal load through the dMTF1 transcription factor. The significance of this polymorphism has been analyzed in depth both at genomic and proteomic levels. Singularly, a fifth MT isoform was recently annotated and named MtnE. The analysis of the MtnE expression pattern revealed some differential traits with regard to the other MTs. We analyze here the peculiarities of the metal binding abilities of the MtnE polypeptide and compare them with those of the other Drosophila MTs determined through the same rationale. Characterization by ESI-MS spectrometry and CD and UV-visible spectrophotometry of the Zn(II)-, Cd(II)- and Cu(I)-MtnE complexes obtained by recombinant synthesis demonstrates that MtnE is the least metal-specific isoform of the Drosophila MTs, and therefore it could play a role when/where a broad spectrum of metal coordination abilities are advantageous in terms of physiological needs.


Texas Children's Medication Algorithm Project: update from Texas Consensus Conference Panel on Medication Treatment of Childhood Major Depressive Disorder.
To revise and update consensus guidelines for medication treatment algorithms for childhood major depressive disorder based on new scientific evidence and expert clinical consensus when evidence is lacking. A consensus conference was held January 13-14, 2005, that included academic clinicians and researchers, practicing clinicians, administrators, consumers, and families. The focus was to review, update, and incorporate the most current data to inform and recommend specific pharmacological approaches and clinical guidance for treatment of major depressive disorder in children and adolescents. Consensually agreed on medication algorithms for major depression (with and without psychosis) and comorbid attention-deficit disorders were updated. These revised algorithms also incorporated approaches to address issues of suicidality, aggression, and irritability. Stages 1, 2, and 3 of the algorithm consist of selective serotonin reuptake inhibitor and norepinephrine serotonin reuptake inhibitor medications whose use is supported by controlled, acute clinical trials and clinical experience. Recent studies provide support that selective serotonin reuptake inhibitors in addition to fluoxetine are still encouraged as first-line interventions. The need for additional assessments, precautions, and monitoring is emphasized, as well as continuation and maintenance treatment. Evidence and expert clinical consensus support the use of selected antidepressants in the treatment of depression in youths. The use of the recommended antidepressant medications requires appropriate monitoring of suicidality and potential adverse effects and consideration of other evidence-based treatment alternatives such as cognitive behavioral therapies.


Trends in hip fracture rates in Canada.
Hip fractures are a public health concern because they are associated with significant morbidity, excess mortality, and the majority of the costs directly attributable to osteoporosis. To examine trends in hip fracture rates in Canada. Ecologic trend study using nationwide hospitalization data for 1985 to 2005 from a database at the Canadian Institute for Health Information. Data for all patients with a hospitalization for which the primary reason was a hip fracture (570,872 hospitalizations) were analyzed. Age-specific and age-standardized hip fracture rates. There was a decrease in age-specific hip fracture rates (all P for trend <.001). Over the 21-year period of the study, age-adjusted hip fracture rates decreased by 31.8% in females (from 118.6 to 80.9 fractures per 100,000 person-years) and by 25.0% in males (from 68.2 to 51.1 fractures per 100,000 person-years). Joinpoint regression analysis identified a change in the linear slope around 1996. For the overall population, the average age-adjusted annual percentage decrease in hip fracture rates was 1.2% (95% confidence interval, 1.0%-1.3%) per year from 1985 to 1996 and 2.4% (95% confidence interval, 2.1%-2.6%) per year from 1996 to 2005 (P < .001 for difference in slopes). Similar changes were seen in both females and males with greater slope reductions after 1996 (P < .001 for difference in slopes for each sex). Age-standardized rates of hip fracture have steadily declined in Canada since 1985 and more rapidly during the later study period. The factors primarily responsible for the earlier reduction in hip fractures are unknown.


Infections in hairy-cell leukemia.
In order to determine the nature of infectious complications in hairy-cell leukemia we studied 20 consecutive patients seen at UCLA and analyzed the available literature. The incidence of serious infection in our series was 40%, and pneumonia and septicemia due to Pseudomonas and E. coli organisms were the leading types of infections. Fungal infections with Cryptococci and Histoplasma organisms were documented, and a single case of Pneumocystis carinii pneumonia was observed. Noninfectious fever occurred in 30% of our patients. There was a clear relationship between fungal disease and corticosteroid therapy, and the overall incidence of infection was correlated with the degree of neutropenia and corticosteroid treatment. No relationship was found between age, duration of disease, or the use of cytotoxic chemotherapy and infectious complications. Of the 13 infectious episodes, 11 occurred in patients prior to splenectomy. Only two episodes were seen in splenectomized patients, both occurring in the immediate postoperative period. We conclude that splenectomy has a beneficial effect in reducing the incidence of infections in hairy-cell leukemia and that corticosteroids should be used cautiously, since they predispose to opportunistic infection in this disease.


Biodiesel from mixed culture algae via a wet lipid extraction procedure.
Microalgae are a source of renewable oil for liquid fuels. However, costs for dewatering/drying, extraction, and processing have limited commercial scale production of biodiesel from algal biomass. A wet lipid extraction procedure was developed that was capable of extracting 79% of transesterifiable lipids from wet algal biomass (84% moisture) via acid and base hydrolysis (90 °C and ambient pressures), and 76% of those extracted lipids were isolated, by further processing, and converted to FAMEs. Furthermore, the procedure was capable of removing chlorophyll contamination of the algal lipid extract through precipitation. In addition, the procedure generated side streams that serve as feedstocks for microbial conversion to additional bioproducts. The capability of the procedure to extract lipids from wet algal biomass, to reduce/remove chlorophyll contamination, to potentially reduce organic solvent demand, and to generate feedstocks for high-value bioproducts presents opportunities to reduce costs of scaling up algal lipid extraction for biodiesel production.


Female nutritional status determines the magnitude and sign of responses to a male ejaculate signal in Drosophila melanogaster.
Ejaculate chemicals transferred from males to females during mating cause significant changes in female behaviour and physiology, but the causes of phenotypic variation in these responses is little understood. We tested here the effect of adult female nutrition on the response of female Drosophila melanogaster to a specific ejaculate component, the sex peptide (SP), which is of interest because of its effects on female egg laying, sexual receptivity, feeding rate, immune responses and potential role in mediating sexual conflict. We exposed adult females to five different diets and kept them continuously with males that did or did not transfer SP. Diet altered the presence, magnitude and sign of the effects of SP on different phenotypic traits (egg laying, receptivity and lifespan) and different traits responded in different ways. This showed that the set of responses to mating can be uncoupled and can vary independently in different environments. Importantly, diet also significantly affected whether exposure to SP transferring males was beneficial or costly to females, with beneficial effects occurring more often than expected. Hence, the food environment can also shape significantly the strength and direction of selection on mating responses.


[Study on anticoagulant material basis and mechanism of Trichosanthis Semen and its shell and kernel based on spectrum-effect relationship integrated molecular docking].
This study explored the anticoagulant material basis and mechanism of Trichosanthis Semen and its shell and kernel based on spectrum-effect relationship-integrated molecular docking. High performance liquid chromatography(HPLC) fingerprints of Trichosanthis Semen and its shell and kernel were established. Prothrombin time(PT) and activated partial thromboplastin time(APTT) in mice in the low-and high-dose(5, 30 g·kg~(-1), respectively) Trichosanthis Semen, the shell, and kernel groups were determined as the coagulation markers. The spectrum-effect relationship and anticoagulant material basis of Trichosanthis Semen and its shell and kernel were analyzed with mean value calculation method of Deng's correlation degree(MATLAB) and the common effective component cluster was obtained. Then the common targets of the component cluster and coagulation were retrieved from TCMSP, Swiss-TargetPrediction, GenCLiP3, GeneCards, and DAVID, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the targets. The main anticoagulant molecular mechanism of the component cluster was verified by SYBYL-X 2.1.1. The spectrum-effect relationship of Trichosanthis Semen and its shell and kernel was in positive correlation with the dosage. The contribution of each component to anticoagulation was not the same, suggesting that the material basis for anticoagulation was different, but they have common effective components(i.e. common material basis), such as adenine(peak 3), uracil(peak 4), hypoxanthine(peak 6), xanthine(peak 9), and adenosine(peak 11). Network pharmacology showed that these components can act on multiple target proteins such as NOS3, KDR, and PTGS2, and exert anticoagulant effect through multiple pathways such as VEGF signaling pathway. They involved the biological functions such as proteolysis, cell component such as cytosol, and molecular functions. The results of molecular docking showed that the binding free energy of these components with NOS3(PDB ID: 1 D0 C), KDR(PDB ID: 5 AMN), and PTGS2(PDB ID: 4 COX) was ≤-5 kJ·mol~(-1), and the docking conformations were stable. Spectrum-effect relationship-integrated molecular docking can be used for the optimization, virtual screening, and verification of complex chemical and biological information of Chinese medicine. Trichosanthis Semen and its shell and kernel have the common material basis for anticoagulation and they exert the anticoagulant through multiple targets and pathways.


[Infestation by Pediculus humanus (Anoplura: Pediculidae) in a metropolitan area of southeast Brazil].
Adults, nymphs and nits of the Pediculus humanus were found in a frame-house infesting the clothes and bedding of three inhabitants in a shanty town in S. Paulo county, S. Paulo State. A total of 198 nits were found glued in 15.0 cm2 of fibres of the infested clothing, giving a ratio of 13.2 nits/cm2. Having in view that the precarious living conditions, promiscuity, absence of basic sanitation and negligence of the sanitary authorities in various Brazilian cities, this situation should not be underestimated. This find should be more frequent than an isolated observation.


