Visualization of cellulose synthases in Arabidopsis secondary cell walls.
Cellulose biosynthesis in plant secondary cell walls forms the basis of vascular development in land plants, with xylem tissues constituting the vast majority of terrestrial biomass. We used plant lines that contained an inducible master transcription factor controlling xylem cell fate to quantitatively image fluorescently tagged cellulose synthase enzymes during cellulose deposition in living protoxylem cells. The formation of secondary cell wall thickenings was associated with a redistribution and enrichment of CESA7-containing cellulose synthase complexes (CSCs) into narrow membrane domains. The velocities of secondary cell wall-specific CSCs were faster than those of primary cell wall CSCs during abundant cellulose production. Dynamic intracellular of endomembranes, in combination with increased velocity and high density of CSCs, enables cellulose to be synthesized rapidly in secondary cell walls.


Age-dependent facilitation of taste-footshock conditioning by prior exposure to the training context.
Among adult rats, gustatory stimuli are easily associated with illness, but not with external unconditioned stimuli such as footshock. Recent evidence indicates, however, that this cue-to-consequence specificity may vary ontongenetically. The present studies examined the acquisition of an aversion to a taste paired with footshock in 5- and 15-day-old rats. Consistent with previous reports, 5-day-old rats avoided the taste that preceded footshock, while 15-day-old subjects did not express an aversion to the taste paired with footshock. Exposure to the training context for either 1 or 5 h prior to conditioning disrupted taste-footshock conditioning in the 5-day-old subjects. For the 15-day-old subjects, 1 h of pre-conditioning exposure to the training context had no effect on conditioning, whereas a longer duration of preexposure promoted conditioning to the taste cue. The results suggest ontogenetic differences in stimulus selection.


Conditional loss of TGF-beta signalling leads to increased susceptibility to gastrointestinal carcinogenesis in mice.
Downregulation of TGF-beta receptors is implicated in colon cancer development. Inactivation of either of the two transmembrane serine/threonine kinases, TGF-beta1 types I/II receptors, is now implicated in carcinogenesis, especially gastrointestinal carcinogenesis. We generated transgenic mice, called pS2-dnRII or ITF-dnRII, of which the dominant negative mutant of the TGF-beta type II receptor was expressed under the control of tissue-specific promoters, the pS2 promoter for stomach and ITF for intestine. They were either infected with H.pylori (ATCC 43504 strain, CagA+ and VacA+) or administered with azoxymethane to determine the significance of loss of TGF-beta signalling in gastrointestinal carcinogenesis. Gastric adenocarcinoma developed in pS2-dnRII mice, whereas only chronic active gastritis was noted in wild-type littermates after 36 weeks of H.pylori infection. Mice lacking in TGF-beta signalling specifically in the stomach showed a significantly higher proliferation cell nuclear antigen-labelling index when infected with H.pylori than wild-type littermates (P < 0.01). Development of colonic aberrant crypt foci was provoked in mice by intraperitoneal injections of azoxymethane, and ITF-dnRII mice showed significantly higher incidences of ACF and colon cancers than wild-type littermates. Maintaining normal TGF-beta signalling in the gastrointestinal tract seems to be important either for preventing abnormal mucosal proliferation, or for suppressing or retarding carcinogenesis.


The influence of race and hospital environment on the care of patients with cervical spine fractures.
The influence of non-white race on outcomes following orthopedic injury has been described in the past. The impact of such factors on hospital processes and quality of care after spinal trauma is less well understood. A cohort control study using the Massachusetts Statewide Inpatient Dataset (2003-2010) was used as the study design. This study aimed to determine whether (1) hospital processes and quality of care associated with the treatment of cervical spine fractures was significantly altered by non-white race and (2) whether findings were different among those treated at academic medical centers (AMCs). The study comprised 10,841 patients. Surgical rate, postoperative morbidity, mortality, and length of stay (LOS) were the outcome measures. Baseline differences between cohorts were evaluated using chi-square or Wilcoxon rank sum tests. Logistic and negative binomial regression techniques were used to adjust for confounders, including whether a surgical intervention was performed. Subset analyses were performed to evaluate whether findings were different for individuals treated at AMCs. The rate of surgical intervention was not significantly different between non-whites and whites (odds ratio [OR] 0.92, 95% confidence interval [CI] 0.82-1.04). LOS (regression coefficient [RC] 0.18, 95% CI 0.13-0.23), mortality (OR 1.49, 95% CI 1.20-1.85), and complications (OR 1.17, 95% CI 1.02-1.33) were significantly increased among non-white patients. These findings were largely preserved among those treated at AMCs. Our results reinforce the fact that efforts at universal access to care may be insufficient to reduce differences in care among minority patients following cervical trauma. Future mixed-methods research is necessary to more effectively evaluate the etiologies behind health-care disparities associated with race in different health-care environments. The level of evidence is Level III, prognostic study.


An exclusive human milk-based diet in extremely premature infants reduces the probability of remaining on total parenteral nutrition: a reanalysis of the data.
We have previously shown that an exclusively human milk-based diet is beneficial for extremely premature infants who are at risk for necrotizing enterocolitis (NEC). However, no significant difference in the other primary study endpoint, the length of time on total parenteral nutrition (TPN), was found. The current analysis re-evaluates these data from a different statistical perspective considering the probability or likelihood of needing TPN on any given day rather than the number of days on TPN. This study consisted of 207 premature infants randomized into three groups: one group receiving a control diet of human milk, formula and bovine-based fortifier ("control diet"), and the other two groups receiving only human milk and human milk-based fortifier starting at different times in the enteral feeding process (at feeding volumes of 40 or 100 mL/kg/day; "HM40" and "HM100", respectively). The counting process Cox proportional hazards survival model was used to determine the likelihood of needing TPN in each group. The two groups on the completely human-based diet had an 11-14 % reduction in the likelihood of needing nutrition via TPN when compared to infants on the control diet (p = 0.0001 and p = 0.001, respectively for the HM40 and HM100 groups, respectively). This was even more pronounced if the initial period of TPN was excluded (p < 0.0001 for both the HM40 and HM100 groups). A completely human milk-based diet significantly reduces the likelihood of TPN use for extremely premature infants when compared to a diet including cow-based products. This likelihood may be reduced even further when the human milk fortifier is initiated earlier in the feeding process. This study was registered at http://www.clinicaltrials.gov reg. # NCT00506584.


Probing the pigment binding sites in LHCII with resonance Raman spectroscopy: The effect of mutations at S123.
Resonance Raman spectroscopy was used to evaluate the structure of light-harvesting chlorophyll (Chl) a/b complexes of photosystem II (LHCII), reconstituted from wild-type (WT) and mutant apoproteins over-expressed in Escherichia coli. The point mutations involved residue S123, exchanged for either P (S123P) or G (S123G). In all reconstituted proteins, lutein 2 displayed a distorted conformation, as it does in purified LHCII trimers. Reconstituted WT and S123G also exhibited a conformation of bound neoxanthin (Nx) molecules identical to the native protein, while the S123P mutation was found to induce a change in Nx conformation. This structural change of neoxanthin is accompanied by a blue shift of the absorption of this carotenoid molecule. The interactions assumed by (and thus the structure of the binding sites of) the bound Chls b were found identical in all the reconstituted proteins, and only marginally perturbed as compared to purified LHCII. The interactions assumed by bound Chls a were also identical in purified LHCII and the reconstituted WT. However, the keto carbonyl group of one Chl a, originally free-from-interactions in WT LHCII, becomes involved in a strong H-bond with its environment in LHCII reconstituted from the S123P apoprotein. As the absorption in the Qy region of this protein is identical to that of the LHCII reconstituted from the WT apoprotein, we conclude that the interaction state of the keto carbonyl of Chl a does not play a significant role in tuning the binding site energy of these molecules.


Vasculitis after blood pressure monitoring.
Description of appearance of ecchymosis on an arm, simultaneously with a classical Leukocytoclastic Vasculitis, the proposal of alternative utilities of measuring blood pressure, and the study of side effects to that measure. Case 80-year-old male came to ER with dyspnea, heart failure, predialysis renal failure with hyperkalemia and hemodynamic instability. During his stay he developed a skin lesion that looks like palpable purpura, from the lower limit of the blood pressure cuff to the distal area of the hand that not disappeared with vitropression, and pruritus. During admission the arm injury was extended to all members, both upper and lower. The study concluded with diagnosis of Leukocytoclastic Vasculitis given the presence of eosinophils, that which suggested probable drug etiology to an antibiotic that had been taken since seven days prior to admission to ER. The need for serial monitoring of blood pressure, and the duration of such monitoring in unstable patients considering the side effects of those techniques was questioned. In addition, the study of other utilities of measuring blood pressure.


Purification, characterization, and gene cloning of thermopsin, a thermostable acid protease from Sulfolobus acidocaldarius.
A thermostable, acid proteolytic activity has been found to be associated with the cells and in the culture medium of Sulfolobus acidocaldarius, an archaebacterium. This acid protease, which has been named thermopsin, was purified to homogeneity from the culture medium by a five-step procedure including column chromatographies on DEAE-Sepharose CL-6B, phenyl-Sepharose CL-4B, Sephadex G-100, monoQ (fast protein liquid chromatography), and gel filtration (high pressure liquid chromatography). The purified thermopsin produced a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the proteolytic activity was associated with the band. Thermopsin is a single-chain protein as indicated by gel electrophoresis and by a single NH2-terminal sequence. It has maximal proteolytic activity at pH 2 and 90 degrees C. A genomic library of S. acidocaldarius was prepared and screened by an oligonucleotide probe designed from the NH2-terminal sequence of thermopsin. Five positive clones were isolated. From these clones the thermopsin gene was mapped and sequenced. The nucleotide sequence showed that the thermopsin structure is encoded in 1020 bases. In the deduced protein sequence, there are 41 amino acid residues (including the initiation Met) preceding the NH2-terminal position of thermopsin. Most of these residues appear to be characteristic of a leader sequence. However, the presence in this region of a short pro sequence cannot be ruled out. Thermopsin contains a single cysteine at residue 237 that is not essential for activity (Fusek, M., Lin, X.-L., Tang, J. (1990) J. Biol. Chem. 265, 1496-1501. Thermopsin has no apparent sequence similarity to aspartic proteases of the pepsin family nor to pepstatin-insensitive acid protease (Maita, T., Nagata, S., Matsuda, G., Murata, S., Oda, K., Murao, S., and Tsura, D. (1984) J. Biochem. 95, 465-475) and thus may represent a new class of acid proteases. Also absent is the characteristic active site aspartyl sequence of aspartic proteases. There are 11 potential N-glycosylation sites on each thermopsin molecule. The molecular weight estimated from gel filtration (45,000) is larger than that calculated from the sequence (32,651), suggesting that thermopsin is the sequence (32,651), suggesting that thermopsin is glycosylated at at least some of these 11 sites.


