Construction of Nef-positive doxycycline-dependent HIV-1 variants using bicistronic expression elements.
Conditionally replicating HIV-1 variants that can be switched on and off at will are attractive tools for HIV research. We previously developed a genetically modified HIV-1 variant that replicates exclusively when doxycycline (dox) is administered. The nef gene in this HIV-rtTA variant was replaced with the gene encoding the dox-dependent rtTA transcriptional activator. Because loss of Nef expression compromises virus replication in primary cells and precludes studies on Nef function, we tested different approaches to restore Nef production in HIV-rtTA. Strategies that involved translation via an EMCV or synthetic internal ribosome entry site (IRES) failed because these elements were incompatible with efficient virus replication. Fusion protein approaches with the FMDV 2A peptide and human ubiquitin were successful and resulted in genetically-stable Nef-expressing HIV-rtTA strains that replicate more efficiently in primary T-cells and human immune system (HIS) mice than Nef-deficient variants, thus confirming the positive effect of Nef on in vivo virus replication.


Radiological-histopathological discordance in patients transplanted for HCC and its impact on post-transplant outcomes.
Contrast-enhanced cross-sectional imaging is the cornerstone in the diagnosis, staging, and management of HCC, including eligibility for liver transplantation (LT). Radiological-histopathological discordance may lead to improper staging and may impact patient outcomes. We aimed to assess the radiological-histopathological discordance at the time of LT in HCC patients and its impact on the post-LT outcomes. We analyzed further the effect of 6-month waiting policy on the discordance. Using United Network for Organ Sharing-Organ Procurement and Transplantation Network (UNOS-OPTN) database, we examined the discordance between pre-LT imaging and explant histopathology for all adult HCC patients who received liver transplants from deceased donors between April 2012 and December 2017. Kaplan-Meier methods and Cox regression analyses were used to evaluate the impact of discordance on 3-year HCC recurrence and mortality. Of 6842 patients included in the study, 66.7% were within Milan criteria on both imaging and explant histopathology, and 33.3% were within the Milan based on imaging but extended beyond Milan on explant histopathology. Male gender, increasing numbers of tumors, bilobar distribution, larger tumor size, and increasing AFP are associated with increased discordance. Post-LT HCC recurrence and death were significantly higher in patients who were discordant, with histopathology beyond Milan (adj HR 1.86, 95% CI 1.32-2.63 for mortality and 1.32, 95% CI 1.03-1.70 for recurrence). Graft allocation policy with 6-month waiting time led to increased discordance (OR 1.19, CI 1.01-1.41), although it did not impact post-LT outcome. Current practice for staging of HCC based on radiological imaging features alone results in underestimation of HCC burden in one out of three patients with HCC. This discordance is associated with a higher risk of post-LT HCC recurrence and mortality. These patients will need enhanced surveillance to optimize patient selection and aggressive LRT to reduce post-LT recurrence and increase survival.


Induction chemotherapy response of HPV-related multiphenotypic sinonasal carcinoma: First-ever reported therapeutic outcome and a brief literature review.
A 54-year-old male patient complained of nasal obstruction and epistaxis for 2 years, with worsening of the symptoms in the preceding year. Physical examination revealed a friable, irregular mass, with yellowish secretion, in the left nasal fossa. Magnetic resonance imaging revealed an expansive lesion in the left nasal cavity, extending into the nasopharynx, ethmoid, right nasal cavity, and cortical bone of the hard palate. An incisional biopsy was then performed. Morphologically, a cellular malignant proliferation with a solid basaloid appearance admixed with adenoid cystic-like areas was observed. Immunohistochemistry revealed positivity for AE1/AE3, CK7, p63, and calponin, with focal labeling for CD117 and α-SMA. p16 had diffuse cytoplasmic and nuclear positivity. Ki-67 index was >80%. Given the morphological and immunohistochemical aspects, the diagnosis was conclusive for HPV-related multiphenotypic sinonasal carcinoma. The tumor was considered irresectable, and the patient was submitted to induction chemotherapy with docetaxel, cisplatin, and infusional 5-fluorouracil, with significant regression after therapy, followed by chemoradiotherapy with carboplatin, without limiting toxicities. The patient is currently under regular follow-up, with complete clinical and radiological response. To date, there are no reports in the literature of induction chemotherapy use or its complete therapeutic responsiveness related to this lesion. A brief literature review was included with the main epidemiological, clinical, therapeutic, and prognostic aspects regarding the 85 cases reported in the literature, including ours.


Successful cardiopulmonary resuscitation following minimally invasive pectus excavatum repair: A case report.
Pectus excavatum is the most common congenital chest wall deformity. The minimally invasive repair of pectus excavatum (MIRPE) is the most commonly practiced method of surgical treatment and there is concern that the pectus bar will prevent effective CPR. There are no recorded cases of successful cardiopulmonary resuscitation with pectus bars in place. A 17-year-old male with pectus excavatum underwent MIRPE. Two years later, he experienced out-of-hospital cardiac arrest and underwent successful cardiopulmonary resuscitation (CPR) in the field with a pectus bar in place. Successful CPR is possible after MIRPE. Clear identification of patients who have undergone MIRPE and education of CPR providers in providing effective chest compressions and defibrillation for this patient population is necessary. This is the first documented case of successful CPR in a patient with a pectus bar in place who experienced out-of-hospital cardiac arrest.


Microengineered Multi-Organoid System from hiPSCs to Recapitulate Human Liver-Islet Axis in Normal and Type 2 Diabetes.
Type 2 diabetes mellitus (T2DM) is a systematic multi-organ metabolic disease, which is characterized by the dynamic interplay among different organs. The increasing incidence of T2DM reflects an urgent need for the development of in vitro human-relevant models for disease study and drug therapy. Here, a new microfluidic multi-organoid system is developed that recapitulates the human liver-pancreatic islet axis in normal and disease states. The system contains two compartmentalized regions connected by a microchannel network, enabling 3D co-culture of human induced pluripotent stem cells (hiPSC)-derived liver and islet organoids for up to 30 days under circulatory perfusion conditions. The co-cultured liver and islet organoids exhibit favorable growth and improved tissue-specific functions. Transcriptional analyses reveal the activation of metabolically relevant signaling pathways in the co-cultured organoids. Notably, the co-culture system facilitates sensitive glucose-stimulated insulin secretion from islet organoids and increased glucose utilization in liver organoids by glucose tolerance tests. Both liver and islet organoids display mitochondrial dysfunction and decreased glucose transport capacity under high glucose conditions, which can be alleviated by metformin treatment. This novel multi-organoid system can recapitulate human-relevant liver-islet axis under both physiological and pathological conditions, providing a unique platform for future T2DM research and drug development.


Red blood cell count and risk of adverse outcomes in patients with mildly reduced left ventricular ejection fraction.
Anemia is associated with increased rates of heart failure (HF)-related mortality and hospitalization. No studies have focused on the association between the red blood cell (RBC) count and the prognosis of patients with HF with mildly reduced left ventricular ejection fraction (HFmrEF). We retrospectively analyzed the effect of the RBC count on outcome events in patients with HFmrEF. We investigated the association of the RBC count with outcome events in 1691 patients with HFmrEF (mean age: 68 years; 35% female) in Xiangtan Central Hospital. Using Cox proportional hazards models, the RBC count was assessed as both a continuous and categorical variable. During follow-up (median: 33 months), cardiovascular death occurred in 168 patients (114 men and 54 women). After adjusting for established risk factors, each 1.0 × 1012 cell/L increase in the RBC count was associated with a 28% lower risk of cardiovascular death in men and a 43% lower risk in women. Patients with low RBC counts had a 0.5-fold higher risk of cardiovascular death than those with normal RBC counts. The hazard ratio for men was 1.42 (95% confidence interval [CI]: 1.07-1.89), and the hazard ratio for women was 1.79 (95% CI: 1.20-2.67). The RBC count was not significantly associated with the composite endpoint of cardiovascular death and HF readmission (cardiovascular events) (p > .05). A decreased RBC count is associated with increased cardiovascular mortality in patients with HFmrEF. Correcting a low RBC count might potentially reduce the risk of cardiovascular death in patients with HFmrEF.


Kurt Semm and an automatic insufflator.
Work on tubal insufflation marked the beginning of Kurt Semm's (b. 1927) scientific career. In the early 1960s, he directed his attention to the fact that, from a technical standpoint, tubal insufflation was similar to creating pneumoperitoneum. In the mid-1960s, Semm--himself a gynecologist--investiged his time and financial resources and risked his university career to develop an automatic abdominal insufflation device. Later he tried it out in the Clinic for Internal Medicine. Since, at that time, the term "laparoscopy" had negative connotations associated with it, Semm formulated a new term "pelviscopy." In 1967, Semm presented his invention to Melvin Cohen, an American pioneer of gynecological laparoscopy, at the meeting of the American Fertility Society, held in Washington.


Gratitude, depression and PTSD: Assessment of structural relationships.
Gratitude, the tendency to appreciate positive occurrences in one's life that can be partially attributed to another person, has been shown to be a robust predictor of greater well-being. Researchers have also found gratitude to be inversely related to several emotional disorders, including major depressive disorder (MDD) and posttraumatic stress disorder (PTSD). Both of these emotional disorders are highly comorbid and share dysphoric symptoms (e.g., restricted affect, detachment, anhedonia) that could account for deficits in the experience and expression of gratitude. We used confirmatory factor analysis to test the relationships between gratitude and the symptom factors of PTSD (using the DSM-5 model) and MDD in a sample of trauma-exposed college students (N=202). Results indicated that gratitude is more strongly related to PTSD's negative alterations in mood and cognition (NAMC) factor than to other PTSD factors. Implications of these findings for the study of gratitude and trauma are discussed, including whether gratitude and gratitude-based interventions might prove particularly suited to targeting depressive symptoms.


Regulation of connexin degradation as a mechanism to increase gap junction assembly and function.
Connexins, the integral membrane protein constituents of gap junctions, are degraded at a rate (t(12) = 1.5-5 h) much faster than most other cell surface proteins. Although the turnover of connexins has been shown to be sensitive to inhibitors of either the lysosome or of the proteasome, how connexins are targeted for degradation and whether this process can be regulated to affect intercellular communication is unknown. We show here that reducing connexin degradation with inhibitors of the proteasome (but not with lysosomal blockers) is associated with a striking increase in gap junction assembly and intercellular dye transfer in cells inefficient in both processes under basal conditions. The effect of proteasome inhibitors on wild-type connexin stability, assembly, and function was mimicked by treatment of assembly-inefficient cells with inhibitors of protein synthesis such as cycloheximide. Sensitivity of connexin degradation to cycloheximide, but not to proteasome inhibitors, was abolished when connexins were rendered structurally abnormal by perturbation of essential disulfide bonds or by mutation. Our findings provide the first evidence that intercellular communication can be up-regulated at the level of connexin turnover and that a short-lived protein may be required for conformationally mature connexins to become substrates of proteasomal degradation.


