Effect of somatostatin on kidney function and vasoactive hormone systems in health subjects.
The acute effects of i.v. somatostatin (250 mcg bolus followed by 250 mcg/h continuous infusion for two hours) on renal hemodynamics, renal electrolyte and water handling, and urinary excretion of catecholamines and prostaglandins, as well as on plasma concentrations of arginine vasopressin, atrial natriuretic factor, norepinephrine, epinephrine, dopamine, glucagon, and plasma renin activity were studied in seven normal subjects. Somatostatin decreased effective renal plasma flow and glomerular filtration rate, osmotic and free water clearances, urine volume, and sodium and potassium excretion, while urinary osmolality, fractional excretion of sodium, and phosphate excretion increased significantly. Plasma concentrations of arginine vasopressin, atrial natriuretic factor, norepinephrine, epinephrine, and dopamine remained unchanged, while plasma renin activity (3.0 +/- 0.25 vs 2.4 +/- 0.2 ng AngI/ml/h; p less than 0.01) and glucagon levels (40 +/- 11 vs 20 +/- 16 pg/ml; p less than 0.01) decreased. Urinary excretion of norepinephrine, epinephrine, dopamine, PGE2, and PGF2 alpha was suppressed under somatostatin. A significant positive correlation was found between urinary dopamine and sodium excretion (r = 0.7; p less than 0.001) and urinary prostaglandin E2 and glomerular filtration (r = 0.52; p less than 0.01). Without accompanying changes in plasma osmolality and vasopressin concentration significant antidiuresis occurred, suggesting a direct tubular effect of somatostatin. However, the hormone-induced changes are due mainly to the decrease in renal plasma flow. The results demonstrate that somatostatin at supraphysiological doses exerts significant effects on the kidney.


Analysis of BCLI, N363S and ER22/23EK Polymorphisms of the Glucocorticoid Receptor Gene in Adrenal Incidentalomas.
Patients with adrenal incidentalomas (AI) may experience detrimental consequences due to a minimal cortisol excess sustained by adrenal adenoma. SNPs of the glucocorticoid receptor gene (NR3C1) modulate individual sensitivity to glucocorticoids and may interfere with the clinical presentation. To compare the frequency of N363S, ER22/23EK and BclI SNPs in patients with AI with the general population and to evaluate whether these SNPs are linked to consequences of cortisol excess. Multicentric, retrospective analysis of patients referred from 2010 to 2014 to 4 centers (Orbassano, Milano, Messina [Italy] and Zagreb [Croatia]). 411 patients with AI; 153 males and 258 females and 186 from blood donors. All patients and controls were genotyped for BclI, N363S and ER22/23EK and SNPs frequency was associated with clinical and hormonal features. SNP frequency was: SNP frequency was: N363S 5.4% (MAF 0.027), BclI 54.7% (MAF 0.328), ER22/23EK 4.4% (MAF 0.022), without any significant difference between patients and controls. N363S was more frequent in hypertensive patients (p = 0.03) and was associated with hypertension (p = 0.015) in patients with suppressed cortisol after the 1-mg DST. Our results demonstrate that SNPs of the glucocorticoid receptor gene do not play a pathogenetic role for AI. The impact of any single SNP on the phenotypic expression of minimal cortisol excess is limited and their analysis does not provide additional data that may be exploited for patient management.


Children in pickup trucks. American Academy of Pediatrics. Committee on Injury and Poison Prevention.
Pickup trucks have become increasingly popular in the United States. A recent study found that in crashes involving fatalities, cargo area passengers were 3 times more likely to die than were occupants in the cab. Compared with restrained cab occupants, the risk of death for those in the cargo area was 8 times higher. Furthermore, the increased use of extended-cab pickup trucks and air bag-equipped front passenger compartments creates concerns about the safe transport of children. The most effective preventive strategies are the legislative prohibition of travel in the cargo area and requirements for age-appropriate restraint use and seat selection in the cab. Parents should select vehicles that are appropriate for the safe transportation needs of the family. Physicians have an important role in counseling families and advocating public policy measures to reduce the number of deaths and injuries to occupants of pickup trucks.


