Cost-effectiveness of nilotinib, dasatinib and imatinib as first-line treatment for chronic myeloid leukemia in Colombia, 2012.
New tyrosine kinase inhibitor treatments for chronic myeloid leukemia based on nilotinib, dasatinib and imatinib have improved patient quality of life and have turned chronic myeloid leukemia from a fatal disease into a chronic disease. To evaluate the cost-effectiveness of nilotinib, 600 mg, and dasatinib, 100 mg, each compared to imatinib, 400 mg, as first-line therapy in chronic myeloid leukemia in Colombia from a third-party payer's perspective. A cost-effectiveness analysis was performed using a Markov model to evaluate a hypothetical cohort of one hundred 55 year-old patients with newly diagnosed chronic myeloid leukemia in the chronic phase, and the time horizon for the baseline case was established as being until the end of life. Progression-free life-years saved were considered the primary outcome. Transition probabilities for major molecular response, disease progression to accelerated phase or blast crisis, and chronic myeloid leukemia related deaths were analyzed in the model for each arm. A 3% discount rate was applied to all costs and patient outcomes. Model robustness was evaluated using both univariate and multivariate Montecarlo sensitivity analysis. Nilotinib was higher in expected progression-free life-years saved (15.21 vs. 12.64 for imatinib), followed by dasatinib (14.91 vs. 14.54 for imatinib). Imatinib had lower total lifetime costs. The incremental cost-effectiveness ratio was US$ 33,120.36 in the nilotinib arm and US$ 514,939.08 in the dasatinib arm per progression-free life-years (PF-LYs) saved, each compared to imatinib. When analyzing nilotinib versus dasatinib indirectly, nilotinib was found to be dominant due to higher efficacy (2.25 PF-LYs) and lower costs (US$ 44,674) in the baseline case. The average estimated cost to manage disease progression per year was US$ 101,978.78, considered to be the threshold. In Colombia, using PF-LYs as the efficacy outcome, nilotinib is highly cost-effective when compared to imatinib and dominant vs. dasatinib in first-line therapy for CML in chronic phase.


DEAD-box RNA helicase protein DDX21 as a prognosis marker for early stage colorectal cancer with microsatellite instability.
DEAD-box RNA helicase DDX21 (also named nucleolar RNA helicase 2) is a nuclear autoantigen with undefined roles in cancer. To explore possible roles of autoimmune recognition in cancer immunity, we examined DDX21 protein expression in colorectal cancer tissue and its association with patient clinical outcomes. Unbiased deep proteomic profiling of two independent colorectal cancer cohorts using mass spectrometry showed that DDX21 protein was significantly upregulated in cancer relative to benign mucosa. We then examined DDX21 protein expression in a validation group of 710 patients, 619 of whom with early stage and 91 with late stage colorectal cancers. DDX21 was detected mostly in the tumor cell nuclei, with high expression in some mitotic cells. High levels of DDX21 protein were found in 28% of stage I, 21% of stage II, 30% of stage III, and 32% of stage IV colorectal cancer cases. DDX21 expression levels correlated with non-mucinous histology in early stage cancers but not with other clinicopathological features such as patient gender, age, tumor location, tumor grade, or mismatch repair status in any cancer stage. Kaplan-Meier analyses revealed that high DDX21 protein levels was associated with longer survival in patients with early stage colorectal cancer, especially longer disease-free survival in patients with microsatellite instability (MSI) cancers, but no such correlations were found for the microsatellite stable subtype or late stage colorectal cancer. Univariate and multivariate analyses also identified high DDX21 protein expression as an independent favorable prognostic marker for early stage MSI colorectal cancer.


Current permissible levels of metal pollutants harm terrestrial invertebrates.
The current decline of invertebrates worldwide is alarming. Several potential causes have been proposed but metal pollutants, while being widespread in the air, soils and water, have so far been largely overlooked. Here, we reviewed the results of 527 observations of the effects of arsenic, cadmium, lead and mercury on terrestrial invertebrates. These four well-studied metals are considered as priorities for public health and for which international regulatory guidelines exist. We found that they all significantly impact the physiology and behavior of invertebrates, even at levels below those recommended as 'safe' for humans. Our results call for a revision of the regulatory thresholds to better protect terrestrial invertebrates, which appear to be more sensitive to metal pollution than vertebrates. More fundamental research on a broader range of compounds and species is needed to improve international guidelines for metal pollutants, and to develop conservation plans to protect invertebrates and ecosystem services.


Diet-induced obesity aggravates NK cell-mediated contact hypersensitivity reaction in Rag1-/- mice.
Previous studies showed that natural killer (NK) cells mediate contact hypersensitivity (CHS) reaction. Many reports are showing that obesity promotes several inflammatory diseases. It was shown that diet-induced obesity (DIO) aggravates classical T cell-mediated CHS in mice. To determine whether the high-fat diet (HFD)-induced obesity modulates antigen-specific NK cell-mediated response. We evaluated the effect of DIO on NK cell-mediated CHS reaction using a model of dinitrofluorobenzene (DNFB)-induced CHS in Rag1-/- mice. Rag1-/- mice fed HFD for 8 but not for 4 weeks developed aggravated CHS reaction determined by ear swelling measurement when compared to animals kept on normal diet (ND) prior to DNFB sensitization. The obese Rag1-/- mice presented the adipose tissue inflammation. Furthermore, in vitro analysis showed that feeding with HFD significantly increases interferon γ (IFN-γ) and interleukin (IL)-12p70 and decreases adiponectin concentration in liver mononuclear cell (LMNC) culture supernatants. The flow cytometry analysis of LMNC revealed that HFD treatment prior to DNFB sensitization increases the percentage of NK1.1+ IFN-γ+ cell population and affects the development and maturation of NK1.1+ cells. In summary, current results suggest that the DIO significantly modulates the local and systemic inflammatory response, contributing to exacerbation of the CHS response mediated by liver NK cells.


Determination and Quantitation of Cytotoxic T Cell-Mediated Cell Death.
Cytotoxic T cell-induced cell death is well documented. Cytotoxic T cell releases various cytolytic proteins. The cytolytic proteins induce target cell death. T cell-induced cell death can be measured by the lytic assay. One of the well-known lytic assays uses radioactive tracer, Chromium-51 (51Cr), and detects the amount of 51Cr released from target cells. This assay can detect cell death and the efficiency of the T cell-induced cell death by coculture effector cells (T cells) and target cells. This assay can determine the kinetics of the cell lysis. The issue of this approach is the use of radioactive material. This chapter describes measuring T cell-induced cell death by determining the epigenetic remodeling and the release of cytolytic proteins. Determine the efficiency of T cell-induced cell death by using a flow cytometry-based detection method.


The protein composition of bovine myelin-free axons.
The proteins of axons prepared from myelinated axons and isolated as myelin-free entities were separated by sodium dodecyl sulfate polyacrylamide electrophoresis and found to consist of more than 10 different molecular weight species. The molecular weights range from 13 000 to over 200 000 with a prominent protein of molecular weight 47 000. The amino acid composition of the seven major proteins showed that the protein with a molecular weigt of 47 000 is distinct from all the other proteins analyzed. A group of three low molecular weight proteins have amino acid compositions which are similar to each other as do a group of three high molecular weight proteins although the two groups are distinctly different from each other and the major axonal protein. Histones, DNA, myelin basic protein and glycoprotein were absent from the proteins but neurotubule protein was present as indicated by cochicine binding activity in the axonal preparations. The cellular origin of these proteins and their relationship to other central nervous system proteins are discussed.


Crystal structure of barium dinickel(II) iron(III) tris-[orthophosphate(V)], BaNi2Fe(PO4)3.
The orthophosphate BaNi2Fe(PO4)3 has been synthesized by a solid-state reaction route and characterized by single-crystal X-ray diffraction and energy-dispersive X-ray spectroscopy. The crystal structure comprises (100) sheets made up of [Ni2O10] dimers that are linked to two PO4 tetra-hedra via common edges and vertices and of linear infinite [010] chains of corner-sharing [FeO6] octa-hedra and [PO4] tetra-hedra. The linkage of the sheets and chains into a framework is accomplished through common vertices of PO4 tetra-hedra and [FeO6] octa-hedra. The framework is perforated by channels in which positionally disordered Ba2+ cations are located.


Factors Affecting Patient Radiation Exposure During Prone and Supine Percutaneous Nephrolithotomy.
Purpose: Radiation exposure from fluoroscopy poses risks to patients and surgeons. Percutaneous nephrolithotomy (PCNL) has traditionally required fluoroscopy, however, the use of ultrasound (US) has decreased radiation exposure. US guidance in supine PCNL (S-PCNL) may further reduce radiation exposure. In this study, we investigate patient and operative factors affecting fluoroscopy time (second), total radiation dose (mGy), and effective dose (ED, mSv) in patients undergoing US-guided S-PCNL or prone PCNL (P-PCNL). Methods: We performed a retrospective study of patients undergoing US-guided PCNL in prone and supine positions. Patients with multiple access tracts, pre-existing renal access, or fluoroscopic renal access were excluded. Patient demographic and radiologic and operative data were collected, and compared between the two groups. Results: Ninety-nine patients were included: 45 P-PCNL and 54 S-PCNL. There were no significant demographic differences between the two groups. Operative time, access location, tract length, and total radiation dose (mGy) also did not differ. S-PCNL was associated with lower ED (2.92 ± 0.32 mSv vs 5.3 ± 0.7 mSv, p = 0.0014) despite increased fluoroscopy time (86.32 ± 7.7 seconds vs 51.00 ± 5.1 seconds, p = 0.004), and was more likely a mini-PCNL (35.2% vs 15.9%, p = 0.032). In multivariate analysis, S-PCNL remained associated with reduced ED compared with P-PCNL (p = 0.002), whereas body mass index (p < 0.001) and staghorn calculi (p < 0.001) were independently associated with increased ED. Conclusions: We demonstrated that ED in US-guided PCNL is increased in the prone position compared with supine position, and in overweight patients regardless of position. US-guided S-PCNL may decrease radiation exposure to patients and surgeons compared with US-guided P-PCNL.


The axonal glycoprotein TAG-1 is an immunoglobulin superfamily member with neurite outgrowth-promoting activity.
Pathfinding of axons in the developing nervous system is thought to be mediated by glycoproteins expressed on the surface of embryonic axons and growth cones. One molecule suggested to play a role in axonal growth is TAG-1, a 135 kd glycoprotein expressed transiently on the surface of subsets of neurons in the developing mammalian nervous system. We isolated a full-length cDNA clone encoding rat TAG-1. TAG-1 has six immunoglobulin-like domains and four fibronectin type III-like repeats and is structurally similar to other immunoglobulin-like proteins expressed on developing axons. Neurons maintained in vitro on a substrate of TAG-1 extend long neurites, suggesting that this protein plays a role in the initial growth and guidance of axons in vivo. TAG-1 is anchored to the neuronal membrane via a glycosyl phosphatidylinositol linkage and is also released from neurons, suggesting that TAG-1 also functions as a substrate adhesion molecule when released into the extracellular environment.