Bioaccumulation and detoxification of trivalent arsenic by Achromobacter xylosoxidans BHW-15 and electrochemical detection of its transformation efficiency.
Arsenotrophic bacteria play an essential role in lowering arsenic contamination by converting toxic arsenite [As (III)] to less toxic and less bio-accumulative arsenate [As (V)]. The current study focused on the qualitative and electrocatalytic detection of the arsenite oxidation potential of an arsenite-oxidizing bacteria A. xylosoxidans BHW-15 (retrieved from As-contaminated tube well water), which could significantly contribute to arsenic detoxification, accumulation, and immobilization while also providing a scientific foundation for future electrochemical sensor development. The minimum inhibitory concentration (MIC) value for the bacteria was 15 mM As (III). Scanning Electron Microscopy (SEM) investigation validated its intracellular As uptake capacity and demonstrated a substantial association with the MIC value. During the stationary phase, the strain's As (III) transformation efficiency was 0.0224 mM/h. Molecular analysis by real-time qPCR showed arsenite oxidase (aioA) gene expression increased 1.6-fold in the presence of As (III) compared to the untreated cells. The immobilized whole-cell also showed As (III) conversion up to 18 days. To analyze the electrochemical oxidation in water, we developed a modified GCE/P-Arg/ErGO-AuNPs electrode, which successfully sensed and quantified conversion of As (III) into As (V) by accepting electrons; implying a functional As oxidase enzyme activity in the cells. To the best of our knowledge, this is the first report on the electrochemical observation of the As-transformation mechanism with Achromobacter sp. Furthermore, the current work highlighted that our isolate might be employed as a promising candidate for arsenic bioremediation, and information acquired from this study may be helpful to open a new window for the development of a cost-effective, eco-friendly biosensor for arsenic species detection in the future.


Double-outlet left atrium with intact ventricular septum.
The clinical course and surgical repair of double-outlet left atrium with intact ventricular septum in a 13-year-old girl are presented. The only outlet of the right atrium was a secundum atrial septal defect, and the left atrium drained into both ventricles through two atrioventricular valves. To our knowledge, there has been only one other published report of repair of double-outlet left atrium.


Serial cytogenetic studies of human colonic tumour xenografts.
Chromosome studies have been made of 2 human colonic tumour lines maintained as xenografts in immune-deprived mice. In both tumours human karyotypes were retained, although progressive changes occurred during serial passage. In one tumour, independent gain of a chromosome 19 was found in the stemline and 2 sidelines. In the other tumour there was selection for a sideline containing a particular deleted marker chromosome. The advantages of chromosome analysis in a xenograft system, both for the study of human solid tumour karyotypes and for monitoring the continued presence of the human genome, are discussed.


Bicyclo[4.4.0]decane oxygenated sesquiterpenes from Eryngium maritimum essential oil.
Investigation of the essential oil of the aerial parts of Eryngium maritimum L. from Corsica led to the isolation of one known sesquiterpene (1) and three new oxygenated sesquiterpenes with a muurolane or cadinane skeleton (2-4). Structure assignments of 4 βH -muurol-9-en-15-al (1), 4 βH -cadin-9-en-15-al (2), 4 βH -muurol-9-en-15-ol (3) and 4 βH -cadin-9-en-15-ol (4) were carried out by GC/MS (EI and CI) and comprehensive 1D and 2D NMR spectroscopy. Antibacterial activity of Eryngium maritimum L. oil and column chromatography fractions have been investigated for the first time. The oxygenated fraction, which contains the four sesquiterpenes, was efficient against Listeria monocytogenes and Echerichia coli.


Flexibility of the mitral annulus with the Duran ring at six years post-implantation.
Although, in theory, the mitral annulus with the Duran flexible ring should change size and shape during the cardiac cycle, no clinical reports are available of such flexibility several years after surgery. The study aim was to evaluate mitral annulus flexibility at medium-term follow up after surgery. Eleven patients (mean age 60+/-8 years) were enrolled for this study. All were in sinus rhythm and had less than mild residual mitral regurgitation. The mean time between study and mitral valve repair was 6.0+/-2.4 years. Three sizes of Duran ring were used (three size 27, six size 29, and two size 31). All patients were studied with transesophageal echocardiography. The mitral annulus was reconstructed 10 times around a single axis of rotation during the cardiac cycle, using published methods. The area, circumferential length and shape of the mitral annulus were analyzed at various times during the cardiac cycle. A significant change in mitral annular area occurred during the cardiac cycle; the maximum to minimum reduction was 23+/-8% (p <0.001). The area and circumferential length of the mitral annulus in patients with size 29 Duran rings changed from 8.2+/-1.3 to 6.2+/-1.3 cm2 (p <0.001) and from 11.8+/-1.1 to 9.9+/-0.8 cm (p <0.001), respectively. Three-dimensional reconstruction of the mitral annulus showed it to retain its non-planar shape, but the shape to change significantly during the cardiac cycle. The mitral annulus with a Duran ring remained flexible and retained its non-planar configuration at six years after implantation.


Multiple Systems for the Motivational Control of Behavior and Associated Neural Substrates in Humans.
In this chapter, we will review evidence about the role of multiple distinct systems in driving the motivation to perform actions in humans. Specifically, we will consider the contribution of goal-directed action selection mechanisms, habitual action selection mechanisms and the influence of Pavlovian predictors on instrumental action selection. We will further evaluate evidence for the contribution of multiple brain areas including ventral frontal and dorsal cortical areas and several distinct parts of the striatum in these processes. Furthermore, we will consider circumstances in which adverse interactions between these systems can result in the decoupling of motivation from incentive valuation and performance.


Prevalence and Antifungal Susceptibility of Sporothrix species in Guangzhou, Southern China.
Sporotrichosis is a subcutaneous and chronic infection caused by traumatic inoculation of pathogenic sporothrix species, usually infecting the skins and subcutaneous tissues of humans and animals. However, the lack of epidemiological data required further molecular identification to describe the distribution of this fungus in our region. In this study, forty-eight clinical sporothrix isolated from Sun Yat-Sen Memorial Hospital were classified, and the susceptibility of each strain to seven antifungal agents was determined. Forty strains of S. globosa and eight strains of S. shenkshii were identified via colony morphology and PCR sequencing of calmodulin gene. Antifungal susceptibility tests of the mycelial phase in vitro showed terbinafine (TRB) and luliconazole (LULI) were the most effective, followed by itraconazole (ITZ) and amphotericin B (AMB). By contrast, voriconazole (VCZ), 5-flucytosine (5FC) and fluconazole (FCZ) have low efficacy with high MIC. Our results showed a predominantly S. globosa infection trend in southern China. Simultaneously, sporothrix is sensitive to TRB, LULI, ITZ and AMB whereas resistant to FCZ. This study firstly reports antifungal sensitivity test in vitro and epidemiological correlation analysis of sporothrix in southern China, and also the first time to find that sporothrix is sensitive to LULI.


Feyerabend on politics, education, and scientific culture.
The purpose of this paper is to offer a sympathetic reconstruction of the political thought of Paul Feyerabend. Using a critical discussion of the idea of the 'free society' it is suggested that his political thought is best understood in terms of three thematic concerns-liberation, hegemony, and the authority of science-and that the political significance of those claims become clear when they are considered in the context of his educational views. It emerges that Feyerabend is best understood as calling for the grounding of cognitive and cultural authorities-like the sciences-in informed deliberation, rather than the uncritical embrace of prevailing convictions. It therefore emerges that a free society is best understood as one of epistemically responsible citizenship rather than epistemically anarchistic relativism of the 'anything goes' sort-a striking anticipation of current debates about philosophy of science in society.


Oak Acorn Poisoning in Cattle during Autumn 2022: A Case Series and Review of the Current Knowledge.
Oak poisoning is a known intoxication in grazing animals, but is slightly described in the literature. This case report describes 7 cattle from 3 different farms admitted to the clinic for ruminants of the University of Liège for suspected acorn poisoning in the autumn of 2022. The clinical signs were, anorexia, apathy with polyuria with low density. Further investigations led to the diagnosis of renal failure (blood urea 162 ± 88 mg/dL; blood creatinine 12 ± 4 mg/L). Supportive treatment, based on infusions (NaCl 0.9%) and electrolyte rebalancing, was administered and renal values were assessed every 24-48 h. Of these animals, 5/7 were euthanized. At necropsy, digestive erosions and ulcerations, oedema and renal hemorrhages, between the pyloric/caliceal cavity and the medulla were observed. Histopathological examination revealed necrosis of the renal tubules. The renal values of the two remaining animals were reduced, their general condition improved, and they were discharged. Acorn poisoning is a serious disease with no specific antidote or characteristic symptoms. Animals are identified as sick too late, when renal failure is already established. Farmers should be made more aware in order to prevent exposure, especially in years when acorns are abundant. Furthermore, there is no antidote for this intoxication.


Debittering effect of Monascus carboxypeptidase during the hydrolysis of soybean protein.
The actions of pepsin and the admixture of pepsin and Monascus pilosus carboxypeptidase 1 (MpiCP-1) on the hydrolysis of soybean protein were studied. The results showed that the pepsin hydrolyzate of soybean protein was much more bitter and contained relatively smaller amounts of total free amino acids than the hydrolyzate obtained with the admixture of pepsin and MpiCP-1. In addition, hydrophilic and hydrophobic amino acids were present in almost equal proportions in the pepsin hydrolyzate, while mainly hydrophobic amino acids made up the hydrolyzate obtained with the admixture of pepsin and MpiCP-1. These results suggest that MpiCP-1 suppresses and reverses the development of the bitterness taste that results from the pepsin hydrolysis of soybean protein by releasing mainly hydrophobic amino acids from the C-termini of the bitter components.


