Attraction basins as gauges of robustness against boundary conditions in biological complex systems.
One fundamental concept in the context of biological systems on which researches have flourished in the past decade is that of the apparent robustness of these systems, i.e., their ability to resist to perturbations or constraints induced by external or boundary elements such as electromagnetic fields acting on neural networks, micro-RNAs acting on genetic networks and even hormone flows acting both on neural and genetic networks. Recent studies have shown the importance of addressing the question of the environmental robustness of biological networks such as neural and genetic networks. In some cases, external regulatory elements can be given a relevant formal representation by assimilating them to or modeling them by boundary conditions. This article presents a generic mathematical approach to understand the influence of boundary elements on the dynamics of regulation networks, considering their attraction basins as gauges of their robustness. The application of this method on a real genetic regulation network will point out a mathematical explanation of a biological phenomenon which has only been observed experimentally until now, namely the necessity of the presence of gibberellin for the flower of the plant Arabidopsis thaliana to develop normally.


The shark-tuna dichotomy: why tuna lay tiny eggs but sharks produce large offspring.
Teleosts such as tunas and billfish lay millions of tiny eggs weighing on the order of 0.001 g, whereas chondrichthyes such as sharks and rays produce a few eggs or live offspring weighing about 2% of adult body mass, as much as 10 000 g in some species. Why are the strategies so extreme, and why are intermediate ones absent? Building on previous work, we show quantitatively how offspring size reflects the relationship between growth and death rates. We construct fitness contours as functions of offspring size and number, and show how these can be derived from juvenile growth and survivorship curves. Convex contours, corresponding to Pearl Type 1 and 2 survivorship curves, select for extremes, either miniscule or large offspring; concave contours select for offspring of intermediate size. Of particular interest are what we call critical straight-line fitness contours, corresponding to log-linear Pearl Type 3 survivorship curves, which separate regimes that select for opposite optimal offspring sizes.


Combined electroencephalography-functional magnetic resonance imaging and electrical source imaging improves localization of pediatric focal epilepsy.
Surgical treatment in epilepsy is effective if the epileptogenic zone (EZ) can be correctly localized and characterized. Here we use simultaneous electroencephalography-functional magnetic resonance imaging (EEG-fMRI) data to derive EEG-fMRI and electrical source imaging (ESI) maps. Their yield and their individual and combined ability to (1) localize the EZ and (2) predict seizure outcome were then evaluated. Fifty-three children with drug-resistant epilepsy underwent EEG-fMRI. Interictal discharges were mapped using both EEG-fMRI hemodynamic responses and ESI. A single localization was derived from each individual test (EEG-fMRI global maxima [GM]/ESI maximum) and from the combination of both maps (EEG-fMRI/ESI spatial intersection). To determine the localization accuracy and its predictive performance, the individual and combined test localizations were compared to the presumed EZ and to the postsurgical outcome. Fifty-two of 53 patients had significant maps: 47 of 53 for EEG-fMRI, 44 of 53 for ESI, and 34 of 53 for both. The EZ was well characterized in 29 patients; 26 had an EEG-fMRI GM localization that was correct in 11, 22 patients had ESI localization that was correct in 17, and 12 patients had combined EEG-fMRI and ESI that was correct in 11. Seizure outcome following resection was correctly predicted by EEG-fMRI GM in 8 of 20 patients, and by the ESI maximum in 13 of 16. The combined EEG-fMRI/ESI region entirely predicted outcome in 9 of 9 patients, including 3 with no lesion visible on MRI. EEG-fMRI combined with ESI provides a simple unbiased localization that may predict surgery better than each individual test, including in MRI-negative patients. Ann Neurol 2017;82:278-287.


DUSP4 is associated with increased resistance against anti-HER2 therapy in breast cancer.
The majority of patients develop resistance against suppression of HER2-signaling mediated by trastuzumab in HER2 positive breast cancer (BC). HER2 overexpression activates multiple signaling pathways, including the mitogen-activated protein kinase (MAPK) cascade. MAPK phosphatases (MKPs) are essential regulators of MAPKs and participate in many facets of cellular regulation, including proliferation and apoptosis. We aimed to identify whether differential MKPs are associated with resistance to targeted therapy in patients previously treated with trastuzumab. Using gene chip data of 88 HER2-positive, trastuzumab treated BC patients, candidate MKPs were identified by Receiver Operator Characteristics analysis performed in R. Genes were ranked using their achieved area under the curve (AUC) values and were further restricted to markers significantly associated with worse survival. Functional significance of the two strongest predictive markers was evaluated in vitro by gene silencing in HER2 overexpressing, trastuzumab resistant BC cell lines SKTR and JIMT-1. The strongest predictive MKPs were DUSP4/MKP-2 (AUC=0.75, p=0.0096) and DUSP6/MKP-3 (AUC=0.77, p=5.29E-05). Higher expression for these correlated to worse survival (DUSP4: HR=2.05, p=0.009 and DUSP6: HR=2, p=0.0015). Silencing of DUSP4 had significant sensitization effects - viability of DUSP4 siRNA transfected, trastuzumab treated cells decreased significantly compared to scramble-siRNA transfected controls (SKTR: p=0.016; JIMT-1: p=0.016). In contrast, simultaneous treatment with DUSP6 siRNA and trastuzumab did not alter cell proliferation. Our findings suggest that DUSP4 may represent a new potential target to overcome trastuzumab resistance.


Safety of subcutaneous octreotide in patients with sulfonylurea-induced hypoglycemia and congestive heart failure.
To report the safe use of subcutaneous octreotide in the treatment of sulfonylurea-induced hypoglycemia in 2 patients with severe congestive heart failure. Two patients with congestive heart failure with systolic dysfunction presented with severe hypoglycemia (blood glucose level: patient 1, 31 mg/dL; patient 2, 36 mg/dL) secondary to sulfonylurea agents. The mechanism of hypoglycemia was poor oral intake and prolonged half-life of the drugs due to renal failure. Hypoglycemia was refractory to glucose supplementation. Patient 1 received 2 doses of octreotide 50 microg subcutaneously 12 hours apart, with resolution of hypoglycemia (blood glucose level: <33 mg/dL before administration of octreotide, 62 mg/dL after first dose, 121 mg/dL after second dose). Patient 2 received an initial dose of octreotide 25 microg subcutaneously; hypoglycemia persisted after this dose, and the patient was given 2 doses of 50 microg subcutaneously 12 hours apart, leading to resolution of hypoglycemia (blood glucose level: 57 mg/dL before administration of octreotide, 80 mg/dL after first dose, 85 mg/dL after second dose, 146 mg/dL after third dose). Resolution of hypoglycemia with octreotide treatment obviated the need for prolonged intravenous dextrose infusions. No local or systemic adverse effects, especially cardiac adverse events, were noted during the course of treatment. Octreotide is considered the antidote for sulfonylurea-induced hypoglycemia. It may be especially useful in patients with congestive heart failure who may not tolerate intravenous infusions of dextrose. The package insert cautions of the occurrence of cardiac adverse effects such as bradycardia, heart block, and worsening heart failure. Patients with heart failure may theoretically be at a higher risk of these adverse effects due to the concurrent use of atrioventricular blocking agents. However, a review of current literature reveals that the adverse effect profile depends on the route, dose, and formulation of the octreotide used along with the clinical indication. These adverse effects may not be clinically significant with the doses used for treatment of sulfonylurea-induced hypoglycemia and the benefits of the drug may outweigh the risks. In these cases, octreotide was safely and effectively used in the treatment of sulfonylurea-induced hypoglycemia in patients with congestive heart failure by adhering to dosing guidelines and close monitoring.


Growth hormone transgenesis does not influence territorial dominance or growth and survival of first-feeding Atlantic salmon Salmo salar in food-limited stream microcosms.
This study explored the relative competitive ability and performance of first-feeding growth hormone (GH) transgenic and non-transgenic Atlantic salmon Salmo salar fry under low food conditions. Pair-wise dominance trials indicated a strong competitive advantage for residents of a contested foraging territory. Transgenic and non-transgenic individuals, however, were equally likely to be dominant. Similarly, in stream environments with limited food, the transgene did not influence the growth in mass or survival at high or low fry densities. Fry in low-density treatments, however, performed better than fry in high-density treatments. These results indicate that, under the environment examined, the growth performance of GH-transgenic and non-transgenic S. salar may be similar during first feeding, an intense period of selection in their life history. Similarities in competitive ability and growth performance with wild-type fish suggest that the capacity of transgenic S. salar to establish in natural streams may not be inhibited during early life history.


Established gastric cancer cell lines transplantable into C57BL/6 mice show fibroblast growth factor receptor 4 promotion of tumor growth.
Previously no mouse gastric cancer cell lines have been available for transplantation into C57BL/6 mice. However, a gastric cancer model in immunocompetent mice would be useful for analyzing putative therapies. N-Methyl-N-nitrosourea (MNU) was given in drinking water to C57BL/6 mice and p53 heterozygous knockout mice. Only 1 tumor from a p53 knockout mouse could be cultured and the cells s.c. transplanted into a C57BL/6 mouse. We cultured this s.c. tumor, and subcloned it. mRNA expression in the most aggressive YTN16 subline was compared to the less aggressive YTN2 subline by microarray analysis, and fibroblast growth factor receptor 4 (FGFR4) in YTN16 cells was knocked out with a CRISPR/Cas9 system and inhibited by an FGFR4 selective inhibitor, BLU9931. These transplanted cell lines formed s.c. tumors in C57BL/6 mice. Four cell lines (YTN2, YTN3, YTN5, YTN16) were subcloned and established. Their in vitro growth rates were similar. However, s.c. tumor establishment rates, metastatic rates, and peritoneal dissemination rates of YTN2 and YTN3 were lower than for YTN5 and YTN16. YTN16 established 8/8 s.c. tumors, 7/8 with lung metastases, 3/8 with lymph node metastases and 5/5 with peritoneal dissemination. FGFR4 expression by YTN16 was 121-fold higher than YTN2. FGFR4-deleted YTN16 cells failed to form s.c. tumors and showed lower rates of peritoneal dissemination. BLU9931 significantly inhibited the growth of peritoneal dissemination of YTN16. These studies present the first transplantable mouse gastric cancer lines. Our results further indicate that FGFR4 is an important growth signal receptor in gastric cancer cells with high FGFR4 expression.


Nursing informatics educational needs.
A survey was conducted among 497 respondents in a northeastern metropolitan area to determine the informatics educational needs of professional nurses. All subjects were asked to indicate on a four-point scale their current knowledge and their desired knowledge in 23 content areas. The population was subdivided into three subsamples based on job classification. There were differences in the informatics educational needs of nurse educators, nurse managers, and informatics nurses. There was interest in returning to school for a graduate degree or certificate in nursing informatics by a majority of the sample, although 71% of the respondents already possess a higher degree.