A modified Robertson's equation of animal growth.
A growth equation is presented, differing from Robertson's equation only in its constnats. It is shown that this equation satisfactorily describes the growth of birds and mammals.


Processing of prothrombin in the secretory pathway.
Antibodies raised against plasma prothrombin and the prothrombin propeptide were used to identify prothrombin precursors in rough and smooth microsomes and in the Golgi apparatus. The data demonstrate that the propeptide is part of the prothrombin molecule when undergoing a variety of modifications in the Golgi apparatus. It is shown that these modifications result in an increase in the apparent molecular mass of the prothrombin precursor from 78 kDa in early processing to 83 kDa in late processing. The 83 kDa prothrombin precursor was not recognized by the anti-propeptide antiserum and most likely represents the final product of the precursor in the secretory pathway. Evidence is presented that the propeptide is released from the parent molecule in the Golgi apparatus by a membrane-bound Ca(2+)-dependent serine proteinase(s) with characteristics similar to those of the proalbumin-to-albumin-converting enzyme. Vitamin K-dependent carboxylase activity was measured in membrane fragments obtained from the Golgi apparatus preparation. Sucrose-density-gradient centrifugation and the use of marker enzymes showed that carboxylase activity was highest in fractions enriched in cis-Golgi cisternae. Two different synthetic peptides were used as substrates for the carboxylase. These peptides were from the N-terminal and the C-terminal part of the gamma-carboxyglutamic acid (Gla) region of prothrombin. It is shown that the N-terminal and the C-terminal peptides were preferred as substrates for the carboxylase in the microsomal and the Golgi apparatus preparations respectively. It is also shown that the prothrombin precursor acquires negative charges in the Golgi apparatus that do not result from addition of sugars in late processing. These negative charges could be eliminated by thermal decarboxylation, suggesting that Gla residues may also be synthesized in late processing.


Human kidney methoxyflurane and sevoflurane metabolism. Intrarenal fluoride production as a possible mechanism of methoxyflurane nephrotoxicity.
Methoxyflurane nephrotoxicity is mediated by cytochrome P450-catalyzed metabolism to toxic metabolites. It is historically accepted that one of the metabolites, fluoride, is the nephrotoxin, and that methoxyflurane nephrotoxicity is caused by plasma fluoride concentrations in excess of 50 microM. Sevoflurane also is metabolized to fluoride ion, and plasma concentrations may exceed 50 microM, yet sevoflurane nephrotoxicity has not been observed. It is possible that in situ renal metabolism of methoxyflurane, rather than hepatic metabolism, is a critical event leading to nephrotoxicity. We tested whether there was a metabolic basis for this hypothesis by examining the relative rates of methoxyflurane and sevoflurane defluorination by human kidney microsomes. Microsomes and cytosol were prepared from kidneys of organ donors. Methoxyflurane and sevoflurane metabolism were measured with a fluoride-selective electrode. Human cytochrome P450 isoforms contributing to renal anesthetic metabolism were identified by using isoform-selective inhibitors and by Western blot analysis of renal P450s in conjunction with metabolism by individual P450s expressed from a human hepatic complementary deoxyribonucleic acid library. Sevoflurane and methoxyflurane did undergo defluorination by human kidney microsomes. Fluoride production was dependent on time, reduced nicotinamide adenine dinucleotide phosphate, protein concentration, and anesthetic concentration. In seven human kidneys studied, enzymatic sevoflurane defluorination was minima, whereas methoxyflurane defluorination rates were substantially greater and exhibited large interindividual variability. Kidney cytosol did not catalyze anesthetic defluorination. Chemical inhibitors of the P450 isoforms 2E1, 2A6, and 3A diminished methoxyflurane and sevoflurane defluorination. Complementary deoxyribonucleic acid-expressed P450s 2E1, 2A6, and 3A4 catalyzed methoxyflurane and sevoflurane metabolism, in diminishing order of activity. These three P450s catalyzed the defluorination of methoxyflurane three to ten times faster than they did that of sevoflurane. Expressed P450 2B6 also catalyzed methoxyflurane defluorination, but 2B6 appeared not to contribute to renal microsomal methoxyflurane defluorination because the P450 2B6-selective inhibitor had no effect. Human kidney microsomes metabolize methoxyflurane, and to a much lesser extent sevoflurane, to fluoride ion. P450s 2E1 and/or 2A6 and P450 3A are implicated in the defluorination. If intrarenally generated fluoride or other metabolites are nephrotoxic, then renal metabolism may contribute to methoxyflurane nephrotoxicity. The relative paucity of renal sevoflurane defluorination may explain the absence of clinical sevoflurane nephrotoxicity to date, despite plasma fluoride concentrations that may exceed 50 microM.


The optimal treatment approaches for stage I small cell lung cancer.
Although small cell lung cancer (SCLC) represents less than one-sixth of all lung cancer cases, it is an aggressive disease with a high metastatic potential to various sites including the brain. Most landmark trials assessing individual therapy benefits for SCLC dichotomized patients as having either limited or extensive disease. Over the last decade, however, there has been a clear shift towards categorizing and analyzing survival patterns using a more thorough staging system that accounts for tumor size and the degree of nodal or metastatic disease burden. For the 5% of patients who present with clinical stage I (cT1-2aN0M0) SCLC, extrapolating treatment recommendations from a rather heterogeneous population that constitutes limited disease SCLC remains controversial and has led to numerous investigations of alternative therapies such as stereotactic body radiation therapy (SBRT) for the management of this highly select group of patients. Given the changing landscape of multimodal therapy, this review will summarize relevant data pertaining to and outline optimal treatment algorithms for stage I SCLC, with a particular focus on SBRT as a primary mode of local therapy.


Liver-specific expression of human insulin-like growth factor binding protein-1 in transgenic mice: repercussions on reproduction, ante- and perinatal mortality and postnatal growth.
Study of the in vivo functions of the insulin-like growth factor binding proteins (IGFBPs) is complicated by their variety (six molecular species) and the differences in their expression related to tissue of origin and stage of development. To investigate the physiological role of IGFBP-1 in the bloodstream, we induced hepatic overexpression of IGFBP-1 in transgenic mice, placing human IGFBP-1 (hIGFBP-1) cDNA under the control of the alpha1-antitrypsin promoter so as to obtain liver-specific expression. Five transgenic founder mice were raised, only two of which (lines 124 and 149) produced transgenic offspring. Northern blotting revealed transgene expression exclusively in the liver during fetal life and unchanged through to adulthood, whereas expression of the endogenous gene was undetectable beyond 10-15 days postnatally. hIGFBP-1 was detected by western immunoblotting in the plasma of transgenic mice and IRMAs yielded mean concentrations of 2.41 +/- 0.33 ng/ml and 13.69 +/- 1.42 ng/ml in homozygous animals of lines 124 and 149, respectively. In the latter, IGFBP-1 levels were distinctly higher than in heterozygotes (2.99 +/- 0.39 ng/ml), P < 0.0001. These levels remained stable in each given animal and did not change with age. Plasma concentrations of IGF-I measured in line 149 exhibited the well-known profile of an increase from birth up to puberty. Values for heterozygotes were similar to those for wild-type mice, with adult levels (544 +/- 98 ng/ml) slightly below those of controls (630 +/- 56 ng/ml), P < 0.05. In homozygotes they were distinctly lower, with adult levels of 370 +/- 75 ng/ml, P = 0.001. In heterozygous and homozygous adults, there was a negative correlation between IGF-I and IGFBP-1 concentrations (r = 0.8, P < 0.0001), suggesting a link between transgene expression and IGF-I levels. Study of body weight gain in line 149 revealed growth retardation within the first weeks after birth, which was marked in homozygous males and females (P < 0.001) but also present in heterozygous males (P = 0.002), indicating some relationship with transgene expression. In addition, body weight in adult mice was negatively correlated to plasma concentrations of IGFBP-1 (r = 0.7, P < 0.0001). Reproductive function also appeared to be severely affected, especially in homozygous females: mating that failed to result in pregnancy in half of the homozygous females crossed with nontransgenic males, suggestive of impaired fertilization or implantation; interrupted or prolonged pregnancies with fetal and neonatal death. Litter size was reduced in transgenic females (by about half in homozygotes) and in nontransgenic females mated with homozygous males, resulting from pre- or neonatal mortality. Moreover, deaths occurred within the first 5 days of life, with an incidence of approximately 50% in the litters of homozygous females, 12-18% among heterozygotes mated with nontransgenic or heterozygous males, respectively, and 30% among those mated with homozygous males. These results, suggesting that fetal transgene expression largely accounted for ante- and perinatal mortality, were confirmed by the predominance of homozygotes among those that could be analyzed genetically. Similarly impaired reproductive function was seen in line 124, but to a lesser degree. Although the mechanisms responsible for these disorders remain to be determined, our results indicate that permanent and uncontrolled hepatic expression of IGFBP-1, even at low levels, affects fertility in females and both ante- and postnatal development.