Molecular characterization of inter-telomere and intra-telomere mutations in human ALT cells.
Telomeres in most immortal cells are maintained by the enzyme telomerase, allowing cells to divide indefinitely. Some telomerase-negative tumors and immortal cell lines maintain long heterogeneous telomeres by the ALT (alternative lengthening of telomeres) mechanism; such tumors are expected to be resistant to anti-telomerase drug therapies. Occasionally telomerase-negative Saccharomyces cerevisiae mutants survive, and 10% of them (type II survivors) have unstable telomeres. As in human ALT+ cells, short telomeres in yeast type II survivors lengthen abruptly; in yeast, this is dependent on the recombination proteins Rad52p and Rad50p. In human cells, ALT involves copying of sequence from a donor to a recipient telomere. We have characterized for the first time a class of complex telomere mutations seen only in ALT+ cells. The mutant telomeres are defined by the replacement of the progenitor telomere at a discrete point (fusion point) with a different telomere repeat array. Among 19 characterized fusion points, one occurred within the first six repeats of the telomere, indicating that these recombination-like events can occur anywhere within the telomere. One mutant telomere may have been involved in a secondary recombination-like mutation event, suggesting that these mutations are sporadic but ongoing in ALT+ cells. We also identified simple intra-allelic mutations at high frequency, which evidently contribute to telomere instability in ALT+ cells.


Hypertonic lactate for the treatment of intracranial hypertension in patients with acute brain injury.
Hypertonic lactate (HL) is emerging as alternative treatment of intracranial hypertension following acute brain injury (ABI), but comparative studies are limited. Here, we examined the effectiveness of HL on main cerebral and systemic physiologic variables, and further compared it to that of standard hypertonic saline (HS). Retrospective cohort analysis of ABI subjects who received sequential osmotherapy with 7.5% HS followed by HL-given at equi-osmolar (2400 mOsmol/L) and isovolumic (1.5 mL/kg) bolus doses-to reduce sustained elevations of ICP (> 20 mmHg). The effect of HL on brain (intracranial pressure [ICP], brain tissue PO2 [PbtO2], cerebral microdialysis [CMD] glucose and lactate/pyruvate ratio [LPR]) and blood (chloride, pH) variables was examined at different time-points (30, 60, 90, 120 min vs. baseline), and compared to that of HS. A total of 34 treatments among 17 consecutive subjects (13 traumatic brain injury [TBI], 4 non-TBI) were studied. Both agents significantly reduced ICP (p < 0.001, at all time-points tested): when comparing treatment effectiveness, absolute ICP decrease in mmHg and the duration of treatment effect (median time with ICP < 20 mmHg following osmotherapy 183 [108-257] vs. 150 [111-419] min) did not differ significantly between HL and HS (all p > 0.2). None of the treatment had statistically significant effects on PbtO2 and CMD biomarkers. Treatment with HL did not cause hyperchloremia and resulted in a more favourable systemic chloride balance than HS (Δ blood chloride - 1 ± 2.5 vs. + 4 ± 3 mmol/L; p < 0.001). This is the first clinical study showing that HL has comparative effectiveness than HS for the treatment of intracranial hypertension, while at the same time avoiding hyperchloremic acidosis. Both agents had no significant effect on cerebral oxygenation and metabolism.


Synthetic generation of DSC-MRI-derived relative CBV maps from DCE MRI of brain tumors.
Perfusion MRI with gadolinium-based contrast agents is useful for diagnosis and treatment response evaluation of brain tumors. Dynamic susceptibility contrast (DSC) MRI and dynamic contrast enhanced (DCE) MRI are two gadolinium-based contrast agent perfusion imaging techniques that provide complementary information about the tumor vasculature. However, each requires a separate administration of a gadolinium-based contrast agent. The purpose of this retrospective study was to determine the feasibility of synthesizing relative cerebral blood volume (rCBV) maps, as computed from DSC MRI, from DCE MRI of brain tumors. One hundred nine brain-tumor patients underwent both DCE and DSC MRI. Relative CBV maps were computed from the DSC MRI, and blood plasma volume fraction maps were computed from the DCE MRIs. Conditional generative adversarial networks were developed to synthesize rCBV maps from the DCE MRIs. Tumor-to-white matter ratios were calculated from real rCBV, synthetic rCBV, and plasma volume fraction maps and compared using correlation analysis. Real and synthetic rCBV in white and gray matter regions were also compared. Pearson correlation analysis showed that both the tumor rCBV and tumor-to-white matter ratios in the synthetic and real rCBV maps were strongly correlated (ρ = 0.87, P < .05 and ρ = 0.86, P < .05, respectively). Tumor plasma volume fraction and real rCBV were not strongly correlated (ρ = 0.47). Bland-Altman analysis showed a mean difference between the synthetic and real rCBV tumor-to-white matter ratios of 0.20 with a 95% confidence interval of ±0.47. Realistic rCBV maps can be synthesized from DCE MRI and contain quantitative information, enabling robust brain-tumor perfusion imaging of DSC and DCE parameters with a single gadolinium-based contrast agent administration.


Hydrodynamic Kelvin-Helmholtz instability on metallic surface.
Kelvin-Helmholtz instability on metallic surface is relevant to intense oblique impact in many physical processes such as explosive welding, Inertial Confinement Fusion and planetary impact events. Evolution of instability results in the formation of wavy morphology leading to material bonding or even mixing. However, mostly due to lack method to describe the dynamic behavior, instability mechanism controlled by elastoplastic properties of metal remains elusive. Here, we introduce a theory to reveal the evolution characteristics aroused by tangential velocity. Our simulations find that the unstable metallic surfaces exhibit amplitude growth and tangential motion by overcoming the depression of yield strength to generate wavy morphology. For diverse loading velocities, corrugated surfaces and material properties, an instability boundary distinguishes all unstable evolutions. Our analytical method with scale-independent variables reproducing numerical findings reveals plentiful characteristics of instability in strength materials. For designed loading velocities and material in oblique impact experiment in laboratory, the property of corrugated surfaces becomes an important factor to determine instability evolution.


Assessment of medication self-administration using artificial intelligence.
Errors in medication self-administration (MSA) lead to poor treatment adherence, increased hospitalizations and higher healthcare costs. These errors are particularly common when medication delivery involves devices such as inhalers or insulin pens. We present a contactless and unobtrusive artificial intelligence (AI) framework that can detect and monitor MSA errors by analyzing the wireless signals in the patient's home, without the need for physical contact. The system was developed by observing self-administration conducted by volunteers and evaluated by comparing its prediction with human annotations. Findings from this study demonstrate that our approach can automatically detect when patients use their inhalers (area under the curve (AUC) = 0.992) or insulin pens (AUC = 0.967), and assess whether patients follow the appropriate steps for using these devices (AUC = 0.952). The work shows the potential of leveraging AI-based solutions to improve medication safety with minimal overhead for patients and health professionals.


Reporter-based assays for analyzing RNA interference in mammalian cells.
RNA interference (RNAi) is a process whereby small RNAs serve as effectors to direct posttranscriptional regulation of gene expression. The effector small RNAs can arise from various sources including plasmids that express short-hairpin RNAs (shRNAs) or microRNA (miRNAs), or alternatively, from synthetic small-interfering RNAs (siRNAs). These small RNAs enter a protein complex that binds directly to mRNA targets and this results in transcript-specific inhibition of protein expression. Though the key core components of the mammalian RNAi processing and effector complexes have been identified, accessory and regulatory factors are less well-defined. Reporter assays that can quantitatively assess RNAi activity can be used to identify modulators of RNAi. We present two methods to quantitatively analyze RNAi activity that have overlapping and distinct utility. The first method uses an eGFP reporter in transiently transfected cells to identify RNAi modulators. The second method uses cells that express luciferase-based reporters in a stable fashion. This assay can easily be conducted in 96-well plate format. Both methods can be used to identify novel proteins or small molecules that modulate RNAi activity.


Reduction of the increased portal vascular resistance of the isolated perfused cirrhotic rat liver by vasodilators.
This study was undertaken to compare the in vitro responses of the portal vascular bed of normal and cirrhotic rat livers to a variety of vasodilator agents. Using carbon tetrachloride-induced cirrhosis in the rat as a model, isolated liver preparations were perfused via the portal vein with a synthetic medium (2.5% bovine serum albumin in Krebs-Henseleit buffer) to eliminate extrahepatic neural and humoral influences. Under these experimental conditions the mean perfusion resistance of the cirrhotic livers was approximately 117% higher than in controls (P less than 0.001). The vascular tone of the normal liver was minimal as assessed by the response to a variety of vasodilator agents, including sodium nitroprusside (3.0 X 10(-3) M), magnesium sulphate (6.0 X 10(-2) M), papaverine hydrochloride (6.4 X 10(-4) M), and cytochalasin B (6.3 X 10(-5) M). In contrast, these agents reduced the perfusion resistance of the cirrhotic livers by approximately 15%. Prostaglandin E1 (3.0 X 10(-6) M) and isoprenaline hydrochloride (2.4 X 10(-6) M) produced a lesser fall in resistance which nevertheless was greater in cirrhotic livers than controls. Cirrhotic livers, unlike the controls, were found to contain large numbers of myofibroblasts in perivenous and perisinusoidal locations. Previous studies have shown that myofibroblasts are capable of sustaining a high level of intrinsic tone and relax in response to vasodilator agents. It is concluded that part of the increased resistance to flow through the portal vascular bed of the cirrhotic rat liver in vitro is due to an increase in intrinsic vascular tone, possibly mediated via myofibroblasts, and can be reversed by pharmacological agents.


Novel approaches for glioblastoma treatment: Focus on tumor heterogeneity, treatment resistance, and computational tools.
Glioblastoma (GBM) is a highly aggressive primary brain tumor. Currently, the suggested line of action is the surgical resection followed by radiotherapy and treatment with the adjuvant temozolomide, a DNA alkylating agent. However, the ability of tumor cells to deeply infiltrate the surrounding tissue makes complete resection quite impossible, and, in consequence, the probability of tumor recurrence is high, and the prognosis is not positive. GBM is highly heterogeneous and adapts to treatment in most individuals. Nevertheless, these mechanisms of adaption are unknown. In this review, we will discuss the recent discoveries in molecular and cellular heterogeneity, mechanisms of therapeutic resistance, and new technological approaches to identify new treatments for GBM. The combination of biology and computer resources allow the use of algorithms to apply artificial intelligence and machine learning approaches to identify potential therapeutic pathways and to identify new drug candidates. These new approaches will generate a better understanding of GBM pathogenesis and will result in novel treatments to reduce or block the devastating consequences of brain cancers.