The transcription factor EMISSION OF BENZENOIDS II activates the MYB ODORANT1 promoter at a MYB binding site specific for fragrant petunias.
Fragrance production in petunia flowers is highly regulated. Two transcription factors, ODORANT1 (ODO1) and EMISSION OF BENZENOIDS II (EOBII) have recently been identified as regulators of the volatile benzenoid/phenylpropanoid pathway in petals. Unlike the non-fragrant Petunia hybrida cultivar R27, the fragrant cultivar Mitchell highly expresses ODO1. Using stable reporter lines, we identified the 1.2-kbp ODO1 promoter from Mitchell that is sufficient for tissue-specific, developmental and rhythmic expression. This promoter fragment can be activated in non-fragrant R27 petals, indicating that the set of trans-acting factors driving ODO1 expression is conserved in these two petunias. Conversely, the 1.2-kbp ODO1 promoter of R27 is much less active in Mitchell petals. Transient transformation of 5' deletion and chimeric Mitchell and R27 ODO1 promoter reporter constructs in petunia petals identified an enhancer region, which is specific for the fragrant Mitchell cultivar and contains a putative MYB binding site (MBS). Mutations in the MBS of the Mitchell promoter decreased overall promoter activity by 50%, highlighting the importance of the enhancer region. We show that EOBII binds and activates the ODO1 promoter via this MBS, establishing a molecular link between these two regulators of floral fragrance biosynthesis in petunia.


Strong Father-Child Relationships and Other Positive Childhood Experiences, Adverse Childhood Experiences, and Sexual Risk Factors for HIV among Young Adults Aged 19-24 Years, Namibia, 2019: A Cross-Sectional Study.
Using cross-sectional data from the 2019 Namibia Violence Against Children and Youth Survey and sex-stratified multivariable models, we assessed the associations between four different positive childhood experiences (PCEs) and having ≥3 adverse childhood experiences (ACEs), including ≥3 ACE-PCE interaction terms, and seven sexual risk factors for HIV acquisition among young adults aged 19-24 years. One PCE, having a strong father-child relationship, was inversely associated with two risk factors among women (lifetime transactional sex (OR, 0.4; 95% CI, 0.2-0.7) and recent age-disparate sexual relationships (OR, 0.3; 95% CI, 0.2-0.5)), and significantly interacted with having ≥3 ACEs for three risk factors among women (not knowing a partner's HIV status, infrequently using condoms, and ever having an STI) and one among men (having multiple sexual partners in the past year). The other PCEs were significantly associated with ≤1 HIV risk factor and had no significant interaction terms. Strong father-child relationships may reduce HIV acquisition risk and mitigate the effect of childhood adversity on HIV risk among young adults in Namibia.


Continuous mixed venous oxygen saturation, not mean blood pressure, is associated with early bupivacaine cardiotoxicity in dogs.
To investigate changes of continuous mixed venous oxygen saturation (cSvO(2)) and mean arterial blood pressure (MBP) in dogs with bupivacaine-induced cardiac depression. Bupivacaine was infused into pentobarbital-anesthetized mongrel dogs (n = 8) at a rate of 0.5 mg.kg(-1).min(-1) until the MBP was 40 mmHg or less (end of bupivacaine infusion; BIE). The infusion time was divided into the early period, first 30 min of bupivacaine infusion and the late period, which was from 30 min of bupivacaine infusion until BIE. cSvO(2) was monitored using a fibreoptic pulmonary artery catheter, and MBP and cardiac output (CO) were measured every ten minutes after the initiation of bupivacaine infusion. Arterial blood gas, serum electrolyte and bupivacaine concentration were measured simultaneously. The relationships between CO and cSvO(2), and of CO vs MBP were compared by regression analysis in the early and late periods. The Pearson's correlation coefficients between CO and cSvO(2) were 0.782 (P = 2.1 x 10(-7)) in the early period and 0.824 (P = 1.3 x 10(-6)) in the late period. The correlation coefficients between CO and MBP were 0.019 (P = 0.921) in the early period and 0.799 (P = 4.8 x 10(-6)) in the late period. cSvO(2), but not MBP, is associated with CO changes in bupivacaine-induced cardiac depression during the early period of bupivacaine intoxication. Decrease of MBP with low cSvO(2) observed during the late period might imply severe cardiac depression induced by bupivacaine infusion.


Unique suites of trabecular bone features characterize locomotor behavior in human and non-human anthropoid primates.
Understanding the mechanically-mediated response of trabecular bone to locomotion-specific loading patterns would be of great benefit to comparative mammalian evolutionary morphology. Unfortunately, assessments of the correspondence between individual trabecular bone features and inferred behavior patterns have failed to reveal a strong locomotion-specific signal. This study assesses the relationship between inferred locomotor activity and a suite of trabecular bone structural features that characterize bone architecture. High-resolution computed tomography images were collected from the humeral and femoral heads of 115 individuals from eight anthropoid primate genera (Alouatta, Homo, Macaca, Pan, Papio, Pongo, Trachypithecus, Symphalangus). Discriminant function analyses reveal that subarticular trabecular bone in the femoral and humeral heads is significantly different among most locomotor groups. The results indicate that when a suite of femoral head trabecular features is considered, trabecular number and connectivity density, together with fabric anisotropy and the relative proportion of rods and plates, differentiate locomotor groups reasonably well. A similar, yet weaker, relationship is also evident in the trabecular architecture of the humeral head. The application of this multivariate approach to analyses of trabecular bone morphology in recent and fossil primates may enhance our ability to reconstruct locomotor behavior in the fossil record.


Substrate specificity of human deoxycytidine kinase toward antiviral 2',3'-dideoxynucleoside analogs.
Deoxycytidine (dCyd) kinase has been purified to homogeneity from human leukemic spleen, and the capacity of the enzyme to phosphorylate 2',3'-dideoxynucleoside (ddN) analogs that are clinically effective inhibitors of human immunodeficiency virus (HIV) replication was evaluated. Cytosine-containing ddN analogs, such as 2',3'-dideoxycytidine, 2',3'-dideoxy-2',3'-dehydrocytidine, and cytallene, were efficiently phosphorylated by dCyd kinase, while no phosphorylation of purine-containing ddN analogs was detected. dCyd kinase was completely inactive toward 2',3'-dideoxyadenosine (ddAdo), 2',3'-dideoxyinosine, 2',3'-dideoxyguanosine, and adenallene, although it was capable of phosphorylating both 2'-deoxyadenosine (dAdo) and 2'-deoxyguanosine (dGuo). The abilities of wild type and mutant human T lymphoblastoid CEM cells to accumulate ddAdo in situ and in vitro were also ascertained. Comparison of the abilities of intact wild type CEM cells and derivatives deficient in nucleoside transport, dCyd kinase, and/or adenosine (Ado) kinase to accumulate [3H]ddAdo-derived radioactivity revealed no significant differences among the wild type and mutant strains. However, ddAdo phosphorylating activity was decreased in extracts from Ado kinase-deficient cells but not in lysates prepared from cells genetically deficient in dCyd kinase activity. In comparative growth rate experiments, wild type, nucleoside transport-deficient, and dCyd kinase-deficient CEM cells were equally sensitive to ddAdo toxicity, while, interestingly, a deficiency in Ado kinase correlated with a 5-fold decreased growth sensitivity to the purine ddN. Insertion of an adenine phosphoribosyltransferase deficiency into the CEM cell lines did not influence ddAdo toxicity or incorporation rate. These results imply that Ado kinase may be an important factor in ddAdo phosphorylation by CEM cells. Furthermore, these studies demonstrate that cytosine- and purine-containing ddNs are transported and activated by independent pathways and, therefore, have important implications for anti-HIV therapy in that pyrimidine and purine ddNs might be used in combination for the treatment of acquired immunodeficiency syndrome.


3D-SHOTGUN: a novel, cooperative, fold-recognition meta-predictor.
To gain a better understanding of the biological role of proteins encoded in genome sequences, knowledge of their three-dimensional (3D) structure and function is required. The computational assignment of folds is becoming an increasingly important complement to experimental structure determination. In particular, fold-recognition methods aim to predict approximate 3D models for proteins bearing no sequence similarity to any protein of known structure. However, fully automated structure-prediction methods can currently produce reliable models for only a fraction of these sequences. Using a number of semiautomated procedures, human expert predictors are often able to produce more and better predictions than automated methods. We describe a novel, fully automatic, fold-recognition meta-predictor, named 3D-SHOTGUN, which incorporates some of the strategies human predictors have successfully applied. This new method is reminiscent of the so-called cooperative algorithms of Computer Vision. The input to 3D-SHOTGUN are the top models predicted by a number of independent fold-recognition servers. The meta-predictor consists of three steps: (i) assembly of hybrid models, (ii) confidence assignment, and (iii) selection. We have applied 3D-SHOTGUN to an unbiased test set of 77 newly released protein structures sharing no sequence similarity to proteins previously released. Forty-six correct rank-1 predictions were obtained, 30 of which had scores higher than that of the first incorrect prediction-a significant improvement over the performance of all individual servers. Furthermore, the predicted hybrid models were, on average, more similar to their corresponding native structures than those produced by the individual servers. This opens the possibility of generating more accurate, full-atom homology models for proteins with no sequence similarity to proteins of known structure. These improvements represent a step forward toward the wider applicability of fully automated structure-prediction methods at genome scales.


[Clinical-epidemiological characteristics of a group of HIV/AIDS infected women in Salvador-Bahia].
The increasing number of HIV/AIDS infected women is one of the main characteristics of the AIDS epidemic, besides the wide expansion of the disease among the poor. This study focused on a group of 82 women with AIDS from Salvador-Bahia. The mean age was 32 +/- 7.2 years, 75% had children, 82.5% were jobless, and 77.8% were illiterate or only had basic education. The number of sexual partners was five or fewer in 71% of cases and 25% had only one sexual partner. Forty-six (55.4%) women reported up to 77 episodes of opportunist infections. The majority of the patients were from the lower classes, with no good education, a low income and few sexual partners. The main via of transmission was heterosexual, and many of the women were infected by their steady partner/husband.


Physiological adaptations to concurrent endurance training and low velocity resistance training.
This study investigated the effects of concurrent endurances and low velocity resistance training (LVR) on measures of strength and aerobic endurance. One group (ES) performed concurrent endurance training 3 days a week and LVR training on alternate days, 3 days a week for 12 weeks. The other group (S) performed only LVR training 3 days a week for 12 weeks without any endurance training. Measurements and increases in training volume were made every three weeks in both groups. Group ES exhibited increases in submaximal exercise responses after 3, 9 and 12 weeks (p less than 0.05). Knee extension peak torque and total work as well as cross-sectional area of quadriceps femoris were significantly increased after 6 and 9 weeks of training in both groups. These findings indicate that no significant differences in strength gains were observed between subjects performing concurrent endurance and resistance training or resistance training only. However, the time-course of adaptations between groups was somewhat different.