Renal cell carcinoma--angioinfarction.
In our experience the mortality rate in 46 patients who underwent angioinfarction for renal cell carcinoma was 4 per cent. Fever, ileus and leukocytosis were noted in 86 to 90 per cent of our patients. The use of absolute ethanol as a medium for renal infarction was associated with a significant incidence of damage to other organs. A 30 per cent decrease in tumor volume following angioinfarction using absorbable gelatin sponge and Gianturco coils was noted in 75 per cent of the patients. There was no evidence of metastatic tumor reduction and we could not document any significant decrease in operative time or blood loss. It would appear that there is some increased survival rate in patients with metastasis who are given adjuvant immune ribonucleic acid therapy. However, the numbers in our series are not significant to draw any definite conclusion. It is apparent that patients treated with infarction, delayed nephrectomy and medroxyprogesterone acetate did not have any significant survival over those treated with palliative nephrectomy or chemotherapy. The macrophage maturation assay may be useful during clinical followup.


Surgical treatment of the cancer of the optic nerve and orbit. An historical overview.
Introduction: Aim of the present study is to present an historical overview of the surgical techniques introduced for the treatment of the cancer of the optic nerve and orbit.Methods: From the late 19th and early twentieth century there were introduced many novel surgical techniques for the treatment of the cancer of the optic nerve and orbit. The majority of these techniques concerned for the cure of orbital malignance, while optic nerve cancer many times was treated by enucleation of the eyeball.Results: Except of the extraction of the malignant tumors the surgeons of the time tried to present the best aesthetic result, fact which points the progress of the surgery those years.Conclusions: Many of the techniques introduced those years are still in use.


NK cell marker gene-based model shows good predictive ability in prognosis and response to immunotherapies in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is the fourth leading cause of malignancy worldwide, and its progression is influenced by the immune microenvironment. Natural killer (NK) cells are essential in the anti-tumor response and have been linked to immunotherapies for cancers. Therefore, it is important to unify and validate the role of NK cell-related gene signatures in HCC. In this study, we used RNA-seq analysis on HCC samples from public databases. We applied the ConsensusClusterPlus tool to construct the consensus matrix and cluster the samples based on their NK cell-related expression profile data. We employed the least absolute shrinkage and selection operator regression analysis to identify the hub genes. Additionally, we utilized the CIBERSORT and ESTIMATE web-based methods to perform immune-related evaluations. Our results showed that the NK cell-related gene-based classification divided HCC patients into three clusters. The C3 cluster was activated in immune activation signaling pathways and showed better prognosis and good clinical features. In contrast, the C1 cluster was remarkably enriched in cell cycle pathways. The stromal score, immune score, and ESTIMATE score in C3 were much higher than those in C2 and C1. Furthermore, we identified six hub genes: CDC20, HMOX1, S100A9, CFHR3, PCN1, and GZMA. The NK cell-related genes-based risk score subgroups demonstrated that a higher risk score subgroup showed poorer prognosis. In summary, our findings suggest that NK cell-related genes play an essential role in HCC prognosis prediction and have therapeutic potential in promoting NK cell antitumor immunity. The six identified hub genes may serve as useful biomarkers for novel therapeutic targets.


Linking inflammation and neuroendocrine differentiation: the role of macrophage migration inhibitory factor-mediated signaling in prostate cancer.
A new paper by Tawadros et al. in Endocrine-Related Cancer demonstrates a link between macrophage migration inhibitory factor and neuroendocrine differentiation in prostate cancer. This paper may have implications in explaining the effect of prostatitis and chronic inflammation on the development of aggressive prostate cancer.


Effect of sport training on forearm bone sites in female handball and soccer players.
The objective of this study was to evaluate the bone mineral density (BMD) and Z-score of a skeletal region, considered as weight-bearing site in trained handball players (HP), but as non-weight-bearing site in trained soccer players (SP). The bone health status of the same site was also analyzed in an untrained group (CG). BMD and Z-score at distal forearm regions (dominant, D; non-dominant, ND) were evaluated in 30 female HP and in 30 female SP, who have been training for 7.7±3.8 years, 17 hours per week, as well as in 30 females CG. Playing handball was associated with higher BMD of the skeleton at both measured sites than in CG. Also in comparison with SP, HPs' arms showed a significant increase in BMD. On the other hand, female SP have been reported to exhibit an enhanced ND arm BMD compared with controls. The benefits of exercise appeared to be significantly improved only in SP sub-group who started sport activity before or at menarche. These athletes showed at ND forearm a BMD 4% greater than those SP who started later, reaching a BMD of 11.6% higher than CG, a value similar to the corresponding in HP sub-group. Moreover, their D arm BMD was 7.1% higher compared with CG. This study indicates that, compared with non-trained subjects, long-term high-impact sport participation is associated with an higher bone health state, especially if the playing careers were started before or at menarche. This effect was observed at level of both forearms in HP, which are considered as weight-bearing sites for this discipline but also on the same skeletal regions of SP, which are not directly loaded by sport-related regular training.


Hormonal and ultrasound markers of ovarian function in a woman with a balanced 1;11 translocation.
To present a case of a poor responder woman with a rare balanced autosomal translocation. Case report. Infertility center in a university teaching hospital. A 34-year-old white woman with primary infertility. Laboratory and ultrasound evaluation, karyotype analysis, ovarian stimulation with gonadotropins. Markers of ovarian reserve and ovarian response to gonadotropin stimulation. The patient was a carrier of a balanced translocation (1;11) (q23;p11.2). The hormone ovarian reserve markers (FSH, anti-Mullerian hormone, inhibin B) were within the normal range. The antral follicle count (AFC) was measured twice and only in one occasion did it reach the lower limit of the normal range. Furthermore, the patient had regular menstrual cycles. However, ovarian response to the administration of a maximal dose FSH was extremely poor. Patients with balanced autosomal translocations should be counseled regarding their increased risk for poor response even if markers of ovarian reserve are in the range of normality.


The distribution and taxonomic value of fatty acids and eicosanoids in the Lipomycetaceae and Dipodascaceae.
Using radioimmunoassay, blood platelet aggregation studies and GC-MS the existence of prostaglandins in the endomycetalean yeast Dipodascopsis uninucleata was confirmed by our group. These findings triggered the search for similar eicosanoids in the rest of the Endomycetales. We commenced by scanning for the easily detectable precursors of eicosanoids, linoleic- and linolenic acid. We selected two families (i.e. Lipomycetaceae and Dipodascaceae), both producing these precursors, for further investigation. Representative strains of the two families were tested for their ability to grow in the presence of 1 mM aspirin, a specific inhibitor of prostaglandin biosynthesis. In contrast to the lipomycetaceous species the dipodascaceous species were insensitive to this drug. These results were verified when representative strains of both families were investigated for their ability to produce eicosanoids from externally fed radio-labeled arachidonic acid along an aspirin sensitive pathway. Thin layer chromatography of culture extracts, followed by autoradiography, showed that while none of the Dipodascaceae produced aspirin sensitive arachidonic acid metabolites, the members of the Lipomycetaceae tested positive for these metabolites. These findings supported the separation of the lipomycetaceous yeast Dipodascopsis from the Dipodascaceae. The findings also correlate with the delimitation of these yeasts in two families (i.e. Dipodascaceae and Lipomycetaceae). Further investigation indicated that prostaglandin production by the genus Dipodascopsis is mainly associated with ascosporogenesis. Thin layer chromatography of cell extracts from Dipodascopsis tóthii, followed by scintillation counting, indicated the presence of PGF2 alpha and PGE2 during ascosporogenesis.


MicroRNA-141 inhibits glioma cells growth and metastasis by targeting TGF-β2 [Retraction].
[This retracts the article on p. 3513 in vol. 8, PMID: 27648141.].


Impact of Extracorporeal Membrane Oxygenation on Mortality in Adults With Cardiac Arrest.
Previous studies regarding benefit of extracorporeal membrane oxygenation (ECMO) in adults with cardiac arrest have yielded conflicting results. We aimed to determine whether ECMO in adults hospitalized with cardiac arrest is associated with improved survival compared with conventional cardiopulmonary resuscitation in a nationally representative sample. The U.S. Healthcare Cost and Utilization Project's Nationwide Inpatient Sample (2006 to 2014) was utilized to identify a cohort of adults hospitalized with cardiac arrest at ECMO-capable facilities (defined by the presence of at least 1 ECMO procedure at the facility during the study period of 2006 to 2014). In-hospital mortality was compared between the ECMO and non-ECMO groups using generalized estimating equations with and without coarsened exact matching analysis. Of 273,690 hospitalizations for adults with cardiac arrest, 33,274 occurred at 363 ECMO-capable facilities, of which 775 (2.3%) involved the use of ECMO. There was no significant difference in in-hospital mortality between patients who received ECMO versus those who did not (60.1% vs 57.2%, p = 0.106). In the risk-adjusted analysis, the presence of ECMO was associated with higher rates of in-hospital mortality in the overall sample [odds ratio 1.59, 95% confidence interval 1.37 to 1.85] and the coarsened exact matching sample (n = 1,068 with 534 adults in each group; odds ratio 1.47, 95% confidence interval 1.14 to 1.88). In this cohort of hospitalizations for cardiac arrest at ECMO-capable centers in the U.S., adults who received ECMO had significantly higher mortality than those who did not receive ECMO. Large scale, adequately powered, randomized controlled trials are warranted to assess the benefit of ECMO in cardiac arrest.


Curcumin induces brown fat-like phenotype in 3T3-L1 and primary white adipocytes.
Recent advances have been made in the understanding of pharmacological and dietary agents that contribute to browning of white adipose tissue in order to combat obesity by promoting energy expenditure. Here, we show that curcumin induces browning of 3T3-L1 and primary white adipocytes via enhanced expression of brown fat-specific genes. Curcumin-induced browning in white adipocytes was investigated by determining expression levels of brown adipocyte-specific genes/proteins by real-time reverse transcriptase polymerase chain reaction, immunoblot analysis and immunocytochemical staining. Curcumin increased mitochondrial biogenesis, as evidenced by transmission electronic microscopic detection and enhanced expression of proteins involved in fat oxidation. Cucurmin also increased protein levels of hormone-sensitive lipase and p-acyl-CoA carboxylase, suggesting its possible role in augmentation of lipolysis and suppression of lipogenesis. Increased expression of UCP1 and other brown adipocyte-specific markers was possibly mediated by curcumin-induced activation of AMP-activated protein kinase (AMPK) based on the fact that inhibition of AMPK by dorsomorphin abolished expression of PRDM16, UCP1 and peroxisome proliferator-activated receptor gamma co-activator 1-alpha while the activator 5-Aminoimidazole-4-carboxamide ribonucleotide elevated expression of these brown marker proteins. Our findings suggest that curcumin plays a dual modulatory role in inhibition of adipogenesis as well as induction of the brown fat-like phenotype and thus may have potential therapeutic implications for treatment of obesity.