18F-FDG-PET in Finnish patients with clinical suspicion of cardiac sarcoidosis: Female sex and history of atrioventricular block increase the prevalence of positive PET findings.
Fluorodeoxyglucose positron emission tomography (FDG-PET) is a non-invasive imaging modality that has been shown to be a feasible method to demonstrate myocardial inflammation. The aim of this study was to identify the patients suspected of having cardiac sarcoidosis (CS), who are most likely to benefit from PET imaging. 137 patients suspected of having CS underwent a dedicated cardiac FDG-PET examination at Tampere University Hospital between August 2012 and September 2015. These examinations were retrospectively analyzed. 33 and 12 of the 137 patients had abnormal left and right ventricular (LV and RV) FDG-uptake, respectively. Abnormal LV-uptake and RV-uptake were significantly associated with female sex and a history of advanced AV-block (P < 0.05). Abnormal RV-uptake was also associated with ventricular tachycardia and atrial fibrillation (P < 0.05). 56% of the 27 female patients with a history of AV-block had a pathological PET finding compared to only 6% of the 49 male patients without a history of AV-block. There were 17 female patients with history of both AV-block and ventricular tachycardia, 71% of them had abnormal PET finding. Abnormal FDG-PET findings were associated with female sex, AV-block, and arrhythmias in this clinical cohort.


Clinical guidelines and the fate of medical autonomy in Ontario.
Conceptually, clinical guidelines and professional autonomy have a paradoxical relationship. Despite being the quintessence of medical knowledge at the corporate level, guidelines diminish the clinical autonomy of individual practitioners, and therefore threaten medicine's justification for its autonomy. Theorists have argued that professional autonomy will be retained through elite dominance of practitioners, while comparative research suggests that economic autonomy can be traded off to retain clinical autonomy. Under government pressure to regulate the growth of Ontario physicians' fee-for-service public expenditure, the profession's representative organization, the Ontario Medical Association (OMA), promoted voluntary clinical guidelines, hoping to both constrain costs and preserve professional control over the content of medical care. The OMA collaborated with the Ministry of Health in developing guidelines and establishing a provincial centre for health service research. Ontario's practitioners disregarded the OMA's exhortations to implement clinical guidelines, suggesting that in the absence of external constraints, practitioners can subvert elite dominance. However, practitioners' unchecked clinical and economic autonomy, combined with evidence of wide provincial variations in medical care, served to legitimize the government's increasingly unilateral control over the schedule of insured medical services, and, in 1993, their imposition of a global cap on physicians' fee-for-service income pool. When analysed in the context of ongoing Ministry-OMA relations, the failure of the OMA's guidelines strategy to constrain medical service costs has expedited an overall decline in medical autonomy in Ontario. The emergence and course of Ontario's clinical guidelines movement is consistent with the view that medical autonomy is contingent upon broad class forces, and the conceptualization of professional organizations as instruments for mediated occupational control.


Effect of amiodarone on the plasma levels of metoprolol.
On average, metoprolol plasma concentration is doubled after an amiodarone loading dose (1.2 g/day over a period of 6 days). However, the individual amount of this drug interaction depends on the CYP2D6 genotype.


Regional neuromuscular regulation within human rectus femoris muscle during gait.
The spatial distribution pattern of neuromuscular activation within the human rectus femoris (RF) muscle was investigated during gait by multi-channel surface electromyography (surface EMG). Eleven healthy men walked on a treadmill with three gait speeds (4, 5, and 6 km/h) and gradients (0°, 12.5°, and 25°). The spatial distribution of surface EMG was tested by central locus activation (CLA), which is calculated from 2-D multi-channel surface EMG with 46 surface electrodes. For all conditions, CLA was around the middle regions during the swing-to-stance transition and moved in a proximal direction during the stance phase and stance-to-swing transition (p<0.05). CLA during the stance-to-swing transition and early swing phase significantly moved to proximal site with increasing gait speed (p<0.05). During the early stance and swing phases, with increasing grade, CLA significantly moved distally (p<0.05). These results suggest that the RF muscle is regionally activated during a gait cycle and is non-uniformly regulated longitudinally.


Shift of Host Range for the Immature Stages of the Lanternfly, Pyrops watanabei (Matsumura) (Hemiptera, Fulgoridae) Native to Taiwan.
Although Pyrops watanabei is a species native to Taiwan, many fundamental aspects of the species are still poorly documented. Populations of the lanternfly in locations of northern Taiwan were found in abundance and were suitable for the conduction of an investigation from 1 July 2021 to 30 June 2022. We established the shift in the main host plants with different developmental stages. The occurrence of immature individuals on Heptapleurum heptaphyllum increased with age from eggs to nymphs in the third instar; however, it declined from the fourth instar onwards due to a shift in preference to Triadica sebifera. In 2021, the earliest detection of an egg mass was on 1 July. More eggs were recorded in August, and some could be found in September and October of the same year. In 2022, we found an egg mass on 28 June. In August 2021, nymphs in the first and second instars were detected. Then, nymphs in the third and fourth instars appeared after September and October 2021, respectively. Furthermore, nymphs in the fifth instar were sighted as late as March 2022. Finally, this study will provide a basis for further evaluation of the impact of invasion of Pyrops candelaria on the ecology of Pyrops watanabei.


Lymphocytes play the music but the macrophage calls the tune.
Researchers interested in immunological aspects of bacterial, fungal, protozoan and helminthic infection are too often kept apart by artificial subject boundaries. These barriers were temporarily breached by a recent workshop* in which the complex interplay between microbes and their mammalian hosts were examined from a global viewpoint. The role of T-cell subsets and their products came under close scrutiny but the most forceful image was that of the macrophage. As host for infective agents, as modulator of specific immune activity and as ultimate mediator of the host response, the macrophage plays a virtuoso's role in the host-parasite drama.


Effect of moderate maternal malnutrition on the placenta.
The results of two studies of the influence of moderate maternal malnutrition on the weight and chemical characteristics of the placenta are discussed. In the first study, two groups of pregnant women of high and low socioeconomic status from Guatemala City were studied. Socioeconomic status was defined by family income, educational level of the mother, and environmental sanitary conditions. Both groups were very similar with respect to age, parity, gestational age, and absence of severe disease during pregnancy. The average placental weight in the low socioeconomic group was 15 per cent below that of the high socioeconomic group and there was a consistent association between the postpartum maternal weight and placental weight. There were no differences between the two groups regarding placental concentration of fat, protein, water, ash, hemoglobin, and DNA; hydroxyproline and fat concentration were significantly lower in the low socioeconomic group. The hypothesis that the difference in placental weight observed between the two groups was primarily due to maternal nutritional status was tested in the second study by means of nutritional intervention in four rural villages in Guatemala. Two of the villages received a protein-calorie preparation while the other two received a calorie supplement. Placental weight was higher among women with high levels of supplemented calories during pregnancy, independently of the type of food supplement ingested. On the average, the groups with low caloric supplementation (smaller than 20,000 calories) had placental weight 11 per cent below those with high caloric supplementation (larger than or equal to 20,000 calories), In contrast to placental weight, the concentration of placental chemical components studied was not associated with caloric supplementation. It was concluded that moderate protein-calorie malnutrition during pregnancy leads to lower placental weight without significantly changing the concentration of the biochemical components studied. The reduction of placental weight may be the mechanism by which maternal malnutrition is associated with high prevalence of low-birth-weight babies in these populations.


SARS-CoV-2 RT-PCR positivity and antibody prevalence among asymptomatic hospital-based health care workers.
The level of asymptomatic infection with SARS-CoV-2 could be substantial and among health care workers (HCWs) a source of continuing transmission of the virus to patients and co-workers. Measure the period prevalence of SARS-CoV-2 PCR positivity and seroprevalence of SARS-CoV-2 IgG antibodies among a random sample of asymptomatic health system hospital-based health care workers (HCWs) 6½ -15½ weeks after 4/5/2020, the peak of the first surge of COVID-19 admissions. Of 524 eligible and consented participants from four metropolitan hospitals, nasopharyngeal swabs were obtained from 439 (83.8 %) and blood from 374 (71.4 %). Using PCR nucleic acid-based amplification (NAAT) methods, the period prevalence of SARS-CoV-2 infection was 0.23 % (95 % confidence interval (CI) 0.01 %-1.28 %; 1/439) from 5/21/20-7/16/20. The seroprevalence of SARS-CoV-2 IgG antibodies from June 17-July 24, 2020 was 2.41 % (95 % CI 1.27 %-4.51 %; 9/374). Those who were reactive were younger (median age 36 versus 44 years; p = 0.050), and those with self-reported Hispanic/Latino ethnicity had a higher seroprevalence (2/12 = 16.7 % versus 7/352 = 2.0 %; p = 0.051). There were no significant differences by sex, race, residence, hospital, unit or job type. The one employee who was found to be PCR test positive in this study was also reactive for IgG antibodies, tested 27 days later. The period prevalence of PCR positivity to SARS-CoV-2 and IgG seroprevalence was unexpectedly low in asymptomatic HCWs after a peak in COVID-19 admissions and the establishment of state and institutional infection control policies, suggesting that routine screening tests while community prevalence is relatively low would produce a minimal yield.


Fat regulatory mechanisms of pine nut oil based on protein interaction network analysis.
Pine nut oil (PNO), a standardized and well-defined extract of Pinus koraiensis (Korean pine), has beneficial effects on wound healing, inflammatory diseases, and cancer. However, the explanation for the mechanism by which PNO reduces body fat remains uncertain. We performed a protein-protein interaction network (PPIN) analysis to explore the genes associated with pinolenic acid using the MEDILINE database from PubChem and PubMed. It was concluded through the PPIN analysis that PNO was involved in a neutral lipid biosynthetic process. This study evaluated the effects of PNO predicted by the network analysis of fat accumulation in chronic obesity mouse models established by feeding a high fat diet (HFD) to C57BL/6J mice and explored potential mechanisms. HFD mice were fed only HFD or HFD with PNO at 822 and 1644 mg/kg. After an oral administration of 7 weeks, several body weight and body fat-related parameters were examined, including the following: adipose weight, adipocyte size, serum lipid profiles, adipocyte expression of PPAR-γ, sterol regulatory element binding protein (SREBP)-1c, lipoprotein lipase (LPL) and leptin. We showed that oral administration of PNO to HFD mice reduces body fat weight, fat in tissue, white adipose tissue weight, and adipocyte size. The serum cholesterol was improved in the HFD mice treated with PNO. Additionally, PNO has significantly attenuated the HFD-induced changes in the adipose tissue expression of PPAR-γ, SREBP-1c, LPL, and leptin. The findings from this study based on the PPIN analysis suggest that PNO has potential as drug to reduce body fat through fat regulatory mechanisms by PPAR-γ and SREBP-1c.