Intestinal and Hepatocellular Transporters: Therapeutic Effects and Drug Interactions of Herbal Supplements.
Herbal supplements are generally considered safe; however, drug disposition is influenced by the interactions of herbal supplements and food constituents with transport and metabolic processes. Although the interference of herbal supplements with drug metabolism has been studied extensively, knowledge of how they interact with the drug transport processes is less advanced. Therefore, we describe here specific examples of experimental and human interaction studies of herbal supplement components with drug transporters addressing, for example, organic anion transporting polypeptides or P-glycoprotein, as such interactions may lead to severe side effects and altered drug efficacy. Hence, it is clearly necessary to increase the awareness of the clinical relevance of the interference of herbal supplements with the drug transport processes.


Longitudinal Associations Among Older Adults' Neurocognitive Performance, Psychological Distress, and Self-Reported Cognitive Function.
Subjective cognitive complaints increase with age. Although subjective cognitive difficulties have been linked to cognitive impairment and psychological distress, some studies have failed to establish a link between subjective cognitive complaints and present or future cognitive impairment. The present study examined the interactive, longitudinal effects of age, psychological distress, and objective cognitive performance on subjective cognitive function. Older adults (N=147, Mage = 74.17) were assessed biannually for up to six years. Subjective cognitive function, psychological distress, and neuropsychological testing were obtained at each assessment. In multilevel models with single predictors, age, poorer average task-switching and poorer memory predicted worse subjective cognitive functioning. Both average levels and within-person deviations in distress predicted worse subjective cognitive function. There were two significant interactions: one between average distress and chronological age, and the other between average memory and within-person distress. Task switching performance and distress had an additive effect on subjective cognitive function. Both individual differences (i.e., between-person differences) and fluctuations over time (i.e., within-person changes) contributed to worse subjective cognitive function. Psychological distress may help explain the relationship between objective cognitive performance and subjective cognitive function and should be assessed when patient concerns about cognitive functioning arise.


A retrospective review of oral low-dose sirolimus (rapamycin) for the treatment of active uveitis.
The purpose of this study is to elicit the role of oral low-dose sirolimus as a corticosteriod-sparing agent for active uveitis. A retrospective, interventional case series was performed by reviewing the clinical records of all patients treated with oral, low-dose sirolimus (1-4 mg daily) for severe uveitis. Data reviewed included symptomatic improvement, Snellen best-corrected visual acuity, corticosteroid requirement, sirolimus levels, intraocular inflammation, spectral-domain optical coherence tomography, and fluorescein angiogram. Primary outcome measures were determined by the ability to decrease the intraocular inflammation, corticosteroid requirement, and frequency of flares. Eight patients with varied diagnoses were treated with oral low-dose sirolimus for severe, chronic uveitis between 2008 and 2010. In four of the eight patients, there was an improvement of all primary outcome measures. While sirolimus monotherapy was successful in only one patient, a sirolimus/methotrexate combination was successful in three patients. Although there was a good initial response in three patients, treatment was a failure after serious side effects forced the cessation of sirolimus therapy. One patient was lost to follow-up. Sirolimus may have a limited role in severe uveitis as an adjunct corticosteroid-sparing agent in combination with more standard immunosuppressive agents. Oral low-dose sirolimus appeared to be better tolerated than higher doses, but there were a significant number of adverse events, requiring therapy to be stopped.


Magnetic Resonance Neurography of the Lumbosacral Plexus in Failed Back Surgery Syndrome.
Retrospective clinical case series. To study the role of magnetic resonance neurography (MRN) of the lumbosacral plexus in management of patients with failed back surgery syndrome (FBSS). FBSS is one of the major problems in health care, affecting up to 40% of patients after spine surgery. To date, no imaging modality has been used to effectively classify nerve compression, because nerve injuries are challenging to detect on conventional lumbar spine magnetic resonance imaging (MRI). To our knowledge, no previous studies have addressed the use of MRN in FBSS or compared it to lumbar spine MRI. From 203 consecutive 3 T MRN studies of lumbosacral plexus in 1 year, 12% (25/203) presented as FBSS. Demographic data, number of previous lumbar MRIs and their findings, MRN findings, interval between MRI and MRN, pre-and post-MRN diagnosis, pain levels, and treatments were recorded. Changes in diagnosis, treatment, and outcomes after MRN were determined. The final sample of 25 patients had a mean age 62 ± 15 and male to female ratio 1:1.08. Approximately 88% (22/25) had previous lumbar MRI, of which 27% had 3 or more. Most common imaging findings were neuroforaminal stenosis 22.6% (7/31) on MRI and neuropathy 22.9% (19/83) on MRN. Mean interval between MRI and MRN was 13.9 ± 28.3 months. Lumbar MRIs were inconclusive in 36% (8/22). MRN detected 63% (52/83) more findings and changed the diagnosis and treatment in 12% and 48% of FBSS cases, respectively. Favorable outcomes were recorded in 40% to 67% of patients following MRN-guided treatments. FBSS is a complex problem and MRN of lumbosacral plexus impacts its management by better directing source of symptoms. 4.


CBL is frequently altered in lung cancers: its relationship to mutations in MET and EGFR tyrosine kinases.
Non-small cell lung cancer (NSCLC) is a heterogeneous group of disorders with a number of genetic and proteomic alterations. c-CBL is an E3 ubiquitin ligase and adaptor molecule important in normal homeostasis and cancer. We determined the genetic variations of c-CBL, relationship to receptor tyrosine kinases (EGFR and MET), and functionality in NSCLC. Using archival formalin-fixed paraffin embedded (FFPE) extracted genomic DNA, we show that c-CBL mutations occur in somatic fashion for lung cancers. c-CBL mutations were not mutually exclusive of MET or EGFR mutations; however they were independent of p53 and KRAS mutations. In normal/tumor pairwise analysis, there was significant loss of heterozygosity (LOH) for the c-CBL locus (22%, n = 8/37) and none of these samples revealed any mutation in the remaining copy of c-CBL. The c-CBL LOH also positively correlated with EGFR and MET mutations observed in the same samples. Using select c-CBL somatic mutations such as S80N/H94Y, Q249E and W802* (obtained from Caucasian, Taiwanese and African-American samples, respectively) transfected in NSCLC cell lines, there was increased cell viability and cell motility. Taking the overall mutation rate of c-CBL to be a combination as somatic missense mutation and LOH, it is clear that c-CBL is highly mutated in lung cancers and may play an essential role in lung tumorigenesis and metastasis.


Transplantation effects of dental pulp-derived cells on peripheral nerve regeneration in crushed sciatic nerve injury.
The effects of transplanted human dental pulp-derived cells (DPCs) on peripheral nerve regeneration were studied in a rat model of sciatic nerve crush injury. In one group, DPCs were transplanted into the compression site (cell transplantation group); the control group underwent no transplantation (crushed group). Sciatic nerve regeneration was determined based on the recovery of motor function and histological and immunohistochemical analyses. The cell transplantation group showed improved motor function compared with the crushed group using the CatWalk XT system, which corresponded to a higher ratio of tibialis to anterior muscle weight 14 days after surgery. Histological analysis revealed a smaller interspace area and few vacuoles in the sciatic nerve after cell transplantation compared with the crushed group. The myelin sheath was visualized with Luxol Fast Blue (LFB) staining and anti-myelin basic protein (anti-MBP) antibody labeling; the percentages of LFB- and MBP-positive areas were higher in the cell transplantation group than in the crushed group. Human mitochondria-positive cells were also identified in the sciatic nerve at the transplantation site 14 days after surgery. Taken together, the observed correlation between morphological findings and functional outcomes following DPC transplantation indicates that DPCs promote peripheral nerve regeneration in rats.


Organisation of care for pregnancy in patients with congenital heart disease.
Improvements in surgery have resulted in more women with repaired congenital heart disease (CHD) surviving to adulthood. Women with CHD, who wish to embark on pregnancy require prepregnancy counselling. This consultation should cover several issues such as the long-term prognosis of the mother, fertility and miscarriage rates, recurrence risk of CHD in the baby, drug therapy during pregnancy, estimated maternal risk and outcome, expected fetal outcomes and plans for pregnancy. Prenatal genetic testing is available for those patients with an identified genetic defect using pregestational diagnosis or prenatal diagnosis chorionic villus sampling or amniocentesis. Centralisation of care is needed for high-risk patients. Finally, currently there are no recommendations addressing the issue of the delivery. It is crucial that a dedicated plan for delivery should be available for all cardiac patients. The maternal mortality in low-income to middle-income countries is 14 times higher than in high-income countries and needs additional aspects and dedicated care.


Response of the rat intestinal microcirculation to experimental endotoxemia is attenuated by ampicillin but not by its derivative KKP723.
KKP723 (KKP), a derivative of ampicillin, is a newly developed beta-lactam antibiotic. Using an experimental endotoxemia model, the intestinal microcirculation in four groups of animals were evaluated using intravital microscopy (IVM). The groups included were a control group, an endotoxemic group (15 mg/kg i.v. LPS from E. coli), an ampicillin (50 mg/kg i.v.) treated endotoxemic group and an endotoxemic group treated with KKP (67.4 mg/kg i.v.). Ampicillin treatment resulted in a significant reduced number of firmly adhering leukocytes in intestinal submucosal venules. KKP treatment did not show this effect on leukocyte activation. We found no changes of the functional capillary density (FCD) of the intestinal wall by treatment with ampicillin or its derivative KKP. The increased leukocyte adherence in the KKP treated LPS animals may be explained by a loss of a possible ampicillin-related anti-inflammatory effect by the biotransformation process. The endotoxemia IVM model is useful to detect effects of antibiotics in an impaired microcirculation.