C-Raf-1 protein kinase is not essential for Ras transformation of mouse embryo fibroblasts.
Transfection of primary cells with mutated oncogenic ras plus a cooperating oncogene such as myc results in the acquisition of the transformed cell phenotype. The pathways downstream of Ras that are required for transformation are an active topic of research. The Raf-MEKK-MAP kinase pathway is triggered by activation of Ras and thought to be important in Ras transformation of rodent fibroblasts. To further explore the involvement of this pathway, fibroblasts from homozygous knock out c-Raf-1 mouse embryos (20 KO) and wild-type c-Raf-1 mouse embryos (16 WT) were transfected with H-ras and myc(v). The resulting cell line derived from the knock out cells grew slower both in tissue culture and had a longer latency period as tumors than the transformed cell line from the wild-type cells. Both cell lines were however able to form tumors in nude mice. These results suggest that c-Raf-1 is not required for Ras transformation in this system.


Vitamin D levels and mortality in type 2 diabetes.
To evaluate vitamin D as a predictor of all-cause and cardiovascular mortality and risk of progression to micro- or macroalbuminuria in type 2 diabetic patients. In a longitudinal observational follow-up study, 289 type 2 diabetic patients with normoalbuminuria (n = 172), microalbuminuria (n = 73), and macroalbuminuria (n = 44) at baseline were followed for a median (range) of 15.0 (0.2-23) years. Mean ± SD age was 54 ± 9 years. Plasma 25-hydroxyvitamin D(3) levels were determined by high-performance liquid chromatography/tandem mass spectrometry on baseline samples. Severe vitamin D deficiency was defined as the lower 10th percentile (<13.9 nmol/l). Median (range) vitamin D level was 35.7 (5-136.7) nmol/l. Vitamin D levels were not associated with age, sex, estimated glomerular filtration rate, urinary albumin excretion rate (UAER), or A1C at baseline, but low levels were weakly associated with elevated systolic blood pressure (R = 0.13, P = 0.03). During follow-up, 196 (68%) patients died. All-cause mortality was increased in patients with severe vitamin D deficiency (hazard ratio 1.96 [95% CI 1.29-2.98]). The association persisted after adjustment for UAER, A1C, diabetes duration, and conventional cardiovascular risk factors (2.03 [1.31-3.13]). Severe vitamin D deficiency was associated with increased cardiovascular mortality (1.95 [1.11-3.44]), and the association persisted after adjustment (1.90 [1.15-3.10]). Severe vitamin D deficiency at baseline did not predict progression to micro- or macroalbuminuria. In type 2 diabetic patients, severe vitamin D deficiency predicts increased risk of all-cause and cardiovascular mortality, independent of UAER and conventional cardiovascular risk factors. Whether vitamin D substitution improves prognosis remains to be investigated.


Concurrent reduction in the sulfation of heparan sulfate and basement membrane assembly in a cell model system.
Basement membranes (BMs) are specialized extracellular matrices that have important roles in cell attachment, migration, growth and differentiation. The murine teratocarcinoma cell line, M1536-B3, has been shown to produce a model BM composed of laminin, entactin and heparan sulfate proteoglycans but lacking collagen. Therefore, M1536-B3 cells are an excellent model system in which to study the role of non-collagenous components in BM assembly. We have used these cells to test for a requirement of mature heparan sulfate (HS) chains in BM assembly. Growth of M1536-B3 cells in the presence of chlorate, an inhibitor of activated sulfate synthesis, resulted in a dose-dependent decrease in the sulfation of glycosaminoglycans and reduction in the charge density of the isolated HS. The undersulfated HS from chlorate-treated cells had a decreased binding capacity for laminin when compared with control HS. Concurrent with these changes in sulfation, chlorate treatment of M1536-B3 cells resulted in the failure of BM assembly, which was restored upon removal of the chlorate from the growth medium. These results were not due to major alterations in cell attachment, spreading, growth, protein synthesis, or to an inability of the cells to synthesize and secrete laminin. These data suggest that the sulfation of HS and its subsequent ability to interact with other BM components play major roles in the assembly and structure of BMs.