Acupuncture for patients with Maigne's syndrome: A case series.
Diagnosing the precise etiology of low back pain (LBP) is crucial for facilitating speedy recovery in patients. Maigne's syndrome (MS), commonly referred to as thoracolumbar junction syndrome, is a condition characterized by pain resulting from nerve entrapment, yet its underlying mechanisms remain poorly understood. This study presents a series of six case reports wherein patients diagnosed with MS received acupuncture treatment. Six individuals with LBP were included in the study, and all were diagnosed with MS. The diagnosis was confirmed in all six patients through pinch-roll and thoracic vertebrae compression tests, indicating the presence of thoracolumbar junction syndrome. Acupuncture treatment was administered to all patients, primarily targeting the T11-L2 facet joints, with additional acupoints selected based on the specific nerve entrapment of MS including the superior cluneal, subcostal, and iliohypogastric nerves. Following acupuncture therapy, all patients reported improvements in their LBP symptoms, while four patients also exhibited amelioration in their thoracic vertebrae compression test results. These findings underscore the significance of promptly diagnosing the underlying cause of LBP and suggest that acupuncture may be an effective approach in alleviating MS-related pain.


Acute and prolonged hindlimb exercise elicits different gene expression in motoneurons than sensory neurons after spinal cord injury.
We examined gene expression in the lumbar spinal cord and the specific response of motoneurons, intermediate gray and proprioceptive sensory neurons after spinal cord injury and exercise of hindlimbs to identify potential molecular processes involved in activity dependent plasticity. Adult female rats received a low thoracic transection and passive cycling exercise for 1 or 4weeks. Gene expression analysis focused on the neurotrophic factors: brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), neurotrophin-3 (NT-3), neurotrophin-4 (NT-4), and their receptors because of their potential roles in neural plasticity. We also examined expression of genes involved in the cellular response to injury: heat shock proteins (HSP) -27 and -70, glial fibrillary acidic protein (GFAP) and caspases -3, -7, and -9. In lumbar cord samples, injury increased the expression of mRNA for TrkB, all three caspases and the HSPs. Acute and prolonged exercise increased expression of mRNA for the neurotrophic factors BDNF and GDNF, but not their receptors. It also increased HSP expression and decreased caspase-7 expression, with changes in protein levels complimentary to these changes in mRNA expression. Motoneurons and intermediate gray displayed little change in mRNA expression following injury, but acute and prolonged exercise increased levels of mRNA for BDNF, GDNF and NT-4. In large DRG neurons, mRNA for neurotrophic factors and their receptors were largely unaffected by either injury or exercise. However, caspase mRNA expression was increased by injury and decreased by exercise. Our results demonstrate that exercise affects expression of genes involved in plasticity and apoptosis in a cell specific manner and that these change with increased post-injury intervals and/or prolonged periods of exercise.


A novel hybrid transcatheter ablation system that combines radiofrequency and cryoenergy.
Radiofrequency (RF) and cryoenergy are largely considered independent modalities for the transcatheter ablation of cardiac arrhythmias. There are numerous theoretical advantages to engineering a system capable of delivering both energy forms. We designed a hybrid steerable catheter capable of delivering RF and cryoenergy independently, sequentially, and simultaneously. The novel catheter system was tested pre-clinically by creating a total of 180 ablation lesions in 20 mongrel dogs. Right atrial and right and left ventricular sites were preselected by a randomized factorial design devised to compare sequential and simultaneous RF and cryoenergy applications to standard RF, irrigated RF, and standard cryoablation. A steerable 4-mm electrode-tip hybrid catheter ("Fire and Ice") was created by modifying a 7 F cryocatheter (Freezor, CryoCath Technologies, Montreal, Canada). RF energy was injected via a copper wire, thermocouples were isolated to reduce RF interference, and 100 KHz band pass filters and RF chokes were added. Sequential low-dose RF (20 W, 60 seconds) preceding or following cryoablation resulted in larger lesions (P = 0.0010). The addition of RF energy did, however, produce more thrombus than cryoenergy alone, with clot detected on 82.4% versus 12.1% of ablation lesions, P < 0.0001. Simultaneously applying the two energy modalities (45 W, 10 or 30 degrees C, 60 seconds) created more voluminous lesions than standard RF ablation (median 288.1 vs 126.1 mm(3), P = 0.0333) of similar dimension to irrigated RF ablation. A versatile catheter system was fashioned capable of creating standard cryoablation lesions, standard RF lesions, and simultaneous lesions of similar dimension to irrigated RF.


Erratum: 10 Years of Toxicogenomics section in Frontiers in Genetics: Past discoveries and Future Perspectives.
[This corrects the article DOI: 10.3389/fgene.2022.979761.].


Developing LC-MS/MS methods to quantify rivaroxabanin human plasma and urine: Application to therapeutic drug monitoring.
Rivaroxaban is an oral anticoagulant directly inhibiting the activity of Factor Xa, which is widely used for the prophylaxis of thromboembolic disorders. Therapeutic drug monitoring (TDM) is required during therapy for individual dosage adjustment. This study aimed at developing a liquid chromatography/tandem mass spectrometry method that was suitable for rivaroxaban TDM in human plasma and urine and exploring the feasibility of urine drug monitoring in medical care. A 3 min run time of the LC-MS/MS methods was established by employing an Acquity UPLC BEH C18 (2.1 × 50 mm, 1.7 μm) column using gradient elution of 10 mmol/L ammonium acetate containing 0.1% formic acid-0.1% formic acid acetonitrile as a mobile phase at a flow rate of 0.4 ml/min with calibration ranges of 0.5-400 and 10-10,000 ng/ml for human plasma and urine, respectively. Rivaroxaban was detected on a triple quadrupole tandem mass spectrometer with an electrospray ionization source in positive ion mode. The methods showed good linearity within the calibration range. The precision and accuracy, matrix effect, extraction recovery and stability in both human matrices were all validated and meet the international guideline requirements. These validated methods were successfully applied to support the TDM of an aged patient receiving rivaroxaban for therapy.


Basal cell carcinoma in farmers: an occupation group at high risk.
Skin cancer is the most commonly diagnosed cancer type worldwide, and 80 % of skin cancers are basal cell carcinoma (BCC). The main risk factor for developing BCC is exposure to ultraviolet radiation (UVR), particularly high-dose exposure at a young age. Outdoor workers, particularly farmers, are at high risk of developing BCC. However, studies of BCC in this population are scant. To comprehensively evaluate all cases of BCC of the head and neck region treated during the years 2007-2013 at our hospital in Poland, and to compare the tumour characteristics in farmers to non-farmers. Retrospective analysis of 312 patients treated for head and neck BCC during the study period (2007-2013). Most patients (198 cases; 63 %) were males, with 114 females (37 %). Median age was 73 years (range 32-96 years). The most common tumour location was the nose and cheek (114 pts; 37 %) followed by the auricle (82 pts; 26 %), lips (54 pts; 18 %), scalp (26 pts; 8 %), and eye (36 pts; 12 %). The most common disease stage on presentation was stage T2 (104 pts, 33 %), followed by stage T1 (79 pts; 25 %), stage T3 (89 pts; 28 %), and stage T4 (40 pts; 14 %). By occupation, farmers accounted for 33 % of all patients (102 of 312 pts). The most common tumour localisations in the farmer subgroup were the nose and cheek (50 pts; 49 %; p < 0.001; odds ratio [OR] 2.19; 95 % confidence interval [CI] 1.35-3.57), followed by the auricle (32 pts; 31 %), scalp (16 pts; 16 %), ocular region (3 pts; 3 %), and lips (1 pt; 1 %). Patients in the farmer group were significantly younger than non-farmers (62 vs. 73 years; p < 0.001; OR 0.90, 95 % CI 0.88-0.93). Farmers were significantly more likely to present disease recurrence (27 vs. 12 % of cases; p < 0.001; OR 5.94; 95 % CI 2.86-12.33). The results highlight the increased incidence and risk of recurrence of BCC in farmers. It is therefore necessary to consider enhancing educational programmes and other preventative measures in this occupational group and to evaluate the effectiveness of such programmes.


Microglial NADPH oxidase mediates leucine enkephalin dopaminergic neuroprotection.
Here, we report that leucine enkephalin (LE) is neuroprotective to dopaminergic (DA) neurons at femtomolar concentrations through anti-inflammatory properties. Mesencephalic neuron-glia cultures pretreated with femtomolar concentrations of LE (10(-15)-10(-13) M) protected DA neurons from lipopolysaccharide (LPS)-induced DA neurotoxicity, as determined by DA uptake assay and tyrosine hydroxylase (TH) immunocytochemistry (ICC). However, des-tyrosine leucine enkephalin (DTLE), an LE analogue that is missing the tyrosine residue required for binding to the kappa opioid receptor, was also neuroprotective (10(-15)-10(-13) M), as determined by DA uptake assay and TH ICC. Both LE and DTLE (10(-15)-10(-13) M) reduced LPS-induced superoxide production from microglia-enriched cultures. Further, both LE and DTLE (10(-14), 10(-13) M) reduced the LPS-induced tumor necrosis factor-alpha (TNFalpha) mRNA and TNFalpha protein from PHOX+/+ microglia, as determined by quantitative real-time RT-PCR and ELISA analysis in mesencephalic neuron-glia cultures, respectively. However, both peptides failed to inhibit TNFalpha expression in PHOX-/- cultures, which are unable to produce extracellular superoxide in response to LPS. Additionally, LE and DTLE (10(-14), 10(-13) M) failed to show any neuroprotection against LPS in PHOX-/- cultures. Together, these data indicate that LE and DTLE are neuroprotective at femtomolar concentrations through the inhibition of oxidative insult associated with microglial NADPH oxidase and the attenuation of the ROS-mediated amplification of TNFalpha gene expression in microglia.


Roles, responsibilities, and outcome expectations of endowed chairs in nursing.
This study, using a modified Delphi approach, examines the purpose, roles, responsibilities, and outcomes of endowed chairholders in nursing. Deans and faculty who hold endowed chairs participated in this 3-phased study. While the ranking of importance varied between these 2 groups of experts, common themes emerged relative to the phenomenon under investigation. Findings from this preliminary work can be used by deans and chairholders to develop job descriptions and evaluate the impact of endowments in a program of nursing.