Formulation of the biological control yeast Meyerozyma caribbica by electrospraying process: effect on postharvest control of anthracnose in mango (Mangifera indica L.) and papaya (Carica papaya L.).
Microorganism for biological control of fruit diseases is an eco-friendly alternative to the use of chemical fungicides. This is the first study evaluating the electrospraying process to encapsulate the biocontrol yeast Meyerozyma caribbica. The effect of encapsulating material [Wey protein concentrate (WPC), Fibersol® and Trehalose], its concentration and storage temperature on the cell viability of M. caribbica, and in vitro and in vivo control of Colletotrichum gloeosporioides was evaluated. The processing with commercial resistant maltodextrin (Fibersol®) 30% (w/v) as encapsulating material showed the highest initial cell viability (95.97 ± 1.01%). The storage at 4 ± 1 °C showed lower losses of viability compared to 25 ± 1 °C. Finally, the encapsulated yeast with Fibersol 30% w/v showed inhibitory activity against anthracnose in the in vitro and in vivo tests, similar to yeast fresh cells. Electrospraying was a highly efficient process due to the high cell viability, and consequently, a low quantity of capsules is required for the postharvest treatment of fruits. Additionally, the yeast retained its antagonistic power during storage. © 2021 Society of Chemical Industry.


Species abundance information improves sequence taxonomy classification accuracy.
Popular naive Bayes taxonomic classifiers for amplicon sequences assume that all species in the reference database are equally likely to be observed. We demonstrate that classification accuracy degrades linearly with the degree to which that assumption is violated, and in practice it is always violated. By incorporating environment-specific taxonomic abundance information, we demonstrate a significant increase in the species-level classification accuracy across common sample types. At the species level, overall average error rates decline from 25% to 14%, which is favourably comparable to the error rates that existing classifiers achieve at the genus level (16%). Our findings indicate that for most practical purposes, the assumption that reference species are equally likely to be observed is untenable. q2-clawback provides a straightforward alternative for samples from common environments.


The response to selection in Glycoside Hydrolase Family 13 structures: A comparative quantitative genetics approach.
The Glycoside Hydrolase Family 13 (GH13) is both evolutionarily diverse and relevant to many industrial applications. Its members hydrolyze starch into smaller carbohydrates and members of the family have been bioengineered to improve catalytic function under industrial environments. We introduce a framework to analyze the response to selection of GH13 protein structures given some phylogenetic and simulated dynamic information. We find that the TIM-barrel (a conserved protein fold consisting of eight α-helices and eight parallel β-strands that alternate along the peptide backbone, common to all amylases) is not selectable since it is under purifying selection. We also show a method to rank important residues with higher inferred response to selection. These residues can be altered to effect change in properties. In this work, we define fitness as inferred thermodynamic stability. We show that under the developed framework, residues 112Y, 122K, 124D, 125W, and 126P are good candidates to increase the stability of the truncated α-amylase protein from Geobacillus thermoleovorans (PDB code: 4E2O; α-1,4-glucan-4-glucanohydrolase; EC 184.108.40.206). Overall, this paper demonstrates the feasibility of a framework for the analysis of protein structures for any other fitness landscape.


Esophagus sparing with IMRT in lung tumor irradiation: an EUD-based optimization technique.
The aim of this study was to evaluate (1) the use of generalized equivalent uniform dose (gEUD) to optimize dose escalation of lung tumors when the esophagus overlaps the planning target volume (PTV) and (2) the potential benefit of further dose escalation in only the part of the PTV that does not overlap the esophagus. The treatment-planning computed tomography (CT) scans of patients with primary lung tumors located in different regions of the left and right lung were used for the optimization of beamlet intensity modulated radiation therapy (IMRT) plans. In all cases, the PTV overlapped part of the esophagus. The dose in the PTV was maximized according to 7 different primary cost functions: 2 plans that made use of mean dose (MD) (the reference plan, in which the 95% isodose surface covered the PTV and a second plan that had no constraint on the minimum isodose), 3 plans based on maximizing gEUD for the whole PTV with ever increasing assumptions for tumor aggressiveness, and 2 plans that used different gEUD values in 2 simultaneous, overlapping target volumes (the whole PTV and the PTV minus esophagus). Beam arrangements and NTCP-based costlets for the organs at risk (OARs) were kept identical to the original conformal plan for each case. Regardless of optimization method, the relative ranking of the resulting plans was evaluated in terms of the absence of cold spots within the PTV and the final gEUD computed for the whole PTV. Because the MD-optimized plans lacked a constraint on minimum PTV coverage, they resulted in cold spots that affected approximately 5% of the PTV volume. When optimizing over the whole PTV volume, gEUD-optimized plans resulted in higher equivalent uniform PTV doses than did the reference plan while still maintaining normal-tissue constraints. However, only under the assumption of extremely aggressive tumors could cold spots in the PTV be avoided. Generally, high-level overall results are obtained when optimization in the whole PTV is also associated with a second simultaneous optimization in the PTV minus overlapping portions of the esophagus. Intensity modulated radiation therapy optimizations that utilize gEUD-based cost functions for the PTV and NTCP-based constraints for the OARs result in increased doses to large portions of the PTV in cases where the PTV overlaps the esophagus, while still maintaining (and confining to the overlap region) minimum dose coverage equivalent to the homogeneous PTV optimization cases.


Genomes of surface isolates of Alteromonas macleodii: the life of a widespread marine opportunistic copiotroph.
Alteromonas macleodii is a marine gammaproteobacterium with widespread distribution in temperate or tropical waters. We describe three genomes of isolates from surface waters around Europe (Atlantic, Mediterranean and Black Sea) and compare them with a previously described deep Mediterranean isolate (AltDE) that belongs to a widely divergent clade. The surface isolates are quite similar, the most divergent being the Black Sea (BS11) isolate. The genomes contain several genomic islands with different gene content. The recruitment of very similar genomic fragments from metagenomes in different locations indicates that the surface clade is globally abundant with little effect of geography, even the AltDE and the BS11 genomes recruiting from surface samples in open ocean locations. The finding of CRISPR protospacers of AltDE in a lysogenic phage in the Atlantic (English Channel) isolate illustrates a flow of genetic material among these clades and a remarkably wide distribution of this phage.


Flexible carbon fiber cloth supports decorated with cerium metal- organic frameworks and multi-walled carbon nanotubes for simultaneous on-site detection of Cd2+ and Pb2+ in food and water samples.
The widespread heavy metal pollution endangers human health; hence, accurate on-site detection and quantification of heavy metal content in the surroundings is a vital step in reversing the harmful effect. Herein, an electrochemical sensor based on flexible cerium metal-organic framework@multi-walled carbon nanotubes/carbon cloth (CeMOF@MWCNTs/CC) was constructed for simultaneous on-site detection of Cd2+ and Pb2+ in food and water samples. The rich carboxyl groups of MWCNTs provided abundant sites for the adsorption of Cd2+ and Pb2+, and the mutual conversion of Ce3+ and Ce4+ in CeMOF facilitated the reduction and reoxidation of metal ions. The prepared electrode showed excellent performance in the simultaneous measurement of Cd2+ and Pb2+, with detection limits of 2.2 ppb and 0.64 ppb, respectively. More importantly, the sensing platform has been successfully used to detect simultaneously Cd2+ and Pb2+ in grain and water samples, and the detection results were consistent with the standard methods, showing great potential in environmental monitoring and food safety.


Developmental dynamics of functionally specific primary sensory afferent projections in the chicken embryo.
The central projections of specific subpopulations of lumbar primary afferents were selectively labeled with the lipophilic tracer DiI in fixed preparations of the chicken embryo. Muscle or cutaneous afferents were selectively labeled by applying DiI to identified peripheral nerves. Medial or lateral afferent populations were selectively labeled by partially lesioning the dorsal root. Muscle and cutaneous afferent populations each contribute to both the medial and the lateral afferent populations. Medial muscle afferents terminate in the intermediate zone and lateral motor column proximally, but only in the intermediate zone distally. Lateral muscle afferents terminate in a ventrolateral region of the dorsal horn both proximally and distally. Medial cutaneous afferents terminate predominantly in lamina III, but a few terminate in the medial region of the intermediate zone. Lateral cutaneous afferents terminate in lamina II and in a ventrolateral region of the dorsal horn. On the basis of the principle termination patterns, specific termination fields were defined and related to the classical cytoarchitectonics of the spinal gray matter. Differential retrograde tracing from the spinal cord with fluorescent dextran-amines demonstrated that the medial afferents originate from the earlier-generated ventrolateral population of large sensory cell bodies, while the lateral afferents originate from the later-generated dorsomedial population of small sensory cell bodies. The medial afferents establish their central projections earlier than the lateral afferents, but for each subpopulation the initial pattern of termination prefigures the mature pattern, throughout the segmental range of the collaterals. Birthdating with 3H-thymidine showed that potential target neurons in the different terminal fields within the dorsal horn are born at different times. In particular, interneurons in lamina II are born after those in lamina III, paralleling the early and late termination of cutaneous afferents in these laminae. Our observations support the notion that primary afferents recognize specific cues in the spinal cord, but also implicate the relative timing of afferent and target differentiation as an important determinant of primary afferent termination patterns.