Ability of ICU Health-Care Professionals to Identify Patient-Ventilator Asynchrony Using Waveform Analysis.
Waveform analysis by visual inspection can be a reliable, noninvasive, and useful tool for detecting patient-ventilator asynchrony. However, it is a skill that requires a properly trained professional. This observational study was conducted in 17 urban ICUs. Health-care professionals (HCPs) working in these ICUs were asked to recognize different types of asynchrony shown in 3 evaluation videos. The health-care professionals were categorized according to years of experience, prior training in mechanical ventilation, profession, and number of asynchronies identified correctly. A total of 366 HCPs were evaluated. Statistically significant differences were found when HCPs with and without prior training in mechanical ventilation (trained vs non-trained HCPs) were compared according to the number of asynchronies detected correctly (of the HCPs who identified 3 asynchronies, 63 [81%] trained vs 15 [19%] non-trained, P < .001; 2 asynchronies, 72 [65%] trained vs 39 [35%] non-trained, P = .034; 1 asynchrony, 55 [47%] trained vs 61 [53%] non-trained, P = .02; 0 asynchronies, 17 [28%] trained vs 44 [72%] non-trained, P < .001). HCPs who had prior training in mechanical ventilation also increased, nearly 4-fold, their odds of identifying ≥2 asynchronies correctly (odds ratio 3.67, 95% CI 1.93-6.96, P < .001). However, neither years of experience nor profession were associated with the ability of HCPs to identify asynchrony. HCPs who have specific training in mechanical ventilation increase their ability to identify asynchrony using waveform analysis. Neither experience nor profession proved to be a relevant factor to identify asynchrony correctly using waveform analysis.


Enzymatic crosslinking to fabricate antioxidant peptide-based supramolecular hydrogel for improving cutaneous wound healing.
Peptide-based supramolecular hydrogels are promising scaffold materials and have been utilized in many fields. The mechanical properties of peptide hydrogels are usually enhanced by synthetic or natural polymers to expand their application scope. In this study, antioxidant supramolecular hydrogels based on feruloyl-modified peptide and glycol chitosan were fabricated via a mild laccase-mediated crosslinking reaction. A natural polysaccharide derivative, feruloyl glycol chitosan (GC-Fer), was used to enhance the mechanical properties of peptide hydrogels. Feruloyl groups were introduced into the gel matrix via covalent bonds, which endowed the hydrogels with inherent antioxidant properties. This was beneficial for their in vivo application via scavenging harmful free radicals existing in a cutaneous wound. Further in vivo experiments demonstrated that the feruloyl-containing antioxidant hydrogel can improve the cutaneous wound healing process. The regeneration process of mature epithelium and connective tissues was accelerated in a full-thickness skin defect model.


Unfavorable social determinants of health and mortality risk by cardiovascular disease status: Findings from a National Study of United States Adults.
The association between cumulative burden of unfavorable social determinants of health (SDoH) and all-cause mortality has not been assessed by atherosclerotic cardiovascular disease (ASCVD) status on a population level in the United States. We assessed the association between cumulative social disadvantage and all-cause mortality by ASCVD status in the National Health Interview Survey, linked to the National Death Index. In models adjusted for established clinical risk factors, individuals experiencing the highest level of social disadvantage (SDoH-Q4) had over 1.5 (aHR = 1.55; 95%CI = 1.22, 1.96) and 2-fold (aHR = 2.21; 95% CI = 1.91, 2.56) fold increased risk of mortality relative to those with the most favorable social profile (SDoH-Q1), respectively for adults with and without ASCVD; those experiencing co-occurring ASCVD and high social disadvantage had up to four-fold higher risk of mortality (aHR = 3.81; 95%CI = 3.36, 4.32). These findings emphasize the importance of a healthcare model that prioritizes efforts to identify and address key social and environmental barriers to health and wellbeing, particularly in individuals experiencing the double jeopardy of clinical and social risk.