Tongue-muscle-controlling motoneurons in the Japanese toad: neural inputs from the thalamus.
The anuran tongue is an effector organ specialized for snapping up prey during visually guided prey-catching behavior. As a step toward elucidating the control mechanisms of the tongue movement and overall organization of visually guided behavior, properties of neural inputs from the thalamus (of which electrical stimulation elicited a behavior very similar to the visually guided predator-avoidance behavior under freely behaving conditions) were investigated in paralyzed Japanese toads. Tongue-muscle-controlling motoneurons (tongue-protractor motoneurons (PMNs) and tongue-retractor motoneurons (RMNs)) were identified antidromically, and synaptic inputs in response to electrical stimuli applied to various points in the thalamus (mainly the posterocentral thalamic nucleus) were examined. Hyperpolarizing potentials were evoked in both PMNs and RMNs in response to single electrical stimuli applied to the thalamus contralateral or ipsilateral to the recording side. Since these potentials reversed to depolarizing ones after injecting Cl- ions into the cell interior, these hyperpolarizing potentials were concluded to be the usual fast type of inhibitory postsynaptic potentials (IPSPs). On the other hand, depolarizing potentials which were superimposed on the underlying IPSPs were evoked when repetitive electrical stimuli were applied to the thalamus. The amplitude of these depolarizing potentials was decreased when depolarizing currents were injected intracellularly, while it was increased when hyperpolarizing currents were injected, indicating that these depolarizing potentials are excitatory postsynaptic potentials (EPSPs).(ABSTRACT TRUNCATED AT 250 WORDS)


Pharmacokinetics and Pharmacokinetic/Pharmacodynamic Properties of Rozibafusp Alfa, a Bispecific Inhibitor of BAFF and ICOSL: Analyses of Phase I Clinical Trials.
Rozibafusp alfa (AMG 570) is a first-in-class bispecific IgG2-peptide fusion designed to inhibit inducible T-cell costimulator ligand (ICOSL) and B-cell activating factor (BAFF). The pharmacokinetics (PK) and pharmacodynamics (PD) of rozibafusp alfa were investigated in two randomized, placebo-controlled clinical studies: a phase Ia single ascending-dose study (7-700 mg subcutaneously (s.c.)) in healthy subjects and a phase Ib multiple ascending-dose study (70-420 mg s.c. every 2 weeks (q2w)) in patients with rheumatoid arthritis. Rozibafusp alfa exhibited nonlinear PK and dose-related and reversible dual-target engagement. Maximal reduction of naïve B cells from baseline (> 40%), reflective of BAFF inhibition, was achieved with rozibafusp alfa exposure (area under the concentration-time curve from time 0 to time infinity (AUCinf ) and AUC within a dosing interval from day 0 to day 14 (AUCtau )) above 51 and 57 days•μg/mL for the single-dose (≥ 70 mg) and multiple-dose studies (≥ 70 mg q2w), respectively. ICOSL receptor occupancy on circulating B cells, a surrogate PD end point for ICOSL inhibition, was directly related to drug concentration. PK/PD analysis showed > 90% RO at rozibafusp alfa ≥ 22.2 μg/mL (≥ 420-mg single dose or ≥ 210 mg q2w multiple dose), with saturation occurring at higher drug concentrations. These results informed the design and dose selection of a phase IIb study assessing the safety and efficacy of rozibafusp alfa in patients with active systemic lupus erythematosus.


[Research Progress of HGF/c-MET Inhibitor in the Treatment of Non-small Cell Lung Cancer].
Molecular targeted therapy has become more and more important in the treatment of non-small cell lung cancer (NSCLC). HGF/c-MET plays the pivotal role in the growth, development and tolerance to epidermal growth factor receptor tyrosine kinase inhibitor of NSCLC. Moreover it has become another heat point in the molecular targeted therapy of NSCLC. c-MET amplification or high expression was deemed to another significant gene modification beyond EGFR and ALK. In the preclinical studies, HGF/c-MET inhibitors have showed the promising anti-tumor effect. Recently, some phase II/III clinical trials have proved that these inhibitors could improve the survival of patients with NSCLC. Hence we performed this review to elaborate the research progress of c-MET inhibitor in the treatment of NSCLC.


Stimulatory effect of PG-KII, an NK3 tachykinin receptor agonist, on isolated pancreatic acini: species-related differences.
More information is needed on the physiological role of the tachykinins (TKs), especially neurokinin3-receptor (NK3) agonists, in the pancreas. In this paper we investigated and compared the effect of PG-KII (10(-9) to 10(-6) M), a natural NK3-receptor agonist, with that of the known secretagogues substance P (10(-9) to 10(-6)M), caerulein (10(-11) to 10(-8) M) and carbachol (10(-8) to 10(-5) M), on amylase secretion from dispersed pancreatic acini of the guinea pig and rat. PG-KII (10(-7) M) significantly increased basal amylase release from guinea pig pancreatic acini (from 5.4+/-0.9% to 11.3+/-0.5%, P < 0.05) but left basal release in the rat unchanged (6.5+/-0.5%). The stimulant effect of PG-KII on guinea pig acini was significantly reduced by the NK3-receptor antagonist, SR 142801 (5 x 10(-7) M), and left unchanged by the NK1-receptor antagonist, SR 140333 (5 x 10(-7) M). Conversely, substance P (10(-7) M) significantly stimulated amylase secretion from rat and guinea pig acini (12.6+/-0.6% and 12.1+/-0.7%, P < 0.05). This stimulated effect of substance P was antagonized by the NK1--receptor antagonist (5 x 10(-7) M), but not by the NK3-receptor antagonist (5 x 10(-7) M). The PG-KII- and substance P-evoked maximal responses were lower than those evoked by caerulein (10(-9) M) (guinea pig, 19.1+/-1.3%; rat, 1802+/-0.9%, P < 0.01) and carbachol (10(-5) M) (guinea pig, 23.3+/-1.2%; rat, 24.0+/-1.1%, P < 0.01). The inhibitors of phospholipase C U-73122 (10(-5) M), phospholipase A2 quinacrine (10(-5)M), and protein tyrosine kinase genistein (10(-4) M), partly but significantly inhibited PG-KII, as well as carbachol-stimulated amylase release. Coincubation of PG-KII 10(-7) M with submaximal doses of caerulein (10(-11) to 10(-10) M) and carbachol (10(-7) to 10(-6) M) had an additive effect on amylase release. Pre-incubation with PG-KII (10(-7) M) for 30 min significantly reduced the subsequent amylase response to PG-KII, whereas pre-incubation with caerulein 10(-10) M or carbachol 10(-6) M did not. These findings suggest that PG-KII directly contributes to pancreatic exocrine secretion by interacting with acinar NK3 receptors of the guinea pig but not of the rat. PG-KII signal transduction involves the intracellular phospholipase C, phospholipase A2 and protein tyrosine kinase pathways. The NK3 receptor system cooperates with the other known secretagogues in regulating guinea pig exocrine pancreatic secretion and undergoes rapid homologous desensitization.


The Neurobiological Basis of Adolescent-onset Borderline Personality Disorder.
Over the past two decades, neurobiological studies in adult onset borderline personality disorder have made important strides, but inquiry into adolescent-onset BPD is still in its infancy and our understanding of the neurobiology of adolescent BPD remains highly tentative. This paper highlights recent findings in genetics, neuroendocrinology and neuroimaging for adult and adolescent-onset BPD. Neurobiological studies of adolescent-onset BPD to date have focused mainly on volumetric studies of various brain regions and measurements of HPA axis components, with comparatively few publications on brain functioning. Such information is essential to developing more effective screening, treatment and preventive strategies.


Modeling Excitable Dynamics of Chemotactic Networks.
The study of chemotaxis has benefited greatly from computational models that describe the response of cells to chemoattractant stimuli. These models must keep track of spatially and temporally varying distributions of numerous intracellular species. Moreover, recent evidence suggests that these are not deterministic interactions, but also include the effect of stochastic variations that trigger an excitable network. In this chapter we illustrate how to create simulations of excitable networks using the Virtual Cell modeling environment.


Role of transglutaminase in immune defense against bacterial pathogens via regulation of antimicrobial peptides.
Transglutaminase (TGase) is critical for blood coagulation, a conserved immunological defense mechanism among invertebrates. Here, a 3248-bp (full-length) TGase cDNA in Eriocheir sinensis (EsTGase) was cloned, with a 2274-bp open reading frame (ORF) encoding a 757 amino acid protein containing two transglut domains, one TGase/protease-like homolog domain and a KGD (Lys-Gly-Asp) motif. Phylogenetic analysis demonstrated that EsTGase appeared earlier in evolution compared with TGases of other crustaceans and mammals. EsTGase mRNA was mainly detected in hemocytes and up-regulated post-challenge with bacteria (Vibrio parahaemolyticus and Staphylococcus aureus), suggesting an immune function for this gene. Moreover, the EsTGase activity in hemocytes challenged with V. parahaemolyticus and S. aureus was decreased significantly. RNA interference of EsTGase down-regulated expression of immune-related genes CrusEs2, EsLecG and Es-DWD1 with or without bacteria stimulation in vitro. Furthermore, absence of EsTGase led to higher bacterial counts in the hemocyte culture medium. Thus, EsTGase is an important component of the crab immune response and is involved in the regulation of certain immune-related genes, particularly those encoding anti-microbial peptides.


Modeling heat loss from the udder of a dairy cow.
A mechanistic model that predicts sensible and latent heat fluxes from the udder of a dairy cow was developed. The prediction of the model was spot validated against measured data from the literature, and the result agreed within 7% of the measured value for the same ambient temperature. A dairy cow can lose a significant amount of heat (388W/m(2)) from the udder. This suggests that the udder could be considered as a heat sink. The temperature profile through the udder tissue (core to skin) approached the core temperature for an air temperature ≥37°C whereas the profile decreased linearly from the core to skin surface for an air temperature less than 37°C. Sensible heat loss was dominant when ambient air temperature was less than 37.5°C but latent heat loss was greater than sensible heat loss when air temperature was ≥37.5°C. The udder could lose a total (sensible + latent) heat flux of 338W/m(2) at an ambient temperature of 35°C and blood-flow rate of 3.2×10(-3)m(3)/(sm(3) tissue). The results of this study suggests that, in time of heat stress, a dairy cow could be cooled by cooling the udder only (e.g., using an evaporative cooling jacket).


1-Isoindolinone scaffold-based natural products with a promising diverse bioactivity.
Isoindolin-1-one or 1-isoindolinone framework is referred to phthalimidines or benzo fused γ-lactams of the corresponding γ-amino carboxylic acids and has been of prime interest for scientists for last several decades. 1-Isoindolinone framework is found in a wide range of naturally occurring compounds with diverse biological activities and therapeutic potential for various chronic diseases. Recent developments in synthetic methods for their procurement have opened a new era of 1-isoindolinone chemistry. This review aims to provide an alphabetical quick reference guide to only 1-isoindolinone based natural products and its variable fused, oxidized and reduced state skeleton with information for advanced chemotaxonomic analyses, cellular targets/pathways and diverse biological activities and future use for medicinal chemistry.


Changes in corneal sensitivity and tear physiology after phacoemulsification.
To identify any changes in corneal sensitivity and tear physiology after phacoemulsification and to evaluate the effectiveness of the use of ophthalmic lubricants. The design of the study was longitudinal, parallel, randomised and double masked. Eighteen patients (average age 70.83 +/- 10.66 years, 7 males and 11 females) undergoing phacoemulsification were recruited and divided into three groups of six. In addition to the prednisolone acetate 1% and chloramphenicol 0.5% routinely given to patients postoperatively, the first two groups were given a tear lubricant (Refresh Soothe and Protect, Allergan Inc., Irvine, CA, USA) and saline (Chauvin Pharmaceuticals Ltd, Essex, UK), respectively, and the third was a control group and did not receive any additional eye drops. Tear production, evaporation, lipid layer interferometry and osmolarity along with corneal sensitivity were measured before and 3 days, 2 weeks, 1 month and 3 months after the surgery. Statistically significant detrimental changes were seen in all parameters of corneal sensitivity and tear physiology immediately after phacoemulsification. Tear physiology recovered within 1 month. Corneal sensitivity did not return to normal levels in 3 months, but a trend towards full recovery was seen. Saline and the tear lubricant were not found to have an effect on the improvement of tear physiology and corneal sensitivity post-surgically. Deterioration in corneal sensitivity and tear physiology is seen immediately after phacoemulsification. Corneal sensitivity does not return to preoperative levels until 3 months postoperatively whereas the tear functions recover within 1 month.