Electrochemiluminescence immunoassay for the N-terminal pro-B-type natriuretic peptide based on resonance energy transfer between a self-enhanced luminophore composed of silver nanocubes on gold nanoparticles and a metal-organic framework of type MIL-125.
The N-terminal pro-B-type natriuretic peptide (NT-proBNP) is a marker of heart failure. A novel sandwich type electrochemiluminescence (ECL) immunoassay is described for the NT-proBNP. The method is based on ECL resonance energy transfer (RET) between silver nanocubes that were covered with semicarbazide-modified gold nanoparticles (AgNC-sem@AuNPs) as the donor, and a Ti(IV)-based metal-organic framework of type MIL-125 as the acceptor. The ECL signal was strongly amplified by increasing the luminous efficiency. ECL-RET occurs due to the partial overlap between the ECL emission of the AgNC-sem@AuNPs (emission wavelength at 470 nm to 900 nm) and the visible absorption spectrum of MIL-125 (absorption wavelength at 406 nm to 900 nm). This results in the quenching of ECL. The AgNC-sem@AuNPs were placed on the electrode. The antibody was immobilized on AgNC-sem@AuNPs via Au-NH2 bond, and MIL-125 was utilized as a label for the secondary antibody. The assay works in the 0.25 pg mL-1 to 100 ng mL-1 concentration range and has a 0.11 pg mL-1 lower detection limit (at S/N = 3). Graphical abstract Schematic representation of self-enhanced luminescence mechanism (semicarbazide (Sem) as co-reaction accelerator) and Electrochemiluminescence resonance energy transfer (ECL-RET): silver nanocubes (AgNCs) as the energy donor and MIL-125 as the energy acceptor.


Impact of pre-exposure and post-exposure prophylaxes prevention programme on HIV burden and services in a low-resource setting: a simulation modelling approach.
sub-Saharan African countries contribute substantially to the global HIV disease burden. Despite this burden, and the promises that prevention could deliver, the implementation and uptake of HIV prevention programmes are still low. The study used the decision support system model to explore the potential impacts of prevention implementation on HIV burden (incidence) and service delivery. an operational research technique known as discrete event simulation model was used to capture an individual patient´s pathways through the HIV care process from diagnosis to treatment and monitoring. The regular monitoring, over a 5-year period, including all the activities and resources utilized at each stage of the pathway were analysed, and the impact of increasing prevention measures for an HIV treatment service in a treatment centre in Nigeria was tested using the simulation model. forty-three patients currently access the Pre-Exposure Prophylaxis (PrEP) and Post Exposure Prophylaxis (PEP) annually, with a 20% and 80% split in the number of patients offered PrEP and PEP, respectively. Scenarios-based on increasing the number of people offered PrEP and PEP from 43 to 250 with a 50/50 split were tested. The outputs revealed improved preventive care by averting new HIV cases, reduction in service demand and utilization, but an increase in the required human resource as well as financial burden. In the next 5 years, the cumulative averted HIV cases are expected to increase from 2 and 5 people (baseline) to 24 and 20 people for PrEP and PEP, respectively. The potentially averted 2 cases per infected persons based on the basic reproductive number of HIV. the effective implementation of PrEP/PEP programme offers an additional safety measure to prevent HIV transmission in at-risk individuals and possibility of ending HIV epidemic.


Corrigendum: Chromatin modification and NBS1: their relationship in DNA double-strand break repair [Genes Genet. Syst. (2015) 90, p. 195-208].
Table 1 on p. 203 should be replaced with the corrected Table 1 shown bellow. <img align="middle" src="./Graphics/abst-91_241_a.jpg">.


[End-of life decisions in intensive care units. Influence of patient wishes on therapeutic decision making].
Among the ethical principles in medicine, respect for patient autonomy has gained the highest revaluation in recent decades. In Germany this was fostered by new legal regulations which came into effect in 2009 and clarified issues regarding end-of-life (EOL) decisions. In this study the influence of direct or mediated wishes of patient wills on EOL decisions in a medical intensive care unit (ICU) were investigated. A retrospective analysis of all patients who died in the years 2009-2010 while being treated in the medical ICU of a large German university hospital was carried out. During the observation period 3,401 patients were treated in the ICU of whom 19 % (n=658) died in hospital. Of the 658 patients who died 126 (19 %) had received unlimited therapy, life support was withheld in 241 patients (37 %) and life support was withdrawn in 245 patients (37 %). In 46 patients (7 %) palliative care was instituted from the beginning of the ICU stay. In 104 cases (16 %) the patients themselves made the EOL decision and in 78 cases (12 %) an advance directive was given. A legal healthcare proxy was designated in 8 %. In 541 cases (82 %) the relatives were involved in the EOL decisions. No serious or unsolvable conflicts with relatives were experienced. Involvement of a court of law was not necessary in any of the cases. In a high percentage of the patients (81 %) who died during the course of intensive care treatment EOL policies were in place. The patients or their relatives were almost always involved in the decision making process. The current German law is in concordance with the established EOL practice in this intensive care unit.


Hemodynamic benefits of atrioventricular sequential pacing after cardiac surgery.
The hemodynamic effects of atrioventricular (A-V) sequential pacing were assessed and compared with those of ventricular and of atrial pacing in 10 patients with and without heart block after cardiac surgery. Ventricular pacing alone was either hemodynamically detrimental or of no benefit in six of the eight patients who initially had sinus or accelerated junctional rhythms. Atrial pacing alone produced significant improvement in cardiac output in all patients who were not pacemaker-dependent. However, five of the eight patients with intact A-V conduction had further increases in cardiac output through A-V sequential pacing at shorter than intrinsic A-V intervals. Optimal A-V intervals for maximal cardiac output could be identified in all patients and varied widely. Significant changes in cardiac output occurred with relatively small diviations in the A-V interval. In selected patients after cardiac surgery, temporary A-V sequential pacing is a workable and valuable adjunctive form of hemodynamic support and is preferable to ventricular or atrial pacing.


The effect of probiotics and organic acids on Shiga-toxin 2 gene expression in enterohemorrhagic Escherichia coli O157:H7.
Probiotics are known to have an inhibitory effect against the growth of various foodborne pathogens, however, the specific role of probiotics in Shiga-toxin-producing Escherichia coli (STEC) virulence gene expression has not been well defined. Shiga toxins are members of a family of highly potent bacterial toxins and are the main virulence marker for STEC. Shiga toxins inhibit protein synthesis in eukaryotic cells and play a role in hemorrhagic colitis and hemolytic uremic syndrome. STEC possesses Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2), both of which have A and B subunits. Although STEC containing both Stx1 and Stx2 has been isolated from patients with hemorrhagic colitis, Stx2 is more frequently associated with human disease complications. Thus, the effect of Lactobacillus, Pediococcus, and Bifidobacterium strains on stx2A expression levels in STEC was investigated. Lactic acid bacteria and bifidobacteria were isolated from farm animals, dairy, and human sources and included L. rhamnosus GG, L. curvatus, L. plantarum, L. jensenii, L. acidophilus, L. casei, L. reuteri, P. acidilactici, P. cerevisiae, P. pentosaceus, B. thermophilum, B. boum, B. suis and B. animalis. E. coli O157:H7 (EDL 933) was coincubated with sub-lethal concentrations of each probiotic strain. Following RNA extraction and cDNA synthesis, relative stx2A mRNA levels were determined according to a comparative critical threshold (Ct) real-time PCR. Data were normalized to the endogenous control glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the level of stx2A expression between treated and untreated STEC was compared. Observed for all probiotic strains tested, stx2A was down-regulated, when compared to the control culture. Probiotic production of organic acids, as demonstrated by a decrease in pH, influenced stx2A gene expression.


Interpersonal violence: secondary analysis of the keep your children/yourself safe and secure (KySS) data.
The purpose of the secondary analysis was to determine agreement of parents and children to the interpersonal violence (IPV) items on the Keep Your Children/Yourself Safe and Secure (KySS) survey to assess mental health concerns. A comparative, descriptive design was used to determine the level of agreement between parent/child dyads on the 15 attitude/knowledge and worry items related to IPV (n = 563 dyads). Single mother/son and 10- to 12-year-olds had less agreement than other dyad combinations and child age groups. Six of the 10 knowledge/attitude items had significant differences, including multigenerational occurrence of sexual abuse, the effect of harsh parenting, witnessing domestic violence, and incidence of physical abuse. The least agreement was on sexual abuse, incidence of physical abuse, and outcome of strict parenting (p < .001). Parents had greater worry about the parent-child relationship than did the children (p < .001). Parents and clinicians may not realize how different their understanding of IPV is from that of children. Identifying items of least agreement can inform parents and clinicians in targeting violence education and prevention.


Hungarian consensus recommendation on the role of vitamin D in disease prevention and treatment
Nine Hungarian medical societies have developed a consensus recommendation on the preferred normal range of vitamin D, the dose of vitamin D supplementation and the method of administration. They summarized the clinical conditions and diseases the development of which may be associated with vitamin D deficiency (VDD). VDD is extremely common in Hungary, especially in late winter. The lower limit of the recommended normal range is 75 nmol/l, although the clinical significance of deficiency is evident mainly at values below 50 nmol/l, but since vitamin D supplementation at the recommended dose is safe, it is worthwhile for everyone to reduce the health risk associated with VDD. The aim of vitamin D supplementation is to prevent deficiency. The recommended normal range is 75–125 nmol/l, above which there is no clear benefit of vitamin D supplementation. To maintain the normal range, a daily intake of 2000 IU in adults is recommended during the UV-B radiation-free period. Vitamin D supplementation is also recommended for children during the same periods and conditions as for adults, but the dose varies with age. In adults, vitamin D3 supplementation at daily, weekly and monthly intervals is equally effective and safe. In severe deficiency, a loading dose is recommended, followed by maintenance supplementation. In addition to the wellknown skeletal, immunological and oncological effects of VDD, more and more data support unfavorable gyneco- logical and obstetric effects. The process of building the consensus has met the requirements of the latest Delphi criteria.


Lipoprotein(a) levels are associated with coronary severity but not with outcomes in Chinese patients underwent percutaneous coronary intervention.
The association between lipoprotein(a) [Lp(a)] levels and the risk of cardiovascular disease is of great interest but still controversial. This study sought to investigate the impact of Lp(a) on coronary severity and long-term outcomes of patients who undergo percutaneous coronary intervention (PCI). A total of 6714 consecutive patients who received PCI were enrolled to analyze the association between Lp(a) and coronary severity and major adverse cardiovascular and cerebrovascular events (MACCE). Patients were divided into tertiles according to Lp(a) levels on admission. Coronary severity was evaluated by SYNTAX scoring system. The MACCE included recurrent myocardial infarction, unplanned target vessel revascularization, stent thrombosis, ischemic stroke and all-cause mortality. Significantly, Lp(a) levels were positively associated with coronary severity (p < 0.001). Multivariate logistic regression analyses showed Lp(a) was an independent predictor of intermediate to high SYNTAX score. During an average of 874 days follow-up, 755 patients presented with MACCE (11.25%) were reported. The incidence rates of MACCE, all-cause mortality, cardiac death, target vessel revascularization, recurrent myocardial infarction, stent thrombosis, stroke and bleeding were not statistically different among the Lp(a) tertile groups. Furthermore, both Kaplan-Meier and Cox regression analyses found no relationship between Lp(a) and cardiovascular outcomes (p > 0.05). Lp(a) is an independent predictor of the prevalence of more complex coronary artery lesions (SYNTAX score ≥ 23) in patients with PCI. In addition, our study has shown that Lp(a) has no relationship with long-term cardiovascular outcomes in Chinese patients with PCI.