A probable Na+(K+)/H+ exchanger on the chloroplast envelope functions in pH homeostasis and chloroplast development in Arabidopsis thaliana.
Electroneutral monovalent cation/proton antiport across the chloroplast envelope has been shown previously to have an important regulatory effect on stromal pH and thereby on photosynthetic carbon reduction. Here we report that an Arabidopsis nuclear gene, AtCHX23, encodes a putative Na(+)(K(+))/H(+) exchanger and functions in the adjustment of pH in the cytosol and possibly in maintaining a high pH level in the chloroplast stroma. The AtCHX23 protein is localized in the chloroplast envelope. Plastids from chx23 mutants had straight thylakoids but lacked grana lamellae. chx23 mutant leaves were pale yellow and had a much reduced chlorophyll content. The chlorophyll content of chx23 was increased by growing in medium at low (4.0) pH and decreased by growing at high (7.0) pH. The cytosolic pH in the leaves of the mutant was significantly higher than that in the wild type. chx23 mutants displayed a high sensitivity to NaCl. Together, these data indicate that CHX23 is a probable chloroplast Na(+)(K(+))/H(+) exchanger important for pH homeostasis and chloroplast development and function.


High doses of olive leaf extract induce liver changes in mice.
Virtually ever since it was first commercialized in 1995, there have been several studies focusing on the use of olive leaf extract (OLE) as a natural therapy and its medical properties. The aim of this study was to investigate the effects of three different concentrations of OLE on the function of mice livers over the course of 14 weeks. Female ICR mice were divided into four groups, depending on OLE concentration used: 0%, 0.25%, 0.5%, and 0.75%. Alanine aminotransferase, alkaline phosphatase, total bilirubin and albumin serum concentrations were all measured. Histopathological changes of the liver were observed after haematoxylin and eosin, reticulin, and Masson's trichrome staining was carried out while liver mitochondrial bioenergetics were also evaluated. Alanine aminotransferase and alkaline phosphatase serum enzyme activities increased significantly in the groups in which 0.5% and 0.75% OLE concentrations were used. Histologically, all the groups exposed to OLE exhibited hyperplasia of the bile ducts, cholestasis, hepatocyte necrosis and inflammatory infiltrated. Hepatic fibrosis was observed in the groups featuring 0.5% and 0.75% OLE concentrations. The mitochondrial membrane potential, respiratory control ratio and ADP/O of samples from animals fed the higher OLE concentration was significantly decreased when compared to the control group.