Dynamic Magnetic Resonance Vascular Fingerprinting During Hypercapnia for Quantitative and Multiparametric Cerebrovascular Reactivity Measures.
Magnetic resonance fingerprinting (MRF) represents a potential paradigm shift in MR image acquisition, reconstruction, and analysis using computational biophysical modelling in parallel to image acquisition. Its flexibility allows for examination of cerebrovascular metrics through MR vascular fingerprinting (MRvF), and this has been extended even further to produce quantitative cerebral blood volume (CBV), microvascular vessel radius, and tissue oxygen saturation (SO2) maps of the whole brain simultaneously every few seconds. This allows for observation of rapid physiological changes like cerebrovascular reactivity (CVR), which is the ability of vessels to dilate in response to a vasoactive stimulus. Here we demonstrated a novel protocol in which a rapid, spin- and gradient-echo pulse sequence allowed for dynamic, and simultaneous acquisition of MRvF and blood oxygen level dependent (BOLD) measures. By combining this with a tailored hypercapnic (5% CO2) breathing paradigm we were able to show how these quantitative CBV, radius, and SO2 parameters changed in response to a stimulus and directly compare those to a colocalized, traditionally used BOLD CVR. We also compared these measures to another traditionally utilized technique in cerebral blood flow CVR from an arterial spin labelling sequence. These imaging, processing, and analysis techniques will allow for further investigation into the magnitude and rate of CVR based on BOLD and MRvF-based metrics and enable investigations to better understand vascular function in healthy aging and cerebrovascular diseases.Clinical Relevance- The development of dynamic magnetic resonance vascular fingerprinting has the potential to enable rapid, quantitative, and multiparametric functional imaging biomarkers of cerebrovascular diseases like vascular cognitive impairment, dementia, and Alzheimer's disease.


Formation of isoprostane bicyclic endoperoxides from the autoxidation of cholesteryl arachidonate.
Autoxidation of polyunsaturated fatty acids and esters leads to a complex mixture containing hydroperoxides and cyclic peroxides. Prostaglandin bicyclic endoperoxides have been detected from the autoxidation of cholesteryl arachidonate by LC-MS and GC-MS techniques. All four possible types (I-IV) of bicyclic endoperoxides have been found starting from different regioisomeric hydroperoxides of cholesteryl arachidonate. Furthermore, the stereochemistry of Type IV bicyclic endoperoxides has been determined by conversion to pentafluorobenzyl (PFB) ester, trimethylsilyl (TMS) derivatives, and comparison with synthetic standards by the use of GC-MS. All eight possible diastereomers of the derivatized isoprostanes were observed and were separated by gas chromatography. The bicyclic endoperoxides with the two alkyl chains syn on the cyclopentane ring were formed preferentially to those with anti configuration, a result anticipated from earlier work. Substantial amounts of the anti-substituted isoprostanes, including PGF(2)(alpha), were, however, observed in the product mixture.


Characterizing local dose perturbations due to gas cavities in magnetic resonance-guided radiotherapy.
Due to differences in attenuation and the electron return effect (ERE), the presence of gas can increase the risk of toxicity in organs at risk (OAR) during magnetic resonance-guided radiotherapy (MRgRT). Current adaptive MRgRT workflows using density overrides negate gas from the dose calculation, meaning that the effects of ERE around gas are not taken into account. In order to achieve an accurate adaptive MRgRT treatment, we should be able to quickly evaluate whether gas present during treatment causes dose constraint violation during an MRgRT fraction. We propose an analytic method for predicting dose perturbations caused by air cavities in OARs during MRgRT. Ten virtual water phantoms were created: nine containing a centrally located spherical air cavity and a reference phantom without an air cavity. Monte Carlo dose calculations were produced to irradiate the phantoms with a single 7 MV photon beam under the influence of a 1.5 T transverse magnetic field (Monaco 5.19.02 Treatment Panning System (TPS) (Elekta AB, Stockholm, Sweden)). Dose distributions of the phantoms with and without air cavities were compared. We used a spherical coordinate system originating in the center of the cavity to sample the dose distributions and calculate the dose perturbation as a result of the presence of each air cavity, ∆D%(θ,Φ)calc . . Dose effects due to ERE and differences in attenuation due to density changes were considered separately. Least squared analysis was used to fit the calculated dose perturbations to mathematical functions. Effects due to ERE were fit to a modulated sinusoidal function and those due to attenuation differences were fit to a 2D Gaussian function. We used the fits to derive a single equation describing dose perturbations around spherical air cavities as a function of angles, θ, Φ, distance from cavity surface, d, and cavity radius, r. We measured the fitting error by calculating the residual error (RE); the difference between the calculated and fitted dose perturbation. Both ERE and differences in attenuation contribute toward the total dose effects of air cavities in MRgRT. Whereas ERE dominates close to the surface of the cavities, attenuation effects dominate at distances >0.5 cm from the cavities. We showed that dose effects around a spherical air cavity (≤1 cm from the surface) due to ERE fit a modulated sinusoidal function with mean (RE) ≤-1.4E-5% and root mean square error (rms) (RE) ≤4.1%. Effects due to attenuation differences fit a Gaussian function with mean (RE) ≤0.7% and rms (RE) ≤1.8%. Our general equation, which we verified using multiple sizes of spherical and cylindrical air cavity, fits Monte Carlo simulated data with mean (RE) ≤±0.9% and rms (RE) ≤6.9%. We show that local dose perturbations around unplanned spherical air cavities during MRgRT can be well characterized analytically. We present an equation that can be incorporated into the clinical workflow to allow for fast evaluation of dose effects of unplanned gas. We also envision this method contributing to the clinical implementation of real time adaptive radiotherapy (ART) for MRgRT using MRI planning.


Granular cell tumor of the stomach: a case report and review of the literature.
A case of granular cell tumor of the stomach in a 39-yr-old black female with peptic ulcer symptoms is presented. This lesion, a soft tissue tumor of disputed origin, has been reported in the stomach only 17 times before. In each case, the tumor was benign. Most of the patients were black, middle aged, and manifested symptoms of peptic ulcer disease. For this reason, we suggest that granular cell tumor be given serious consideration in the differential diagnosis should an upper gastrointestinal contrast study reveal a submucosal, benign mass lesion in a patient with these characteristics.


Programmed cell death in the developing epithelium of the mouse inner ear.
Programmed cell death is as essential to development as is proliferation. Thus, the objective of this study was to elucidate the spatiotemporal involvement of programmed cell death in the development of the inner ear epithelia. Programmed cell death is seen in situ as apoptosis. A time-sequence study was performed on the distribution of apoptosis during the development of the inner ear epithelia of the mouse using the TdT-mediated dUTP-biotin nick labeling (TUNEL) method to detect apoptosis histochemically. Apoptosis occurred during the early stages of development of the inner ear and took place earlier in the organs of equilibrium than in the cochlea. These periods corresponded to those of active proliferation of epithelial cells in the inner ear. Since cell-cell interactions change after the removal of neighboring cells by apoptosis, apoptosis may influence cytodifferentiation.


In-depth Analysis of the Lid Subunits Assembly Mechanism in Mammals.
The 26S proteasome is a key player in the degradation of ubiquitinated proteins, comprising a 20S core particle (CP) and a 19S regulatory particle (RP). The RP is further divided into base and lid subcomplexes, which are assembled independently from each other. We have previously demonstrated the assembly pathway of the CP and the base by observing assembly intermediates resulting from knockdowns of each proteasome subunit and the assembly chaperones. In this study, we examine the assembly pathway of the mammalian lid, which remains to be elucidated. We show that the lid assembly pathway is conserved between humans and yeast. The final step is the incorporation of Rpn12 into the assembly intermediate consisting of two modular complexes, Rpn3-7-15 and Rpn5-6-8-9-11, in both humans and yeast. Furthermore, we dissect the assembly pathways of the two modular complexes by the knockdown of each lid subunit.


Visual function and fitness to drive.
Driving is recognized to be a visually intensive task and accordingly there is a legal minimum standard of vision required for all motorists. The purpose of this paper is to review the current United Kingdom (UK) visual requirements for driving and discuss the evidence base behind these legal rules. The role of newer, alternative tests of visual function that may be better indicators of driving safety will also be considered. Finally, the implications of ageing on driving ability are discussed. A search of Medline and PubMed databases was performed using the following keywords: driving, vision, visual function, fitness to drive and ageing. In addition, papers from the Department of Transport website and UK Royal College of Ophthalmologists guidelines were studied. Current UK visual standards for driving are based upon historical concepts, but recent advances in technology have brought about more sophisticated methods for assessing the status of the binocular visual field and examining visual attention. These tests appear to be better predictors of driving performance. Further work is required to establish whether these newer tests should be incorporated in the current UK visual standards when examining an individual's fitness to drive.


Loss of the female advantage with cardiovascular disease for women with diabetes.
The relative risk of cardiovascular disease (CVD) associated with diabetes is greater for women than men, and diabetic women did not experience temporal declines in CVD mortality observed for the general population and diabetic men. To examine sex differences in CVD risk factors for persons with diabetes over time. Population-based historical cohort study. The provider-linked medical records of all Rochester, MN, residents assigned a clinical diagnosis of diabetes were reviewed to confirm case status and assign diagnosis date. Data on fasting glucose, obesity, persistent proteinuria, smoking, hypertension, and dyslipidemia were obtained at diagnosis for confirmed incidence cases. There were 1330 diabetes cases 1970- 1989. Compared to men, women at diagnosis were older and more likely hypertensive, had similar levels of fasting glucose and persistent proteinuria, and less likely to smoke. Among persons diagnosed at younger ages, women were more likely than men to be obese. Comparison with published data for the Rochester population revealed the excess obesity and hypertension associated with diabetes were highest for women < age 55 y. Temporal trends in CVD risk factors did not differ between diabetic men and women.


Increasing use of linezolid in a tertiary NICU during a 10-year period: reasons and concerns for the future.
Linezolid has been increasingly used in tertiary NICUs. The objectives of this study were to explore the indications of these linezolid prescriptions, to analyze a possible misuse and to provide solutions to avoid such misuse. A monocentric retrospective cohort study included all neonates hospitalized in one tertiary NICU between January 1st, 2010 and December 31st, 2019 and who received at least one administration of linezolid. These data were confronted to epidemiological and antibiotic use data from the same NICU. Two independent pediatricians secondarily classified linezolid uses as adequate or not. During the study period, 66 infections in 57 patients led to linezolid use. Most patients were pre-term and 21 patients (37%) died. Infections were mainly related to methicillin-resistant coagulase negative staphylococci and were frequently either pneumoniae (35%) or isolated bacteremia (48%), including 25 persistent bacteremia (64% of the 39 bacteremia). Need for a better tissue distribution or first-line treatment failure were the main reasons to initiate linezolid. Linezolid was administered for a median duration of 7 [3;10] days. No side effects were reported. Twenty-two (33%) of the 66 linezolid prescriptions were retrospectively classified as inadequate. A rapid increase in linezolid prescriptions has been observed in our tertiary NICU, from 2014 to 2019, with 33% inadequate uses. This worrisome trend should lead to search for therapeutic alternatives and to work on antibiotic stewardship to prevent the emergence of new antimicrobial bacterial resistance.