Outward and inward directed aggressiveness: the interaction between violence and suicidality.
This paper examines the relations and interactions between suicidality and violence. It reviews the recent literature indicating that more than 40 variables have been identified as risk factors for suicide. Evidence is presented to indicate that the differences between suicide attempters and suicide completers are a matter of degree, and that most conclusions that apply to one also apply to the other. A review of the recent literature on violence revealed that 37 variables are risk factors for violence and that 23 of them are risk factors for suicide as well. Evidence indicates that at least 17 variables have been identified as protective factors that decrease the risk of both suicide and violence. These considerations and others have led to the development of a Two-Stage Model of Countervailing Forces that systematically relates suicide to violence.


Intramural duodenal hematoma secondary to pancreatitis: case report and review of the literature.
Spontaneous intramural duodenal hematoma is uncommon and is usually associated with coagulopathy, anticoagulant therapy and endoscopic procedures. The aim here was to describe a case of intramural duodenal hematoma caused by chronic exacerbation of pancreatitis. A 46-year-old male with chronic alcoholic pancreatitis was admitted to hospital due to abdominal pain, melena and low hemoglobin. An intramural duodenal hematoma with active bleeding was detected and selective angioembolization was warranted. The patient evolved with a perforated duodenum and underwent laparotomy with exclusion of the pylorus and Roux-en-Y gastrojejunostomy. He was discharged nine days later. Intramural duodenal hematoma is a rare complication of pancreatitis. Selective embolization is the preferred treatment for hemorrhagic complications of pancreatitis. However, the risk of visceral ischemia and perforation should be considered.


Surgical Trial In Traumatic intraCerebral Haemorrhage (STITCH): a randomised controlled trial of Early Surgery compared with Initial Conservative Treatment.
While it is accepted practice to remove extradural (EDH) and subdural haematomas (SDH) following traumatic brain injury, the role of surgery in parenchymal traumatic intracerebral haemorrhage (TICH) is controversial. There is no evidence to support Early Surgery in this condition. There have been a number of trials investigating surgery for spontaneous intracerebral haemorrhage but none for TICH. This study aimed to establish whether or not a policy of Early Surgery for TICH improves outcome compared with a policy of Initial Conservative Treatment. This was an international multicentre pragmatic parallel group trial. Patients were randomised via an independent telephone/web-based randomisation service. Neurosurgical units in 59 hospitals in 20 countries registered to take part in the study. The study planned to recruit 840 adult patients. Patients had to be within 48 hours of head injury with no more than two intracerebral haematomas greater than 10 ml. They did not have a SDH or EDH that required evacuation or any severe comorbidity that would mean they could not achieve a favourable outcome if they made a complete recovery from their head injury. Patients were randomised to Early Surgery within 12 hours or to Initial Conservative Treatment with delayed evacuation if it became clinically appropriate. The Extended Glasgow Outcome Scale (GOSE) was measured at 6 months via a postal questionnaire. The primary outcome was the traditional dichotomised split into favourable outcome (good recovery or moderate disability) and unfavourable outcome (severe disability, vegetative, dead). Secondary outcomes included mortality and an ordinal assessment of Glasgow Outcome Scale and Rankin Scale. Patient recruitment began in December 2009 but was halted by the funding body because of low UK recruitment in September 2012. In total, 170 patients were randomised from 31 centres in 13 countries: 83 to Early Surgery and 87 to Initial Conservative Treatment. Six-month outcomes were obtained for 99% of 168 eligible patients (82 Early Surgery and 85 Initial Conservative Treatment patients). Patients in the Early Surgery group were 10.5% more likely to have a favourable outcome (absolute benefit), but this difference did not quite reach statistical significance because of the reduced sample size. Fifty-two (63%) had a favourable outcome with Early Surgery, compared with 45 (53%) with Initial Conservative Treatment [odds ratio 0.65; 95% confidence interval (CI) 0.35 to 1.21; p = 0.17]. Mortality was significantly higher in the Initial Conservative Treatment group (33% vs. 15%; absolute difference 18.3%; 95% CI 5.7% to 30.9%; p = 0.006). The Rankin Scale and GOSE were significantly improved with Early Surgery using a trend analysis (p = 0.047 and p = 0.043 respectively). This is the first ever trial of surgery for TICH and indicates that Early Surgery may be a valuable tool in the treatment of TICH, especially if the Glasgow Coma Score is between 9 and 12, as was also found in Surgical Trial In spontaneous intraCerebral Haemorrhage (STICH) and Surgical Trial In spontaneous lobar intraCerebral Haemorrhage (STICH II). Further research is clearly warranted. Current Controlled Trials ISRCTN 19321911. This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 19, No. 70. See the NIHR Journals Library website for further project information.


Multi-scale network imaging in a mouse model of amyloidosis.
The adult neocortex is not hard-wired but instead retains the capacity to reorganise across multiple spatial scales long into adulthood. Plastic reorganisation occurs at the level of mesoscopic sensory maps, functional neuronal assemblies and synaptic ensembles and is thought to be a critical feature of neuronal network function. Here, we describe a series of approaches that use calcium imaging to measure network reorganisation across multiple spatial scales in vivo. At the mesoscopic level, we demonstrate that sensory activity can be measured in animals undergoing longitudinal behavioural assessment involving automated touchscreen tasks. At the cellular level, we show that network dynamics can be longitudinally measured at both stable and transient functional assemblies. At the level of single synapses, we show that functional subcellular calcium imaging approaches can be used to measure synaptic ensembles of dendritic spines in vivo. Finally, we demonstrate that all three levels of imaging can be spatially related to local pathology in a preclinical rodent model of amyloidosis. We propose that multi-scale in vivo calcium imaging can be used to measure parallel plasticity processes operating across multiple spatial scales in both the healthy brain and preclinical models of disease.


Rapid GAL gene switch of Saccharomyces cerevisiae depends on nuclear Gal3, not nucleocytoplasmic trafficking of Gal3 and Gal80.
The yeast transcriptional activator Gal4 localizes to UAS(GAL) sites even in the absence of galactose but cannot activate transcription due to an association with the Gal80 protein. By 4 min after galactose addition, Gal4-activated gene transcription ensues. It is well established that this rapid induction arises through a galactose-triggered association between the Gal80 and Gal3 proteins that decreases the association of Gal80 and Gal4. How this happens mechanistically remains unclear. Strikingly different hypotheses prevail concerning the possible roles of nucleocytoplasmic distribution and trafficking of Gal3 and Gal80 and where in the cell the initial Gal3-Gal80 association occurs. Here we tested two conflicting hypotheses by evaluating the subcellular distribution and dynamics of Gal3 and Gal80 with reference to induction kinetics. We determined that the rates of nucleocytoplasmic trafficking for both Gal80 and Gal3 are slow relative to the rate of induction. We find that depletion of the nuclear pool of Gal3 slows the induction kinetics. Thus, nuclear Gal3 is critical for rapid induction. Fluorescence-recovery-after-photobleaching experiments provided data suggesting that the Gal80-Gal4 complex exhibits kinetic stability in the absence of galactose. Finally, we detect Gal3 at the UAS(GAL) only if Gal80 is covalently linked to the DNA-binding domain. Taken altogether, these new findings lead us to propose that a transient interaction of Gal3 with Gal4-associated Gal80 could explain the rapid response of this system. This notion could also explain earlier observations.


Linkage analysis of rheumatoid arthritis in US and UK families reveals interactions between HLA-DRB1 and loci on chromosomes 6q and 16p.
HLA is the most strongly associated locus in rheumatoid arthritis (RA), accounting for up to one-third of the genetic contribution. Conditioning on the effect of true disease loci such as HLA can lead to increased power to detect effects at other loci and, in addition, allows investigation of the underlying disease models, including interactions. The aim of this study was to detect susceptibility loci for RA by conditioning on HLA in a large sample of affected sibling pairs (ASPs) and to test for evidence of interaction between novel loci and HLA. Genotype data from 3 whole-genome linkage scans for RA in a US population and a UK population were pooled, resulting in a combined data set of 886 ASPs. This pooling of data increased the power to detect loci showing low levels of heterogeneity. Nonparametric linkage analysis was performed to identify regions of interest. Joint 2-locus analysis was then performed for HLA and each of the loci that demonstrated evidence of linkage in the 886 ASPs. Evidence for linkage was most significant at HLA (P = 4 x 10(-16)), with 7 non-HLA loci showing some evidence for linkage (P = 0.05-0.003). Joint modeling of these loci with HLA provided evidence for linkage at a genome-wide significance level for loci on 6q (P = 2.7 x 10(-6)) and 16p (P = 2 x 10(-4)). These data provide the most convincing evidence to date that 6q and 16p harbor susceptibility genes. In addition, these loci may interact with HLA, facilitating the search for candidate genes within this region.


Validity of the Novel Radiological Classification System of the Distal Femur.
Aseptic loosening (AL) is among the most important causes of failure after total knee arthroplasty (TKA). However, while there are numerous underlying causes of AL, the morphometry of the distal femur and intramedullary canal has not been sufficiently demonstrated. This study aimed to show the interobserver and intraobserver reliability and validity of the Citak classification, which has been recently defined according to the morphometry of the distal femur and provides a risk factor definition for AL. A total of 200 patients whose standardized anteroposterior (AP) and lateral images of the knee joint were obtained between October 2019 and April 2020 were retrospectively evaluated in this study. Patients with a history of extra-articular deformity and knee surgery were excluded from the study. For AL, morphologies of the distal femur were identified by two observers using the new radiological classification system of the distal femur. Mean pairwise Cronbach's alpha coefficient was used to assess the intra- and interobserver agreement of the classification. There was excellent interobserver agreement for the 20 cm proximal and 2 cm proximal to the lateral joint line (PLJL) and adductor tubercle (PAD), respectively. The mean Cronbach's alpha coefficient was 0.96 (range 0.764-0.944) for the PAD and 0.98 (range 0.734-0.929) for the PLJL. There was also an excellent intraobserver agreement, with 93% average pairwise percent agreement for the index group and 95.5% average pairwise percent agreement for the anatomical classification group. The level of inter- and intraobserver agreement for the morphology of the distal femur was excellent in the new radiological classification system, which was shown to be beneficial in the planning of revision knee arthroplasty for AL. However, there is a need for further studies in order to make a correlation of the classification with specific intraoperative findings.