Neurotrophin receptor structure and interactions.
Although ligand-induced dimerization or oligomerization of receptors is a well established mechanism of growth factor signaling, increasing evidence indicates that biological responses are often mediated by receptor trans-signaling mechanisms involving two or more receptor systems. These include G protein-coupled receptors, cytokine, growth factor and trophic factor receptors. Greater flexibility is provided when different signaling pathways are merged through multiple receptor signaling systems. Trophic factors exemplified by NGF and its family members, ciliary neurotrophic factor (CNTF) and glial derived neurotrophic factor (GDNF) all utilize increased tyrosine phosphorylation of cellular substrates to mediate neuronal cell survival. Actions of the NGF family of neurotrophins are not only dictated by ras activation through the Trk family of receptor tyrosine kinases, but also a survival pathway defined by phosphatidylinositol-3-kinase activity (Yao and Cooper, 1995), which gives rise to phosphoinositide intermediates that activate the serine/threonine kinase Akt/PKB (Dudek et al., 1997). Induction of the serine-threonine kinase activity is critical for cell survival, as well as cell proliferation. Hence, for many trophic factors, multiple proteins constitute a functional multisubunit receptor complex that activates ras-dependent and ras-independent intracellular signaling. The NGF receptors provide an example of bidirectional crosstalk. In the presence of TrkA receptors, p75 can participate in the formation of high affinity binding sites and enhanced neurotrophin responsiveness leading to a survival or differentiation signal. In the absence of TrkA receptors, p75 can generate, in only specific cell populations, a death signal. These activities include the induction of NF kappa B (Carter et al., 1996); the hydrolysis of sphingomyelin to ceramide (Dobrowsky et al., 1995); and the pro-apoptotic functions attributed to p75. Receptors are generally drawn and viewed as isolated integral membrane proteins which span the lipid bilayer, with signal transduction proceeding in a linear step-wise fashion. There are now numerous examples which indicate that each receptor acts not only in a linear, independent manner, but can also influence the activity of other cell surface receptors, either directly or through signaling intermediates. Which step and which intermediates are utilized for crosstalk between the receptors is a critical question. For neurotrophins, their primary function in sustaining the viability of neurons is counterbalanced by a receptor mechanism to eliminate cells by an apoptotic mechanism. It is conceivable that this bidirectional system may be utilized selectively during development and in neurodegenerative diseases.


Skeletal muscle loss: cachexia, sarcopenia, and inactivity.
Loss of skeletal muscle mass occurs during aging (sarcopenia), disease (cachexia), or inactivity (atrophy). This article contrasts and compares the metabolic causes of loss of muscle resulting from these conditions. An understanding of the underlying causes of muscle loss is critical for the development of strategies and therapies to preserve muscle mass and function. Loss of skeletal muscle protein results from an imbalance between the rate of muscle protein synthesis and degradation. Cachexia, sarcopenia, and atrophy due to inactivity are characterized by a loss of muscle mass. Each of these conditions results in a metabolic adaptation of increased protein degradation (cachexia), decreased rate of muscle protein synthesis (inactivity), or an alteration in both (sarcopenia). The clinical consequences of bedrest may mimic those of cachexia, including rapid loss of muscle, insulin resistance, and weakness. Prophylaxis against bedrest-induced atrophy includes nutrition support with an emphasis on high-quality protein. Nutritional supplementation alone may not prevent muscle loss secondary to cachexia, but, in combination with the use of an anabolic agent, it may slow or prevent muscle loss.


High-dose chemotherapy and hematopoietic stem cell transplantation for patients with nasopharyngeal cancer: a feasibility study.
Nasopharyngeal cancer (NPC) is a highly chemosensitive malignancy. The purpose of this study was to evaluate the clinical efficacy of high-dose chemotherapy (HDCT) in combination with hematopoietic stem cell transplantation in patients with locally advanced or metastatic NPC. Nine patients with locally advanced or metastatic NPC were recruited after three to four courses of cisplatin-based chemotherapy followed by a single course of cyclophosphamide 1600 mg/m(2) day 1-4, carboplatin 400 mg/m(2) day 1-3, and thiotepa 120 mg/m(2) day 1-4 or melphalan 120 mg/m(2) day 5. Chemoirradiation was administered after HDCT only if the patient had never received radiotherapy or had residual cervical nodes. A median of 8.32 x 10(6) CD34+ cells/kg was collected. Two patients were rendered disease-free before HDCT, one by massectomy and one by cisplatin-based chemotherapy. All patients recovered rapidly after peripheral blood stem cell transplantation (PBSCT). Among seven assessable patients, response to HDCT was observed in four patients. Only one patient achieved complete response after HDCT. The median time to failure and median survival after HDCT was eight and 18 months, respectively. One patient died of pulmonary hemorrhage two months after transplantation. No long-term disease-free survival was noted. HDCT with autologous PBSCT is feasible with an acceptable toxicity, and can convert partial remission into complete remission. While no long-term disease-free survival was observed in this study, further investigations are needed to establish the role of HDCT in the treatment of NPC.