Relationship between neck circumference and overactive bladder in women with metabolic syndrome: a preliminary study.
To investigate relationship between overactive bladder (OAB) and metabolic syndrome (MtS) by using neck circumference (NC). In retrospective view of prospective collected data, 204 women with or without OAB were enrolled into study, between August 2012 and December 2013. All patients were administered OAB validated questionnaires (OAB-V8) and whose score was > 8 were accepted as OAB. Patients were divided into two groups and group 1 consisted of patients with OAB, group 2 consisted of patients without OAB. Demographic data with anthropometric measurements and blood analyses were recorded. Statistical analyses including receiver operating characteristic (ROC) curves were performed; statistically significant p was < 0.05. Mean age was 41.06 ± 9.78 years. There were 115 (56.4 %) patients in group 1, and 89 (43.6 %) patients in group 2. OAB-V8 scores were significant higher in group 1 than group 2 (p < 0.001). Waist circumference (WC) and NC measurements were statistical significant longer in group 1 than group 2 (p < 0.001). In multivariate logistic regression analyses age, body mass index, MtS, WC, and NC were statistical significant associated with OAB. In ROC curves, area under the curve (AUC) was 0.72 cm2 for relationship between OAB and WC (p < 0.001), and AUC was 0.73 cm2 for relationship between OAB and NC (p = 0.004). Cut-off NC and WC values for OAB were determined as 35.25 cm and 98.5 cm, respectively. OAB with metabolic syndrome seems like more common in women than in those without. NC may be a novel indicator for OAB in selected female patients with MtS.


IL-2 responsiveness of CD4 and CD8 lymphocytes: further investigations with human IL-2Rbeta transgenic mice.
Responsiveness to IL-2 varies from one lympho-mononuclear subset to another. NK lymphocytes and monocytes spontaneously respond to IL-2 whereas it is generally accepted that T and B lymphocytes need to be activated to fully acquire this competence. To further investigate this phenomenon, we studied human IL-2Rbeta (hIL-2Rbeta) transgenic mice constitutively expressing heterospecific, intermediate-affinity IL-2R (hIL-2Rbeta/mouse IL-2Rgamma(c)). We noted that the B lymphocytes and monocytes from spleens of these hIL-2Rbeta transgenic animals failed to grow when cultured in IL-2-containing medium. Under the same experimental conditions, CD4 lymphocytes survived, again without growth, whereas CD8 lymphocytes and NK cells were able to proliferate and develop potent LAK cytotoxicity. The properties of these CD4 and CD8 lymphocytes were then compared after purification. Both subsets expressed functional IL-2R able to induce global protein phosphorylation and, more precisely, signal transducer and activation of transcription 5 and Erk phosphorylation. Therefore, the differential growth potential of these CD4 and CD8 lymphocytes cannot be explained by the lack of IL-2R-dependent early signaling events. When the entrance of purified CD4 and CD8 lymphocytes into the cell cycle was analyzed, we found that the CD4 lymphocytes were unable to enter the G1 phase in the absence of anti-CD3 stimulation. This correlates with the effect of IL-2 on cyclin-dependent kinase inhibitor p27(kip1). In CD4 lymphocytes, IL-2 does not affect p27(kip1) expression. But in CD8 lymphocytes, IL-2 down-modulates p27(kip1). These results indicate that, aside from IL-2R expression and function, IL-2 responsiveness is also controlled by lineage-specific mechanisms.


Current trends in tobacco prevention and cessation in Nebraska physicians' offices.
Despite years of intervention, few studies describe the extent to which recommended tobacco use prevention and cessation activities occur in community-based family practices. This study was designed to discover current practice patterns in these areas and to describe physician outcome and efficacy expectations. An exploratory comparative case study of 11 family practices used direct observation of practices and clinical encounters, chart reviews, and in-depth interviews. Qualitative and quantitative information was gathered on (1) intensity of tobacco use prevention and cessation; (2) physicians' attitudes and beliefs regarding outcome expectations; and (3) physicians' perceptions of their ability to counsel. Qualitative content analysis and descriptive statistics were used to construct case studies for comparisons. Themes common to most practices included the "provision of little prevention" and "a lack of perceived need to address smokeless tobacco." Responsibility for tobacco activities fell almost solely to physicians. Although physicians felt confident in their counseling skills, the skills they identified were fairly basic. Most physicians were pessimistic about the positive effects of these activities. None of the practices was using any specifically developed "package," and pharmaceutical companies provided almost all patient education material. There was considerable variation in intensity of activities because of differences in attitudes, expectation, and background. To increase tobacco control activities, practice systems need to be individually evaluated to identify what is needed, how it will fit within the practice culture, and how it can best be implemented in this specific practice. One-size-fits-all interventions probably will not be widely implemented.


Molecular characterization of Malassezia isolates from dogs using three distinct genetic markers in nuclear DNA.
Little precise information is available on the systematics, genetics, ecology and epidemiology of yeasts of the genus Malassezia from different animal species. In the present study, one hundred and four isolates of Malassezia (lipid dependent or non-lipid dependent) from dogs were characterized by their chitin synthase 2 gene (CHS2), and the large subunit (LSU) and the first internal transcribed spacer (ITS-1) of nuclear ribosomal DNA sequences, and compared genetically with well-defined reference strains of Malassezia pachydermatis and heterologous species, including Malassezia furfur and Candida albicans. For each locus examined, three main sequence types (i.e. A, B and C) represented all of the 104 isolates, which were designated as genotypes A, B and C, respectively. A fourth, minor sequence type was also defined for the ITS-1. The nucleotide differences among genotypes was consistent with the magnitudes of intraspecific variability reported in previous studies. The genetic analysis of the sequence data sets (for individual loci) showed that all Malassezia genotypes clustered (with moderate to strong support) with the reference sequences of M. pachydermatis to the exclusion of the outgroups M. furfur and C. albicans. The present study reveals that multiple genetic variants of M. pachydermatis occur on dogs. The multilocus approach employed herein provides a foundation for future investigations of M. pachydermatis from other animals and humans, and their ecology and epidemiology.


Lipoxygenase-catalyzed transformation of epoxy fatty acids to hydroxy-endoperoxides: a potential P450 and lipoxygenase interaction.
Herein, we characterize a generally applicable transformation of fatty acid epoxides by lipoxygenase (LOX) enzymes that results in the formation of a five-membered endoperoxide ring in the end product. We demonstrated this transformation using soybean LOX-1 in the metabolism of 15,16-epoxy-α-linolenic acid, and murine platelet-type 12-LOX and human 15-LOX-1 in the metabolism of 14,15-epoxyeicosatrienoic acid (14,15-EET). A detailed examination of the transformation of the two enantiomers of 15,16-epoxy-α-linolenic acid by soybean LOX-1 revealed that the expected primary product, a 13S-hydroperoxy-15,16-epoxide, underwent a nonenzymatic transformation in buffer into a new derivative that was purified by HPLC and identified by UV, LC-MS, and ¹H-NMR as a 13,15-endoperoxy-16-hydroxy-octadeca-9,11-dienoic acid. The configuration of the endoperoxide (cis or trans side chains) depended on the steric relationship of the new hydroperoxy moiety to the enantiomeric configuration of the fatty acid epoxide. The reaction mechanism involves intramolecular nucleophilic substitution (SNi) between the hydroperoxy (nucleophile) and epoxy group (electrophile). Equivalent transformations were documented in metabolism of the enantiomers of 14,15-EET by the two mammalian LOX enzymes, 15-LOX-1 and platelet-type 12-LOX. We conclude that this type of transformation could occur naturally with the co-occurrence of LOX and cytochrome P450 or peroxygenase enzymes, and it could also contribute to the complexity of products formed in the autoxidation reactions of polyunsaturated fatty acids.


Activity of the novel BCR kinase inhibitor IQS019 in preclinical models of B-cell non-Hodgkin lymphoma.
Pharmacological inhibition of B cell receptor (BCR) signaling has recently emerged as an effective approach in a wide range of B lymphoid neoplasms. However, despite promising clinical activity of the first Bruton's kinase (Btk) and spleen tyrosine kinase (Syk) inhibitors, a small fraction of patients tend to develop progressive disease after initial response to these agents. We evaluated the antitumor activity of IQS019, a new BCR kinase inhibitor with increased affinity for Btk, Syk, and Lck/Yes novel tyrosine kinase (Lyn), in a set of 34 B lymphoid cell lines and primary cultures, including samples with acquired resistance to the first-in-class Btk inhibitor ibrutinib. Safety and efficacy of the compound were then evaluated in two xenograft mouse models of B cell lymphoma. IQS019 simultaneously engaged a rapid and dose-dependent de-phosphorylation of both constitutive and IgM-activated Syk, Lyn, and Btk, leading to impaired cell proliferation, reduced CXCL12-dependent cell migration, and induction of caspase-dependent apoptosis. Accordingly, B cell lymphoma-bearing mice receiving IQS019 presented a reduced tumor outgrowth characterized by a decreased mitotic index and a lower infiltration of malignant cells in the spleen, in tight correlation with downregulation of phospho-Syk, phospho-Lyn, and phospho-Btk. More interestingly, IQS019 showed improved efficacy in vitro and in vivo when compared to the first-in-class Btk inhibitor ibrutinib, and was active in cells with acquired resistance to this latest. These results define IQS019 as a potential drug candidate for a variety of B lymphoid neoplasms, including cases with acquired resistance to current BCR-targeting therapies.


From the Cover: Segmented spiral waves in a reaction-diffusion system.
Pattern formation in reaction-diffusion systems is often invoked as a mechanism for biological morphogenesis. Patterns in chemical systems typically occur either as propagating waves or as stationary, spatially periodic, Turing structures. The spiral and concentric (target) waves found to date in spatially extended chemical or physical systems are smooth and continuous; only living systems, such as seashells, lichens, pine cones, or flowers, have been shown to demonstrate segmentation of these patterns. Here, we report observations of segmented spiral and target waves in the Belousov-Zhabotinsky reaction dispersed in water nanodroplets of a water-in-oil microemulsion. These highly ordered chemical patterns, consisting of short wave segments regularly separated by gaps, form a link between Turing and trigger wave patterns and narrow the disparity between chemistry and biology. They exhibit aspects of such fundamental biological behavior as self-replication of structural elements and preservation of morphology during evolutionary development from a simpler precursor to a more complex structure.


12-O-Tetradecanoylphorbol-13-acetate increases cardiomyogenesis through PKC/ERK signaling.
12-O-Tetradecanoylphorbol-13-acetate (TPA) is the most widely used diacylglycerol (DAG) mimetic agent and inducer of protein kinase C (PKC)-mediated cellular response in biomedical studies. TPA has been proposed as a pluripotent cell differentiation factor, but results obtained have been inconsistent. In the present study we show that TPA can be applied as a cardiomyogenesis-promoting factor for the differentiation of mouse embryonic stem (mES) cells in vitro. The mechanism of TPA action is mediated by the induction of extracellular signal-regulated kinase (ERK) activity and the subsequent phosphorylation of GATA4 transcription factor. Interestingly, general mitogens (FGF, EGF, VEGF and serum) or canonical WNT signalling did not mimic the effect of TPA. Moreover, on the basis of our results, we postulate that a TPA-sensitive population of cardiac progenitor cells exists at a certain time point (after days 6-8 of the differentiation protocol) and that the proposed treatment can be used to increase the multiplication of ES cell-derived cardiomyocytes.