Mutation analysis of the phenylalanine hydroxylase gene and its clinical implications in two Japanese patients with non-phenylketonuria hyperphenylalaninemia.
We describe a mutation analysis for the phenylalanine hydroxylase gene and the clinical outcome of two Japanese patients with non-phenylketonuria (PKU) hyperphenylalaninemia (serum phenylalanine level below 600 mumol/l under a free diet) detected by a mass-screening program. Single strand conformation polynorphism analysis and direct sequencing of their genomic DNAs revealed that non-PKU hyperphenylalaninemia resulted from compound heterozygosity for a mutation causing classical PKU and a mutation with a milder effect on phenylalanine hydroxylase activity. The mutations were R241C and R243Q in exon 7, and R413P in exon 12. The mutation genotypes of the two patients were R241C/R243Q and R241C/R413P. It has been demonstrated that homozygosity for the R243Q or R413P mutation is associated with a severe phenotype of PKU and low in vitro expression activity. In contrast, the R241C mutation has much less effect on phenylalanine hydroxylase activity. The metabolic consequence of each variant allele was confirmed by a phenylalanine loading test in the patients and their parents. The patients achieved normal results in all intellectual and neurologic tests. Brain magnetic resonance imaging revealed no abnormalities. The dietary restriction was continued until 10 years of age in order to maintain the serum phenylalanine level below 400 mumol/l. The genetic analysis to distinguish non-PKU hyper-phenylalaninemia from classical PKU helps to determine the principles of dietary management in the early infantile period.


Dietary fat saturation produces lipid modifications in peritoneal macrophages of mouse.
We investigated the effects of a saturated fat diet on mice lipid metabolism in resident peritoneal macrophages. Male C57BL/6 mice were weaned at 21 days of age and assigned to either the experimental diet, containing coconut oil (COCO diet), or the control diet, containing soybean oil as fat source. Fat content of each diet was 15% (w/w). Mice were fed for 6 weeks until sacrifice. In plasma of mice fed the COCO diet, the concentration of triglyceride, total cholesterol, HLD- and (LDL+VLDL)-cholesterol, and thiobarbituric acid-reactive substances (TBARS) increased, without changes in phospholipid concentration, compared with the controls. In macrophages of COCO-fed mice, the concentration of total (TC), free and esterified cholesterol, triglyceride, phospholipid (P) and TBARS increased, while the TC/P ratio did not change. The phospholipid compositions showed an increase of phosphatidylcholine and phosphatidylserine + phosphadytilinositol, a decrease of phosphatidylethanolamine, and no change in phosphatidylglycerol. (3)H(2)O incorporation into triglyceride and phospholipid fractions of macrophages increased, while its incorporation into free cholesterol decreased. Incorporation of [(3)H]cholesterol into macrophages of COCO-fed mice and the fraction of [(3)H]cholesterol ester increased. COCO diet produced an increase in myrystic, palmitic and palmitoleic acids proportion, a decrease in linoleic and arachidonic acids and no changes in stearic and oleic acids, compared with the control. Also, a higher relative percentage of saturated fatty acid and a decrease in unsaturation index (p <0.001) were observed in macrophages of COCO-fed mice. These results indicate that the COCO-diet, high in saturated fatty acids, alters the lipid metabolism and fatty acid composition of macrophages and produces a significant degree of oxidative stress.


Correction: Rivera-Picón et al. Factors Associated with Adherence to Treatment in Patients with HIV and Diabetes Mellitus. J. Pers. Med. 2023, 13, 269.
There was an error in the original publication [...].


Synthesis, structural characterization and biological properties of phosphorescent iridium(III) complexes.
Two phosphorescent cyclometalated iridium(III)-triptycenyl-1,10-phenanthroline complexes [Ir(ppy)2(tpt-phen)]+ (1) and [Ir(bhq)2(tpt-phen)]+ (2) {ppy=2-phenylpyridine, bhq=Benzo[h]quinoline, tpt-phen=triptycenyl-1,10-phenanthroline} have been synthesized and structurally characterized. The structure of complex 2 has been studied by single crystal X-ray crystallography. The photophysical properties of complexes in a different solvent have also been investigated. The binding of complexes to the double stranded calf thymus (CT-DNA) has been investigated by spectroscopic techniques. These complexes condense originally circular plasmid DNA into particulate structures. The DNA-condensation induced by these complexes have been investigated by electrophoretic mobilty shift assay, dynamic light scattering, and fluorescence microscopy. Furthermore, the cytotoxicity of these complexes towards HeLa cells have been studied and their cellular localisation properties have been investigated by fluorescence microscopy.


Effect of P to A mutation of the N-terminal residue adjacent to the Rgd motif on rhodostomin: importance of dynamics in integrin recognition.
Rhodostomin (Rho) is an RGD protein that specifically inhibits integrins. We found that Rho mutants with the P48A mutation 4.4-11.5 times more actively inhibited integrin α5β1. Structural analysis showed that they have a similar 3D conformation for the RGD loop. Docking analysis also showed no difference between their interactions with integrin α5β1. However, the backbone dynamics of RGD residues were different. The values of the R(2) relaxation parameter for Rho residues R49 and D51 were 39% and 54% higher than those of the P48A mutant, which caused differences in S(2), R(ex), and τ(e). The S(2) values of the P48A mutant residues R49, G50, and D51 were 29%, 14%, and 28% lower than those of Rho. The R(ex) values of Rho residues R49 and D51 were 0.91 s(-1) and 1.42 s(-1); however, no R(ex) was found for those of the P48A mutant. The τ(e) values of Rho residues R49 and D51 were 9.5 and 5.1 times lower than those of P48A mutant. Mutational study showed that integrin α5β1 prefers its ligands to contain (G/A)RGD but not PRGD sequences for binding. These results demonstrate that the N-terminal proline residue adjacent to the RGD motif affect its function and dynamics, which suggests that the dynamic properties of the RGD motif may be important in Rho's interaction with integrin α5β1.


Increase in myeloid-derived suppressor cells (MDSCs) associated with minimal residual disease (MRD) detection in adult acute myeloid leukemia.
Myeloid-derived suppressor cells (MDSCs) are thought to help provide a cellular microenvironments in many solid tumors, in which transformed cells proliferate, acquire new mutations, and evade host immunosurveillance. In the present study, we found that MDSCs (CD33 + CD11b + HLA-DR(low/neg)) in bone marrow were significantly increased in adult acute myeloid leukemia (AML) patients. MDSCs levels in newly diagnosed AML patients correlated well with extramedullary infiltration and plasma D-dimer levels. Remission rates in the MDSCs > 1500 group and MDSCs < 1500 group were 72.73 and 81.25 %, respectively. No significant differences were found between the two groups. MDSC levels in the complete remission group were significantly decreased after chemotherapy, while in the partial remission and non-remission groups, there were no significant differences. The level of MDSCs in the high minimal residual disease (MRD) group was significantly higher than that in the middle and low MRD groups. High levels of Wilms' Tumor-1 (WT-1) protein were strongly correlated with higher bone marrow MDSC levels. In conclusion, we report here a population of immunosuppressive monocytes in the bone marrow of patients with AML characterized by the CD33(high)CD11b + HLA-DR(low/neg) phenotype. These cells appear to impact the clinical course and prognosis of AML. This data may provide potentially important targets for novel therapies.


Scientific reports controlling droplet splashing and bouncing by dielectrowetting.
Stopping droplets from bouncing or splashing after impacting a surface is fundamental in preventing cross-contamination, and the spreading of germs and harmful substances. Here we demonstrate that dielectrowetting can be applied to actively control the dynamics of droplet impact. Moreover, we demonstrate that dielectrowetting can be used to prevent droplet bouncing and suppress splashing. In our experiments, the dielectrowetting effect is produced on a flat substrate by two thin interdigitated electrodes connected to an alternating current potential. Our findings show that the strength of the electric potential can affect the dynamic contact angle and regulate the spreading, splashing and receding dynamics at the right time-scales.


[Spermatic cord liposarcoma: a case report and review of the literature].
To explore the clinical features and treatment of spermatic cord liposarcoma (SCL). We retrospectively analyzed the clinical data of a case of SCL, reviewed the related literature and investigated the diagnosis and treatment of the disease. The patient underwent tumor resection and left inguinal orchidectomy. Postoperative pathology confirmed the case to be s SCL. Neither recurrence nor metastasis was found during the five-month follow-up. SCL is a rare medical condition with no specific imaging and laboratory features. Radical orchidectomy with wide local excision of the mass is recommended for its treatment, and adjuvant radiotherapy can be considered in intermediately or highly differentiated tumors and recurrent liposarcomas, while the role of chemotherapy is not well-defined.


[Fiber emissions from weathered asbestos cement products. 1. Fiber release in ambient air].
Emissions of fibrous aerosols were measured on buildings with weathered and corroded asbestos-cement-plates (roofing and facade shingles) by means of an already published equipment and procedure. The measured emission factors for asbestos fibers longer than 5 microns were in the range of 10(6) to 10(8) fibers/m2.h. They depended on the type of the AC-plates as well as on their age and corrosion intensity.


The academic consequences of early childhood problem behaviors.
Social/emotional skills in early childhood are associated with education, labor market, and family formation outcomes throughout the life course. One explanation for these associations is that poor social/emotional skills in early childhood interfere with the development of cognitive skills. In this paper, we use data from the Fragile Families and Child Wellbeing Study (N=2302) to examine how the timing of social/emotional skills-measured as internalizing, externalizing, and attention problem behaviors in early childhood-is associated with cognitive test scores in middle childhood. Results show that externalizing problems at age 3 and attention problems at age 5, as well as externalizing and attention problems at both ages 3 and 5, are associated with poor cognitive development in middle childhood, net of a wide array of control variables and prior test scores. Surprisingly, maternal engagement at age five does not mediate these associations.