Laboratory evaluation of four techniques of stapled gastroplasty.
Four techniques of stapled gastroplasty including unreinforced horizontal gastroplasty (UHG), suture-reinforced horizontal gastroplasty (SHG), Marlex banded horizontal gastroplasty (HBG), and Marlex banded vertical gastroplasty (VBG) were studied in mongrel dogs. Twelve dogs survived operations by each technique and were killed in groups of four at 3, 6, and 12 weeks after operation. Five deaths caused by leaks occurred after UHG (one), SHG (one), and HBG (three). Disruption of the staple line occurred after UHG (four), HBG (one), and VBG (three). Mean stomal diameter at death for UHG (2.1 cm) and SHG (1.5 cm) was substantially greater than the initial diameter of 0.9 cm. Mean stomal diameter at death for HBG (1.4 cm) and VBG (1.3 cm) was only slightly larger than the initial diameter of 1.2 cm. There was no correlation between the degree of stomal dilatation and time of death. There was no evidence of erosion of Marlex through the gastric wall in dogs that survived to the time of death. These data suggest that stomal reinforcement with Marlex mesh effectively limits dilatation and is safe when sutured to itself rather than to the stomach. Reinforcement of the stapled partition with sutures may be necessary to eliminate staple-line disruption.


Is overexpression of HER-2 a predictor of prognosis in colorectal cancer?
The development of novel chemotherapeutic agents in colorectal cancer has improved survival. Following initial response to chemotherapeutic strategies many patients develop refractory disease. This poses a significant challenge common to many cancer subtypes. Newer agents such as Bevacizumab have successfully targeted the tyrosine kinase receptor epidermal growth factor receptor in metastatic colorectal cancer. Human epidermal growth factor receptor-2 is another member of the tyrosine kinase receptor family which has been successfully targeted in breast cancer. This may play a role in colorectal cancer. We conducted a clinicopathological study to determine if overexpression of human epidermal growth factor receptor-2 is a predictor of outcome in a cohort of patients with colorectal cancer. Clinicopathological data and paraffin-embedded specimens were collected on 132 consecutive patients who underwent colorectal resections over a 24-month period at Mayo General Hospital. Twenty-six contained non-malignant disease. Her-2/neu protein overexpression was detected using immunohistochemistry (IHC). The HER-2 4B5 Ventana monoclonal antibody was used. Fluorescent insitu hybridisation (FISH) was performed using INFORM HER-2/Neu Plus. Results were correlated with established clinical and pathological predictors of outcome including TNM stage. Statistical analysis was performed using SPSS version 11.5. 114 were HER-2/Neu negative using IHC, 7 showed barely perceptible positivity (1+), 9 showed moderate staining (2+) and 2 were strongly positive (3+). There was no correlation with gender, age, grade, Dukes' stage, TNM stage, time to recurrence and 5-year survival (p > 0.05). FISH was applied to all 2+ and 3+ cases as well as some negative cases selected at random. Three were amplified (2 were 3+ and 1 was 2+). Similarly, HER-2 gene overexpression did not correlate with established prognostic indicators. HER-2 protein is over expressed in 11% of colorectal cancer patients. The gene encoding HER-2 is amplified in 3% of cases. Overexpression of HER-2 is not a predictor of outcome. However, patients who over express HER-2 may respond to Herceptin therapy.


[Fullerenol - properties and applications in biomedical sciences].
Fullerenols, the water-soluble derivatives of fullerenes, are currently being recently intensively studied in the context of the possibility of their application in the biomedicine. Due to their hydrophilic properties and the ability to eliminate free radicals, fullerenols may in the future provide a solid alternative to currently used pharmacological methods in chemotherapy, treatment of neurodegenerative diseases and radiobiology. Depending on the research protocol applied, fullerenols may also act as pro oxidants. The dualistic nature of fullerenols may contribute to finding new biomedical applications of these agents in the future, by exerting a cytotoxic or protective effect respectively against cancer cells or healthy cells. Because of the encapsulated structure of fullerenols, there exists the possibility of their application in medical diagnostics in the transfer of contrast agents or in the drug transport. During the planning of an experiment designed to investigate the effects of radiation in combination with derivatives of water-soluble fullerenes, the possibility of appearance of the "dose-response effect" should be taken into consideration since it significantly contributes to one of the two possible effects: protection or sensitization. The same applies to the possibility of using these compounds as potential neuroprotectors. Fullerenol may protect neurons in the particular areas of the brain but in the definedcertain doses it may also induce cell death. A giant leap in the field of nanotechnology not only leads scientists to search for new applications of nanomaterials such as fullerenols, but also raises the question about their harmful effect on the environment. High utilization of hardly biodegradable fullerenols increases the likelihood of their accidental release into natural systems and their bioaccumulation. Despite convincing evidences about the potential applications of fullerenols in biomedicine, we still have insufficient knowledge about the mechanism of action of these molecules and their possible side effects.


Left ventricular M-mode prediction intervals in 7651 dogs: Population-wide and selected breed-specific values.
Echocardiography is a common method to measure heart size in dogs. The heart dimensions are influenced by body weight (BW) and potentially by breed. To establish BW-dependent prediction intervals (PIs) of the left ventricular (LV) linear dimensions in a population of dogs of many breeds in multicenter environment, and to identify breeds deviating from these intervals. DOGS: Seven thousand six hundred and fifty-one dogs. Retrospectively, data from heart screens conducted between 2009 and 2016 were included. Cardiac dimensional PIs were generated using allometric scaling including all nonsighthound dogs and values were compared to previously published PIs. The values measured in dogs of respective breeds, including sighthounds, were then compared to the overall nonsighthound PIs to identify deviant breeds. The interobserver-variability of the measurements was determined using the explained residual variance. Prediction intervals for the nonsighthound dogs were in agreement with previously published cardiac PIs, although the upper limits of the generated PIs of our study were slightly below those currently applied (except the interventricular septum in systole and the left ventricular free wall in diastole below 10.0 kg and 15.0 kg, respectively). Values measured in the nonsighthound breed Newfoundland deviated for most dimensions. Most of the sighthound breeds analyzed had greater cardiac dimensions, with the exception of the Irish Wolfhound. Findings of our study reinforces the value of BW-dependent PIs for cardiac dimensions in dogs and suggest that these PIs are valid for most nonsighthound breeds, but not the sighthound breeds.


Inhibition of humoral immunity and mitogen responsiveness of lymphoid cells following oral administration of the heterocyclic food mutagen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) to B6C3F1 mice.
In these studies, the food promutagen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) was evaluated for its immunotoxicity in B6C3F1 mice following oral 5-day dosing at total doses of 50 and 150 mg/kg. Results indicated that PhIP produced a dose-dependent suppression of the humoral immune response of spleen cells to sheep erythrocytes, with a 50% decrease in the number of PFC detected at the 150 mg/kg dose of PhIP. A 40-90% inhibition of the phytohemagglutinin (PHA) response of spleen cells, mesenteric lymph nodes (MLNs), and Peyer's patch (PP) lymphocytes was seen in the treatment groups. The lipopolysaccharide (LPS) response was somewhat more variable and less affected with 20-30% inhibition observed in the spleen and PPs, whereas PhIP increased the LPS response in the MLNs. There was no effect of PhIP on cell recovery or viability in any of the treatment groups. Flow cytometry analysis revealed a depletion of T cells (Thy 1.2+ cells) and a slight increase in B cells (Ly5+ cells) in the PPs. The percentage of B and T cells present in the spleen and MLNs was unaffected by PhIP. These results demonstrate that the oral administration of PhIP produces immunotoxicity to mice, especially to lymphoid tissues present in the GI tract (i.e., PPs), and demonstrates that T cell mitogen (PHA) responses in PPs are the most sensitive indicator of PhIP-induced immunotoxicity.


Management of thumb duplication.
Radial polydactyly, a common congenital problem, most often occurs as thumb duplication. It appears in pentadactylous, polydactylous, and hypoplastic hands and is simulated by complex syndactyly. Classically, there are three categories for radial polydactyly: severe hypoplasia, partial duplication, and total duplication (easily confused with pseudoduplication). The common second category is subcategorized by Wassel's seven types. Management techniques vary with these classifications. Until a decade ago, the standard recommendation was simple excision of one of the duplicates. Problems of deformity, instability, and weakness became apparent, and reconstructive options were developed. Review of a series of 54 duplicate thumbs, all surgically treated, reflects the changing pattern of reconstruction. With current techniques, good results are usual, although secondary reconstructive procedures may be required. Simple excision is seldom indicated. Combination procedures, involving core tissues of bone, joint, and tendon or peripheral tissues--neurovascular, subcutaneous, and skin--or sometimes all of these, are preferred for reconstruction. The major current problem is to achieve the maximal good result with a minimal number of surgical operations.