Synthesis and biological activity of tuftsin and of [O = C Thr1]-tuftsin. A novel synthetic route to peptides containing N-terminal L -O = C Thr and L - O = C Ser residues.
The cyclization, under alkaline conditions, of N-benzyloxycarbonyl-L-threonine and of N-benzyloxycarbonyl-L-serine to produce 5-methyl-2-oxo-oxazolidine-4-carboxylic acid (O = C Thr1 and 2-oxo-oxazolidine-4-carboxylic acid (O = C Ser), respectively, was investigated and found to be efficient and racemization-free. Similar was the cyclization which accompanied the basic hydrolysis of N-benzyl-oxycarbonyl-L-threonyl-L-phenylalanine methyl ester and of N-benzyloxy-carbonyl-L-seryl-DL-valine ethyl ester, and which resulted in the formation of L - O = C Thr-L-Phe and L - O = C Ser-DL-Val, respectively. The reaction was applied to the synthesis of [O = C Thr1] tuftsin, an active analog of the phagocytosis stimulating peptide tuftsin. A new synthetic route to tuftsin is also described.


Protein kinase networks regulating glucocorticoid-induced apoptosis of hematopoietic cancer cells: fundamental aspects and practical considerations.
Glucocorticoids (GCs) are integral components in the treatment protocols of acute lymphoblastic leukemia, multiple myeloma, and non-Hodgkin lymphoma owing to their ability to induce apoptosis of these malignant cells. Resistance to GC therapy is associated with poor prognosis. Although they have been used in clinics for decades, the signal transduction pathways involved in GC-induced apoptosis have only partly been resolved. Accumulating evidence shows that this cell death process is mediated by a communication between nuclear GR affecting gene transcription of pro-apoptotic genes such as Bim, mitochondrial GR affecting the physiology of the mitochondria, and the protein kinase glycogen synthase kinase-3 (GSK3), which interacts with Bim following exposure to GCs. Prevention of Bim up-regulation, mitochondrial GR translocation, and/or GSK3 activation are common causes leading to GC therapy failure. Various protein kinases positively regulating the pro-survival Src-PI3K-Akt-mTOR and Raf-Ras-MEK-ERK signal cascades have been shown to be activated in malignant leukemic cells and antagonize GC-induced apoptosis by inhibiting GSK3 activation and Bim expression. Targeting these protein kinases has proven effective in sensitizing GR-positive malignant lymphoid cells to GC-induced apoptosis. Thus, intervening with the pro-survival kinase network in GC-resistant cells should be a good means of improving GC therapy of hematopoietic malignancies.


Coordinate, biphasic activation of p44 mitogen-activated protein kinase and S6 kinase by growth factors in hamster fibroblasts. Evidence for thrombin-induced signals different from phosphoinositide turnover and adenylylcyclase inhibition.
In CCL39 cells transfected with m1-muscarinic receptors, carbachol stimulates phosphoinositide turnover and early events associated with mitogenesis as efficiently as thrombin but, in contrast to thrombin, fails to induce cell proliferation (Seuwen, K., Kahan, C., Hartmann, T., and Pouysségur, J. (1990) J. Biol. Chem. 265, 22292-22299). We show here that the action of the two agents can be dissociated at the level of S6 kinase and mitogen-activated protein kinase (MAP kinase) activation. Mitogenic concentrations of thrombin and basic FGF were found to stimulate S6 kinase activity, measured in whole cell lysates, with a biphasic time course; an early peak of activity is induced 10 min following stimulation and a sustained phase of activity can be measured over several hours. A very similar profile emerged for p44 MAP kinase (p44mapk), assayed in immunoprecipitates. In this case, the activity first peaks at 6-8 min, preceding S6 kinase. In contrast to thrombin and FGF, carbachol stimulates S6 kinase and MAP kinase only transiently, corresponding to the first peak of activity, but the sustained phase is not observed. Similarly, phorbol dibutyrate induces an early phase of activity only. Pertussis toxin (PTX), which is known to block thrombin mitogenicity efficiently, inhibited the first peak of thrombin-induced S6 kinase and MAP kinase activity only partially, but totally blocked the sustained phase. The toxin had no effect on FGF-induced kinase activities. The cAMP elevating hormone PGE1 did not inhibit p44mapk or S6 kinase activation by thrombin or FGF, demonstrating that the PTX-sensitive signal generated by thrombin does not depend on a Gi-mediated sustained inhibition of adenylylcyclase. Surprisingly, PGE1 was found to stimulate sustained phase S6 kinase activity both alone and in synergy with FGF or thrombin. This result, as well as the biphasic activation of S6 kinase by thrombin, could be qualitatively reproduced in immunocomplex kinase assays using an antiserum immunoprecipitating p70 S6 kinase (p70S6k). Our data show that activation of phosphoinositide turnover and PKC does not quantitatively explain thrombin action, in particular the sustained phase of kinase activities, which critically depends on a PTX-sensitive signal different from adenylylcyclase inhibition. We postulate that this signal does not exclusively originate from the recently identified G protein-coupled thrombin receptor.


Audit feedback interventions to address high-risk prescriptions in long-term care homes: a costing study and return on investment analysis.
Audit and feedback is a common implementation strategy, but few studies describe its costs. 'MyPractice' is a province-wide audit and feedback initiative to improve prescribing in nursing homes. This study sought to estimate the costs of 'MyPractice' and assess whether the financial benefit of 'MyPractice' offsets those costs. We conducted a costing study from the perspective of the Ontario government. Total cost of 'MyPractice' was calculated as the sum of the costs of producing and disseminating the reports (covering three report releases) which were obtained from Ontario Health staff interviews and document reviews. Return on investment (ROI) was calculated as the ratio of net cost-savings and the intervention cost. Cost savings were based on the effectiveness of 'MyPractice' derived from a published cohort study. Cost-savings attributable to 'MyPractice' were estimated from the changes in the rates of antipsychotics over time between physicians who signed up and viewed the reports and those who did not sign up to the reports. Total intervention costs were C$223,691 (C$838 per physician and C$74,564 per release). Costs incurred during the development phase accounted for 74% of the total cost (C$166,117), while implementation costs for three report releases were responsible for 26% of the total costs (C$57,575). The ROI for every C$1 spent on the 'MyPractice' intervention was 1.02 (95% CI 0.51, 1.93) for three report releases. 'MyPractice' report offers a good return on investment and the value for money could improve with greater number of report releases.


Modeling of hydrogen-assisted cracking in iron crystal using a quasi-Newton method.
A Quasi-Newton method was applied in the context of a molecular statics approach to simulate the phenomenon of hydrogen embrittlement of an iron lattice. The atomic system is treated as a truss-type structure. The interatomic forces between the hydrogen-iron and the iron-iron atoms are defined by Morse and modified Morse potential functions, respectively. Two-dimensional hexagonal and 3D bcc crystal structures were subjected to tensile numerical tests. It was shown that the Inverse Broyden's Algorithm-a quasi-Newton method-provides a computationally efficient technique for modeling of the hydrogen-assisted cracking in iron crystal. Simulation results demonstrate that atoms of hydrogen placed near the crack tip produce a strong deformation and crack propagation effect in iron lattice, leading to a decrease in the residual strength of numerically tested samples.


Sclerosis of the intestinal tract with severe malabsorption.
The clinical findings and pathological changes in a 14 year old boy who died from severe malabsorption are presented. Early signs of the illness were attacks of abdominal pain, diarrhoea, and intermittent periods of constipation. Later, severe malabsorption together with chronic ileus were apparent but no organic obstruction was found at several laparotomies. X-ray examination revealed extremly distended intestinal loops and a very slow transit time of barium. Malnutrition could not be improved by dietary, medical, or surgical therapy. Microscopic examination of the tissues obtained at autopsy showed changes in the small vessels of the lungs, heart, kidneys, and intestine. Increased fibrous tissue combined with atrophy of the muscular coat was observed in the wall of the entire intestine. Although cutaneous changes of scleroderma were not evident the deposition of collagenous material and the vascular changes seem typical of systemic sclerosis.


Investigation of seismicity after the initiation of a Seismic Electric Signal activity until the main shock.
The behavior of seismicity in the area candidate to suffer a main shock is investigated after the observation of the Seismic Electric Signal activity until the impending main shock. This is based on the view that the occurrence of earthquakes is a critical phenomenon to which statistical dynamics may be applied. In the present work, analysing the time series of small earthquakes, the concept of natural time chi was used and the results revealed that the approach to criticality itself can be manifested by the probability density function (PDF) of kappa(1) calculated over an appropriate statistical ensemble. Here, kappa(1) is the variance kappa(1)(=<chi(2)>-<chi>(2)) resulting from the power spectrum of a function defined as Phi(omega)= summation operator(k=1)(N) p(k) exp(iomegachi(k)), where p(k) is the normalized energy of the k-th small earthquake and omega the natural frequency. This PDF exhibits a maximum at kappa(1) asymptotically equal to 0.070 a few days before the main shock. Examples are presented, referring to the magnitude 6 approximately 7 class earthquakes that occurred in Greece.


Study of stiffness effects of poly(amidoamine)-poly(n-isopropyl acrylamide) hydrogel on wound healing.
The mechanical miss-match between the host and an implanted foreign body is one of the primary causes for implantation failure. To enhance the efficacy in wound repair, we developed stiffness-tunable temperature-sensitive hydrogels composed of poly(amidoamine) (PAA)-based poly(n-isopropyl acrylamide) (PNIPAM). PNIPAM-PAA hydrogels with three different stiffness fabricated by varying the concentrations of poly(amidoamine) were chosen for morphology and rheology tests. The degradation rate and cell compatibility of gels were also characterized. The PAA-PNIPAM hydrogels were then tested in a wound healing model of mice with full-thickness skin loss. We found that the stiffness of hydrogels has an impact on the wound healing process mainly by regulating the cell activities in the proliferation phase. PNIPAM-PAA hydrogels with appropriate stiffness reduce scar formation and improve wound healing by promoting myofibroblast transformation, keratinocytes proliferation, extracellular matrix synthesis and remodeling. Moreover, the stiffness of hydrogels impact on the secretion of TGF-β1 and bFGF, which play an important role in skin wound healing. These results suggest that the therapeutic effects of hydrogels in skin wound healing can by regulated by hydrogels' stiffness.