Efficacy and safety of low-dose sirolimus for treatment of lymphangioleiomyomatosis.
Lymphangioleiomyomatosis (LAM) is a rare disease caused by dysregulated activation of the mammalian target of rapamycin (mTOR). Sirolimus, an inhibitor of mTOR, has been reported to decrease the size of angiomyolipomas and stabilize pulmonary function in patients with LAM. However, the optimal dose for the treatment of LAM remains unclear. We conducted a retrospective, observational study of 15 patients with LAM who underwent sirolimus therapy for more than 6 months. The efficacy was evaluated by reviewing the patients' clinical courses, pulmonary function and chest radiologic findings before and after the initiation of sirolimus treatment. All patients had blood trough levels of sirolimus lower than 5ng/mL. Sirolimus treatment improved the annual rates of change in FVC and FEV1 in the 9 patients who were free from chylous effusion (FVC, -101.0 vs. +190.0mL/y, p=0.046 and FEV1, -115.4 vs. +127.8mL/y, p=0.015). The remaining 7 patients had chylous effusion at the start of sirolimus treatment; the chylothorax resolved completely within 1-5 months of treatment in 6 of these cases. These results resembled those of previous studies in which blood trough levels of sirolimus ranged from 5 to 15ng/mL. Low-dose sirolimus (trough level, 5ng/mL or less) performed as well as the higher doses used previously for improving pulmonary function and decreasing chylous effusion in patients with LAM.


An educational panopticon? New technology, nurse education and surveillance.
This papers raises concerns about the use of new technology in nurse education. It is possible that new forms of computer-based learning and teaching carry with them the possibility of new kinds of 'panoptic' surveillance and control of students. We discuss how this is possible, and set this development within a wider social context. These forms of panoptic surveillance are, we believe, inimical to the underlying values and philosophy of nurse education. New technology does not necessarily have to be used in this way, and can be part of a liberating pedagogy. We would not propose that surveillance of students should never be used, but rather that its use should be considered and debated.


The prediction of hydrophobicity gradients within membrane interactive protein alpha-helices using a novel graphical technique.
Here, it is shown that amphiphilicity profiling based on the mean hydrophobic moment provides a simple visual guide for the identification of oblique orientated alpha-helices. The methodology has an efficiency of circa 70% and predicts that approximately 40% of transmembrane alpha-helices may possess these structures.


Cardiovascular events in patients with coronary artery disease with and without myocardial ischemia: Long-term follow-up.
After the results of the ISCHEMIA Trial, the role of myocardial ischemia in the prognosis of coronary artery disease (CAD) was under debate. We sought to comparatively evaluate the long-term prognosis of patients with multivessel CAD with or without documented myocardial ischemia. This is a single-center, retrospective, observational cohort study that included patients with CAD obtained from the research protocols database of "The Medicine, Angioplasty or Surgery Study," the MASS Study Group. Patients were stratified according to the presence or absence of myocardial ischemia. Cardiovascular events (overall mortality and myocardial infarction) were tracked from the registry entry up to a median follow-up of 8.7 years. Myocardial ischemia was assessed at baseline by a functional test with or without imaging. From 1995 to 2018, 2015 patients with multivessel CAD were included. Of these, 1001 presented with conclusive tests at registry entry, 790 (79%) presenting with ischemia and 211 (21%) without ischemia. The median follow-up was 8.7 years (IQR 4.04 to 10.07). The primary outcome occurred in 228 (28.9%) patients with ischemia and in 64 (30.3%) patients without ischemia (plog-rank=0.60). No significant interaction was observed with the presence of myocardial ischemia and treatment strategies in the occurrence of the combined primary outcome (pinteration=0.14). In this sample, myocardial ischemia was not associated with a worse prognosis compared with no ischemia in patients with multivessel CAD. These results refer to debates about the role of myocardial ischemia in the occurrence of cardiovascular events.


A model of the role of natural killer cells in immune surveillance--II.
The functioning of natural killer (NK) cells as immune surveillance effector cells against tumors is explored. In part I (J. Math. Biol. 12, 363-373 (1981], it was predicted that susceptible tumors would be eliminated if they have parameter lambda 0 value negative. They would not be eliminated if lambda 0 greater than 0. As the lambda 0 less than 0 result was local, one expected either that tumors of all sizes with lambda 0 less than 0 will be eliminated (global stability) or that tumor population will go to zero if in a domain of attraction of the critical point which is not all of the positive orthant. In this paper, the second is shown to be true. The general results are illustrated by a specific model.


Maternal acetaminophen use during pregnancy and childhood behavioural problems: Discrepancies between mother- and teacher-reported outcomes.
Maternal acetaminophen use during pregnancy is common and has been associated with childhood behavioural problems among offspring, specifically hyperactivity and conduct problems. Assessments of child behaviour in many previous studies have relied on maternal or parent report. Acknowledging that results of behavioural assessments vary between informants, we examined the association between maternal acetaminophen use during pregnancy and behaviour problems in childhood based on mother- and teacher-report. A longitudinal study of 560 mother-child pairs with data on illnesses and medication use during pregnancy and neurodevelopmental assessments during childhood was conducted. Acetaminophen use during pregnancy was captured using a standardised maternal interview, completed 1 year after delivery on average. Measures of childhood (6-12 years of age) behaviour were obtained via mother- and teacher-report, using the Child Behaviour Checklist and Teacher Report Form. Linear and log-binomial models were used to calculate adjusted mean differences (MD) and risk ratios (RR), respectively and 95% confidence intervals (CI) for internalising, externalising, and total behaviour problems comparing acetaminophen users to non-users. Stabilized inverse probability weights were used to account for loss to follow-up, and adjustments for indication were made. Approximately 60% (n = 354) of women reported use of acetaminophen during pregnancy. Acetaminophen use during pregnancy was associated with an increase in total behaviour problem score and risk of clinical behaviour problems according to mother report (MD 2.2, 95% CI 0.3, 4.1 and RR 1.93, 95% CI 0.99, 3.76) but not according to teacher report. Weighting to account for participation did not alter results, while adjustment for indications of acetaminophen use greatly attenuated the associations with mother-reported total behaviour problem score and risk of clinical behaviour problems (MD 0.1, 95% CI -2.1, 2.3 and RR 1.31, 95% CI 0.67, 2.58). Acetaminophen use during pregnancy was weakly associated with mother-reported behaviour problems and not associated with teacher-reported problems.


Nuclear magnetic resonance-determined lipoprotein profile and risk of breast cancer: a Mendelian randomization study.
While crudely quantified lipoproteins have been reported to affect the risk of breast cancer, the effects of subclass lipoproteins characterized by particle size, particle number, and lipidomes remain unknown. Utilizing nuclear magnetic resonance-based GWAS of 85 lipoprotein traits, we performed two-sample univariable Mendelian randomization (MR) to evaluate the causal relationship between each trait with breast cancer (Ncase/control = 133,384/113,789) and with its estrogen receptor (ER) subtypes. Then, we applied multivariable MR to investigate the independent effects considering both general and central obesity. In univariable MR, a heterogeneous effect of subclass high-density lipoproteins (HDL) was observed, in which small HDL traits (ORs ranged from 0.89 to 0.94) were associated with a decreased risk of breast cancer while non-small HDLs traits (OR ranged from 1.04 to 1.08) were associated with an increased risk of breast cancer. Very-low-density lipoproteins (VLDL) traits and serum total triglycerides (TG) were associated with a decreased risk of breast cancer (ORs ranged from 0.88 to 0.94). Similar association patterns were found for ER + subtype. In multivariable MR, only the protective effects of small HDL, VLDL and TG on ER + subtype remained significant. We identified a heterogeneous effect of subclass HDLs and a consistent protective effect of VLDL on breast cancer. Only the effects of small HDL and VLDL on ER + subtype remained robust after controlling for obesity. These findings provide new insight into the causal pathway underlying lipoproteins and breast cancer.


Risk assessment of the National Institute of Standards and Technology petroleum crude oil standard water accommodated fraction: further application of a copepod-based, full life-cycle bioassay.
The U.S. National Institute of Standards and Technology (NIST) petroleum crude oil was used to generate NIST water-accommodated hydrocarbon fractions (WAFs) for standardized assessment of crude oil effects on the copepod Amphiascus tenuiremis. Effects were assessed using a 96-well microplate, full life-cycle test. Briefly, nauplii (age, 24 h) were reared individually to adults (n > or =120 nauplii/treatment) in microplate wells containing 200 microl of treatment solution (seawater control [0%] or 10, 30, 50, or 100% NIST-WAF). Nauplii were monitored through development to adulthood, and mature virgin male:female pairs mated in wells containing original treatments (<30 d). A second bioassay using 0, 10, 30, and 50% WAFs (n > or =60 nauplii/treatment) was conducted to assess the effects of ultraviolet (UV) light on naupliar endpoints (<16 d). In the first experiment, nauplius-to-copepodite survival in exposures to 100% WAF was 27% +/- 6% lower than in controls (92% +/- 1%), but copepodite-to-adult survival was greater than 90% across all treatments. Analysis of development curves showed that nauplii in the 10% WAF developed into copepodites 25% faster, whereas nauplii in the 50 and 100% WAFs developed 17% slower, than controls. Copepodite development into male and female copepods was significantly delayed (2 and 4 d, respectively) in the 100% WAF compared to controls. Although none of the WAF exposures had significant effects on fertilization success or total viable production (p > 0.05), embryo hatching in the 100% WAF was significantly less (70.0% +/- 21.2%) than that in controls (87.0% +/- 19.4%). Results from the UV bioassay showed that relatively short exposures (<14 d) to 30 and 50% WAFs in the presence of UV light caused negative effects on copepod survival and development. Naupliar-stage survival and developmental endpoints were the most sensitive indicators of exposure to the NIST crude oil WAF


Harmonic to anharmonic tuning of moiré potential leading to unconventional Stark effect and giant dipolar repulsion in WS2/WSe2 heterobilayer.
Excitonic states trapped in harmonic moiré wells of twisted heterobilayers is an intriguing testbed for exploring many-body physics. However, the moiré potential is primarily governed by the twist angle, and its dynamic tuning remains a challenge. Here we demonstrate anharmonic tuning of moiré potential in a WS2/WSe2 heterobilayer through gate voltage and optical power. A gate voltage can result in a local in-plane perturbing field with odd parity around the high-symmetry points. This allows us to simultaneously observe the first (linear) and second (parabolic) order Stark shift for the ground state and first excited state, respectively, of the moiré trapped exciton - an effect opposite to conventional quantum-confined Stark shift. Depending on the degree of confinement, these excitons exhibit up to twenty-fold gate-tunability in the lifetime (100 to 5 ns). Also, exciton localization dependent dipolar repulsion leads to an optical power-induced blueshift of ~ 1 meV/μW - a five-fold enhancement over previous reports.