Development and characterization of a bacteriophage cocktail with high lytic efficacy against field-isolated Salmonella enterica.
Salmonella spp. is a prevalent pathogen that causes great public health concern worldwide. Bacteriophage-based cocktails have arisen as an alternative to antibiotics to inhibit the growth of Salmonella. However, the bactericidal effect of bacteriophage cocktails in vivo largely differs from their observed effect in vitro. This is partly because in vitro developments of cocktails do not always consider the bacterial diversity nor the environmental conditions where bacteriophages will have to replicate. Here, we isolated and sequenced 47 bacteriophages that showed variable degrees of lytic activity against 258 Salmonella isolates from a commercial broiler company in Brazil. Three of these bacteriophages were characterized and selected to assemble a cocktail. In vitro quantitative assays determined the cocktail to be highly effective against multiple serovars of Salmonella, including Minnesota and Heidelberg. Remarkably, the in vitro lytic activity of the cocktail was retained or improved in conditions that more closely resembled the chicken gut, such as anaerobiosis, 42°C, and Salmonella mono-strain biofilms. Analysis of bacterial cross-resistance between the 3 bacteriophages composing the cocktail revealed limited or no generation of cross-resistance. Our results highlight the relevance of an optimized flux of work to develop bacteriophage cocktails against Salmonella with high lytic efficacy and strong potential to be applied in vivo in commercial broiler farms.


Eruptive Cherry Angiomas and Skin Melanoma: Further Insights into an Intriguing Association.
Some risk factors for malignant melanoma (MM) are recognized. To compare the strength of association between MM and eruptive cherry angiomas (CAs) with that of other well-known associations. This cross-sectional study included all subjects referred to the Outpatient Dermatology-Oncology and Dermoscopy Units of the Universities of Ferrara and Bologna, Italy, over a 5-month period and submitted to total body skin examination. We recorded: age, sex, cutaneous and non-cutaneous malignancies, presence of CAs, arbitrarily considered as "eruptive" when >10, >40 common melanocytic naevi or >2 clinically atypical naevi. The strength of association between the possible risk factors and MM was calculated by odds ratio in both the whole population and age quartiles. Variables associated with MM were included in multiple logistic regression analysis. 1,190 subjects were included; 615 had malignant skin tumours, 462 MM, 85 extracutaneous tumours. Five hundred and eighty-seven subjects had eruptive CAs, 485 subjects >40 melanocytic naevi and 368 more than 2 atypical melanocytic naevi. Eruptive CAs, especially in subjects younger than 70, and >2 atypical melanocytic naevi, mostly in subjects older than 50, were significantly associated with MM. The strength of these 2 associations was similar. The presence of >40 melanocytic naevi was not associated with MM. These findings confirmed an association between MM and eruptive CAs, which was as strong as the one between MM and >2 atypical melanocytic naevi. CAs seem an intriguing model of interaction between heterogeneous variables, like immunocompetence, stimuli inducing endothelial cell proliferation, and oncogenesis, which deserves further investigation.