Effects of nitrogen supply and elevated carbon dioxide on construction cost in leaves of Pinus taeda (L.) seedlings.
Seedlings of loblolly pine (Pinus taeda L.) were grown under varying conditions of soil nitrogen and atmospheric carbon dioxide availability to investigate the interactive effects of these resources on the energetic requirements for leaf growth. Increasing the ambient CO2 partial pressure from 35 to 65 Pa increased seedling growth only when soil nitrogen was high. Biomass increased by 55% and photosynthesis increased by 13% after 100 days of CO2 enrichment. Leaves from seedlings grown in high soil nitrogen were 7.0% more expensive on a g glucose g-1 dry mass basis to produce than those grown in low nitrogen, while elevated CO2 decreased leaf cost by 3.5%. Nitrogen and CO2 availability had an interactive effect on leaf construction cost expressed on an area basis, reflecting source-sink interactions. When both resources were abundant, leaf construction cost on an area basis was relatively high (81.8±3.0 g glucose m-2) compared to leaves from high nitrogen, low CO2 seedlings (56.3±3.0 g glucose m-2) and low nitrogen, low CO2 seedlings (67.1±2.7 g glucose m-2). Leaf construction cost appears to respond to alterations in the utilization of photoassimilates mediated by resource availability.


Weak-acid preservatives: pH and proton movements in the yeast Saccharomyces cerevisiae.
Weak-acid preservatives commonly used to prevent fungal spoilage of low pH foods include sorbic and acetic acids. The "classical weak-acid theory" proposes that weak acids inhibit spoilage organisms by diffusion of undissociated acids through the membrane, dissociation within the cell to protons and anions, and consequent acidification of the cytoplasm. Results from 25 strains of Saccharomyces cerevisiae confirmed inhibition by acetic acid at a molar concentration 42 times higher than sorbic acid, in contradiction of the weak-acid theory where all acids of equal pK(a) should inhibit at equimolar concentrations. Flow cytometry showed that the intracellular pH fell to pH 4.7 at the growth-inhibitory concentration of acetic acid, whereas at the inhibitory concentration of sorbic acid, the pH only fell to pH 6.3. The plasma membrane H⁺-ATPase proton pump (Pma1p) was strongly inhibited by sorbic acid at the growth-inhibitory concentration, but was stimulated by acetic acid. The H⁺-ATPase was also inhibited by lower sorbic acid concentrations, but later showed recovery and elevated activity if the sorbic acid was removed. Levels of PMA1 transcripts increased briefly following sorbic acid addition, but soon returned to normal levels. It was concluded that acetic acid inhibition of S. cerevisiae was due to intracellular acidification, in accord with the "classical weak-acid theory". Sorbic acid, however, appeared to be a membrane-active antimicrobial compound, with the plasma membrane H⁺-ATPase proton pump being a primary target of inhibition. Understanding the mechanism of action of sorbic acid will hopefully lead to improved methods of food preservation.


Value of the HFA-PEFF diagnostic algorithms for heart failure with preserved ejection fraction to the inflammatory myopathy population.
The HFA-PEFF score has been validated to hold great diagnostic and prognostic utility for heart failure with preserved ejection fraction (HFpEF). Idiopathic inflammatory myopathy (IIM) is recognized as one of the potential etiologies underlying HFpEF. Here, we intended to investigate the real prevalence of HFpEF in IIM via the HFA-PEFF score and explore the prognostic value of this score. Two hundred twenty IIM patients were enrolled for assessment. The cohort was divided into low, intermediate and high tertiles of the HFA-PEFF score. Spearman's correlation analysis was used to explore the association between the score and disease activity. Chi-square test was applied to investigate the distribution discrepancy of HFA-PEFF tertiles among patients with different myositis-specific antibodies (MSAs) or myositis-associated antibodies (MAAs). Univariate and multivariate ordinal regression analyses were performed to screen risk factors for high HFA-PEFF scores. Survival curves were obtained using the Kaplan-Meier method and log-rank tests. In total, 79 (35.9%), 107 (48.6%) and 34 (15.5%) patients were rated low, intermediate and high probability of HFpEF, respectively. The HFA-PEFF score correlated well with disease activity. Patients with positive AMA-M2 scored higher in the HFA-PEFF score (p = 0.011). During follow-up, patients with positive AMA-M2 or anti-SRP antibody developed an inclination towards concentric hypertrophy on echocardiography. Additionally, palpitation symptom, AMA-M2 positivity and elevated serum levels of LDH, cTnI were independent risk factors for high HFA-PEFF scores. Finally, a high-tertile HFA-PEFF score was related to lower overall survival rate (p < 0.001). Patients with positive AMA-M2 had poorer outcomes (p = 0.002). HFpEF was prevailing in IIM patients according to the HFA-PEFF score. The HFA-PEFF score correlated well with disease activity and held significant prognostic value. Patients with AMA-M2 antibody were prone to have poor outcomes.


Lateral force-displacement behaviour of the human patella and its variation with knee flexion--a biomechanical study in vitro.
This study measured the patellar lateral force-displacement behaviour at a range of knee flexion angles in normal human cadaver specimens. The knee extensor muscles were loaded in proportion to their physiological cross-sectional areas, the tensions being applied in physiological directions along the separate quadriceps muscles. Knee extension was blocked at a range of knee flexion angles from 0 to 90 degrees, and patellar lateral displacement versus force characteristics were measured. This experiment was repeated with three total muscle forces, 20, 175 and 350 N, which were held constant at all flexion angles. It was shown that similar stability variation was obtained with the different total muscle loads, and also the forces required to produce a range of patellar displacements (1.5, 9 mm) were examined. A 5 mm lateral patellar displacement required a constant displacing force (i.e. the patella had constant lateral stability) up to 60 degrees knee flexion, and then a significant increase at 90 degrees. The results were related to surgical and anatomical observations.


Association of haplotypes of interleukin-10 gene with the risk of cancer.
Polymorphisms of promotor region of IL-8, IL-10, and IL-12 genes were analyzed in cancer patients and subjects without history of cancer. The distribution of alleles of the analyzed polymorphisms in the control group coincided with that in other Caucasian populations. The incidences of three IL-10 gene polymorphisms (G-1082A, C-819T, and C-592A) significantly differed in controls and patients. Of 8 theoretically probable IL-10 gene haplotypes determined by these polymorphisms, 3 variants were revealed. Haplotype ACC was more incident in cancer patients, while ATA haplotype was rarer. The results are in line with the findings of other studies indicating the involvement of the immune system genes in the pathogenesis of cancer.


Research Note: Effects of Bacillus coagulans X26 on the production performance, intestinal structure, short-chain fatty acids and flora composition of laying hens during the peak laying period.
This study investigated the effects of Bacillus coagulans X26 on the production performance, egg quality, intestinal structure, intestinal short-chain fatty acids, and microbial diversity of laying hens during the peak laying period and explored the possibility of using B. coagulans X26 as an alternative to antibiotics. The results showed that adding 1.0 × 106 CFU/g B. coagulans X26 to the basal diet had the best effect: the average laying rate increased by 4.20% (P < 0.05), the survival rate increased by 7.15% (P < 0.05), the feed intake decreased by 3.06% (P < 0.05), the ratio of feed to egg decreased by 7.42% (P < 0.05), the rate of soft-broken eggshell decreased by 73.04% (P < 0.05), the average egg weight increased by 2.94% (P < 0.05), and the content of egg white protein increased by 5.77% (P < 0.05). The production performance and egg quality of laying hens reached the same level as that of hens fed a diet supplemented with chlortetracycline hydrochloride in this study, and there were significant advantages in the average laying rate and feed-egg ratio (P < 0.05). Both chlortetracycline hydrochloride and B. coagulans X26 altered the flora composition and the SCFA content of the intestinal contents; however, B. coagulans X26 also significantly increased the villus height of the ileum and the ratio of villus height to crypt depth (P < 0.05).


Suicidal ideation and related factors in construction industry apprentices.
Suicide rates within the construction industry are disproportionately high and there is accumulating evidence suggesting that young apprentices working in this industry may be particularly vulnerable. This study examined the presence of suicidal ideation and exposure to suicidal behaviours in construction industry apprentices, and explored associations between suicidal ideation and other demographic, workplace, and psychosocial factors. A large sample of apprentices were recruited for the study (N = 1402). The study employed a cross-sectional survey design. In addition to demographic items, the survey asked questions relating to suicidal ideation in the past year, exposure to suicidal behaviours, substance use, stress management, workplace bullying, psychological distress, and well-being. Nearly one third of apprentices reported suicidal ideation in the previous year, and approximately half to two-thirds knew someone who had either attempted or died by suicide. Suicidal ideation was associated with knowing someone who had attempted suicide, greater psychological distress, substance use, and poorer well-being. The response rate was low. There were also portions of missing data. Multiple imputation was used to help overcome this, and results from the sensitivity analyses are presented. The cross-sectional design is another limitation. Suicidal ideation in construction apprentices is common and is associated with poorer overall mental health and well-being, substance use, and knowing others who have attempted suicide. These findings can help inform the development of tailored workplace suicide prevention activities for apprentices at-risk of suicide and poor mental health.


Breakthrough invasive fungal diseases during echinocandin treatment in high-risk hospitalized hematologic patients.
The frequency of breakthrough invasive fungal diseases (IFDs) during echinocandin therapy is unclear. We retrospectively analyzed 534 hematologic patients treated with echinocandin (caspofungin, N = 55; micafungin, N = 306; anidulafungin, N = 173). Four proven IFDs were found, caused by Candida parapsilosis (N = 2), C. parapsilosis and Candida glabrata (N = 1), and Fusarium species (N = 1). Four cases of possible IFDs were observed, all showing pulmonary infection. One case showed features suggestive of hepatosplenic candidiasis. Six of these eight cases had previously received the purine analog clofarabine. Breakthrough IFD during echinocandin treatment occurred infrequently (1.5 %), caused predominantly by Candida species. Clofarabine usage was an important risk factor.


Propyl gallate induces cell death and inhibits invasion of human trophoblasts by blocking the AKT and mitogen-activated protein kinase pathways.
Propyl gallate (PG) is an antioxidant widely used in food additives, cosmetics, adhesives, and lubricants. PG protects the oils in food products from reacting with hydrogen peroxide and oxygen free radicals, thus preventing spoilage. It is known to have both protective and cytotoxic effects on various cells, but its effects on human trophoblasts remain unclear. Therefore, we investigated the effects of PG on proliferation, apoptosis, and invasiveness of human trophoblasts using an immortalized HTR8/SVneo cell line. We found that PG significantly reduced proliferation of and induced apoptosis in HTR8/SVneo cells. In addition, the invasiveness of HTR8/SVneo cells was attenuated in response to PG. Signaling pathways including the PI3K/AKT and MAPK pathways involved in the proliferation and invasiveness of human trophoblasts were regulated by PG treatment in HTR8/SVneo cells. We confirmed that mitochondrial membrane disruption and Ca2+ overload were markedly elevated in response to PG treatment, suggesting that PG-induced apoptosis is closely related to mitochondrial dysfunction and further PG-induced apoptosis in HTR8/SVneo cells is related to endoplasmic reticulum (ER) stress. Collectively, these results indicate that PG exerts harmful effects on human trophoblasts; therefore, exposure to PG in early pregnancy is predicted to cause abnormal implantation and placental development.