A systematic review and meta-analysis comparing everolimus and calcineurin inhibitors (CNIs) to mycophenolate and CNIs in kidney transplant patients.
This study compared everolimus and mycophenolate mofetil, each paired with calcineurin inhibitors (CNIs) and used with or without steroids, for maintaining immunosuppression in kidney transplant (KT) patients. Relevant studies published before August 21, 2022 were retrieved from PubMed, the Cochrane Central Register of Controlled Trials, and the gray literature. The risk of bias was assessed independently using the revised Cochrane risk of bias assessment tool (RoB 2). RevMan ver. 5.4 was used to calculate the risk ratios (RRs) with corresponding 95% confidence intervals (CIs) for biopsy-proven acute rejection, death, and infection. The mean difference (MD) was used to compare the estimated glomerular filtration rate (eGFR) between the groups. Sixteen randomized controlled trials with a total of 5,403 patients were synthesized to compare everolimus (n=2,763) with mycophenolate (n=2,542) for maintaining post-KT immunosuppression. The meta-analysis showed no significant difference in the risk for biopsy-proven acute rejection (RR=1.12; 95% CI, 0.92-1.35; I2=29%) and death (RR=0.85; 95% CI, 0.63-1.16; I2=0%). The eGFR had no significant difference between the two groups (MD=0.93; 95% CI, -2.25 to 4.1; I2=84%). The risk for any infection was significantly higher in the mycophenolate group than in the everolimus group (RR=0.83; 95% CI, 0.73-0.93; I2=66%). Our meta-analysis showed that when paired with a CNI, everolimus and mycophenolate had no difference in risk for biopsy-proven acute rejection, death, or increase in eGFR. However, the mycophenolate group exhibited a significantly higher risk of infection.


PEDOT: PSS/AuNPs-Based Composite as Voltammetric Sensor for the Detection of Pirimicarb.
An electrochemical sensor for the pesticide Pirimicarb (PMC) has been developed. A screen-printed electrode (SPCE) was used and modified with the conducting polymer poly (3,4-ethylenedioxythiophene) (PEDOT) and gold nanoparticles (AuNPs) to enhance electrochemical proprieties. Electrode characterizations were performed using scattering electron microscopy (SEM) and cyclic voltammetry (CV). With the SPCE/PEDOT:PSS/AuNPs modified electrode, a new peak at 1.0 V appeared in the presence of PMC related to the PMC oxidation. To elucidate the mechanism of PMC oxidation, Gas Chromatography-Mass Spectrometry (GC-MS), where two major peaks were identified, evidencing that the device can both detect and degrade PMC by an electro-oxidation process. Exploring this peak signal, it was possible the sensor development, performing detection from 93.81-750 µmol L-1, limits of quantification (LOQ) and detection (LOD) of 93.91 µmol L-1 and 28.34 µmol L-1, respectively. Thus, it was possible to study and optimization of PMC degradation, moreover, to perform detection at low concentrations and with good selectivity against different interferents using a low-cost printed electrode based on graphite modified with conductive polymer and AuNPs.


Comparison of efficacies of haploidentical transplantation and matched sibling donor transplantation in treating T-cell lymphoblastic lymphoma.
To investigate the differences in efficacy and safety between haploidentical donor hematopoietic stem cell transplantation (HID-HSCT) and matched sibling donor HSCT (MSD-HSCT) in patients with T-cell lymphoblastic lymphoma (T-LBL). In this retrospective analysis, we enrolled 38 patients who had undergone allogeneic HSCT at our institution between 2013 and 2021. The study participants included 28 patients who underwent HID-HSCT and 10 patients who underwent MSD-HSCT. We compared the patient characteristics and treatment effectiveness and safety between the two groups and evaluated potential prognostic variables for patients with T-LBL. The median follow-up durations in the HID-HSCT and MSD-HSCT groups were 23.5 (range: 4-111) and 28.5 (range: 13-56) months, respectively. All patients showed full-donor chimerism after hematopoietic stem cell transplantation (HSCT). Except for two patients in the HID-HSCT cohort who developed poor graft function, all patients showed neutrophil and platelet engraftments after HSCT. The cumulative incidences of grades III-IV acute graft-versus-host disease were 37.5% and 28.57% in the HID-HSCT and MSD-HSCT groups, respectively (p = 0.84). The cumulative incidences of limited (34.13% vs. 28.57%, p = 0.82) and extensive (31.22% vs. 37.50%, p = 0.53) chronic graft-versus-host disease did not differ between the two cohorts. In the HID-HSCT and MSD-HSCT cohorts, the estimated 2-year overall survival rates were 70.3% (95% confidence interval [CI]: 54.9%-90.0%) and 56.2% (95% CI: 31.6%-100%), respectively (p = 1.00), and the estimated 2-year progression-free survival (PFS) rates were 48.5% (95% CI: 32.8%-71.6%) and 48.0% (95% CI: 24.6%-93.8%), respectively (p = 0.94). Furthermore, the Cox proportional-hazards model showed that a positive positron emission tomography/computed tomography (PET/CT) status before HSCT in patients who had completed chemotherapy was an independent risk factor for PFS in the multivariate analysis (p = 0.0367). This study showed that HID-HSCT had comparable effectiveness and safety to MSD-HSCT in treating T-LBL. HID-HSCT could serve as an alternate treatment option for T-LBL in patients without an eligible identical donor. Achievement of the PET/CT-negative status before HSCT may contribute to better survival.


Vancomycin-resistant Enterococcus faecium (VREF) from Norwegian poultry cluster with VREF from poultry from the United Kingdom and The Netherlands in an amplified fragment length polymorphism genogroup.
The genetic relationship between 197 vancomycin-resistant Enterococcus faecium (VREF) isolates and 21 vancomycin-susceptible E. faecium isolates from Norwegian poultry was analyzed by amplified fragment length polymorphism (AFLP). The isolates were compared to 255 VREF isolates from various sources and countries. The Norwegian isolates constituted a relatively homogeneous population of E. faecium and clustered in a previously defined poultry AFLP genogroup.


Accuracy of formula-based volume and image segmentation-based volume in calculation of preoperative cystic jaw lesions' volume.
The aim of this study was to assess the accuracy of formula-based volume measurements and the 3D volume analysis with different software packages in the calculation of preoperative cystic jaw lesions' volume. The secondary aim was to assess the reliability and the accuracy of 3 imaging software programs for measuring the cystic jaw lesions' volume in CBCT images. This study consisted of two parts: an in vitro part using 2 dry human mandibles that were used to create simulated osteolytic lesions to assess the accuracy of the volumetric analysis and formula-based volume. As a gold standard, the volume of each bone defect was determined by taking an impression using rapid soft silicone (Vinylight) and then quantifying the volume of the replica. Afterward, each tooth socket was scanned using a high-resolution CBCT. A retrospective part using archived CBCT radiographs that were taken from the database of the outpatient clinic of the oral and maxillofacial radiology department, Faculty of Dentistry, Minia University to assess the reliability of the 3 software packages. The volumetric data set was exported for volume quantification using the 3 software packages (MIMICS-OnDemand and InVesalius software). Also, the three greatest orthogonal diameters of the lesions were calculated, and the volume was assessed using the ellipsoid formula. Dunn's test was used for pair-wise comparisons when Friedman's test was significant. The inter-examiner agreement was assessed using Cronbach's alpha reliability coefficient and intra-class correlation coefficient. Regarding the results of the retrospective part, there was a statistically significant difference between volumetric measurements by equation and different software (P value < 0.001, Effect size = 0.513). The inter-observer reliability of the measurements of the cystic lesions using the different software packages was very good. The highest inter-examiner agreement for volume measurement was found with InVesalius (Cronbach's alpha = 0.992). On the other hand, there was a statistically significant difference between dry mandible volumetric measurements and Gold Standard. All software showed statistically significantly lower dry mandible volumetric measurements than the gold standard. Computer-aided assessment of cystic lesion volume using InVesalius, OnDemand, and MIMICS is a readily available, easy to use, non-invasive option. It confers an advantage over formula-based volume as it gives the exact morphology of the lesion so that potential problems can be detected before surgery. Volume analysis with InVesalius software was accurate in determining the volume of simulated periapical defects in a human cadaver mandible as compared to true volume. InVesalius software proved that open-source software can be robust yet user-friendly with the advantage of minimal cost to use.


Regulation of (+):(-)-strand asymmetry in replication of brome mosaic virus RNA.
Transfection of barley protoplasts with brome mosaic virus (BMV) RNAs 1 + 2 in the absence of RNA-3 yielded a molar ratio for (+):(-)-strand progeny at 24 hr postinoculation near unity, whereas over 100-fold more (+)- than (-)-strand progeny accumulated in its presence. The presence of RNA-3 enhanced total (+)-strand RNA production 205-fold and that of RNAs 1 + 2 by 29-fold. In contrast, total (-)-strand RNA accumulation decreased by 68% and that for (-)RNAs 1 + 2 by 79% in the presence of RNA-3. Transfections containing an RNA-3 mutant (Gsgi----U RNA-3) that is incapable of yielding RNA-4 as a result of a single nucleotide substitution at the subgenomic RNA initiation site yielded only 66% of the (+):(-) asymmetry seen in the presence of wild-type RNA-3. Only 1.8-fold excess of (+)-over (-)-strand production was obtained for transfections that included delta SGP RNA-3, a deletion that includes the subgenomic promoter core and extends 43 nt into the RNA-4 sequence. Transfections containing RNA-3 mutants bearing frameshifts or deletions in the coat protein cistron yielded levels of asymmetry similar to those seen for Gsgi----U RNA-3. These findings implicate the subgenomic promoter and other sequences in the intercistronic region of RNA-3 as the primary determinants of asymmetric replication, although the coat protein may be an additional factor enhancing the accumulation of (+)-strand RNA.


Adjuvant glycerol is not beneficial in experimental pneumococcal meningitis.
Bacterial meningitis in children causes high rates of mortality and morbidity. In a recent clinical trial, oral glycerol significantly reduced severe neurological sequelae in paediatric meningitis caused by Haemophilus influenzae type b, and a tendency towards a benefit of adjunctive glycerol was seen in pneumococcal meningitis. Here we examined the effects of glycerol in pneumococcal meningitis of infant rats and adult mice. All animals received ceftriaxone, and glycerol or placebo. Brain damage, hearing loss, and inflammatory parameters were assessed. Clinically and by histopathology, animals treated with glycerol or placebo did not differ. While both groups showed equally high levels of matrix metalloproteinase-9 at 24 h after infection, a significant difference in favour of glycerol was observed at 40 h after infection. However, this difference in matrix metalloproteinase-9 in late disease did not result in an improvement of histopathologic parameters. No benefit of adjunctive glycerol was found in these models of pneumococcal meningitis.


Motivational techniques of dietitians counseling individuals with type II diabetes.
A survey was conducted to ascertain to what extent dietitians use techniques cited in the literature considered to affect adherence by patients with non-insulin dependent (Type II) diabetes. A survey instrument listing techniques related to changing behavior, the counseling process, and content of teaching was developed and mailed to 500 members of the ADA Diabetes Care and Education Practice Group. Responses were received from 39% of the sample. For a majority of techniques, data analysis showed statistical association between frequency of use, perceived importance, and adequacy of practitioner training. Dietitians regularly used only 40% of the cited techniques related to changing behavior and the counseling process, but they said most of the items were important. Only half of the items related to the content of teaching were rated as important, and dietitians felt prepared to use only 70% of them. Most dietitians did not teach concepts related to glucose utilization, energy metabolism, and the metabolic role of insulin in diabetes. Those findings suggest that dietitians do not regularly use many techniques considered effective in obtaining dietary adherence of patients with diabetes. Since other studies show compliance is directly influenced by skills of the provider, dietitians could improve cost-effective health care by developing further professional expertise in the area of counseling patients with diabetes.