Reversible magnetism between an antiferromagnet and a ferromagnet related to solvation/desolvation in a robust layered [Ru2]2TCNQ charge-transfer system.
The charge-transfer compound [{Ru(2)(O(2)CPh-o-Cl)(4)}(2)TCNQ(MeO)(2)] x CH(2)Cl(2) (1; o-ClPhCO(2)(-) = o-chlorobenzoate; TCNQ(MeO)(2) = 2,5-dimethoxy-7,7,8,8-tetracyanoquinodimethane) was synthesized from the reaction of the neutral precursors [Ru(2)(II,II)(O(2)CPh-o-Cl)(4)] (abbreviated as [Ru(2)(II,II)] or [Ru(2)(4+)]) and TCNQ(MeO)(2) in a CH(2)Cl(2)/nitrobenzene solution. The structure consists of two-dimensional layers consisting of an infinite array in which [Ru(2)(II,II)] units are involved in charge transfer to TCNQ(MeO)(2) to give a formal charge of [{Ru(2)(4.5+)}-TCNQ(MeO)(2)(*-)-{Ru(2)(4.5+)}]. Interstitial CH(2)Cl(2) molecules are located in the void spaces between the layers. Strong intralayer magnetic coupling between the units [Ru(2)(II,II)] with S = 1 or [Ru(2)(II,III)] with S = 3/2 and TCNQ(MeO)(2)(*-) with S = 1/2, as well as long-range ordering due to antiferromagnetic interlayer interactions, was observed. An antiferromagnetic ground state exists below T(N) = 75 K, which undergoes a metamagnetic transition under applied fields less than 2 T to a field-induced canted antiferromagnetic state with large coercivities up to H(c) = 1.6 T at 1.8 K. Compound 1 gradually loses the interstitial CH(2)Cl(2) molecule at room temperature to form a dried sample (1-dry) without loss of crystallinity and converts nearly reversibly back to 1 after being exposed to CH(2)Cl(2) vapor for 72 h (distinguished as 1'). Interestingly, during this process there is no significant change in lattice dimensions and bond distances or angles with a volume change of only 1.2 vol %. The only discernible difference is ordering/disordering of a pendant ligand orientation, but the magnetism is dramatically altered to a ferromagnetic state with T(c) approximately 56 K for 1-dry. The magnetic property changes are gradual and depend on the degree of interstitial CH(2)Cl(2) molecule loss with reversibility in the process of going between 1 and 1-dry. In addition, in the case of partially desolvated crystals that have mixed domains of ferromagnetically and antiferromagnetically ordered domains for desolvated and solvated segments, respectively, the complete change to ferromagnet can also be triggered by magnetic fields even if the desolvated segments are comparatively minor compared to the solvated segments in a crystal. Surprisingly, the information of the existence of ferromagnetically ordered domains is dynamically recorded in the entire crystal after applying significant magnetic fields as if the majority of the antiferromagnetically ordered domains for solvated segments were never present.


Target Screen of Anti-Hyperuricemia Compounds from Cortex Fraxini In Vivo Based on ABCG2 and Bioaffinity Ultrafiltration Mass Spectrometry.
The ATP-binding cassette (ABC) transporter ABCG2 is a significant urate transporter with a high capacity, and it plays a crucial role in the development of hyperuricemia and gout. Therefore, it has the potential to be targeted for therapeutic interventions. Cortex Fraxini, a traditional Chinese medicine (TCM), has been found to possess anti-hyperuricemia properties. However, the specific constituents of Cortex Fraxini responsible for this effect are still unknown, particularly the compound that is responsible for reducing uric acid levels in vivo. In this study, we propose a target screening protocol utilizing bio-affinity ultrafiltration mass spectrometry (BA-UF-MS) to expediently ascertain ABCG2 ligands from the plasma of rats administered with Cortex Fraxini. Our screening protocol successfully identified fraxin as a potential ligand that interacts with ABCG2 when it functions as the target protein. Subsequent investigations substantiated fraxin as an activated ligand of ABCG2. These findings imply that fraxin exhibits promise as a drug candidate for the treatment of hyperuricemia. Furthermore, the utilization of BA-UF-MS demonstrates its efficacy as a valuable methodology for identifying hit compounds that exhibit binding affinity towards ABCG2 within TCMs.