Potential clinical value of catheters impregnated with antimicrobials for the prevention of infections associated with peritoneal dialysis.
Peritoneal dialysis (PD) is a widely used dialysis modality, which offers the advantage of being a home therapy but is associated with a risk of potentially serious infections, including exit site infection, catheter tunnel infection, and peritonitis that may result in morbidity, technique failure, and increased mortality. Catheters impregnated with antimicrobials hold promise as a novel technique to reduce PD associated infections. We describe PD modalities, catheters, technique, complications, and the microbiology of associated infections, as well as standard measures to reduce the risk of infection. A novel technique for the impregnation of silicone devices with antimicrobial agents has been used to produce antimicrobial impregnated ventricular shunt catheters with proven clinical efficacy that have now been adopted as the standard of care to reduce neurosurgical infections. Using the same technology, we have developed PD and urinary catheters impregnated with sparfloxacin, triclosan, and rifampicin. Safety and tolerability have been demonstrated in urinary catheters, and a similar study is planned in PD catheters. Catheters impregnated with antimicrobials offer a simple technique to reduce PD associated infections and thereby enable more people to enjoy the advantages of PD. Clinical trials are needed to establish efficacy.


Distinct and essential roles of bZIP transcription factors in the stress response and pathogenesis in Alternaria alternata.
The ability to cope with environmental abiotic stress and biotic stress is crucial for the survival of plants and microorganisms, which enable them to occupy multiple niches in the environment. Previous studies have shown that transcription factors play crucial roles in regulating various biological processes including multiple stress tolerance and response in eukaryotes. This work identified multiple critical transcription factor genes, metabolic pathways and gene ontology (GO) terms related to abiotic stress response were broadly activated by analyzing the transcriptome of phytopathogenic fungus Alternaria alternata under metal ions stresses, oxidative stress, salt stresses, and host-pathogen interaction. We investigated the biological functions and regulatory roles of the bZIP transcriptional factor (TF) genes in the phytopathogenic fungus A.alternata by analyzing targeted gene disrupted mutants. Morphological analysis provides evidence that the bZIP transcription factors (Gcn4, MeaB, Atf1, the ER stress regulator Hac1, and the all development altered-1 gene Ada1) are required for morphogenesis as the colony morphology of these gene deletion mutants was significantly different from that of the wild-type. In addition, bZIPs are involved in the resistance to multiple stresses such as oxidative stress (Ada1, Yap1, MetR) and virulence (Hac1, MetR, Yap1, Ada1) at varying degrees. Transcriptome data demonstrated that the inactivation of bZIPs (Hac1, Atf1, Ada1 and Yap1) significantly affected many genes in multiple critical metabolism pathways and gene ontology (GO) terms. Moreover,the ΔHac1 mutants displayed reduced aerial hypha and are hypersensitivity to endoplasmic reticulum disruptors such as tunicamycin and dithiothreitol. Transcriptome analysis showed that inactivation of Hac1 significantly affected the proteasome process and its downstream unfolded protein binding, indicating that Hac1 participates in the endoplasmic reticulum stress response through the conserved unfolded protein response. Taken together, our findings reveal that bZIP transcription factors function as key regulators of fungal morphogenesis, abiotic stress response and pathogenesis, and expand our understanding of how microbial pathogens utilize these genes to deal with environmental stresses and achieve successful infection in the host plant.


Running economy of elite surf iron men and male runners, on soft dry beach sand and grass.
The primary aim of this study was to measure the energetics of six elite surf iron men (who participate in regular sand running training), performing steady-state running trials on grass in shoes at 8, 11 and 14 km x h(-1), and on sand bare foot and in shoes, at both 8 and 11 km x h(-1). The net total energy cost (EC, J x kg(-1) x m(-1)) was determined from the net steady-state oxygen consumption and respiratory exchange ratio (net aerobic EC) plus net lactate accumulation (net anaerobic EC). For the sand barefoot and sand in shoes running trials at 8 and 11 km x h(-1), net aerobic EC and total net EC (but not anaerobic EC) were significantly greater (P < 0.001) than the grass running trial values. No differences (P > 0.05) existed between the sand barefoot and sand in shoes trials. These measures were compared with data obtained from eight well-trained male recreational runners who performed the same protocol in a previous study, but who were not accustomed to running on sand. Comparisons of net aerobic EC between the two groups for the surface conditions were not significantly different (P > 0.05). For net anaerobic EC, the iron man values were significantly less (P < 0.02) than the recreational runner values. For net total EC, the iron man values were less than the recreational runner values, but the differences were only significant when both groups ran on sand barefoot (P < 0.03: on grass P = 0.158; on sand in shoes P = 0.103). The lower lactate accumulation values recorded for the iron men on both grass and sand may indicate that running on sand potentially reduces metabolic fatigue when running on firm or soft surfaces.


Evaluation of Gastroesophageal Reflux Disease 1 Year after Esophageal Atresia Repair: Paradigms Lost from a Single Snapshot?
To analyze the findings of both multichannel intraluminal impedance with pH (MII-pH) and endoscopy/histopathology in children with esophageal atresia at age 1 year, according to current recommendations for the evaluation of gastroesophageal reflux disease (GERD) in esophageal atresia. We retrospectively reviewed both MII-pH and endoscopy/histopathology performed in 1-year-old children with esophageal atresia who were followed up in accordance with international recommendations. Demographic data and clinical characteristics were also reviewed to investigate factors associated with abnormal GERD investigations. In our study cohort of 48 children with esophageal atresia, microscopic esophagitis was found in 33 (69%) and pathological esophageal acid exposure on MII-pH was detected in 12 (25%). Among baseline variables, only the presence of long-gap esophageal atresia was associated with abnormal MII-pH. Distal baseline impedance was significantly lower in patients with microscopic esophagitis, and it showed a very good diagnostic performance in predicting histological changes. Histological esophagitis is highly prevalent at 1 year after esophageal atresia repair, but our results do not support a definitive causative role of acid-induced GERD. Instead, they support the hypothesis that chronic stasis in the dysmotile esophagus might lead to histological changes. MII-pH may be a helpful tool in selecting patients who need closer endoscopic surveillance and/or benefit from acid suppression.


JAK1/STAT3 activation directly inhibits IL-12 production in dendritic cells by preventing CDK9/P-TEFb recruitment to the p35 promoter.
Inhibition of Janus-activated kinase-1 (JAK1) is a promising clinical concept for post-transplant immunosuppression and autoimmunity. However, it also raises concerns regarding possible immunosuppressive side effects. Our study investigates JAK1 signalling in the context of CD40L and bacterially activated human MoDC using siRNA and biological inhibitors. We demonstrate that strong stimuli (e.g. intact Escherichia coli or LPS in addition to IL-1β) induce IL-12p70 via a ROS/RELA/CDK9 pathway that is inhibited by simultaneous JAK1/STAT3 signalling. Transcription is effective if RELA recruits the positive transcription elongation factor b (P-TEFb) component CDK9 to a combined RELA/STAT3 binding site -50 to -20bp upstream of the start site of the IL-12p35 promoter. STAT3 simultaneously attaches to this site and inhibits CDK9 binding. In the presence of IFNγ, JAK1/2 inhibitors block STAT1/IRF1/IRF8-dependent activation and simultaneously enhance CDK9-dependent activation signals. This inverse regulation of IFNγ- vs. E. coli-induced cytokine production by JAK inhibitors including Ruxolitinib was similarly observed for IL-6 and TNF-α production, but not for IL-10 production. Thus, JAK1 inhibition enhances IL-12p70 production in this context by increased DNA binding of CDK9. In contrast, weak RELA-activation signals (CD40L, LPS) depended on IFN-γ induced STAT1/IRF1/IRF8 co-signalling, which was completely blocked by JAK inhibitors as reported before. Our results suggest a novel molecular mechanism of how cytokine responses to invading pathogens are separable from IFNγ-dependent autoimmunity by targeting JAK1/STAT3 activation.


Activity standardisation of ²²⁶Ra by 4πα liquid scintillation counting method.
Activity of (226)Ra in radium daughter products free solution was determined by 4πα liquid scintillation counting (LSC) method, where the detection efficiency of radium decay is practically equal to 1. The sources were prepared from solution with known (226)Ra mass concentration, from which, immediately before LS counting, (222)Rn and its daughter nuclides were removed by solvent extraction. LS counting results were corrected practically only for a <0.6% loss of radium from measured sample and for the ingrowth of (222)Rn and (218)Po concentrations in the sample after the separation was completed. The combined relative standard uncertainty was estimated to be lower than 0.34%.


Alterations in striatal neuropeptide Y system activity of rats with haloperidol-induced behavioral supersensitivity.
The study was conducted to determine whether the expression of behavioral supersensitivity induced by haloperidol (HAL) administered once daily (2 mg/kg i.p.) for 14 days is associated with the alterations in activity of neuropeptide Y (NPY) system in the striatum (caudate-putamen) and nucleus accumbens. Dopamine supersensitivity was tested by measurement of locomotor activity and stereotyped behavior after administration of the dopamine D2/D3 receptor agonist quinpirole (1 mg/kg i.p.) on day 1, 3 and 7 after HAL withdrawal. Neuropeptide Y-like immunoreactivity (NPY-LI) was determined in the striatum and nucleus accumbens isolated 6 h after quinpirole injection on day 1, 3 and 7 after the end of HAL treatment. NPY mRNA was quantified in these structures on day 7 after HAL withdrawal. HAL increased spontaneous locomotor activity and prevalence of rearing, grooming and head-down sniffing. At the same time, striatal NPY-LI increased progressively from the reduced level found on day 1 of haloperidol withdrawal. NPY mRNA remained unchanged. In saline-treated rats, quinpirole enhanced locomotion, rearing, and induced intense head-down sniffing and oral activity. These behavioral effects were accompanied by a decrease in striatal NPY-LI. NPY mRNA was slightly increased. HAL treatment altered response to quinpirole, namely it increased locomotion, intensified oral activity and reduced rearing and head-down sniffing. The second and the third quinpirole injection decreased NPY-LI levels. NPY mRNA was unchanged. In the nucleus accumbens, apart from a decrease in NPY-LI on day 1 after the last haloperidol dose, the level of NPY-LI and NPY mRNA in any experimental group did not differ from the control value. The presented results suggest that the alterations in the activity of the striatal but not nucleus accumbens NPY system contribute to adaptive changes induced by long-term haloperidol treatment and may be of significance to the motor hyperactivity induced by intermittent stimulation of postsynaptic dopamine D2 receptors.