Validation of Use of Flipped Classroom for Surgery Core Clerkships.
The flipped classroom (FC) is an integrated learning paradigm that equips students with self-directed study materials before scheduled meeting times allowing for the deeper application of acquired knowledge with an instructor. There is limited data on the application of FC to clinical undergraduate medical education and particularly as it applies to a surgical clerkship. This study is a four-year retrospective study that includes two cohorts of students who matriculated through two training paradigms, traditional classroom (TC) and FC. Information regarding the FC cohort was collected from June 2018 to July 2020 (N=166). A two-year matched historical cohort of students enrolled in the clerkship and taught with the TC paradigm from June 2016 to July 2018 was used for comparison (N=157). The primary aim of this study is to assess the National Board of Medical Examiners (NBME) surgery shelf performance when the FC model is utilized compared to the TC model. This study will validate a prior similar study that had a smaller cohort with different secondary endpoints. Therefore, the secondary aim of this study assesses how teaching style can affect other performance metrics of a rotation (such as clinical performance, quizzes, Objective Structured Clinical Examinations (OSCE), and practicums) and how time was utilized by faculty, staff, and students. There was no difference in overall NBME surgery shelf performance when comparing the FC to the traditional classroom teaching (68.94 vs 70.34, P= 0.1667). Likewise, there was no difference in quiz performance. The FC did allow instructors to spend more time in other clinical components of the curriculum leading to a significant difference in student practicum (84.2 vs 88.26, P = 0.0186) and OSCE grades (87.54 vs 90.58, P <0.0001). The surgery NBME shelf performance is not compromised by FC and therefore can be used as an alternative to traditional classroom setting for teaching medical knowledge to surgery clerkship students. In addition, the FC can improve time management for instructors allowing for improved teaching and development in other components of the surgery curriculum.


Genetics considerations in cerebral palsy.
Cerebral palsy refers to a collective of neurologic conditions that share in common disorders of motor function and posture. Traditionally, and still today in many circles, the term is considered almost synonymous with brain injury. Multiple lines of evidence, however, point to the fact that cerebral palsy is rarely caused by problems with perinatal management. In fact, a mounting body of evidence points to strong genetic influences on the occurrence of cerebral palsy. Like most neurogenetic conditions, cerebral palsy exhibits complex inheritance. The best descriptor of the inheritance of cerebral palsy would be that of "multifactorial inheritance." This implies etiologic and genetic heterogeneity with complex interactions with multiple environmental influences. This article reviews known genetic influences on the origin of cerebral palsy. A proposed scheme for the genetic evaluation in identifying the etiology of cerebral palsy is provided.


Detection and discrimination of pathogenic bacteria with nanomaterials-based optical biosensors: A review.
Pathogenic bacteria can pose a great threat to food safety and human health. It is therefore imperative to develop a rapid, portable, and sensitive determination and discrimination method for pathogenic bacteria. Over the past few years, various nanomaterials (NMs) have been employed as desirable nanoprobes because they possess extraordinary properties that can be used for optical signal enabled detection and identification of bacteria. By means of modification, NMs can, depending on different mechanisms, sense targets directly or indirectly, which then provides an essential support for the detection and differentiation of pathogenic bacteria. In this review, recent application of NMs-based optical biosensors for food safety bacterial detection and discrimination is performed, mainly in but not limited to noble metal NMs, fluorescent NMs, and point-of-care testing (POCT). This review also focuses on future trends in bacterial detection and discrimination, and machine learning in performing intelligent rapid detection and multiple accurate identification of bacteria.


Integrated characterisation of nine species of the Schistorchiinae (Trematoda: Apocreadiidae) from Indo-Pacific fishes: two new species, a new genus, and a resurrected but 'cryptic' genus.
We report nine species of the Schistorchiinae Yamaguti, 1942 (Apocreadiidae Skrjabin, 1942) from Indo-Pacific marine fishes. Molecular data (ITS2 and 28S rDNA and cox1 mtDNA) are provided for all species and the genus-level classification of the subfamily is revised. For Schistorchis Lühe, 1906, we report the type-species Sch. carneus Lühe, 1906 and Sch. skrjabini Parukhin, 1963. For Sphinteristomum Oshmarin, Mamaev & Parukhin, 1961 we report the type-species, Sph. acollum Oshmarin, Mamaev & Parukhin, 1961. We report and re-recognise Lobatotrema Manter, 1963, for the type and only species, L. aniferum Manter, 1963, previously a synonym of Sph. acollum. Lobatotrema aniferum is phylogenetically distant from, but morphologically similar to, Sph. acollum and Lobatotrema is recognised as a 'cryptic genus'. We propose Blendiella n. gen. for B. trigintatestis n. sp. and B. tridecimtestis n. sp. These species are broadly consistent with the present morphological concept of Schistorchis but are phylogenetically distant from the type-species; a larger number of testes and some other subtle morphological characters in species of Blendiella serve to distinguish the two genera. We report three species of Paraschistorchis Blend, Karar & Dronen, 2017: P. stenosoma (Hanson, 1953) Blend, Karar & Dronen, 2017 (type-species), P. seychellesiensis (Toman, 1989) Blend, Karar & Dronen, 2017, and P. zancli (Hanson, 1953) Blend, Karar & Dronen, 2017. Lobatotrema aniferum, P. stenosoma, and Sch. carneus each have two distinct cox1 populations either over geographical range or in sympatry. Available evidence suggests that most of these species, but not all, are widespread in the tropical Indo-Pacific.


Effects of cyclosporine on the production of the reactive oxygen species in the glial cells.
After organ transplantation, some patients suffer mild neurological symptoms such as tremor to severe complications including seizures and encephalopathy. Among the immunosuppressants, cyclosporine (CsA) can induce neurological side effects. However, the mechanisms of encephalopathy by CsA are not fully understood. We measured the production of reactive oxygen species (ROS) in the glioma cells after CsA treatment. CsA (2.5 mmol/L) added to glioma cells was incubated for 60 minutes at 37 degrees C. ROS production was evaluated by measuring the fluorescent product from the oxidation of an oxidant-sensitive 2',7'-dichlorofluorescin using VICTOR3 multilabel counter. CsA resulted in ROS production by glioma cells. The ROS production increased with the time of exposure to CsA. These findings indicated that CsA may contribute to neurological side effects via ROS production.


Inflammation Markers Among Schizophrenia Patients Who Use Cannabis.
The mechanism of inflammation of the immune system, for example, such circulatory markers as the neutrophil-to-lymphocyte ratio (NLR) and mean platelet volume (MPV), has been shown in many studies to be associated with schizophrenia. In addition, it has been shown that the cannabidiol component reduces the activation of the acquired immune system. This study examined the differences in the levels of NLR and MPV among schizophrenia patients with cannabis use versus those without. In 2019 to 2020, a retrospective cross-sectional study was conducted based on digital medical records. Demographic, clinical, and complete blood cell count data were collected from records of rehospitalization of active psychotic schizophrenia inpatients. Data on NLR, MPV values, and demographic and clinical characteristics were compared between the groups and according to the degree of prevalence of cannabis use. No differences were found in the NLR and MPV values between the groups. The results were contrary to our expectations. These results may be explained by the presentation of a "pseudo-balanced" picture created when multiple processes affect inflammatory indices.


Genetic analysis of the interleukin-18 system highlights the role of the interleukin-18 gene in cardiovascular disease.
Interleukin (IL)-18 plays a key role in atherosclerosis and its complications. The present study investigated the genetic variability of 4 genes of the IL-18 system-IL18, IL18R1, IL18RAP, and IL18BP-in relation to circulating IL-18 levels and cardiovascular mortality. Twenty-two polymorphisms were genotyped in 1288 patients with coronary artery disease prospectively followed up during a median period of 5.9 years. The end point was death from cardiovascular causes (n=142). Baseline IL-18 levels were predictive of cardiovascular deaths occurring during < or =4 years of follow-up (HR=2.96, 95% CI 1.54 to 5.70, P=0.001 for the top compared with the bottom quartile) but not of later deaths. Haplotypes of the IL18 gene were associated with IL-18 levels (P=0.002) and cardiovascular mortality (P=0.006) after adjustment for cardiovascular risk factors. The same haplotype was associated with both a 9% lowering effect on IL-18 levels and a protective effect on risk (HR=0.57, 95% CI 0.36 to 0.92). IL18 haplotypes explained only 2% of IL-18 variability. Adjustment for baseline IL-18 levels abolished the association of haplotypes with cardiovascular risk. The haplotype associated with phenotypes was the only one carrying the minor allele of the IL18/A+183G polymorphism located in the 3'untranslated region and potentially affecting mRNA stability. The other genes of the system were not related to IL-18 levels or cardiovascular outcome. Variations of the IL18 gene consistently influence circulating levels of IL-18 and clinical outcome in patients with coronary artery disease, which supports the hypothesis of a causal role of IL-18 in atherosclerosis and its complications.


Assessment of Human Leukocyte Antigen Differences between Smokers with Reinke's Edema and Those with Laryngeal Cancer.
The present study aimed to assess human leukocyte antigen (HLA) typing differences between smokers with Reinke's edema and those with laryngeal squamous cell carcinoma (SCC). The HLA class I, II alleles were examined in 76 unrelated Iranian patients using low-resolution polymerase chain reaction with the sequence-specific primer (PCR-SSP) method. The frequency of the HLA-A*36 allele and HLA-B*35 was significantly higher in patients with SCC. The frequency of HLA-DRB1*01 alleles in Reinke's edema was significantly higher, as compared to that in others. In the volunteer group, HLA-DRB1*13 and HLA-DRB1*15 were significantly higher. As evidenced by the obtained results, HLA-A*36 was significantly higher in SCC, as compared to that in volunteers and Reinke's edema patients. It can be concluded that being positive for HLA-A*36 increases the chance of SCC by three times. This result should be further investigated in cohort studies conducted on larger samples. Furthermore, HLA-A*24 was significantly higher in the volunteer group, as compared to that in other groups. The HLADRB1*01 was remarkably higher in Reinke's edema, as compared to that in SCC.


Recycled concrete aggregate in base course applications: Review of field and laboratory investigations of leachate pH.
The prevalence of construction and demolition (C&D) waste and the concurrent demand for construction aggregate presents the opportunity to recycle C&D waste materials as substitutes for virgin aggregate. Commonly, recycled concrete aggregate (RCA) is used as base course in pavement construction. Environmentally responsible applications of RCA must consider the high pH leachate and trace element leaching risks reported in the literature. This review presents the methodology, results, and limitations of existing laboratory and field investigations of RCA leachate chemistry. Long-term highway field studies of RCA leachate illustrate that an initially high leachate pH approaches neutral within approximately one to two years of construction. Conversely, laboratory investigations of RCA leachate pH using batch reactor leaching tests and column leaching tests measure consistently high leachate pH (pH > 10). The discrepancies between field and laboratory measurements of RCA leachate pH suggest that the current laboratory methodology inadequately describes leachate conditions in the field. The authors recommend that future laboratory investigations consider intermittent wetting and drying cycles, eliminate particle abrasion, employ relevant contact times, and consider additional environmental processes that reduce leachate pH such as soil acidity and carbonation.