Pharmacokinetic interactions between flunixin and sulphadimidine in horses.
The pharmacokinetic aspects of sulphadimidine were studied in clinically healthy (control) and Flunixin-medicated horses after a single intravenous and oral administration of 100 mg/kg body weight. Plasma sulphadimidine concentration were determined by high-performance liquid chromatography (HPLC). Following the intravenous injection, all plasma sulphadimidine data were best approximated by a two-compartment open model using sequential, weight non-linear regression. Flunixin induced a 67% increase in the rate of sulphadimidine return to the central compartment from peripheral tissues (K21) and there were a trend to a 30% increase in K12. The sulphadimidine elimination half-life was decreased 21%, the Vdss was reduced by 18% and MRT was decreased by 20%. Following the oral administration, sulphadimidine was rapidly absorbed in control and Flunixin-medicated horses with absorption half-lives (t1/2 ab) of 0.5 and 0.43 hours respectively. The peak plasma concentration (Cmax) were 93.7 and 109 micrograms/ml attained at (tmax) 2.36 and 1.9 hours respectively. The elimination half-life after oral administration (t1/2 ab) was shorter in flunixin pre-medicated horses than in control ones. The systemic bioavalability percentages (F%) of sulphadimidine after oral administration of 100 mg/kg body weight was 79.3 and 71.2% in control and flunixin medicated horses, respectively. Therefore care should be exercised in the use of sulphadimidine in equine patients concurrently treated with flunixin.


Are superoxide dismutase 2 and nitric oxide synthase polymorphisms associated with idiopathic infertility?
The aim of this study was to investigate in a case-control study the associations between idiopathic infertility and antioxidant gene polymorphisms. One hundred ten infertile subjects (58 women and 52 men) with a history of idiopathic infertility and 69 fertile subjects (35 women and 34 men) with no history of infertility were included by three hospital departments of reproductive biology in the NCT01093378 French government clinical trial. Genotyping was assessed by real-time polymerase chain reaction with TaqMan assay. We examined genetic polymorphisms affecting five antioxidant enzymes: manganese superoxide dismutase (MnSOD), myeloperoxidase (MPO), glutathione peroxidase 1 (GPx1), catalase (CAT), and endothelial nitric oxide synthase (eNOS). The presence of at least 1 Ala-MnSOD allele (rs4880) increased significantly the risk of infertility (odds ratio [OR] 2.94; 95% confidence interval [CI], 1.14, 7.60; p=0.03) in male subjects. Moreover, the presence of 2 G-eNOS allele (rs1799983) increased significantly the risk of infertility in both men and women (OR 1.91; 95% CI, 1.04, 3.54; p=0.04). Our observations lead to the hypothesis that the genetic susceptibility modulating oxidative stress may represent a risk factor for male idiopathic infertility.


Friction-Stir Welding of a Wrought Al-Si-Mg Alloy in As-Fabricated and Heat-Treatment States.
A wrought Al-11.3Si-0.6Mg alloy under hot extrusion (T1), solution treatment (T4), and solution treatment + artificial aging (T6) states were friction stir welded at welding speed of 100 mm/min and rotation rate of 800 rpm. The effect of prior heat-treatment on the microstructure and mechanical properties of the welds were investigated. The results show that the microstructures of the nugget zones have little dependence on the initial states of the base material. In the nugget zones, complete recrystallized structures with equaxied grains in the Al matrix were formed under all conditions. The Si particles in the nugget zones are almost unchanged compared with those of their base materials (BMs) in the three states. In contrast, the joint efficiency of the obtained welds was very sensitive to the initial material condition. The joint efficiency under the T1 state is more than 90% due to the fact that the microstructure is almost unchanged, except for the slight coarsening of the Al matrix grains and some of the Mg2Si phases during the friction stir welding process. However, the joint efficiency in the T4 and T6 conditions is only 77.22% and 62.03%, respectively. The relatively low weld strength in the T4 and T6 conditions is due to the elimination of the solid solution strengthening and age hardening effects during friction stir welding. The hardness distributions along the cross section of joints are all W-shaped under T1, T4, and T6 conditions.


The inflammation-related biomarker CXCR7 independently predicts patient outcome after radical prostatectomy.
The influence of inflammation on prostate tumor carcinogenesis is currently much better known than with its role in prostate cancer (CaP) progression. We evaluated the prognostic value of epigenetic (HDAC1, HDAC4, H3Ac) and inflammation-related (CXCR4, CXCR7, CXCL12) biomarkers immunoexpression, in radical prostatectomy specimens, from 2 cohorts of CaP patients with long term follow-up. Formalin-fixed and paraffin-embedded radical prostatectomy specimens were obtained from the pathology archives of Prof. Doutor Fernando Fonseca Hospital, in Amadora, Portugal and Portuguese Oncology Institute of Porto, in Porto, Portugal, and tissue microarrays were assembled. It was achieved a set of 234 patients submitted to radical retropubic prostatectomy between January 2000 and December 2005. Immunohistochemistry was used for evaluation of protein expression of epigenetic and inflammation-related markers. Nuclear staining was assessed using digital image analysis. Study outcomes included disease-specific survival and disease-free survival (DFS). Statistical analysis was tabulated using SPSS version 23.0. Hazard ratios (HRs) and survival curves were estimated using Cox-regression and Kaplan-Meyer models, respectively. Statistical significance was set at P < 0.05. Complete follow-up data was available for 234 patients and median follow-up time was 164 [11-218] months. Patients with higher CXCR4 immunoexpression experienced significantly worse disease-specific survival compared to patients with low expression (HR = 1.016, 95% CI: 1.002-1.031). The same happened with CXCL12 (HR = 0.546 95% CI: 0.322-0.926) and H3Ac (HR = 1.015, 95% CI: 1.001c1.029). In what concerns to DFS, patients with higher expression of CXCR4 and CXCR7 were significantly more prone to experience disease recurrence (HR = 1.003, 95% CI: 1.000-1.005 and HR = 1.111, 95% CI:1.032-1.196, respectively). When adjusted to pTStage and WHO Grade Groups, CXCR7 maintained independent impact on DFS (HR = 1.119, 95% CI: 1.032-1.214). The interplay between inflammation and epigenetics and its impact in CaP outcome deserves further studies in the future. CXCR7 shows an independent predictor for worse DFS after radical prostatectomy, and could provide important prognostic information for patient management after radical prostatectomy.


Evolutionary consequences of nascent multicellular life cycles.
A key step in the evolutionary transition to multicellularity is the origin of multicellular groups as biological individuals capable of adaptation. Comparative work, supported by theory, suggests clonal development should facilitate this transition, although this hypothesis has never been tested in a single model system. We evolved 20 replicate populations of otherwise isogenic clonally reproducing 'snowflake' yeast (Δace2/∆ace2) and aggregative 'floc' yeast (GAL1p::FLO1 /GAL1p::FLO1) with daily selection for rapid growth in liquid media, which favors faster cell division, followed by selection for rapid sedimentation, which favors larger multicellular groups. While both genotypes adapted to this regime, growing faster and having higher survival during the group-selection phase, there was a stark difference in evolutionary dynamics. Aggregative floc yeast obtained nearly all their increased fitness from faster growth, not improved group survival; indicating that selection acted primarily at the level of cells. In contrast, clonal snowflake yeast mainly benefited from higher group-dependent fitness, indicating a shift in the level of Darwinian individuality from cells to groups. Through genome sequencing and mathematical modeling, we show that the genetic bottlenecks in a clonal life cycle also drive much higher rates of genetic drift-a result with complex implications for this evolutionary transition. Our results highlight the central role that early multicellular life cycles play in the process of multicellular adaptation.


Two-Year Safety and Efficacy Experience in Patients with Methotrexate-Resistant Active Rheumatoid Arthritis Treated with Etanercept and Conventional Disease-Modifying Anti-rheumatic Drugs in the Latin American Region.
Although long-term data are available from biologic studies in North American/European populations with rheumatoid arthritis (RA), long-term findings in Latin American RA populations are limited. To examine long-term safety/efficacy of etanercept, methotrexate, and/or other disease-modifying anti-rheumatic drugs (DMARDs) in Latin American patients with moderate-to-severe active RA. In the first phase of this open-label study, patients were randomized to etanercept 50 mg weekly plus methotrexate or conventional DMARD (hydroxychloroquine or sulfasalazine) plus methotrexate for 24 weeks. At the start of the second phase (week 24), investigators selected a treatment regimen that included any combination/dosage of etanercept, methotrexate, hydroxychloroquine, or sulfasalazine based on previous treatment response, preference, and local product labeling, and was continued for the 104-week extension. In the extension, in the group previously randomized to etanercept-plus-methotrexate therapy, etanercept was continued in 259/260 patients; methotrexate continued in 260/260; and hydroxychloroquine and sulfasalazine added in 8/260 and 3/260, respectively. In the group previously randomized to conventional DMARD-plus-methotrexate therapy, conventional DMARD was discontinued in 86/126 and etanercept added in 105/126. Among etanercept-exposed patients (total exposure, 798.1 patient-year [PY]), rates of adverse events, serious adverse events, and serious infections per PY were 1.7, 0.07, and 0.02 events per PY. In both groups, after treatment modification was permitted, clinical response rates and improvements in clinical/patient-reported outcomes from baseline were sustained to week 128. After investigators were permitted to modify treatment, etanercept was part of the treatment regimen in 95% of patients. Continuation or addition of etanercept in the 2-year extension resulted in a consistently good risk:benefit profile. Open-Label Study Comparing Etanercept to Conventional Disease Modifying Antirheumatic Drug (DMARD) Therapy; ClinicalTrials.gov, number NCT00848354; https://clinicaltrials.gov/ct2/show/NCT00848354.