Neoadjuvant Chemotherapy and Expandable Prosthesis Reconstruction to Treat Osteosarcoma around the Knee in Children.
Survival and reconstruction in osteosarcoma is quite challenging. The study aimed to investigate the prognosis in patients treated with neoadjuvant chemotherapy and determine the clinical outcomes of expandable endoprosthesis reconstruction in children. From January 2009 to December 2014, we retrospectively analyzed 29 skeletally immature children (mean age, 10.5 years; range, 6-15 years) with osteosarcoma around the knee. Of the 29 patients who underwent neoadjuvant chemotherapy and limb salvage surgery, an expandable prosthesis was implanted for reconstruction. No patients were missed during follow-up. The evaluation index involved follow-up time, complication, functional results, and lengthening procedures. The survivorship and recurrence were assessed by GraphPad Software, and the function was evaluated by the Musculoskeletal Tumor Society (MSTS) scoring system. A mean follow-up time was 8.9 years (range, 6-12 years), and the overall 5-year survival was 89.1% based on Kaplan-Meier analysis. Three patients suffered a relapse and one underwent amputation. Lung metastasis developed in one patient. At 6 months after the operation, patients had a mean MSTS score of 27 points (range, 24-29). Two patients underwent revision surgery, one for implant infection and one for aseptic loosening. Prognosis is correlated with alkaline phosphatase change after treatment. Chemotherapy scheme and limb salvage can achieve high survival rates. This expandable prosthesis was associated with good function and low complication rates. The character of expandability could be a method to overcome discrepancies in the growth period.


Bone metabolism and oxidative stress in postmenopausal rats with iron overload.
Osteoporosis is associated with many etiological causes such as nutrition, cytokines, hormones, and aging. Recently, reactive oxygen species (ROS) are considered to be responsible for the aging process and osteoporosis. We investigated the relationship between ROS and bone metabolism in young female and postmenopausal rats, by using dietary iron overload and several indices including bone metabolic markers, oxidative stress and antioxidant markers, and cytokines. Postmenopausal rats exhibited significant decreases in serum alkaline phosphatase activity and the level of osteocalcin as bone formation markers compared with young female rats; however, urinary excretion of deoxypyridinoline, a bone resorption marker, did not change. On the other hand, a 5% iron lactate diet for 4 weeks in postmenopausal rats led to significantly increased excretion of urinary deoxypyridinoline and 8-hydroxy-2'-deoxyguanosine (8-OHdG) but not serum alkaline phosphatase activity. Interestingly, the diet induced significant increases of serum osteopontin and TGF-beta1, augumenting osteoclast-mediated bone resorption through the RANK/RANKL pathway [J. Clin. Invest. 112 (2003) 181]. TGF-beta1 showed a negative correlation with serum glutathione peroxidase (GPx) activity (r = -0.674, P < 0.003), but a positive correlation with the serum iron level (r = 0.836, P < 0.0001). Taken together, these results suggest for the first time that oxidative stress could be involved in the pathogenesis of metabolic bone diseases such as osteoporosis as demonstrated by analysis of the relationship between bone metabolism and oxidative stress.


Bcl-2 family members and disease.
Apoptosis plays an important role during development and in the maintenance of multicellular organisms. Bcl-2 family members affect cell death in either a positive or negative fashion. Although some redundancy exists between family members, expression of certain family members is important during development in an organ-specific manner. The founding family member bcl-2 tends to be highly expressed in the embryo and declines postnatally following differentiation and maturation. Altered expression of bcl-2, as well as other family members, has been observed in disease states potentially affecting treatment modalities. Here we examine the distribution and role death repressors bcl-2, bcl-x(L) and bcl-w as well as death effectors bax and bak play regulating apoptosis in a tissue-specific manner. Understanding the normal role of these proteins during embryogenesis and in the mature organ will give us important insight into what goes awry in various disease states.


The impact of periodontitis in the course of chronic obstructive pulmonary disease: Pulmonary and systemic effects.
The aim of this study was to verify the impact of periodontitis in the course of chronic obstructive pulmonary disease (COPD) in C57Bl/6J mice. The animals were randomly divided into four groups (n = 8): Basal, Periodontitis (P), COPD and COPD+P. COPD was induced by orotracheal instillation of 30 μl of cigarette extract 3 times/week for 7 weeks. Periodontitis was induced by ligation technique for 22 days. Euthanasia was performed on 51st day. The analyzes were total/differential cells and cytokines recovered from bronchoalveolar lavage (BAL), total/differential blood cell count, platelets, total marrow cell count, airway collagen deposition, alveolar enlargement analyzed by mean linear intercept (Lm), mucus and bone crest reabsorption. One-way ANOVA followed by the Student-Newman-Keuls was used. The association COPD+P decreased macrophages (p = 0,0351), TNF-α (p = 0,0071) and INF-γ (p = 0,0004) in BAL, when compared to the COPD group maintaining emphysema levels by alveolar enlargement (p < .05) reorganization of collagen fibers (p = .001) and also mean linear intercept (lm) (p = .001) and mucus (p = .0001). The periodontitis group caused TNF-α increase (p = 0, 0001) in BAL. Periodontitis, per se, does not alter any of the parameters analyzed, except for increased TNF-α in BAL. However, its association with COPD caused macrophages TNF-α and INF-γ alterations, when compared to the COPD group maintaining emphysema levels by alveolar enlargement and reorganization of collagen fibers. It seems that periodontitis is influencing the course of Th1 profile cell, and cytokines and pulmonary alterations. Further studies are needed to clarify the regulatory process underlying these two diseases.


Monitoring human ovarian carcinoma with a combination of CA 125, CA 19-9, and carcinoembryonic antigen.
CA 125 and CA 19-9 are antigenic determinants associated with human epithelial ovarian carcinomas. Murine monoclonal antibodies have been raised against these determinants, and immunoradiometric assays have been developed to monitor antigen levels in the serum of cancer patients. This study was undertaken to determine whether concomitant measurement of CA 125, CA 19-9, and carcinoembryonic antigen would provide a more precise correlation with tumor progression or regression than could be obtained with any single assay. Among 105 patients with surgically demonstrable epithelial ovarian carcinoma, serum CA 125 levels were elevated (greater than 35 U/ml) in 83%, CA 19-9, levels (greater than 37 U/ml) in 17%, and carcinoembryonic antigen levels (greater than or equal to 2.5 ng/ml) in 37%. Within individual samples, no correlation was found among values for the three markers, but patients with elevated CA 19-9 levels also had increased levels of CA 125. At least one of the three markers was elevated in 90% of the subjects. When 41 patients were monitored serially over 2 to 60 months, alterations in CA 125 levels correlated with disease progression or regression in 94% of instances, whereas alterations in CA 19-9 levels correlated in 33% and alterations in carcinoembryonic antigen levels in 25% of instances. Concomitant measurement of CA 125, CA 19-9, and carcinoembryonic antigen did not prove superior to measurement of CA 125 alone in the monitoring of patients with epithelial ovarian carcinoma.


Multi-pumping flow system for the spectrophotometric determination of dipyrone in pharmaceutical preparations.
A novel flow system for the spectrophotometric determination of dipyrone with p-dimethylaminobenzaldehyde exploiting the multi-pumping approach was developed. The proposed methodology utilises several micro-pumps for propelling the involved fluids under improved mixing conditions, introducing sample/reagent aliquots and providing commuting facilities. As a consequence the multi-pumping system presents high versatility and manifold simplicity, as well as a straightforward operational control and enhanced analytical capabilities. Linearity of the analytical curve was observed within 10 and 400 mg l(-1) dipyrone (r=0.9997; n=6), results were precise (r.s.d.<0.12%; n=20) and sampling rate was 50 h(-1). Detection limit was estimated as 1 mg l(-1) dipyrone. The method was applied to pharmaceutical preparations and the results were in agreement with those obtained by the reference procedure with relative deviations within -1.7 and +2.2%.


Pregnancy Outcomes in Women with Arrhythmias following Surgical Repair of Cardiac Defects.
The goal of this study was to investigate whether preexisting cardiac arrhythmias are associated with adverse obstetrical outcomes in women with a history of open cardiac surgery. This was a retrospective cohort study of women with a history of open cardiac surgery who delivered at MedStar Washington Hospital Center (Washington, DC) from January 2007 through December 2018. Women with the isolated percutaneous cardiac surgical repair were excluded. Maternal and neonatal outcomes were compared between patients with preexisting cardiac arrhythmias and patients without preexisting cardiac arrhythmias. Maternal outcomes studied were intensive care unit admission, postpartum blood loss greater than 1,000 mL, congestive heart failure development, preeclampsia with severe features, postpartum readmission, postpartum cardiac events, and postpartum length of stay >5 days. Neonatal outcomes investigated were low birth weight <2,500 g, Apgar's scores <7 at 5 minutes, and neonatal intensive care unit admission. Multivariate logistic regression model was used to calculate the adjusted odds ratio (aOR) and 95% confidence intervals. The outcomes for 69 deliveries from 56 women with a history of open cardiac surgery were examined. Thirty-three women (48%) had arrhythmias after cardiac surgery with fourteen (20%) requiring implantable cardioverted defibrillators. Two women (6%) with preexisting arrhythmias after cardiac surgery developed postpartum volume overload requiring readmission (p = 0.06). After controlling for age, gestational age at delivery, and BMI, preeclampsia with severe features (p = 0.02) and low birth weight neonates (p = 0.02, aOR = 2.26 [0.56-9.03]) remained statistically more like to occur in patients with preexisting cardiac arrhythmias than in patients without preexisting arrhythmias. Women with a history of open cardiac surgery and preexisting cardiac arrhythmias prior to pregnancy are more likely to develop preeclampsia with severe features and have low birth weight neonates compared with women with a history of open cardiac surgery without preexisting cardiac arrhythmias. · Preexisting arrhythmias after cardiac surgery was associated with a risk of preeclampsia.. · Neonates of women with preexisting cardiac arrhythmias are more likely to be low birth weight.. · Forty-seven percent of women with open cardiac surgery developed subsequent arrhythmias..


Designing stable redox-active surfaces: chemical attachment of an osmium complex to glassy carbon electrodes prefunctionalized by electrochemical reduction of an in situ-generated aryldiazonium cation.
The production of stable redox-active layers on electrode surfaces can lead to improvements in electronic device design. Enhanced stability can be achieved by pretreatment of electrode surfaces to provide surface chemical functional groups for covalent tethering of redox complexes. Herein, we describe pretreatment of glassy carbon electrodes to provide surface carboxylic acid groups by electro-reduction of an in situ-generated aryl diazonium salt from 3-(4-aminophenyl)propionic acid. This surface layer is characterized by attenuated total reflection infrared spectroscopy, atomic force microscopy, and electrochemical blocking studies. The surface carboxylic acid generated is then used to tether an osmium complex, [Os(2,2'-bipyridyl)2(4-aminomethylpyridine)Cl]PF6, to provide a covalently bound redox-active monolayer, E(0) ' of 0.29 V (vs Ag/AgCl in phosphate buffer, pH 7.4), on the pretreated glassy carbon electrode. The layer proves stable to pH, temperature, and storage conditions, retaining electroactivity for at least 6 months.