Absence of an hypoxic depression of metabolism in preproenkephalin knockout mice.
Opioids inhibit breathing in mammals, especially in newborns, and are also implicated in the control of hypoxic anapyrexia. We measured breathing patterns and metabolic responses to 12% oxygen in six adult male wildtype C57B/6J mice and six preproenkephalin knockout (PPNK-/-) mice in a flow-through respirometer and barometric plethysmograph with ambient temperature maintained in the thermoneutral zone. Breathing air, there was no significant difference between the two groups of mice in ventilation ((.)V), oxygen consumption ((.)V(O(2)), convection requirement ((.)V/(.)V(O(2)), tidal volume (V(t)), frequency (f), or inspiratory time (T(i)); however, PPNK-/- mice had a significantly shorter expiratory time (T(e)). The breathing pattern response to 5% CO(2) was the same between wildtype and PPNK-/- in terms of absolute values, but the % change in V(t) was greater in the wildtype. Breathing 12% O(2), there was no significant difference in V , V(t), f, T(i), T(e) or body temperature between groups, but there was a significant difference in (.)V(O(2) (PPNK-/- 1.24+/-0.05 ml O(2)min(-1) versus 0.91+/-0.05 for wildtype, P<0.001) and % change in (.)V(O(2), (2.3+/-6.6% for PPNK-/- versus -28+/-3.8% for wildtype); in ((.)V/(.)V(O(2)), (54+/-4 versus 78+/-10, P<0.05) and the % change in (.)V/(.)V(O(2), (37+/-9 versus 131+/-28, P<0.01). These data implicate enkephalin as a signaling molecule in the control of hypoxic depression of metabolism in mice.


Label-Free Identification of Persistent Particles in Association with Primary Immune Cells by Imaging Flow Cytometry.
The frequency of human exposure to persistent particles via consumer products, air pollution, and work environments is a modern-day hazard and an active area of research. Particle density and crystallinity, which often dictate their persistence in biological systems, are associated with strong light absorption and reflectance. These attributes allow several persistent particle types to be identified without the use of additional labels using laser light-based techniques such as microscopy, flow cytometry, and imaging flow cytometry. This form of identification allows the direct analysis of environmental persistent particles in association with biological samples after in vivo studies and real-life exposures. Microscopy and imaging flow cytometry have progressed with computing capabilities and fully quantitative imaging techniques can now plausibly detail the interactions and effects of micron and nano-sized particles with primary cells and tissues. This chapter summarises studies which have utilized the strong light absorption and reflectance characteristics of particles for their detection in biological specimens. This is followed by the description of methods for the analysis of whole blood samples and the use of imaging flow cytometry to identify particles in association with primary peripheral blood phagocytic cells, using brightfield and darkfield parameters.


Factors associated with mortality or transplantation versus Fontan completion after cavopulmonary shunt for patients with tricuspid atresia.
Tricuspid atresia with normally related great vessels (TA) is considered the optimal substrate for the Fontan pathway. The factors associated with death or transplantation after cavopulmonary shunt (CPS) are underappreciated. We aimed to determine factors associated with CPS-Fontan interstage death/transplantation versus transition to Fontan in TA. A total of 417 infants younger than 3 months of age with TA were enrolled (January 1999 to February 2020) from 40 institutions into the Congenital Heart Surgeons' Society TA cohort. Parametric competing risk methodology was used to determine factors associated with the competing end points of death/transplantation without Fontan completion, and transition to Fontan. CPS was performed in 382 patients with TA; of those, 5% died or underwent transplantation without transition to Fontan and 91% transitioned to Fontan by 5 years after CPS. Prenatal diagnosis (hazard ratio [HR], 0.74; P < .001) and pulmonary artery band (PAB) at CPS (HR, 0.50; P < .001) were negatively associated with Fontan completion. Preoperative moderate or greater mitral valve regurgitation (HR, 3.0; P < .001), concomitant mitral valve repair (HR, 11.0; P < .001), PAB at CPS (HR, 3.0; P < .001), postoperative superior vena cava interventions (HR, 9.0; P < .001), and CPS takedown (HR, 40.0; P < .001) were associated with death/transplantation. The mortality rate after CPS in patients with TA is notable. Those with preoperative mitral valve regurgitation remain a high-risk group. PAB at the time of CPS being associated with both increased risk of death and decreased Fontan completion may represent a deleterious effect of antegrade pulmonary blood flow in the CPS circulation.


Size does matter: An exploration of the relationship between licensed venue capacity and on-premise assaults.
Venue capacity has been proposed as a factor associated with increased number of violent incidents on-premises, though no specific research has demonstrated this association, and instead has tended to focus on the relationship between crowding and aggression. The aim of current paper is to investigate the association between venue capacity and the number of violent incidents on-premises. Venue capacity data (the maximum capacity listed on the liquor license) were obtained for all venues in central Melbourne from 2010 until 2016. These data were then matched with police-recorded on-premises assaults that occurred within high-alcohol hours (Friday and Saturday 8 pm-6 am) inside the venue. Analyses were conducted on 5729 venue-years (yearly assault counts per venue, per year) across central Melbourne. Compared with venues that have a maximum capacity of between 0 and 100 patrons, venues with higher capacities have increasingly more recorded assaults. Venues with maximum capacities between 501 and 1000 are 6.1 times more likely to have an assault recorded compared with venues with a maximum capacity between 0 and 100. Further, each additional high-alcohol hour that a venue can be open for is associated with a 72% increase in the number of recorded assaults. Greater venue capacity was found to be strongly associated with an increased risk of violent incidents for any given venue. This was further exacerbated by late-night trading which substantially adds to the risk of assaults inside the venue.


Integrated eco-strategies towards sustainable carbon and nitrogen cycling in agriculture.
To meet the ever-growing human demands for food, fuel, and fiber, agricultural activities have dramatically altered the global carbon (C) and nitrogen (N) cycles. These biogeochemical cycles along with water, phosphorus, and sulfur cycles are fundamental features of life on Earth. Human alteration of the global N cycle has had both positive and negative outcomes. To efficiently feed a growing population, crop-livestock production systems have been developed, however, these systems also contribute significantly to environmental pollution and global climate change. Management of agricultural waste (AW) and the application of N fertilizers are central to the issues of greenhouse gas (GHG) emissions and nutrient runoff that contributes to the eutrophication of water bodies. If managed properly, AW can provide nutrients for plants and contribute to the conservation of soil health. In order to achieve the long-term conservation of agricultural production systems, it is important to promote the proper recycling of AW in agroecosystems and to minimize the reliance on chemical N fertilizers. Composting is one of the sustainable and effective approaches for recycling AW in agriculture. However, the conventional composting process is dilatory and produces compost with low N content compared to chemical N fertilizers. For this reason, comprehensive research is required to improve the composting process and the N content of the soil organic amendments. This work aims to explore the beneficial effects of the integrated application of biochar and specific C and N cycling microorganisms to the composting process and the quality of the composted products. In pursuit of replacing chemical N fertilizers with bio/organic fertilizers, we further discussed the power of the combined application of compost, biochar, and N-fixing bacteria in agricultural production systems. The knowledge of smart integration of AW and microorganisms in agriculture could solve the main agricultural and environmental problems associated with human-induced flows of C and N. Building upon the knowledge disseminated in review to further extensive research will pave the way for better management of agricultural production systems and sustainable C and N cycling in agriculture.


[The surgical treatment of villous tumours of the large bowel (author's transl)].
The treatment of 12 patients with villous tumours of the rectum is described. In benign lesions a sphincter conserving operation should be employed. With the technique of submucosal local excision per anum which was described by A.G. Parks it is possible to remove large villous tumours of the rectum. If there is malignant change with invasion of the muscularis muscosae which is found in about 20% of the cases resection therapy is necessary.


Chylous ascites: an etiology of peritonitis in infancy.
Fifty-six cases of chylous ascites in patients under the age of fifteen have been reported in the last three hundred years. A case of chylous ascites that presented with the signs and symptoms of peritonitis is reviewed. The normal anatomy and physiology of the intraabdominal lymphatic system, pathophysiology of the entity, as well as a mechanism for diagnosis, are addressed and a treatment algorithm is presented.


The need for perforator treatment after VenaSeal and ClosureFast endovenous saphenous vein closure in CEAP 6 patients.
The authors have previously demonstrated that VenaSeal (Medtronic, Inc, Minneapolis, Minn) adhesive, compared with radiofrequency ablation (RFA, ClosureFast; Medtronic, Inc), in treatment of refluxing saphenous veins in CEAP 6 limbs, results in shorter healing times of venous ulcers. The authors hypothesize that the longer treated length possible with VenaSeal's nonthermal modality may affect the number of critical refluxing perforators contributing to the nonhealing wound. This follow-up study compares the need for follow-up treatment of perforator veins after saphenous vein treatment with either radiofrequency ablation (ClosureFast RFA) or adhesive closure (VenaSeal). A multi-institutional retrospective review of CEAP 6 patients who had closure of their saphenous veins from 2015 to 2020 was conducted. Patients who underwent follow-up treatment of perforator veins were grouped according to their method of initial management of their saphenous veins. The primary end point was incidence of a perforator procedure after ClosureFast or VenaSeal ablation. Secondary end points included sclerotherapy to facilitate wound healing. Bivariate analysis used the χ2 test, Fisher exact test, t-test, and Wilcoxon rank sum test. A P value of <.05 defined statistical significance. There were 119 CEAP 6 patients with saphenous closure: 51 limbs treated with VenaSeal and 68 with RFA. Median follow-up was 105 days (interquartile range: 44, 208). All limbs achieved wound healing during the study period. Mean time to wound healing post index procedure was shorter for VenaSeal than RFA (72 vs 293.8 days, P > .0009), as was median time (43 vs 104 days, P = .001). More limbs treated with RFA had previous known deep vein thrombosis (29% vs 10%, P = .009), deep venous insufficiency (82% vs 51%, P = .0003), and perforator reflux (57% vs 29%, P = .002). Limbs with identified follow-up perforator reflux treated with RFA had a higher prevalence of initially treated saphenous veins with RFA compared with those treated with VenaSeal (49% vs 27%, P = .003). There was no difference between the methods of vein closure and use of concurrent sclerotherapy. ClosureFast and VenaSeal are both effective and safe modalities of saphenous ablation, but VenaSeal treatment was associated with less perforator RFA intervention.