Improved brain community structure detection by two-step weighted modularity maximization.
The human brain can be regarded as a complex network with interacting connections between brain regions. Complex brain network analyses have been widely applied to functional magnetic resonance imaging (fMRI) data and have revealed the existence of community structures in brain networks. The identification of communities may provide insight into understanding the topological functions of brain networks. Among various community detection methods, the modularity maximization (MM) method has the advantages of model conciseness, fast convergence and strong adaptability to large-scale networks and has been extended from single-layer networks to multilayer networks to investigate the community structure changes of brain networks. However, the problems of MM, suffering from instability and failing to detect hierarchical community structure in networks, largely limit the application of MM in the community detection of brain networks. In this study, we proposed the weighted modularity maximization (WMM) method by using the weight matrix to weight the adjacency matrix and improve the performance of MM. Moreover, we further proposed the two-step WMM method to detect the hierarchical community structures of networks by utilizing node attributes. The results of the synthetic networks without node attributes demonstrated that WMM showed better partition accuracy than both MM and robust MM and better stability than MM. The two-step WMM method showed better accuracy of community partitioning than WMM for synthetic networks with node attributes. Moreover, the results of resting state fMRI (rs-fMRI) data showed that two-step WMM had the advantage of detecting the hierarchical communities over WMM and was more insensitive to the density of the rs-fMRI networks than WMM.


Metabolism of acetyl-L-carnitine for energy and neurotransmitter synthesis in the immature rat brain.
Acetyl-L-carnitine (ALCAR) is an endogenous metabolic intermediate that facilitates the influx and efflux of acetyl groups across the mitochondrial inner membrane. Exogenously administered ALCAR has been used as a nutritional supplement and also as an experimental drug with reported neuroprotective properties and effects on brain metabolism. The aim of this study was to determine oxidative metabolism of ALCAR in the immature rat forebrain. Metabolism was studied in 21-22 day-old rat brain at 15, 60 and 120 min after an intraperitoneal injection of [2-(13)C]acetyl-L-carnitine. The amount, pattern, and fractional enrichment of (13)C-labeled metabolites were determined by ex vivo(13)C-NMR spectroscopy. Metabolism of the acetyl moiety from [2-(13)C]ALCAR via the tricarboxylic acid cycle led to incorporation of label into the C4, C3 and C2 positions of glutamate (GLU), glutamine (GLN) and GABA. Labeling patterns indicated that [2-(13)C]ALCAR was metabolized by both neurons and glia; however, the percent enrichment was higher in GLN and GABA than in GLU, demonstrating high metabolism in astrocytes and GABAergic neurons. Incorporation of label into the C3 position of alanine, both C3 and C2 positions of lactate, and the C1 and C5 positions of glutamate and glutamine demonstrated that [2-(13)C]ALCAR was actively metabolized via the pyruvate recycling pathway. The enrichment of metabolites with (13)C from metabolism of ALCAR was highest in alanine C3 (11%) and lactate C3 (10%), with considerable enrichment in GABA C4 (8%), GLN C3 (approximately 4%) and GLN C5 (5%). Overall, our (13)C-NMR studies reveal that the acetyl moiety of ALCAR is metabolized for energy in both astrocytes and neurons and the label incorporated into the neurotransmitters glutamate and GABA. Cycling ratios showed prolonged cycling of carbon from the acetyl moiety of ALCAR in the tricarboxylic acid cycle. Labeling of compounds formed from metabolism of [2-(13)C]ALCAR via the pyruvate recycling pathway was higher than values reported for other precursors and may reflect high activity of this pathway in the developing brain. This is, to our knowledge, the first study to determine the extent and pathways of ALCAR metabolism for energy and neurotransmitter biosynthesis in the brain.


Culturing adult human bone marrow stem cells on gelatin scaffold with pNIPAAm as transplanted grafts for skin regeneration.
Skin tissue engineering is a possible solution for the treatment of extensive skin defect. The ultimate goal of skin tissue engineering is to restore the complete functions of native skin, but until now the structures and functions of skins are only partially restored. By negative immunoselection (CD45 and glycophorin A), we isolated and cultivated adult human bone marrow stem cells (hBMSCs) that are of multilineage differentiation potential. In this study, we first demonstrated that by using gelatin/thermo-sensitive poly N-isopropylacrylamide (pNIPAAm) and the immunocompromised mice model, the hBMSCs possess the differentiation potential of epidermis and the capability of healing skin wounds. The in vitro observations and the results of the scanning electron microscope showed that the hBMSCs can attach and proliferate in the gelatin/thermo-sensitive pNIPAAm. To further monitor the in vivo growth effect of the hBMSCs in the skin-defected nude mice, the green fluorescence protein (GFP) gene was transduced into the hBMSCs by the murine stem cell viral vector. The results showed that the rates of cell growth and wound recovery in the hBMSC-treated group were significantly higher than those in the control group, which was only treated with the gelatin/pNIPAAm (p < 0.01). More importantly, the re-epithelialization markers of human pan-cytokeratin and E-cadherin were significantly increased on day 7, day 14, and day 21 after the hBMSC-scaffold with the pNIPAAM in the mice with skin defects (p < 0.05). Moreover, the stem cell markers of human CD13 and CD105 were gradually decreased during the period of wound healing. In sum, this novel method provides a transferring system for cell therapies and maintains its temperature-sensitive property of easy-peeling by lower-temperature treatment. In addition, the in vitro and in vivo GFP imaging systems provide a new imaging modality for understanding the differentiation process and the effective expression of stem cells in wound healing.


Purification of the glycoprotein lectin from the broad bean (Vicia faba) and a comparison of its properties with lectins of similar specificity.
1. The lectin from the broad bean (Vicia faba) was purified by affinity chromatography by using 3-O-methylglucosamine covalently attached through the amino group to CH-Sepharose (an omega-hexanoic acid derivative of agarose). Its composition and the nature of its subunits were compared with concanavalin A and the lectins from pea and lentil. 2. Unlike the other three lectins, broad-bean lectin is a glycoprotein; a glycopeptide containing glucosamine and mannose was isolated from a proteolytic digest. 3. The mol.wt. is about 47500; the glycoprotein consists of two apprently identical subunits, held together by non-covalent forces. Fragments of the subunits, similar to those found in concanavalin A and soya-bean agglutinin, were found in active preparations. 4. Broad-bean lectin was compared with concanavalin A and the lectins from pea and lentil in an investigation of the inhibition of their action by a number of monosaccharides, methyl ethers of monosaccharides, disaccharides and glycopeptides. The most striking differences concern 3-O-substituted monosaccharides, which are strong inhibitors of the action of broad-bean, pea and lentil lectins but not of the action of concanavalin A. There is, however, no strong inhibition of the action of these lectins by 3-Olinked disaccharides.


UV and visible light screening by individual sporopollenin exines derived from Lycopodium clavatum (club moss) and Ambrosia trifida (giant ragweed).
We have investigated the UV-visible light transmission of three types of micrometre-sized sporopollenin exine shells, two derived from Lycopodium clavatum (club moss) spores and one from Ambrosia trifida (giant ragweed) pollen. We have used spectrophotometer measurements of partial monolayers of exines and microscope absorbance imaging to derive the light transmission properties of individual exines. Measurements have been made for exines in air when light transmission losses are due to a combination of absorption, reflection and scattering processes and for exines dispersed in a liquid for which the refractive index (RI) is approximately equal to the RI of the exine such that reflection and scattering effects are negligible. Overall, it found that the light transmission of a single exine wall is approximately 50%. This value of the transmission is due mainly to light absorption, is similar for the three exines studied here and varies only slightly with light wavelength over the range 200-900 nm.


HilD-mediated transcriptional cross-talk between SPI-1 and SPI-2.
The acquisition of new genetic traits by horizontal gene transfer and their incorporation into preexisting regulatory networks have been essential events in the evolution of bacterial pathogens. An example of successful assimilation of virulence traits is Salmonella enterica, which acquired, at distinct evolutionary times, Salmonella pathogenicity island 1 (SPI-1), required for efficient invasion of the intestinal epithelium and intestinal disease, and SPI-2, essential for Salmonella replication and survival within macrophages and the progression of a systemic infection. A positive regulatory cascade mainly composed of HilD, HilA, and InvF, encoded in SPI-1, controls the expression of SPI-1 genes, whereas the two-component regulatory system SsrA/B, encoded in SPI-2, controls expression of SPI-2 genes. In this study, we report a previously undescribed transcriptional cross-talk between SPI-1 and SPI-2, where the SPI-1-encoded regulator HilD is essential for the activation of both the SPI-1 and SPI-2 regulons but at different times during the stationary phase of growth in Luria-Bertani medium. Our data indicate that HilD counteracts the H-NS-mediated repression exerted on the OmpR-dependent activation of the ssrAB operon by specifically interacting with its regulatory region. In contrast, HilD is not required for SPI-2 regulon expression under the in vitro growth conditions that are thought to resemble the intracellular environment. Our results suggest that two independent SPI-2 activation pathways evolved to take advantage of the SPI-2-encoded information at different niches and, in consequence, in response to different growth conditions.


Vibrio litoralis sp. nov., isolated from a Yellow Sea tidal flat in Korea.
Two Gram-negative, facultatively anaerobic bacterial strains, MANO22D(T) and MANO22P, were isolated from a tidal flat area of Dae-Chun, Chung-Nam, Korea. The isolates were rod-shaped and were motile by means of one or more polar flagella. They grew at 1-12 % NaCl, 4-45 degrees C and pH 4.1-8.8 and were oxidase-positive, arginine dihydrolase-negative and sensitive to the vibriostatic agent O/129. The isolates required Na(+) for growth, produced acid, but no gas, from D-glucose under anaerobic conditions and utilized a wide range of compounds as sole carbon and energy sources. A phylogenetic analysis based on 16S rRNA gene sequences revealed that the strains belong to the Gammaproteobacteria and are specifically related to Vibrio species. They were most closely related to Vibrio rumoiensis FERM P-14531(T), with which they were found to share 98.65 % 16S rRNA gene sequence similarity. In the phylogenetic tree, the two novel strains comprised a relatively long subline of descent, sharing a branching point with the outlying species V. rumoiensis, and were found to occupy a phylogenetically distant position on the main Vibrio branch. The levels of DNA-DNA hybridization with respect to V. rumoiensis FERM P-14531(T), which is their most closely related phylogenetically related Vibrio species, were 7.4 % (MANO22D(T)) and 3.9 % (MANO22P). Thus, the two novel isolates appear to represent a novel species within the genus Vibrio, for which the name Vibrio litoralis sp. nov. is proposed. The type strain is MANO22D(T) (=KCTC 12520(T)=DSM 17657(T)).


[Reproduction of the larvivorous fish Poecilia reticulata (Poeciliidae) in natural conditions].
The main reproduction characteristics of the larvivorous fish Poecilia reticulata, were determined under natural conditions. The study was carried out in a ditch of dirty water sink at the Lenin Park (Havana City) in the period comprised between November 1986 and May 1987. The samplings were performed monthly, and the presence of gravid females was demonstrated during the whole period, while the average of embryo/female increased remarkably during April. Such results contribute to the knowledge of the biology of this fish for its better use in the biologic control of mosquito larvae.