Assessing ad-hoc adaptations' alignment with therapeutic goals: a qualitative study of lay counselor-delivered family therapy in Eldoret, Kenya.
A key question in implementation science is how to balance adaptation and fidelity in translating interventions to new settings. There is growing consensus regarding the importance of planned adaptations to deliver interventions in contextually sensitive ways. However, less research has examined ad-hoc adaptations, or those that occur spontaneously in the course of intervention delivery. A key question is whether ad-hoc adaptations ultimately contribute to or detract from intervention goals. This study aimed to (a) identify ad-hoc adaptations made during delivery of a family therapy intervention and (b) assess whether they promoted or interrupted intervention goals. Tuko Pamoja (Swahili: "We are Together") is an evidence-informed family therapy intervention aiming to improve family dynamics and mental health in Kenya. Tuko Pamoja employs a task-shifting model, delivered by lay counselors who are afforded a degree of flexibility in presenting content and in practices they use in sessions. We used transcripts of therapy sessions with 14 families to examine ad-hoc adaptations used by counselors. We first identified and characterized ad-hoc adaptations through a team-based code development, coding, and code description process. Then, we evaluated to what extent ad-hoc adaptations promoted the principles and strategies of the intervention ("TP-promoting"), disrupted them ("TP-interrupting"), or neither ("TP-neutral"). To do this, we first established inter-coder agreement on application of these categories with verification by the intervention developer. Then, coders categorized ad-hoc adaptation text segments as TP-promoting, TP-interrupting, or TP-neutral. Ad-hoc adaptations were frequent and included (in decreasing order): incorporation of religious content, exemplars/role models, community dynamics and resources, self-disclosure, and metaphors/proverbs. Ad-hoc adaptations were largely TP-promoting (49%) or neutral (39%), but practices were TP-interrupting 12% of the time. TP-interrupting practices most often occurred within religious content and exemplars/role models, which were also the most common practices overall. Extra attention is needed during planned adaptation, training, and supervision to promote intervention-aligned use of common ad-hoc adaptation practices. Discussing them in trainings can provide guidance for lay providers on how best to incorporate ad-hoc adaptations during delivery. Future research should evaluate whether well-aligned ad-hoc adaptations improve therapeutic outcomes. Pilot trial registered at clinicaltrials.gov (C0058).


Enhanced bioavailability of tripterine through lipid nanoparticles using broccoli-derived lipids as a carrier material.
Chemotherapy via the oral route remains a considerable challenge due to poor water-solubility and permeability of anticancer agents. This study aimed to construct lipid nanoparticles using broccoli-derived lipids for oral delivery of tripterine (Tri), a natural anticancer candidate, and to enhance its oral bioavailability. Tri-loaded broccoli lipid nanoparticles (Tri-BLNs) were prepared by a solvent-diffusion method. The resulting Tri-BLNs were 75±10 nm in particle size with entrapment efficiency over 98%. In vitro release study indicated that Tri was almost not released from Tri-BLNs (<2%), whereas the lipolytic experiment showed that Tri-BLNs possessed a relatively strong anti-enzymatic degradation ability to Tri-CLNs (Tri-loaded common lipid nanoparticles). In situ single-pass intestinal perfusion manifested that the effective permeability of Tri-BLNs were significantly higher than that of Tri-CLNs. Further, Tri-BLNs exhibited more efficient cellular uptake in MDCK-II cells as evidenced by flow cytometry and confocal microscopy. The relative bioavailability of Tri-BLNs and Tri-CLNs was 494.13% and 281.95% compared with Tri suspensions, respectively. Depending on the ability in enhancement of biomembrane permeability, broccoli-derived lipids as an alternative source should be useful to construct lipid nanoparticles for bettering oral delivery of drugs with low bioavailability.


Chromogranin B (secretogranin I), a putative precursor of two novel pituitary peptides through processing at paired basic residues.
During the course of reversed-phase high-pressure liquid chromatography (RP-HPLC) purification of the 7B2 peptide originally isolated in our laboratory from human pituitary gland extracts, two novel peptides were identified and purified to homogeneity. The complete amino acid sequence of the first one was established in 1985 and recently found to be entirely homologous to positions 420-493 of the just published chromogranin B sequence. This peptide, denoted GAWK, could originate from chromogranin B following specific cleavage at the basic amino acids flanking both termini of GAWK. Moreover, another peptide isolated in our laboratory from the same source and denoted CCB has been discovered and its sequence is also part of the same chromogranin B molecule. Here again, this peptide, occupying positions 597-653 and located at the COOH-terminal region of chromogranin B, could derive from specific processing at basic amino acids, Arg-Lys-Lys, present at positions 594-596. In a manner reminiscent of the relationship between pancreastatin and chromogranin A, it is proposed that both GAWK and CCB are produced from chromogranin B after specific processing at basic amino acids. These data are thus in favor of a putative role of chromogranins as precursors to potentially bioactive peptides.


Immunomodulating pectic polysaccharides from waste rose petals of Rosa damascena Mill.
A water-soluble polysaccharide (RP-1) was obtained from distilled rose petals of Rosa damascena Mill. as an attempt for valorization of the waste. RP-1 showed in vitro intestinal immune system modulating activity through Peyer's patch cells and IL-6 producing activity from macrophages. RP-1 lost most of its immunomodulating activity by degradation of the carbohydrate moiety with periodate. RP-1 was fractionated by anion-exchange and gel filtration chromatography and some of the fractions showed significant intestinal immune system modulating activity. The active fractions were suggested to be pectic polysaccharides and type II arabino-3,6-galactan from the component sugar analyses and the reactivity with Yariv antigen. When some active fractions were digested with endo α-d-(1→4)-polygalacturonase, highest molecular weight fragments which were considered as rhamnogalacturonan I, showed potent immunomodulating activities. To our knowledge, this is a first report which explores the possibility for utilization of waste rose petals as a source of immunomodulating pectic polysaccharides.


Antibacterial activity of the alkaloid-enriched extract from Prosopis juliflora pods and its influence on in vitro ruminal digestion.
The purpose of this study was to assess the in vitro antimicrobial activity of alkaloid-enriched extracts from Prosopis juliflora (Fabaceae) pods in order to evaluate them as feed additives for ruminants. As only the basic chloroformic extract (BCE), whose main constituents were juliprosopine (juliflorine), prosoflorine and juliprosine, showed Gram-positive antibacterial activity against Micrococcus luteus (MIC = 25 μg/mL), Staphylococcus aureus (MIC = 50 μg/mL) and Streptococcus mutans (MIC = 50 μg/mL), its influence on ruminal digestion was evaluated using a semi-automated in vitro gas production technique, with monensin as the positive control. Results showed that BCE has decreased gas production as efficiently as monensin after 36 h of fermentation, revealing its positive influence on gas production during ruminal digestion. Since P. juliflora is a very affordable plant, this study points out this alkaloid enriched extract from the pods of Prosopis juliflora as a potential feed additive to decrease gas production during ruminal digestion.


A TCP-based early regression index predicts the pathological response in neo-adjuvant radio-chemotherapy of rectal cancer.
Introducing a radiobiological index based on early tumor regression during neo-adjuvant radio-chemotherapy (RCT, including oxaliplatin) of rectal adenocarcinoma and testing its discriminative power in predicting the tumor response. Seventy-four patients were treated with Helical Tomotherapy following an adaptive (ART) protocol (41.4 Gy/18 fr, 2.3 Gy/fr) delivering a simultaneous integrated boost on the residual tumor in the last 6 fractions up to 45.6 Gy. T2-weighted MRI were taken before (MRIpre) and at mid (MRImid) therapy and the corresponding tumor volumes were considered (Vpre,Vmid). The "Early Regression Index" [Formula: see text] was introduced and its discriminative power was assessed in terms of AUC, sensitivity/specificity, positive/negative predictive value (PPV/NPV). Two end-points were considered: (a) pathological complete response (pCR) or clinical complete response followed by watch-and-wait, (cCR); (b) limited response (residual vital cells (RVC) in the surgical specimen >10%). Complete data were available for 65 patients: pCR, cCR and RVC >10% were 20, 2 and 19 respectively. The discriminative power of ERITCP was moderately high (AUC = 0.81/0.75 for /pCRorcCR/RVC >10% respectively, p < 0.0005). ERITCP was highly sensitive (86-89%) with very high NPV (90-94%). The discriminative power of ERITCP was confirmed on a subgroup of 44/65 patients when considering tumor volumes delineated by a skilled radiologist. A radiobiologically consistent index based on early regression showed high performances in predicting the pathological response after neo-adjuvant RCT for rectal cancer with relevant potentialities for ART/treatment customization.


Pregnancy heartburn in Nigerians and Caucasians with theories about aetiology based on manometric recordings from the oesophagus and stomach.
A study by questionnaire of the incidence of pregnancy heartburn in two groups of Nigerians and one group of Caucasians showed an incidence of 9-8 per cent in all Nigerians as against 78-8 per cent in Caucasians. The difference was highly significant (chi 2 = 102-75; P less than 0-0001). An oesophageal manometric study of the lower esophageal sphincter in 12 non-pregnant women and in 12 pregnant Nigerians (10 without heartburn and 2 with heartburn) showed that the mean lower oesophageal sphincter pressure (+/- SD) in the pregnant patients without heartburn was 20-1 +/- 7-0 mm Hg as against 17-9 +/- 7-0 in the non-pregnant women. While one pregnant subject without heartburn had a lower oesophageal sphincter which was partially displaced into the thorax, none of the non-pregnant women had such sphincter displacement. The two pregnant women with heartburn had low resting sphincter pressures and in both of them the sphincters were partially in the thorax. It is argued that pregnancy heartburn is due to the displacement of the lower oesophageal sphincter into the negative pressure environment of the thorax where the sphincter is less capable of resisting reflux. It is also argued that pregnancy heartburn is commoner in Caucasians that in Nigerians because the spincter in non-pregnant Caucasians is often partly intrathoracic and is thus more easily displaced completely into the thorax by the pregnant uterus.


Switching costs, price sensitivity and health plan choice.
We investigate the extent to which sensitivity to health plan premiums differs across individuals according to characteristics related to the cost of switching plans. Our results indicate substantial variation in price sensitivity related to expected health care costs: younger, healthier employees are between two and four times more sensitive to price than employees who are older and who have been recently hospitalized or diagnosed with cancer. We also find evidence of status quo bias: estimated premium elasticities are significantly higher for new hires than for incumbent employees. Simulations combining our results with actuarial data illustrate the cost implications of risk-related differences in price elasticity.