Effect of Cognitive Training on Daily Function in Older People without Major Neurocognitive Disorder: A Systematic Review.
There is increasing interest in the effect of non-pharmacological treatments on preserving cognition and function in older adults without major neurocognitive disorder (dementia). However, its effect on everyday function in terms of instrumental activities of daily living (IADL) is unclear. We conducted a systematic review to examine whether cognitive training, independent of other interventions, can improve IADL function in older adults without major neurocognitive disorder. We searched multiple databases including MEDLINE, EMBASE, and PSYCINFO and found thirteen studies that met our inclusion criteria with 7130 participants in total. Six out of thirteen studies reported a significant change on validated IADL assessment. On subgroup analysis, five studies included older adults with normal cognition and one included mild cognitive impairment (MCI). Eleven out of twelve studies showed improvement in measures of cognition. None of the studies described changes in the ability to live independently. While variation in study protocol, outcome measurement, and effect size reporting precluded further inferential statistical analysis, our review found a sizable number of studies showing improvement in IADL. Cognitive training may have some benefit in improving IADL function in older adults without major neurocognitive disorder. Future long-term studies focusing on maintained IADL function and preserved independence are needed.


Why i.p. therapy cannot yet be considered as a standard of care for the first-line treatment of ovarian cancer: a systematic review.
A National Cancer Institute (NCI) clinical announcement recommended i.p. therapy for women with optimally debulked ovarian cancer. Its basis was a summary of eight randomised controlled trials and two systematic reviews, which appear to indicate benefit of i.p. therapy. However, the systematic reviews that inform the recommendations have been inappropriately presented and interpreted. The systematic reviews inappropriately pooled results from 'confounded' trials in which different drugs and different doses of drugs were given in the control and i.p. treatment arms. Therefore, it is not possible to assess which component of treatment is responsible for improving outcome. In addition, none of the trials use a control arm of the internationally accepted standard of care. Using just the unconfounded trials, indirect comparisons show that the magnitude of benefit observed when i.p. regimens are compared with older i.v. regimens [hazard ratio (HR) for overall survival (OS) 0.75; 95% confidence interval (CI) 0.60-0.92, P = 0.006] is smaller than the magnitude of benefit achieved with modern day standard of i.v. treatment compared with the same i.v. regimen used as control in the unconfounded i.p. trials (HR for OS 0.68; 95% CI 0.58-0.80, P < 0.001). A further difficulty is that the reviews cannot recommend an i.p. regimen for standard use. Drug-related toxicity and catheter complications that occur with i.p. therapy are considerable. The NCI recommendations have major implications for the treatment of women with ovarian cancer and for the next generation of clinical trials. We do not believe that the body of evidence currently available supports the recommendation that i.p. therapy should form part of routine care. The choice of treatment of women with newly diagnosed, optimally debulked, ovarian cancer, where therapy has the best chance of influencing OS, is too important to be left with this uncertainty. A clinical trial that investigates a practical and acceptable regimen which gives some or all chemotherapy by the i.p. route and compares this with standard i.v. chemotherapy should be a priority for those who wish to promote its use.


[Purulent pericarditis and pleural empyema due to Streptococcus pneumoniae].
We present the case of a 59 year old woman with no previous clinical history of disease who was admitted in our ICU due to respiratory failure and hemodynamic instability. She was found to have purulent pericarditis due to Streptococcus pneumoniae, complicated by massive pericardial effusion and pleural empyema. A pericardial tap was performed unsuccessfully and finally pericardial effusion was evacuated by surgical drainage. Pleural empyema was also evacuated. She received antibiotic treatment and recovered with normalisation of heart function.


Metastatic intracranial chordoma in a child with massive pulmonary tumor emboli.
A 27-month-old boy of Hispanic background developed multiple cranial nerve palsies, difficulty swallowing, bloody nasal discharge, and irritability. Radiographic evaluations showed extensive destruction of the clivus by a large tumor that invaded the sphenoid bone, left cavernous sinus, ethmoid sinus, nasal cavity, and left orbit. Multiple pulmonary nodules were also noted. The bone marrow and spinal fluid showed no evident tumor cells. Transnasal biopsy revealed a chordoma. Treatment was initiated with a combination of ifosfamide, mesna, and etoposide along with radiation therapy to the cranial tumor. Shifting pulmonary densities were noted on serial films. Despite some clinical improvement, the child developed rapidly progressive hypoxemia 3 weeks after admission and died. Autopsy showed persistent viable tumor in the primary site and massive pulmonary arteriolar tumor emboli, infarcts, and widespread lung parenchymal metastases. No other sites of tumor involvement were discovered. This is the second child reported with intracranial chordoma, pulmonary metastases at diagnosis, and early death attributed to pulmonary tumor emboli.


ER-α36: a novel biomarker and potential therapeutic target in breast cancer.
Estrogen receptor-alpha36 (ER-α36) is a 36-kDa variant of estrogen receptor-alpha (ER-α) firstly identified and cloned by Wang et al in 2005. It lacks both transactivation domains (activation function 1 and activation function 2) and has different biological characteristics compared to traditional ER-α (ER-α66). ER-α36 primarily locates on plasma membrane and cytoplasm and functions as a mediator in the rapid membrane-initiated non-genomic signaling pathway. Additionally, it inhibits the traditional genomic signaling pathway mediated by ER-α66 in a dominant-negative pattern. Accumulating evidence has demonstrated that ER-α36 regulates the physiological function of various tissues. Thus, dysregulation of ER-α36 is closely associated with plenty of diseases including cancers. ER-α36 is recognized as a molecular abnormality which solidly correlates to carcinogenesis, aggressiveness, and therapeutic response of breast cancer. Additionally, special attention has been paid to the role of ER-α36 in endocrine therapy resistance. Therefore, ER-α36 provides a novel biomarker of great value for diagnosis, prognosis, and treatment of breast cancer. It may also be a potential therapeutic target for breast cancer patients, especially for those who are resistant to endocrine therapy. In this review, we will overview and update the biological characteristics, underlying mechanism, and function of ER-α36, focusing on its biological function in breast cancer and endocrine therapy resistance. We will evaluate its application value in clinical practice.


Quadricuspid aortic valve: a rare cause of aortic regurgitation and stenosis.
Quadricuspid aortic valve is a rare congenital condition that occurs not only as an isolated anomaly, but also with other cardiac defects. We describe a 10-year-old boy whose aortic stenosis was diagnosed during infancy. Transthoracic echocardiography revealed dilation of the left ventricle, valvular and subvalvular aortic stenosis, bicuspid aortic valve, aortic regurgitation, and mitral valve prolapse. The results of cardiac catheterization and aortography showed severe aortic regurgitation, an aortic valve gradient of 76 mmHg, a bicuspid aortic valve, a subaortic membrane, and an ascending aortic aneurysm. The patient underwent elective valve replacement with a mechanical prosthesis, and during surgery, the valve was noted to be quadricuspid. The patient was diagnosed as having a quadricuspid aortic valve associated with aortic regurgitation, severe aortic stenosis, and an ascending aortic aneurysm.


Analgesic and sedative effects of perioperative gabapentin in total knee arthroplasty: a randomized, double-blind, placebo-controlled dose-finding study.
Gabapentin has shown acute postoperative analgesic effects, but the optimal dose and procedure-specific benefits vs harm have not been clarified. In this randomized, double-blind, placebo-controlled dose-finding study, 300 opioid-naive patients scheduled for total knee arthroplasty were randomized (1:1:1) to either gabapentin 1300 mg/d (group A), gabapentin 900 mg/d (group B), or placebo (group C) daily from 2 hours preoperatively to postoperative day 6 in addition to a standardized multimodal analgesic regime. The primary outcome was pain upon ambulation 24 hours after surgery, and the secondary outcome was sedation 6 hours after surgery. Other outcomes were overall pain during well-defined mobilizations and at rest and sedation during the first 48 hours and from days 2-6, morphine use, anxiety, depression, sleep quality, and nausea, vomiting, dizziness, concentration difficulty, headache, visual disturbances, and adverse reactions. Pain upon ambulation (visual analog scale, mean [95% confidence interval]) 24 hours after surgery in group A vs B vs C was as follows: 41 [37-46] vs 41 [36-45] vs 42 [37-47], P = 0.93. Sedation (numeric rating scale, median [range]) 6 hours after surgery was as follows: 3.2 [0-10] vs 2.6 [0-9] vs 2.3 [0-9], the mean difference A vs C being 0.9 [0.2-1.7], P = 0.046. No between-group differences were observed in overall pain or morphine use the first 48 hours and from days 2-6. Sleep quality was better during the first 2 nights in group A and B vs C. Dizziness was more pronounced from days 2-6 in A vs C. More severe adverse reactions were observed in group A vs B and C. In conclusion, gabapentin may have a limited if any role in acute postoperative pain management of opioid-naive patients undergoing total knee arthroplasty and should not be recommended as a standard of care.