Dietary fat influences the expression of contractile and metabolic genes in rat skeletal muscle.
Dietary fat plays a major role in obesity, lipid metabolism, and cardiovascular diseases. To determine whether the intake of different types of dietary fats affect the muscle fiber types that govern the metabolic and contractile properties of the skeletal muscle, we fed male Wistar rats with a 15% fat diet derived from different fat sources. Diets composed of soybean oil (n-6 polyunsaturated fatty acids (PUFA)-rich), fish oil (n-3 PUFA-rich), or lard (low in PUFAs) were administered to the rats for 4 weeks. Myosin heavy chain (MyHC) isoforms were used as biomarkers to delineate the skeletal muscle fiber types. Compared with soybean oil intake, fish oil intake showed significantly lower levels of the fast-type MyHC2B and higher levels of the intermediate-type MyHC2X composition in the extensor digitorum longus (EDL) muscle, which is a fast-type dominant muscle. Concomitantly, MyHC2X mRNA levels in fish oil-fed rats were significantly higher than those observed in the soybean oil-fed rats. The MyHC isoform composition in the lard-fed rats was an intermediate between that of the fish oil and soybean oil-fed rats. Mitochondrial uncoupling protein 3, pyruvate dehydrogenase kinase 4, and porin mRNA showed significantly upregulated levels in the EDL of fish oil-fed rats compared to those observed in soybean oil-fed and lard-fed rats, implying an activation of oxidative metabolism. In contrast, no changes in the composition of MyHC isoforms was observed in the soleus muscle, which is a slow-type dominant muscle. Fatty acid composition in the serum and the muscle was significantly influenced by the type of dietary fat consumed. In conclusion, dietary fat affects the expression of genes related to the contractile and metabolic properties in the fast-type dominant skeletal muscle, where the activation of oxidative metabolism is more pronounced after fish oil intake than that after soybean oil intake.


High mobility group box protein 1: an endogenous signal for dendritic cell maturation and Th1 polarization.
High mobility group box protein 1 (HMGB1), a DNA binding nuclear and cytosolic protein, is a proinflammatory cytokine released by monocytes and macrophages. This study addressed the hypothesis that HMGB1 is an immunostimulatory signal that induces dendritic cell (DC) maturation. We show that HMGB1, via its B box domain, induced phenotypic maturation of DCs, as evidenced by increased CD83, CD54, CD80, CD40, CD58, and MHC class II expression and decreased CD206 expression. The B box caused increased secretion of the proinflammatory cytokines IL-12, IL-6, IL-1alpha, IL-8, TNF-alpha, and RANTES. B box up-regulated CD83 expression as well as IL-6 secretion via a p38 MAPK-dependent pathway. In the MLR, B box-activated DCs acted as potent stimulators of allogeneic T cells, and the magnitude of the response was equivalent to DCs activated by exposure to LPS, nonmethylated CpG oligonucleotides, or CD40L. Furthermore, B box induced secretion of IL-12 from DCs as well as IL-2 and IFN-gamma secretion from allogeneic T cells, suggesting a Th1 bias. HMGB1 released by necrotic cells may be a signal of tissue or cellular injury that, when sensed by DCs, induces and/or enhances an immune reaction.


New learning area in laparoscopic gastrectomy for gastric cancer: YouTube® or WebSurg®?
Laparoscopic gastrectomy cannot be performed routinely in many centres, because there is still no standardisation in the centres where it is performed, and therefore, new learning areas are needed for residents working in these centres for surgical training. These areas are various courses, video training sets and video platforms watched over the internet. The most frequently used platforms are YouTube® and WebSurg®. Our aim is to compare these two online video platforms in terms of laparoscopic gastrectomy and to evaluate the contribution of video platforms to surgical training and whether they have sufficient technical quality. We made a search on YouTube® and WebSurg® using the keyword 'laparoscopic gastrectomy' on 13 November 2020. A total of 143 videos were analysed, 111 of them on YouTube® and 32 on WebSurg®. All these videos were examined by two surgical oncologists experienced in laparoscopic gastrectomy and using laparoscopy in their daily practice. The average video duration was 53.54 min in the YouTube® group and 18.20 min in the WebSurg® group, and this difference was found to be statistically significant between the two groups. According to the LGSS based on surgical procedures, the average score of WebSurg® videos was 10.37 and of YouTube® videos was 5.55, and there was a statistically significant difference between the two groups. Today, video platforms have started to play a major role in surgical training. Of these platforms examined, WebSurg® is superior to YouTube® in terms of education and quality, but these platforms still have some deficiencies and need regulation.


Methods to study cartilage and bone development.
In humans, the study of linear growth at the level of growth plate is limited to the use of mainly noninvasive methods. The use of in vitro studies and diverse animal models are alternative applied methods to unravel the underlying mechanisms of endochondral ossification and bone remodeling. In this chapter, we will outline important techniques to study molecular interactions and the regulation of gene expression in chondrocytes as well as bone cells. Microdissection of growth plate provides a powerful approach to study gene expression of individual zones and temporal senescence of growth plate cartilage by using microarray and real-time PCR analysis. IHC gives valuable information about the distribution of proteins of interest throughout distinct layers of the growth plate. The TUNEL assay has been successfully applied to study apoptosis in chondrocytes. In addition, organ(metatarsal) culture, primary and cell line cultures have been widely used to explore the regulation of chondrocyte differentiation under varying experimental conditions. Ultimately, these methods are essential to give new insights into the search for new diagnostic and therapeutic approaches for growth and various metabolic bone disorders.


External quality control program in infectious diseases screening at laboratories and blood banks in Latin America: an analysis of the past 5 years.
To evaluate the screening of blood samples for infectious disease markers at laboratories and blood banks in Latin America per the findings of an External Quality Assessment Program (EQAP). This qualitative analysis used data from the EQAP coordinated by the Fundação Pro Sangue Hemocentro de São Paulo with the support of the Pan American Health Organization to assess the performance of blood screening for infectious diseases from 2014 to 2018 in Latin America. Each participating laboratory or blood bank received an identical blind panel with 24 blood samples with variable reactivity for all the screening parameters. Panels were processed at each participating facility and results were returned to the Fundação Pro Sangue Hemocentro de São Paulo for individual and joint analyses. Two types of discrepant results were potential failures: false positive results (FPRs) and false nonreactive results (FNRRs). A total of 23 136 samples were evaluated. Global rates of FPR, FNRR, and concordant results were 0.3%, 1.0% and 98.7%, respectively. Seven FNRRs were found for HBsAg (1.0%), 12 for syphilis (2.6%), and 21 for Chagas disease (2.9%). No FNRRs were found for the HIV, HCV, and HTLV viruses. The average accuracy of all the laboratories and blood banks participating in the EQAP during the study period was 99.5% (standard deviation, 0.5%). The findings of this qualitative analysis are positive for blood safety in Latin America, with an average accuracy of 99.5% among the participating laboratories and blood banks. This report reflects an important improvement in blood bank serological screening EQAP-PAHO report since the 2003.


Explorations of fungal biosynthesis of reduced polyketides - a personal viewpoint.
This viewpoint on biosynthesis of reduced polyketides in fungi traces evolution of the research area over more than 4 decades. It is a companion to the related articles by two personal and scientific friends with whom there has been free exchange of ideas for over 30 years. Beginning with very rudimentary knowledge about assembly of such natural products, developments using stable isotope labelling and subsequently identification of biosynthetic genes, led to understanding of the processive nature of polyketide formation. Recent expression and isolation of fungal iterative polyketide synthase enzymes has enabled more detailed exploration of the mechanisms of these fascinating molecular machines.


Depressed Physical Performance Outlasts Hormonal Disturbances after Military Training.
The aim of this study was to investigate the effect of an arduous 1-wk military course on measures of physical performance, body composition, and blood biomarkers. Participants were apprentices in an annual selection course for the Norwegian Special Forces. Fifteen soldiers (23 ± 4 yr, 1.81 ± 0.06 m, 78 ± 7 kg) completed a hell week consisting of rigorous activity only interspersed by 2 to 3 h of sleep per day. Testing was conducted before and 0, 1, 3, 7, and 14 d after the hell week. Physical performance was measured as muscle strength and jump performance. Body composition was measured by bioelectrical impedance and blood samples were collected and analyzed for hormones, creatine kinase, and C-reactive protein. Body mass was reduced by 5.3 ± 1.9 kg during the hell week and returned to baseline within 1 wk. Fat mass was reduced by 2.1 ± 1.7 kg and muscle mass by 1.9 ± 0.9 kg. Muscle strength in leg press and bench press was reduced by 20% ± 9% and 9% ± 7%, respectively, and both were approximately 10% lower than baseline after 1 wk of recovery. Jump-height was reduced by 28% ± 13% and was still 14% ± 5% below baseline after 2 wk of recovery. Testosterone was reduced by 70% ± 12% and recovered gradually within a week. Cortisol was increased by 154% ± 74% and did not fully recover during the next week. Insulin-like growth factor 1 was reduced by 51% ± 10% and triiodothyronine and thyroxine by 12% to 30%, all recovered within a week. One-week arduous military exercise resulted in reductions in body mass and performance, as well as considerable hormonal disturbances. Our most important observation was that whereas the hormonal systems was normalized within 1 wk of rest and proper nutrition, lower body strength and jump performance were still depressed after 2 wk.


Olive oil-enriched diet reduces brain oxidative damages and ameliorates neurotrophic factor gene expression in different life stages of rats.
Our aim was to assess the influence of maternal diet rich in monounsaturated fatty acids on oxidative and molecular parameters in brains of mouse pups as well as their body weight during their lifetime. Female rats received a diet containing 20% of olive oil-enriched diet (OOED) and a standard diet control diet (CD) in different periods: pregnancy, lactation and after weaning until pups' adulthood. On the last prenatal day (Group 1), embryos from OOED group showed smaller body weight, brain weight and lower levels of sulphydryl groups glutathione reduced (GSH) in the brain. On postnatal delay-21 (PND21) (Group 2), pups from OOED group showed higher body weight and brain weight, reduced brain weight/body weight ratio and lower brain lipid peroxidation (LP). On PND70 (Group 3), pups from OOED group showed lower brain LP and higher levels of GSH in prefrontal cortex and lower brain levels of reactive species in the hippocampus. Interestingly, the group of animals whose diet was modified from OOED to CD on PND21 showed greater weight gain compared to the group that remained in the same original diet (OOED) until adulthood. Furthermore, OOED consumption during pregnancy and lactation significantly increased BDNF only, as well as its main transcripts exon IV and VI mRNA levels in the prefrontal cortex. In addition, OOED significantly up-regulated FGF-2 mRNA levels in the prefrontal cortex. These findings open a pioneering line of investigation about dietary adjunctive therapeutic strategies and the potential of healthy dietary habits to prevent neonatal conditions and their influence on adulthood.