Increased BMI and late-life mobility dysfunction; overlap of genetic effects in brain regions.
How obesity earlier in life impacts upon mobility dysfunctions in late life is not well understood. Pernicious effects of excess weight on the musculoskeletal system and mobility dysfunctions are well-recognized. However, increasingly more data support the link of obesity to overall motor defects that are regulated in the brain. To assess the causal relationship between body mass index (BMI) at midlife and performance of the Timed Up-and-Go test (TUG) in late life among a population-based longitudinal cohort of Chinese adults living in Singapore. We evaluated genetic predispositions for BMI in 8342 participants who were followed up from measurement of BMI at average 53 years, to TUG test (as a functional mobility measure) 20 years later. A robust 75.83% of genetically determined BMI effects on late-life TUG scores were mediated through midlife BMI (Pindirect-effect = 9.24 × 10-21). Utilizing Mendelian randomization, we demonstrated a causal effect between BMI and functional mobility in late life (βIVW = 0.180, PIVW = 0.001). Secondary gene enrichment evaluations highlighted down-regulation of genes at BMI risk loci that were correlated with poorer functional mobility in the substantia nigra and amygdala regions as compared to all other tissues. These genes also exhibit differential expression patterns during human brain development. We report a causal effect of obesity on mobility dysfunction. Our findings highlight potential neuronal dysfunctions in regulating predispositions on the causal pathway from obesity to mobility dysfunction.


Agranulocytosis secondary to zinc excess: Clinically relevant observations, including response to G-CSF and oral copper.
Anemia and leukopenia because of copper deficiency can be mistaken for myelodysplasia. Key issues, including response to G-CSF and oral copper, are discussed. This case illustrates a significant deleterious effect of excessive zinc consumption.


A staging system for forelimb regeneration in the axolotl, Ambystoma mexicanum.
A staging system has been devised for normal regeneration from the upper arm in the mature axolotl. It consists of seven externally definable stages: (1) Wound healing (WH): (2) Dedifferentiation (DD); (3) Early bud (EB); (4) Medium bud (MB); (5) Late bud (LB); (6) Palette (Pal), and (7) Digital outgrowth (DO). Serial histological sections of 38 regenerating limbs were used to correlate gross stages with microscopic events in the regenerative process.


Controllable proton-reservoir ordered gel towards reversible switching and reliable electromagnetic interference shielding.
Smart and dynamic electromagnetic interference (EMI) shielding materials possess a remarkable capacity to modify their EMI shielding abilities, rendering them invaluable in various civil and military applications. However, the present response mechanism of switch-type EMI shielding materials is slightly restricted, as it primarily depends on continuous pressure induction, thereby resulting in concerns regarding their durability and reliability. Herein, for the first time, we demonstrate a novel method for achieving solvent-responsive, reversible switching, and robust EMI shielding capabilities using a controlled proton-reservoir ordered gel. The gel contains polyaniline (PANI) and sodium alginate (SA). Initially, SA acts as a proton reservoir for PANI in an aqueous system, enhancing the doping level of PANI and improving its electrical conductivity. Additionally, PANI and SA chains respond to diverse polar solvents, such as water, acetonitrile, ethanol, n-hexane, and air, inducing distinct conformations that affect the degree of PANI conjugation and electron migration along the chains. This process is reversible and non-destructive to the polymer chain, ensuring the effective and uncompromised performance of the EMI shielding switch. We can achieve precise and reversible tuning (on/off) of EMI shielding with different effectiveness levels by manipulating the solvents within the framework. This work opens a new solvent-stimuli avenue for the development of EMI shielding materials with reliable and intelligent on/off switching capabilities.


Effect of cocaine on the affinity of -adrenoceptors for noradrenaline.
1. Doses of cocaine which cause specific or unspecific supersensitivity in cat spleen did not alter the blocking effect of phenoxybenzamine on the responses of isolated cat spleen strips to noradrenaline.2. The same doses of cocaine did not increase the protection of alpha-adrenoceptors given by noradrenaline during a standard exposure to phenoxybenzamine.3. It is concluded that cocaine does not change the affinity of the alpha-adrenoceptor for noradrenaline, and therefore changes in affinity are not responsible for the potentiating action of cocaine.


Molecular tectonics based nanopatterning of interfaces with 2D metal-organic frameworks (MOFs).
The nanostructuring of the graphite surface with 2D coordination networks, based on a combination of an acentric porphyrin tecton bearing two divergently oriented monodentate pyridyl units and a CoCl2 metallatecton, behaving as a four connecting node, was achieved at the solid-liquid interface and characterized by scanning tunnelling microscopy.


Help-seeking for child psychopathology: pathways to informal and professional services in the Netherlands.
To devise and test a model describing the process of help-seeking for child psychopathology in professional and informal service settings. Using structural equation modeling, associations between several help-seeking stages, and the influence of child, family, and context characteristics on these stages were investigated in 246 Dutch children (4-11 years old) between April 2000 and July 2002. Children were selected for having emotional or behavioral problems from a representative general practice sample; 68% of the selected children participated. School personnel played an important role in the process of help-seeking for child psychopathology, in both detecting service need and the provision of and the referral for help. Although Dutch general practitioners are supposed to be gatekeepers of mental health care, their role in help-seeking for child psychopathology was limited. Various family characteristics were shown to influence service need and informal or professional help-seeking. The influence of child characteristics on the help-seeking process was limited. Educating parents about child psychopathology and the availability and accessibility of care, improving general practitioners' skills in detecting child psychopathology, and direct contact of mental health professionals with general practitioners and schools may enhance access to care for children in need.


When Does Alzheimer's Disease Really Start? The Role of Biomarkers.
(Appeared originally in Int J Mol Sci 2019, 20 5536).


The analgesic efficacy of anterior femoral cutaneous nerve block in combination with femoral triangle block in total knee arthroplasty: a randomized controlled trial.
Ultrasound-guided femoral triangle block (FTB) can provide motor-sparing anterior knee analgesia. However, it may not completely anesthetize the anterior femoral cutaneous nerve (AFCN). We hypothesized that an AFCN block (AFCNB) in combination with an FTB would decrease pain during movement in the immediate 12 h postoperative period compared with an FTB alone. Eighty patients scheduled to undergo total knee arthroplasty were randomized to receive either FTB alone (FTB group) or AFCNB with FTB (AFCNB + FTB group) as part of the multimodal analgesic regimen. The primary outcome was pain during movement at 12 h postoperatively. Secondary outcomes included numeric rating scale (NRS) pain scores, incidence of surgical incision site pain, intravenous morphine consumption, immediate functional performance, patient satisfaction, and length of hospital stay. The NRS pain scores on movement 12 h postoperatively were significantly lower in the AFCNB + FTB group than in the FTB group (mean difference: -2.02, 95% CI: -3.14, -0.89, P < 0.001). The incidence of pain at the surgical incision site at 24 h postoperatively and morphine consumption within 48 h postoperatively were significantly lower (P < 0.001), and quadriceps muscle strength at 0° immediately after surgery was significantly greater in the AFCNB + FTB group (P = 0.04). The addition of ultrasound-guided AFCNB to FTB provided more effective analgesia and decreased opioid requirement compared to FTB alone after total knee arthroplasty and may enhance immediate functional performance on the day of surgery.


Immunoglobulin treatment reduces atherosclerosis in apolipoprotein E-/- low-density lipoprotein receptor-/- mice via the complement system.
Atherosclerosis is associated with activation of the immune system. Intravenously applied normal polyclonal immunoglobulins (IVIg) have broad therapeutic applications in the treatment of autoimmune and systemic inflammatory diseases. Recently, IVIg have been shown to inhibit atherogenesis in experimental animal models. To investigate the role of the complement system in this process, we used third complement component-deficient (C3(-/-)) and control atherosclerosis-prone apolipoprotein E (ApoE) and low-density lipoprotein receptor (LDLR) double knock-out mice fed a normal diet. IVIg treatment reduced lesion fraction area in the aortic root of complement-sufficient mice whereas the lesion fraction area of C3(-/-) mice was not affected. Thus, complement activation plays a role in the anti-atherosclerotic effects of IVIg, possibly by C3-derived fragments generated through Fc-dependent complement activation.


Differential expression of the GLUT1 and GLUT4 glucose transporters during differentiation of L6 muscle cells.
Skeletal muscle is the main tissue responsible for glucose utilization in the fed state, and it expresses the ubiquitous GLUT1 glucose transporter and the muscle/fat specific GLUT4 glucose transporter. Here we investigated the expression of these transporters during muscle cell differentiation in vitro. Rat L6 muscle cells were grown to the stages of myoblasts, alignment and fused myotubes. Glucose (2-deoxy-D-glucose) transport was higher in myoblasts, decreasing with the progression of alignment and cell fusion. Conversely, insulin-stimulated glucose uptake was negligible in myoblasts, and increased with cell alignment and fusion. The cellular content of GLUT1 transporters decreased and that of GLUT4 transporters increased with cell fusion. Insulin rapidly stimulated glucose uptake in fused myotubes maintained in 2% serum but not in 10% serum. In 10% serum, basal glucose uptake increased as did the cellular content of GLUT1 transporters, while GLUT4 transporter content did not change. These results indicate that both transporters are regulated oppositely during muscle cell differentiation, and that high serum concentrations override the capacity of insulin to regulate transport by inducing overexpression of the GLUT1 transporter.


Thyrocalcitonin: inhibitor of bone resorption in tissue culture.
A partially purified protein from thyroid glands of rats inhibits release of calcium from long bones of embryo rats in tissue culture. Inhibition was more easily detected when resorption of bone was stimulated by parathyroid hormone.


Variations of renal tissue oxygenation during abdominal compartment syndrome and sepsis.
This experimental study was designed to evaluate the renal tissue oxygenation under the coexistence of abdominal compartment syndrome and sepsis. Fourteen non-breed dogs were divided into two groups: the control group (8) and the study group (6). Sepsis was established with intravenous endotoxin infusion at 100μg/kg for over 30min. Insufflation of CO2 in the peritoneal cavity was used for the increase in intra-abdominal pressure (IAP). A special catheter placed and fixed in the renal cortex at a depth of 3mm from the renal capsule was used for the measurement of renal tissue oxygenation. Study parameters were recorded at the starting phase, at IAP of 15mmHg and 30mmHg and after decompression of the abdomen in the control group, and at the same intervals plus the induction of sepsis, prior to increasing abdominal pressure, in the study group. With the elevation of the IAP a reduction of renal tissue oxygenation presents itself, which is more pronounced in the presence of sepsis, especially for IAP over 15mmHg. Like other parameters, after abdominal decompression the renal tissue oxygenation returns to the initial levels, independently of sepsis. The afferent arterioles vasoconstriction, which takes place during sepsis, and the intra-renal shunt, which occurs and leads to blood diversion to the medulla from the renal cortex due to the combination of intra-abdominal hypertension (IAH) and sepsis, seem to explain this finding.