[Health working conditions and environment: conceptual model for remote and rural areas].
This report proposes a conceptual model on workplace environment and working conditions that integrates the available evidence to facilitate the design and evaluation of interventions aimed at improving the attraction, recruitment and retention of health personnel at the first level of care in rural and remote areas. Theoretical, empirical and testimonial evidence was consulted to support the model, and 15 frameworks disseminated in the last 20 years were synthesized. The article shows the diversity of perspectives and the complexity involved in establishing the dimensions to be considered in a proposal that is useful to apply to human resources for health policies.The proposed model includes four categories of components: factors of the external context, organizational factors, employment and work conditions, and individual factors. The boundaries between the components -as well as the weight and influence of each one- vary according to the interrelationship among them and the interaction with the environment, and thus its interpretation must be adapted to the context in which it is intended to be applied.Based on this conceptual model, the design and evaluation of interventions aimed at increasing the availability of health personnel -particularly at the primary care level in rural and remote areas of the Region of the Americas- should emerge from an interaction between health and employment policies, and the realities and expectations of workers and the communities.


Retinoblastoma protein in human renal cell carcinoma in relation to alterations in G1/S regulatory proteins.
The retinoblastoma gene product (pRb) is the main substrate for cyclin-dependent kinases (CDKs) during the G1/S transition. Aberrations in cell cycle regulatory proteins, which have been observed in many malignancies, can theoretically cause increased phosphorylation of pRb due to unbalanced CDK activities. The expression and phosphorylation of pRb and potential associations to cell cycle aberrations in renal cell carcinomas (RCC) has only partly been clarified. We therefore evaluated the presence of pRb and the level of pRb-phosphorylation in 216 RCCs arranged in tissue microarrays by using different pRb-antibodies, including pRb-phosphospecific antibodies. Most RCCs (95%) expressed pRb, while cases with the low pRb levels, potentially indicative for pRb-inactivation, were few. In order to detect secondary alterations to a potential pRb-inactivation, the p16 expression was also monitored. None of the tumors exhibited increased p16 levels, confirming that pRb-inactivation is rare in RCC. Phosphorylated pRb was detected in approximately 50% of the RCCs, using Western blotting or immunohistochemistry. The immunohistochemical ppRb(ser807/811) levels were associated with high proliferation, cyclin D1, cyclin E and p27 protein content. Surprisingly, there was no association between pRb-phosphorylation and clinicopathological data. In summary, pRb seemed to be functional and aberrations in G1/S-regulatory proteins were associated with increased phosphorylation of pRb and proliferation. The data supports that pRb might be one of the main cell cycle regulators in RCC.


The role of (E)-6-chloro-3-(3-methyl-1-phenyl-1H-pyrazol-5-yl)-2-styrylquinazolin-4(3H)-one in the modulation of cannabinoidergic system. A pilot study.
Compounds acting on endocannabinoid system regulate different neuronal processes through the cannabinoid receptors activation. The main aim of this study was determining whether the 2-styrylquinazolin-4(3H)-one 5, a structural analogue of rimonabant, was able to counteract the behavioural signs of the activation of the endocannabinoidergic system induced by CP 55.940. Behavioural assessment was carried out using the tetrad task and the novel object recognition test. The endocannabinoidergic system activation was possible by the administration of CP 55.940 and 30min after rats were tested in the tetrad task for the evaluation of the antinociceptive-, cataleptic-, hypothermic- and locomotor- effects. The evaluation of the declarative memory was carried out through the novel object recognition test. The administration of the new compound was made at three different doses, 30min before CP 55.940 administration on a separate group of animals. Our results demonstrated that compound 5, at the highest dose, was able to counteract the effects exerted by CP 55.940, shown by an increase in body temperature, total distance travelled, latency to fall and decrease in tail flick latency, interfering conjointly in memory impairment. This study shows that compound 5 is able to counteract the cannabinoid activation induced by the agonist CP 55.940. Further investigations on its pharmacological profile are mandatory before considering it as a potential candidate for clinical studies and its possible employment as pharmacological agent for the management of different pathological conditions such as motor incoordination, obesity and brain related disorders.


What is the evidence for heterozygote advantage selection?
Recent genomic data have found that many genes show the signal of selection. How many of these genes are undergoing heterozygote advantage selection is only beginning to be known. Initial genomic surveys have suggested that only a small proportion of loci have polymorphisms maintained by heterozygote advantage and this is consistent with the few examples generated from other approaches within given species. Unless further studies provide large numbers of loci with heterozygote advantage, it appears that loci with heterozygote advantage must be considered only a small minority of all loci in a species. This is not to say that some heterozygote advantage loci do not have important adaptive functions, but that their role in overall evolutionary change might be more of an unusual phenomenon than a major player in adaptation.


Modulation of lentivirus replication by antibodies. Non-neutralizing antibodies to caprine arthritis-encephalitis virus enhance early stages of infection in macrophages, but do not cause increased production of virions.
Non-neutralizing antibodies to caprine arthritis-encephalitis virus (CAEV) enhance the early stages of the virus life cycle but do not potentiate enhanced production of virus particles by macrophages. In primary macrophages used for these studies, there was enhancement in binding, internalization and uncoating of virus pretreated with non-neutralizing sera in comparison to virus pretreated with a non-immune serum. However, this did not lead to enhanced production of virus particles. Failure of non-neutralizing sera to inactivate CAEV may be due in part to low avidity of the antibodies for the virus particles which contain sialic acids on their envelopes, because desialylation of the particles made them neutralizable. The non-neutralizing antibodies probably bound to most of the native virus particles which were then internalized via Fc receptor-mediated endocytosis and degraded. Sialylated particles that failed to bind antibodies probably caused the infection. Thus there was no true enhancement of infection. The previously reported increase in severity of lesions in animals immunized with inactivated CAEV particles prior to challenge with live virus suggested enhancement of infection but in the light of our finding this may have been caused by factors other than an increase in production in the number of infectious virus particles.


Water Security and Nutrition: Current Knowledge and Research Opportunities.
Water is an essential nutrient that has primarily been considered in terms of its physiological necessity. But reliable access to water in sufficient quantities and quality is also critical for many nutrition-related behaviors and activities, including growing and cooking diverse foods. Given growing challenges to water availability and safety, including climate change, pollution, and infrastructure degradation, a broader conceptualization of water and its diverse uses is needed to sustainably achieve global nutrition targets. Therefore, we review empirical and qualitative evidence describing the linkages between water security (the reliable availability, accessibility, and quality of water for all household uses) and nutrition. Primary linkages include water security for drinking, food production and preparation, infant and young child feeding, and limiting exposure to pathogens and environmental toxins. We then identify knowledge gaps within each linkage and propose a research agenda for studying water security and nutrition going forward, including the concurrent quantification of both food and water availability, accessibility, use, and stability. By making explicit the connections between water security and nutritional well-being, we aim to promote greater collaboration between the nutrition and water, sanitation, and hygiene sectors. Interdisciplinary policies and programs that holistically address the water-nutrition nexus, versus those that focus on water and nutrition independently, are likely to significantly advance our ability to ensure equitable access to healthy foods and safe water for all.


Mechanisms of rejection in vascular composite allotransplantation.
For patients with devastating injuries in whom standard reconstruction is not an option, vascularized composite allotransplantation (VCA) has become a viable means of restoring form and function. However, immunological rejection continues to be a problem in VCA and has not yet been fully characterized. As the field is relatively new, much of the data on rejection and immunosuppression have been extrapolated from that of solid organ transplantation. In this review, we cover the basic mechanisms of rejection as they relate specifically to VCA with analysis of recent literature and future directions. Recent clinical studies have supported previously postulated T-cell-mediated mechanism of acute rejection and have also made strides in differentiating rejection from inflammation from other skin conditions and with different treatment regimens. Antibody-mediated rejection has been described in recent cases as well as treatment of presensitized patients receiving VCAs. With more long-term grafts, chronic changes, including vasculopathy, are being reported. Clinically observed types of rejection in VCA include mainly cell-mediated, antibody-mediated and chronic rejection. Advances in diagnosis and treatment of rejection have been made, but there is still much to be learned about VCA-specific rejection.


The Oddity Detection in Diverse Scenes (ODDS) database: Validated real-world scenes for studying anomaly detection.
Many applied screening tasks (e.g., medical image or baggage screening) involve challenging searches for which standard laboratory search is rarely equivalent. For example, whereas laboratory search frequently requires observers to look for precisely defined targets among isolated, non-overlapping images randomly arrayed on clean backgrounds, medical images present unspecified targets in noisy, yet spatially regular scenes. Those unspecified targets are typically oddities, elements that do not belong. To develop a closer laboratory analogue to this, we created a database of scenes containing subtle, ill-specified "oddity" targets. These scenes have similar perceptual densities and spatial regularities to those found in expert search tasks, and each includes 16 variants of the unedited scene wherein an oddity (a subtle deformation of the scene) is hidden. In Experiment 1, eight volunteers searched thousands of scene variants for an oddity. Regardless of their search accuracy, they were then shown the highlighted anomaly and rated its subtlety. Subtlety ratings reliably predicted search performance (accuracy and response times) and did so better than image statistics. In Experiment 2, we conducted a conceptual replication in which a larger group of naïve searchers scanned subsets of the scene variants. Prior subtlety ratings reliably predicted search outcomes. Whereas medical image targets are difficult for naïve searchers to detect, our database contains thousands of interior and exterior scenes that vary in difficulty, but are nevertheless searchable by novices. In this way, the stimuli will be useful for studying visual search as it typically occurs in expert domains: Ill-specified search for anomalies in noisy displays.