From genetics to structure to function: exploring sleep in Drosophila.
Sleep consists of quiescent periods with reduced responsiveness to external stimuli. Despite being maladaptive in that when asleep, animals are less able to respond to dangerous stimuli; sleep behavior is conserved in all animal species studied to date. Thus, sleep must be performing at least one fundamental, conserved function that is necessary, and/or whose benefits outweigh its maladaptive consequences. Currently, there is no consensus on what that function might be. Over the last 10 years, multiple groups have started to characterize the molecular mechanisms and brain structures necessary for normal sleep in Drosophila melanogaster. These researchers are exploiting genetic tools developed in Drosophila over the past century to identify and manipulate gene expression. Forward genetic screens can identify molecular components in complex biological systems and once identified, these genes can be manipulated within specific brain areas to determine which neuronal groups are important to initiate and maintain sleep. Screening for mutations and brain regions necessary for normal sleep has revealed that several genes that affect sleep are involved in synaptic plasticity and have preferential expression in the mushroom bodies (MBs). Moreover, altering MB neuronal activity alters sleep. Previous genetic screens found that the same genes enriched in MB are necessary for learning and memory. Increasing evidence in mammals, including humans, points to a beneficial role for sleep in synaptic plasticity, learning and memory. Thus, results from both flies and mammals suggest a strong link between sleep need and wake plasticity.


A BCWD-Resistant line of rainbow trout is less sensitive to cortisol implant-induced changes in IgM response as compared to a susceptible (control) line.
In salmonids, stress responses increase cortisol levels and disease susceptibility, including to Flavobacterium psychrophilum (Fp), the causative agent of BCWD. A BCWD-resistant line (R-line) of rainbow trout was used here to investigate potential differences in immunoglobulin response after a combined treatment of cortisol and Fp, as compared to a susceptible (S-line) control line. Expression of membrane and secreted immunoglobulin heavy chains mu and tau were determined by RT-qPCR in spleen and anterior kidney. Cortisol treatment did not affect B cell development in the anterior kidney, but delayed IgM responses at the early stage of infection in the spleen of both lines. An earlier IgM response was a determining factor in differential disease progression between resistant- and susceptible fish after Fp-challenge. S-line fish had a delayed and exacerbated IgM response after cortisol implant indicative of a detrimental cycle of sustained IgM responses and high pathogen loads. In contrast, R-line fish had delayed but milder, and protective IgM responses and cleared pathogen successfully. Fp challenge after cortisol implant increased serum cortisol levels on days 3 and 5 compared to mock treatments in S-line fish, but only on day 3 in R-line. Hence, combined cortisol treatment and Fp challenge differentially modulated B cell activation and Fp loads in BCWD-resistant and susceptible lines of rainbow trout. We propose that under conditions of increased stress, BCWD-resistant fish may remain immunologically better equipped to respond to infections compared to BCWD susceptible fish.


Occurrence of Virulence and Resistance Genes in Salmonella in Cloacae of Slaughtered Chickens and Ducks at Pluck Shops in Trinidad.
The study used PCR to determine the molecular basis of the antibiotic resistance and virulence profiles of isolates of Salmonella by targeting genes encoding for carriage and persistence in the poultry. Of a total 1,503 cecal samples collected from poultry, 91 (6.1%) were positive for Salmonella. Ten different serotypes were detected from Salmonella isolates. The study was also conducted to determine the occurrence of 13 virulence and 12 resistance genes in isolates of Salmonella. All 46 isolates of Salmonella tested were positive for one or more of the 12 virulence genes detected, ranging from 0.0% (viaB) to 100.0% (invA, mgtB, pipA, and spi4D) (P < 0.05). Occurrence of virulence genes varied significantly (P < 0.05) by serotype but not by animal species. Only 4 (33.3%) of 12 resistance genes assayed were detected: strA, ampC, cmy2, and qnrB. Overall, the occurrence of detected resistance genes was 71.7% (33 of 46), and 87.1 and 40.0% of the isolates from chickens and ducks, respectively, were positive (P = 0.0009). The occurrence of resistance genes ranged from 2.2% (cmy2) to 50.0% (qnrB) in isolates positive for resistance gene. The findings provide evidence that poultry from "pluck shops" are colonized by Salmonella pathogens that harbor virulence and antimicrobial resistance genes; this may have clinical and therapeutic consequences, if the genes detected are expressed. Although there is a need for prudent use of antimicrobial agents in poultry production systems, there should be constant monitoring for the prevalence of resistance in Salmonella isolates using phenotypic methods. The importance of monitoring the occurrence of resistance genes in the pathogens in Trinidad cannot be ignored.


Women in Surgery Italia: what are the opportunities in the operatory room? Results from a nationwide interdisciplinary survey.
Many studies documented the "glass ceiling" effect for women in surgery: achievements in academic and leadership positions are not consistent with the percentage of female surgeons in practice. A solid surgical case volume and expertise in high-complexity cases are required to pursue leadership positions. The aim of the study was to determine whether part of the difficulties encountered by female surgeons may lie in reduced surgical opportunities. This is the first study to investigate this issue in Italy. An online survey, conceived and promoted by Women in Surgery Italia, was administered through the RedCap platform, between November and December 2020, and female surgeons actively working in Italian academic and non-academic hospitals were invited to answer anonymously. A multivariate analysis was performed to evaluate the role of different factors on two main variables: overall procedures done and a sub-analysis of complex cases performed as first surgeon. 1810 respondents were included; the women who responded participated in 3% fewer cases on average, when compared to the mean staff case volume, and were significantly more often listed as the assistant surgeon and as primary surgeons in low-complexity cases. 36.5% of the respondents declared that at least one female physician on staff had to abandon the operatory room (OR) and does not regularly perform any surgical procedure. 73% female surgeons would like to spend more hours in the OR. While acquiring skills and surgical autonomy, many obstacles still exist for female surgeons. A portion of women are relegated to non-surgical activities, irrespective of their specialty, and these results confirm the theory that gender-biased underemployment still exists in surgical fields.


Identification and characterization of the mediator kinase-dependent myometrial stem cell phosphoproteome.
To identify, in myometrial stem/progenitor cells, the presumptive cell of origin for uterine fibroids, substrates of Mediator-associated cyclin dependent kinase 8/19 (CDK8/19), which is known to be disrupted by uterine fibroid driver mutations in Mediator complex subunit 12 (MED12). Experimental study. Academic research laboratory. Women undergoing hysterectomy for uterine fibroids. Stable isotopic labeling of amino acids in cell culture (SILAC) coupled with chemical inhibition of CDK8/19 and downstream quantitative phosphoproteomics and transcriptomic analyses in myometrial stem/progenitor cells. High-confidence Mediator kinase substrates identified by SILAC-based quantitative phosphoproteomics were determined using an empirical Bayes analysis and validated orthogonally by in vitro kinase assay featuring reconstituted Mediator kinase modules comprising wild-type or G44D mutant MED12 corresponding to the most frequent uterine fibroid driver mutation in MED12. Mediator kinase-regulated transcripts identified by RNA sequencing were linked to Mediator kinase substrates by computational analyses. A total of 296 unique phosphosites in 166 proteins were significantly decreased (≥ twofold) upon CDK8/19 inhibition, including 118 phosphosites in 71 nuclear proteins representing high-confidence Mediator kinase substrates linked to RNA polymerase II transcription, RNA processing and transport, chromatin modification, cytoskeletal architecture, and DNA replication and repair. Orthogonal validation confirmed a subset of these proteins, including Cut Like Homeobox 1 (CUX1) and Forkhead Box K1 (FOXK1), to be direct targets of MED12-dependent CDK8 phosphorylation in a manner abrogated by the most common uterine fibroid driver mutation (G44D) in MED12, implicating these substrates in disease pathogenesis. Transcriptome-wide profiling of Mediator kinase-inhibited myometrial stem/progenitor cells revealed alterations in cell cycle and myogenic gene expression programs to which Mediator kinase substrates could be linked directly. Among these, CUX1 is an established transcriptional regulator of the cell cycle whose corresponding gene on chromosome 7q is the locus for a recurrent breakpoint in uterine fibroids, linking MED12 and Mediator kinase with CUX1 for the first time in uterine fibroid pathogenesis. FOXK1, a transcriptional regulator of myogenic stem cell fate, was found to be coordinately enriched along with kinase, but not core, Mediator subunits in myometrial stem/progenitor cells compared with differentiated uterine smooth muscle cells. These studies identify a new catalog of pathologically and biologically relevant Mediator kinase substrates implicated in the pathogenesis of MED12 mutation-positive uterine fibroids, and further uncover a biochemical basis to link Mediator kinase activity with CUX1 and FOXK1 in the regulation of myometrial stem/progenitor cell fate.


The influence of mass of [11C]-laniquidar and [11C]-N-desmethyl-loperamide on P-glycoprotein blockage at the blood-brain barrier.
An earlier report suggested that mass amount of PET tracers could be an important factor in brain uptake mediated by P-glycoprotein. Thereby, this study investigated the influence of mass dose of laniquidar, desmethyl-loperamide and loperamide on the P-glycoprotein-mediated brain uptake of, respectively, [(11)C]-laniquidar and [(11)C]-N-desmethyl-loperamide ([(11)C]-dLop). Wild-type (WT) mice were injected intravenously with solutions of 5.6 MBq [(11)C]-laniquidar (either no carrier added or 60 mg/kg laniquidar added) or with 5.0-7.4 MBq [(11)C]-dLop (either no carrier added or 3 mg/kg desmethyl loperamide). Mice were killed, and brain and blood were collected, weighted and counted for radioactivity. Mdr1a(-/-) knockout mice were incorporated as the control group. Injection of (11)C-laniquidar (no carrier added) in WT mice resulted in a statistical significant lower brain uptake (0.7±0.2 %ID/g) compared to the carrier-added formulation (60 mg/kg laniquidar) (3.1±0.3 %ID/g) (P=.004), while no statistical difference could be observed between formulations of [(11)C]-dLop. The [(11)C]-laniquidar and [(11)C]-dLop blood concentrations were not significantly different between the tested formulations in WT mice. In control animals, no effect of mass amount on brain uptake of both tracers could be demonstrated. These results demonstrate the bivalent character of laniquidar, acting as a substrate at low doses and as a blocking agent for P-glycoprotein transport in the brain at higher doses. In comparison, no difference was observed in [(11)C]-dLop uptake between carrier- and no-carrier-added formulations, which confirms that desmethyl-loperamide is a substrate of P-glycoprotein at the blood-brain barrier.