Systemic macrophage depletion inhibits Helicobacter bilis-induced proinflammatory cytokine-mediated typhlocolitis and impairs bacterial colonization dynamics in a BALB/c Rag2-/- mouse model of inflammatory bowel disease.
Helicobacter bilis, an enterohepatic helicobacter, is associated with chronic hepatitis in aged immunocompetent inbred mice and inflammatory bowel disease (IBD) in immunodeficient mice. To evaluate the role of macrophages in H. bilis-induced IBD, Rag2(-/-) BALB/c or wild-type (WT) BALB/c mice were either sham dosed or infected with H. bilis Missouri strain under specific-pathogen-free conditions, followed by an intravenous injection of a 0.2-ml suspension of liposomes coated with either phosphate-buffered saline (control) or clodronate (a macrophage depleting drug) at 15 weeks postinfection (wpi). At 16 wpi, the ceca of H. bilis-infected Rag2(-/-) mice treated with control liposomes had significantly higher histopathological lesional scores (for cumulative typhlitis index, inflammation, edema, epithelial defects, and hyperplasia) and higher counts of F4/80(+) macrophages and MPO(+) neutrophils compared to H. bilis-infected Rag2(-/-) mice treated with clodronate liposomes. In addition, cecal quantitative PCR analyses revealed a significant suppression in the expression of macrophage-related cytokine genes, namely, Tnfa, Il-1β, Il-10, Cxcl1, and iNos, in the clodronate-treated H. bilis-infected Rag2(-/-) mice compared to the H. bilis-infected Rag2(-/-) control mice. Finally, cecal quantitative PCR analyses also revealed a significant reduction in bacterial colonization in the clodronate-treated Rag2(-/-) mice. Taken together, our results suggest that macrophages are critical inflammatory cellular mediators for promoting H. bilis-induced typhlocolitis in mice.


Airway hyper-responsiveness in young adults with asthma that remitted either during or before adolescence.
More than 50% of patients with childhood asthma enter clinical remission by puberty, although 40-50% of these people will probably develop asthma symptoms during early adulthood. The mechanism of relapsing asthma in early adulthood remains unclear. This study determined the characteristics of young adults whose asthma remitted either during or before adolescence. A comparative study was performed on 24 students whose childhood asthma had gone into clinical remission by puberty (remission group), 25 atopic students with no history of asthma (atopy group) and 19 non-atopic students without allergic diseases (control group). Examinations included spirometry, levels of serum-specific IgE-antibodies, airway responsiveness to methacholine, exhaled nitric oxide (eNO) and evidence of airway inflammation in induced sputum. Airway responsiveness (P < 0.01), eosinophil counts in sputum (P < 0.05) and the prevalence of sensitization to Dermatophagoides forinae (P < 0.01) were significantly higher, and FEF(25-75%) and FEF(75%) (P < 0.01) were significantly lower in the remission group than in the atopy and control groups. Furthermore, 50% and 33% of the remission group had airway hyper-responsiveness (AHR) and sputum eosinophilia, respectively. The eNO levels in the remission (P < 0.01) and atopy (P < 0.05) groups were significantly higher than in controls. Remission group members with AHR had a significantly longer period of childhood asthma, a shorter period of remission and greater airway eosinophilic inflammation than those without AHR (P < 0.05). One half of young adults with childhood asthma that remitted either during or before adolescence continued to have evidence of AHR and airway eosinophilic inflammation, and might be at risk of future relapse.


Turnip yellow mosaic virus variants produced from DNA clones encoding their genomes.
Full-length dsDNA clones that encode the genomes of two Australian turnip yellow mosaic isolates, TYMV-BL and TYMV-CL have been constructed. These clones were transcribed to give 6.3 kb capped ssRNA which infects Chinese cabbages to give symptoms indistinguishable from those produced by the parental viruses. Extensions of up to 26 nucleotides at the 3' end of the TYMV-BL clone delay infections, but virus particles isolated from these plants 4 weeks after inoculation contain RNA with the original TYMV-BL 3' terminus. A 90 nucleotide-long portion of the virion protein gene of TYMV-BL was replaced by a synthetic 90-mer primer with 16 nucleotide changes to decrease the large cytosine content (34-42%) characteristic of tymovirus genomic RNA. No reversion of any of the mutated nucleotides to cytosine occurred during 7 passages in Chinese cabbage. Hybrids between the TYMV-BL and TYMV-CL clones were also constructed, by exchanging various portions of the genome. However, it was not possible to determine definitively which part of the viral genome is responsible for the more severe symptoms caused by TYMV-BL as the hybrids gave intermediate symptoms.


Laparoscopic Billroth II gastrectomy: a review.
The operation of Billroth II gastrectomy has not changed substantially in concept or technique since it was first performed in 1885 by Theodor Billroth. The advent of stapling has made the procedure faster for the surgeon but the operation has remained as painful and as traumatic for the patient. The totally intra-abdominal laparoscopic Billroth II gastrectomy offers a minimally invasive option that is remarkably less traumatic and more 'patient friendly'. Initial experience in this operation around the world has largely concentrated on resection for benign gastric ulcer but the possibility of resection for cancer is not inconceivable. Experience with a small experimental series of eight cases showed that this operation has many advantages over open surgery in terms of postoperative pain, quicker mobilization, fewer wound problems, better cosmesis and quicker discharge. No anastomotic leak, chest or wound complications were encountered. The main problem remains the cost of disposable stapling devices.


Impact of Lung Parenchymal-Only Failure on Overall Survival in Early-Stage Lung Cancer Patients Treated With Stereotactic Ablative Radiotherapy.
The impact of lung parenchymal-only failure on patient survival after stereotactic ablative body radiotherapy (SABR) for early-stage non-small-cell lung cancer (NSCLC) remains unclear. The study population included 481 patients with early-stage NSCLC who were treated with 3- to 5-fraction SABR between 2000 and 2016. The primary study objective was to assess the impact of out-of-field lung parenchymal-only failure (OLPF) on overall survival (OS). At a median follow-up of 5.9 years, the median OS was 2.7 years for all patients. Patients with OLPF did not have a significantly different OS compared to patients without failure (P = .0952, median OS 4.1 years with failure vs. 2.6 years never failure). Analysis in a 1:1 propensity score-matched cohort for Karnofsky performance status, comorbidity score, and smoking status showed no differences in OS between patients without failure and those with OLPF (P = .8). In subgroup analyses exploring the impact of time of failure on OS, patients with OLPF 6 months or more after diagnosis did not have significantly different OS compared to those without failure, when accounting for immortal time bias (P = .3, median OS 4.3 years vs. 3.5 years never failure). Only 7 patients in our data set experienced failure within 6 months of treatment, of which only 4 were confirmed to be true failures; therefore, limited data are available in our cohort on the impact of OLPF for ≤ 6 months on OS. OLPF after SABR for early-stage NSCLC does not appear to adversely affect OS, especially if occurring at least 6 months after SABR. More studies are needed to understand if OLPF within 6 months of SABR is associated with adverse OS. These data are useful when discussing prognosis of lung parenchymal failures after initial SABR.


Cyclic AMP inducibility of the myelin basic protein gene promoter requires the NF1 site.
In the central nervous system oligodendrocyte differentiation is accompanied by the activation of a specific transcriptional program responsible for the synthesis of myelin genes. One of the signals leading to the expression of myelin components, such as the myelin basic protein (MBP) gene is cyclic AMP (cAMP). Previous work using a cell line in which the endogenous MBP gene can be induced by increased cAMP levels (D6P2T) showed that the region of the MBP gene that was required for induction of the gene by cAMP lay between -248 and -105 in the 5' flanking region. This region contains numerous transcription factor binding sites, including sites for NF1, Sp1, and MEBA. In order to determine if the NF1 site itself was specifically responsible for the cAMP responsiveness of the MBP promoter, stably transfected cells carrying MBP promoter deletion constructs were used. Deletion of just the NF1 site caused loss of responsiveness to cAMP levels. Furthermore, site-specific mutations in the NF1 site that interfere with NF1 protein binding, in the context of the full length promoter, abolished cAMP responsiveness and caused derepression of the promoter. Analysis of protein binding to the NF1 site showed that the mutation resulted in loss of binding to the site and that the proteins binding at the site are modified in the presence of cAMP elevating agents. These results demonstrate that the NF1 site is indispensable for cAMP responsiveness of the MBP promoter and, together with other DNA elements, plays a role in controlling MBP gene expression.


A MT1-MMP/NF-kappaB signaling axis as a checkpoint controller of COX-2 expression in CD133+ U87 glioblastoma cells.
The CD133(+) stem cell population in recurrent gliomas is associated with clinical features such as therapy resistance, blood-brain barrier disruption and, hence, tumor infiltration. Screening of a large panel of glioma samples increasing histological grade demonstrated frequencies of CD133(+) cells which correlated with high expression of cyclooxygenase (COX)-2 and of membrane type-1 matrix metalloproteinase (MT1-MMP). We used qRT-PCR and immunoblotting to examine the molecular interplay between MT1-MMP and COX-2 gene and protein expression in parental, CD133(+), and neurospheres U87 glioma cell cultures. We found that CD133, COX-2 and MT1-MMP expression were enhanced when glioma cells were cultured in neurosphere conditions. A CD133(+)-enriched U87 glioma cell population, isolated from parental U87 cells with magnetic cell sorting technology, also grew as neurospheres and showed enhanced COX-2 expression. MT1-MMP gene silencing antagonized COX-2 expression in neurospheres, while overexpression of recombinant MT1-MMP directly triggered COX-2 expression in U87 cells independent from MT1-MMP's catalytic function. COX-2 induction by MT1-MMP was also validated in wild-type and in NF-kappaB p65-/- mutant mouse embryonic fibroblasts, but was abrogated in NF-kappaB 1 (p50-/-) mutant cells. We provide evidence for enhanced COX-2 expression in CD133(+) glioma cells, and direct cell-based evidence of NF-kappaB-mediated COX-2 regulation by MT1-MMP. The biological significance of such checkpoint control may account for COX-2-dependent mechanisms of inflammatory balance responsible of therapy resistance phenotype of cancer stem cells.