Postmarketing Safety Profile of Subcutaneous Interferon Beta-1a Given 3 Times Weekly: A Retrospective Administrative Claims Analysis.
Health insurance administrative claims databases represent a valuable source of information regarding the safety profile of marketed products as used in actual clinical practice in a broader range of patients than that assessed in clinical trials. Interferon beta-1a administered subcutaneously 3 times weekly (IFN β-1a SC tiw), which was approved in 2002 by the FDA for the treatment of relapsing-remitting multiple sclerosis (MS), has over a decade of postmarketing experience. To date, however, its postmarketing safety profile has not been described using a real-world evidence source such as administrative claims data. To describe the safety profile of IFN β-1a SC tiw as presented in its U.S. prescribing information (PI) for patients with MS initiating IFN β-1a SC tiw therapy using data from U.S. health care administrative claims databases. This study featured an observational and retrospective "new start" cohort design using data from the Truven MarketScan Commercial and Medicare Supplemental health care administrative claims databases. Patients were eligible for inclusion if they were aged ≥ 18 years; had ≥ 1 diagnosis for MS recorded between January 1, 2006, and December 31, 2012; had ≥ 2 prescriptions for IFN β-1a SC tiw; and had ≥ 90 days of continuous eligibility pre-index date and ≥ 180 days of continuous eligibility post-index date. Patients with a prescription for IFN β-1a SC tiw without a MS diagnosis were excluded. Patients were followed from first prescription for IFN β-1a SC tiw (index date) until date of therapy switch or discontinuation, end of insurance eligibility, or end of observation period. Adverse events (AEs) examined were those listed in the Warnings and Precautions, Adverse Reactions, and Postmarketing Experience sections of the 2014 U.S. PI. Outcomes of interest were identified at the Medical Dictionary for Regulatory Activities (version 17.1) Preferred Term level and then coded to the corresponding ICD-9-CM criteria. Descriptive analyses of patient demographic, health status, health care utilization, and adherence status were performed, and incidence rates (IRs) per 100 person-years of labeled AEs with corresponding 95% CIs were calculated. The IR calculation was based on events that presented after therapy initiation and that were not present in the 90-day pre-index period. The top 6 AEs included influenza-like symptoms (IR = 15.65, 95% CI = 14.96-16.36); malaise (IR = 15.33, 95% CI = 14.65-16.04; fatigue (IR = 15.02, 95% CI = 14.35-15.72); abdominal pain (IR = 10.18, 95% CI = 9.67-10.70); chest pain (IR = 8.48, 95% CI = 8.03-8.95); and depression (IR = 7.75, 95% CI = 7.32-8.20). In contrast, the 6 lowest IRs were for maculo-papular rash (IR = 0.01, 95% CI = 0.00-0.04; injection-site necrosis (IR = 0.01, 95% CI = 0.00-0.03); erythema multiforme (IR = 0.01, 95% CI = 0.00-0.04); hypoesthesia (IR = 0.00, 95% CI = 0.00-0.02); Stevens-Johnson Syndrome (IR = 0.00, 95% CI = 0.00-0.02); and xerophthalmia (IR = 0.00, 95% CI = 0.00-0.02). Study results show strong convergence between the real-world safety profile of IFN β-1a SC tiw and its U.S. label. Our findings demonstrate the value of using real-world evidence obtained from administrative claims to complement clinical trial and postmarketing surveillance data in order to characterize the safety profile of established products, such as IFN β-1a SC tiw, in the postmarketing context.


Relationship between caregiving burden and depression in caregivers of individuals with intellectual disabilities in Korea.
Individuals with intellectual disabilities (IWID) need intensive long-term care. Consequently, caregivers experience not only economic burdens but also tension, depression, insecurity, etc. This study aimed to provide basic data and materials for preparing policy alternatives to improve caregiver quality of life of by examining relationships between IWID caregiving burden and depression. This study involved secondary analysis of data from the 2011 Survey on the Actual Conditions of Individuals with Developmental Disabilities, conducted by the Ministry of Health and Welfare of Korea. Effects of caregivers' characteristics, caregiving time and cost, and perceptions of time and cost, on depression were analyzed. All components of caregiving burden and depression were significantly higher in the 1st grade, designating the most severe disabilities. Overall, caregiving burden tended to be higher in female caregivers, the unemployed, parents, co-residents, and people with health problems. Caregivers' characteristics such as spouse relationship, having health problems, and perception of caregiving time and cost were associated with depression. Caregiving burden and depression vary depending on the characteristics of IWID and their caregivers. Services for decreasing caregiving burden may be optimized by focusing on these characteristics.


Abomasal displacement in cattle: influence of concentrates in the ration on fatty acid concentrations in ruminal, abomasal, and duodenal contents.
The primary causative factor in abomasal displacement appears to be atony or hypotony of the abomasum. Seemingly, high-concentrate feeding inhibits abomasal motility by increasing the amount of fatty acids which enter the abomasum. In the present study, cows with ruminal, abomasal, and duodenal fistulas were fed a hay ad libitum ration or a hay and concentration ration and were compared. It was shown that a considerable increase in ruminal volatile free fatty acid (VFFA) concentrations was not followed by a subsequent increase in abomasal VFFA concentrations. Differences in abomasal VFFA levels between the 2 rations could not be found. There was a slight, but insignificant, increase in duodenal VFFA concentrations after cows were fed the hay ration. One cow given the hay and concentrate diet had a small, but significant, increase in duodenal VFFA concentrations during the first 2 hours after feeding. The VFFA concentrations in duodenal fluid were too low to support the hypothesis that changes in duodenal VFFA concentrations could be responsible for abomasal hypotony.


Loss of Oct4 expression during the development of murine embryoid bodies.
We describe the internal organization of murine embryoid bodies (EBs) in terms of the structures and cell types formed as Oct4 expression becomes progressively lost. This is done by making the EBs from iPS cells carrying a novel Oct4 reporter (Oct4-MerCreMer;mTmG) which is inducible, sensitive, and permanent in all cellular progeny. When these EBs are treated with tamoxifen, the Oct4 expressing cells switch from a red to a green fluorescence color, and this is maintained thereafter by all their progeny. We show that there is no specific pattern in which Oct4 is downregulated, rather it appears to be spatially random. Many of the earliest cells to lose Oct4 expression stain positive for markers of visceral endoderm (DAB2, α-fetoprotein (AFP), HNF4). These are randomly located, although if endoderm differentiation is allowed to commence before EB formation then an external layer is formed. This is true both of EBs made from the reporter iPS cells, or from an embryo-derived mouse ES line (R1 cells). Markers of the early body axis, Brachyury (BRA) and FOXA2, usually showed a concentration of positive cells in one region of the EB, but the morphology is not predictable and there are also scattered cells expressing these markers. These patterns are similar in R1 cells. Use of the Oct4 reporter showed a difference between BRA and FOXA2. BRA, which marks the early mesoderm, node and notochord, arises in Oct4 expressing cells on days 3-4. FOXA2, which marks the floor plate of the neural tube and definitive endoderm, as well as the node and notochord, arises at the same time but mostly in cells that have already lost Oct4 expression. Several clumps of cardiomyocytes are visible by days 7-8 of EB development, both in our iPS cells and in R1 cells. Using the Oct4 reporter we show that the cells forming these clumps lose Oct4 expression between days 3 and 5. Overall, our results indicate that EBs recapitulate normal development quite well in terms of the tempo of events and the appearance of specific markers, but they do not resemble embryos in terms of their morphology.


Functional connectivity in elderly controls and AD patients using resting state fMRI: a pilot study.
Conventionally, Alzheimer's disease (AD) and other dementias are diagnosed using clinical assessment, neuropsychology and also structural neuroimaging, showing neuronal degeneration starting in the hippocampal regions. However, there is an increasing need for a new method that is more sensitive to early AD identification than currently possible. A new promising technique that may be used for this is to measure local brain activation using functional magnetic resonance imaging (fMRI), since functional loss predates structural loss of brain tissue. A new method to apply fMRI is to study connectivity between brain regions during a resting state without application of a task. Recent data suggest that connectivity within memory systems during such a resting state is associated with the level of memory function. Here we explain how we will study healthy elderly controls, patients with a mild cognitive impairment (MCI, considered to be a transitional stage between normal condition and AD), and AD patients using resting state connectivity fMRI. If resting state connectivity is sensitive to cognitive decline, this will be of great importance for noninvasive dementia research, offering a tool to easily study functional networks in the brain without the requirement of a memory task, and perhaps offering a tool sensitive for early diagnostics.


Spinal Cord Signal Change on Magnetic Resonance Imaging May Predict Worse Clinical In- and Outpatient Outcomes in Patients with Spinal Cord Injury: A Prospective Multicenter Study in 459 Patients.
Prognostic factors for clinical outcome after spinal cord (SC) injury (SCI) are limited but important in patient management and education. There is a lack of evidence regarding magnetic resonance imaging (MRI) and clinical outcomes in SCI patients. Therefore, we aimed to investigate whether baseline MRI features predicted the clinical course of the disease. This study is an ancillary to the prospective North American Clinical Trials Network (NACTN) registry. Patients were enrolled from 2005-2017. MRI within 72 h of injury and a minimum follow-up of one year were available for 459 patients. Patients with American Spinal Injury Association impairment scale (AIS) E were excluded. Patients were grouped into those with (n = 354) versus without (n = 105) SC signal change on MRI T2-weighted images. Logistic regression analysis adjusted for commonly known a priori confounders (age and baseline AIS). Main outcomes and measures: The primary outcome was any adverse event. Secondary outcomes were AIS at the baseline and final follow-up, length of hospital stay (LOS), and mortality. A regression model adjusted for age and baseline AIS. Patients with intrinsic SC signal change were younger (46.0 (interquartile range (IQR) 29.0 vs. 50.0 (IQR 20.5) years, p = 0.039). There were no significant differences in the other baseline variables, gender, body mass index, comorbidities, and injury location. There were more adverse events in patients with SC signal change (230 (65.0%) vs. 47 (44.8%), p < 0.001; odds ratio (OR) = 2.09 (95% confidence interval (CI) 1.31-3.35), p = 0.002). The most common adverse event was cardiopulmonary (186 (40.5%)). Patients were less likely to be in the AIS D category with SC signal change at baseline (OR = 0.45 (95% CI 0.28-0.72), p = 0.001) and in the AIS D or E category at the final follow-up (OR = 0.36 (95% CI 0.16-0.82), p = 0.015). The length of stay was longer in patients with SC signal change (13.0 (IQR 17.0) vs. 11.0 (IQR 14.0), p = 0.049). There was no difference between the groups in mortality (11 (3.2%) vs. 4 (3.9%)). MRI SC signal change may predict adverse events and overall LOS in the SCI population. If present, patients are more likely to have a worse baseline clinical presentation (i.e., AIS) and in- or outpatient clinical outcome after one year. Patients with SC signal change may benefit from earlier, more aggressive treatment strategies and need to be educated about an unfavorable prognosis.


Same-day batch measurement of glycine betaine, carnitine, and other betaines in biological material.
Glycine betaine, carnitine, carnitine esters, butyrobetaine, and proline betaine (stachydrine) concentrations in biological materials can be reliably measured in 100-microliters samples, with a detection limit below 1 mumol/liter. The procedure is suitable for batches of more than 30 specimens and it is possible to obtain a single result within 2 h. The betaines are extracted into an acetonitrile:methanol mixture, dried with anhydrous disodium hydrogen phosphate containing argentous oxide. The 4-bromophenacyl ester derivatives are formed using 4-bromophenacyl triflate as reagent, in the presence of solid magnesium oxide as base. The derivatives are separated by high-performance chromatography on a silica column, in a mixed partition and ion-exchange mode.


Global analysis of genomic instability caused by DNA replication stress in Saccharomyces cerevisiae.
DNA replication stress (DRS)-induced genomic instability is an important factor driving cancer development. To understand the mechanisms of DRS-associated genomic instability, we measured the rates of genomic alterations throughout the genome in a yeast strain with lowered expression of the replicative DNA polymerase δ. By a genetic test, we showed that most recombinogenic DNA lesions were introduced during S or G2 phase, presumably as a consequence of broken replication forks. We observed a high rate of chromosome loss, likely reflecting a reduced capacity of the low-polymerase strains to repair double-stranded DNA breaks (DSBs). We also observed a high frequency of deletion events within tandemly repeated genes such as the ribosomal RNA genes. By whole-genome sequencing, we found that low levels of DNA polymerase δ elevated mutation rates, both single-base mutations and small insertions/deletions. Finally, we showed that cells with low levels of DNA polymerase δ tended to accumulate small promoter mutations that increased the expression of this polymerase. These deletions conferred a selective growth advantage to cells, demonstrating that DRS can be one factor driving phenotypic evolution.


Aberrant T cell immunity triggered by human Respiratory Syncytial Virus and human Metapneumovirus infection.
Human Respiratory syncytial virus (hRSV) and human metapneumovirus (hMPV) are the two major etiological viral agents of lower respiratory tract diseases, affecting mainly infants, young children and the elderly. Although the infection of both viruses trigger an antiviral immune response that mediate viral clearance and disease resolution in immunocompetent individuals, the promotion of long-term immunity appears to be deficient and reinfection are common throughout life. A possible explanation for this phenomenon is that hRSV and hMPV, can induce aberrant T cell responses, which leads to exacerbated lung inflammation and poor T and B cell memory immunity. The modulation of immune response exerted by both viruses include different strategies such as, impairment of immunological synapse mediated by viral proteins or soluble factors, and the induction of pro-inflammatory cytokines by epithelial cells, among others. All these viral strategies contribute to the alteration of the adaptive immunity in order to increase the susceptibility to reinfections. In this review, we discuss current research related to the mechanisms underlying the impairment of T and B cell immune responses induced by hRSV and hMPV infection. In addition, we described the role each virulence factor involved in immune modulation caused by these viruses.


Evaluating Cost-Effective Methods for Rapid and Repeatable National Scale Detection and Mapping of Invasive Species Spread.
Invasive species can spread rapidly at local and national scales, creating significant environmental and economic impacts. A central problem in mitigation efforts is identifying methods that can rapidly detect invasive species in a cost-effective and repeatable manner. This challenge is particularly acute for species that can spread over large areas (>1 million km2). Wild pigs (Sus scrofa) are one of the most prolific invasive mammals on Earth and cause extensive damage to agricultural crops, native ecosystems, and livestock, and are reservoirs of disease. They have spread from their native range in Eurasia and North Africa into large areas of Australia, Africa, South America, and North America. We show that the range of invasive wild pigs has increased exponentially in Canada over the last 27 years following initial and ongoing releases and escapes from domestic wild boar farms. The cumulative range of wild pigs across Canada is 777,783 km2, with the majority of wild pig distribution occurring in the Prairie Provinces. We evaluate eight different data collection and evaluation/validation methods for mapping invasive species over large areas, and assess their benefits and limitations. Our findings effectively map the spread of a highly invasive large mammal and demonstrate that management efforts should ideally rely on a set of complementary independent monitoring methods. Mapping and evaluating resulting species occurrences provide baseline maps against which future changes can be rapidly evaluated.


A randomized, blinded, clinical investigation of breath odor reduction efficacy of a stabilized chlorine-dioxide containing flavored mouthwash.
To evaluate the efficacy of a flavored, non-fluoridated, alcohol-free mouthwash containing 0.1% chlorine dioxide in reducing oral malodor. This was a randomized, 8-week, single site, double blind, crossover design with a 2-week washout period between crossover phases. Fifty subjects with clinically diagnosed intrinsic oral malodor were enrolled according to inclusion/exclusion criteria and randomized to one of two groups. Washout period initiated at end of Phase I and crossover design implemented prior to Phase II. Calibration for organoleptic judges performed at baseline for both phases. 48 subjects completed the study. No significant differences in intensity scores at baseline were found for both groups during both phases (P> 0.05). Within group comparisons for placebo revealed no significant differences with organoleptic intensity scores for all visits during both phases (P> 0.05). During Phase I, the mean changes in organoleptic scores for the test group were significantly different from the baseline at each visit: Weeks 1 to 3 (P< 0.05). After crossover, significant differences were found for the last two visits: Weeks 7 and 8 (P< 0.05). No adverse effects to oral tissues were observed or reported. This product is safe to use for up to 3 weeks and resulted in a decrease in oral malodor. Results suggested that twice-daily use of a 0.1% chlorine dioxide-containing flavored mouthwash, in conjunction with normal oral hygiene care, provided clinically relevant improvements in oral malodor for up to 3 weeks.


Steroidogenesis during in vitro maturation of bovine cumulus oocyte complexes and possible effects of tri-butyltin on granulosa cells.
Steroids are known as important factors on the route of oocytes development and cumulus oocyte complexes (COC) as well as follicular granulosa cells (GC) are suggested to be themselves involved in steroidogenesis. The aim of this study was to characterize such a local sex steroidogenic system during in vitro maturation (IVM) of bovine COCs according to the production of estradiol (E), testosterone (T) and progesterone (P). The expression of two steroid-converting key-enzymes was measured in parallel by quantitative RT-PCR. Furthermore, possible effects of the environmental pollutant tri-butyltin (TBT) were elucidated for the first time on bovine COC and GC in vitro concerning that steroidogenic system. During IVM of bovine COCs concentrations of P increased continuously, corresponding with steady-state levels of 3-beta-hydroxy-steroid-dehydrogenase (HSD) transcripts. In contrast, E together with P450 aromatase mRNA (ARO) increased in the first hours of IVM but declining thereafter, whereas T reached almost balanced levels. However, TBT showed only slight effects during IVM of COC. In cultured GC, LH caused highest P- and E-production within 24h and treatment with 50pM TBT induced a significant decrease of E in contrast to 100pM TBT and the control. These results indicate, that (1) COCs were able to modulate their steroidogenic environment in vitro and that (2) TBT may possibly influence or disturb steroidogenesis in the cows reproductive tract shown here for GC.


Strain Reversal in Actuated Origami Structures.
Origami in engineering is gaining interest for its potential as deployable or shape-adaptive structures. Practical systems could employ a network of actuators distributed across the structure to induce these deformations. Selecting the actuator locations requires an understanding of how the effect of a single actuator propagates spatially in an origami structure. We combine experimental results, finite element analysis, and reduced-order bar-and-hinge models to show how a localized static actuation decays elastically in Miura-ori tubes and sheets. We observe a strain reversal, before the origami structure springs back to the initial configuration further away from the point of actuation. The strain reversal is the result of bending of the facets, while the spring back requires in-plane facet deformations.