Urinary PGE-M in colorectal cancer: predicting more than risk?
Progress in cancer chemoprevention has been hindered by a lack of validated biomarkers of risk and interventive response. The identification of accurate, reliable, and easily measurable risk and response biomarkers within the field of cancer prevention could dramatically alter our approach to the disease. Colorectal cancer is associated with substantial morbidity and a limited 5-year survival rate for late-stage disease. The identification of biomarkers to predict (i) those most at risk of clinically significant colorectal neoplasia in conjunction with or building upon current risk models and/or (ii) those most likely to respond to potential colorectal chemopreventive agents, such as aspirin and NSAIDs, would significantly advance colorectal cancer risk management. Urinary PGE-M is an established indicator of systemic prostaglandin E2 production and has previously been demonstrated to predict risk of advanced colorectal neoplasia in a handful of studies. In the July 2014 issue, Bezawada and colleagues confirmed those earlier risk associations and demonstrated that PGE-M can also predict responsiveness to aspirin/NSAIDs in a small subset of women undergoing lower endoscopy in the Nurse's Health Study. PGE-M has the potential to define subsets of the population that may derive greater chemopreventive benefit from NSAIDs, as well as the potential to optimize the use of expensive and/or invasive screening tests. Additional larger and more diverse prospective studies meeting the criteria for phase IV biomarker studies are needed to advance the development of PGE-M as a noninvasive biomarker of both risk and chemopreventive response in populations at risk for colorectal cancer.


Machine learning analysis of the T cell receptor repertoire identifies sequence features of self-reactivity.
The T cell receptor (TCR) determines specificity and affinity for both foreign and self-peptides presented by the major histocompatibility complex (MHC). Although the strength of TCR interactions with self-pMHC impacts T cell function, it has been challenging to identify TCR sequence features that predict T cell fate. To discern patterns distinguishing TCRs from naive CD4+ T cells with low versus high self-reactivity, we used data from 42 mice to train a machine learning (ML) algorithm that identifies population-level differences between TCRβ sequence sets. This approach revealed that weakly self-reactive T cell populations were enriched for longer CDR3β regions and acidic amino acids. We tested our ML predictions of self-reactivity using retrogenic mice with fixed TCRβ sequences. Extrapolating our analyses to independent datasets, we predicted high self-reactivity for regulatory T cells and slightly reduced self-reactivity for T cells responding to chronic infections. Our analyses suggest a potential trade-off between TCR repertoire diversity and self-reactivity. A record of this paper's transparent peer review process is included in the supplemental information.


Synthesis of bile acid monosulphates by the isolated perfused rat kidney.
Perfusion of an isolated rat kidney with labelled bile acids, in a protein-free medium, resulted in the urinary excretion of the labelled bile acid, 3% being converted into polar metabolities in 1h. These metabolities were neither glycine nor taurine conjugates, nor bile acid glucuronides, and on solovolysis yielded the free bile acid. On t.l.c. the metabolite of [24-14C]lithocholic acid had the mobility of lithocholate 3-sulphate. The principal metabolite of [24-14C]chenodeoxycholic acid had the mobility of chenodeoxycholate 7-sulphate; trace amounts appeared as chenodeoxycholate 3-sulphate. [35S]sulphate was incorporated in chenodeoxycholic acid by the kidney, resulting in a similar pattern of sulphation. No disulphate salt of chenodeoxycholic acid was detected. These findings lend support to the hypothesis that renal synthesis may account for some of the bile acid sulphates present in urine in the cholestatic syndrome in man.


Model on cell movement, growth, differentiation and de-differentiation: reaction-diffusion equation and wave propagation.
We construct a model for cell proliferation with differentiation into different cell types, allowing backward de-differentiation and cell movement. With different cell types labeled by state variables, the model can be formulated in terms of the associated transition probabilities between various states. The cell population densities can be described by coupled reaction-diffusion partial differential equations, allowing steady wavefront propagation solutions. The wavefront profile is calculated analytically for the simple pure growth case (2-states), and analytic expressions for the steady wavefront propagating speeds and population growth rates are obtained for the simpler cases of 2-, 3- and 4-states systems. These analytic results are verified by direct numerical solutions of the reaction-diffusion PDEs. Furthermore, in the absence of de-differentiation, it is found that, as the mobility and/or self-proliferation rate of the down-lineage descendant cells become sufficiently large, the propagation dynamics can switch from a steady propagating wavefront to the interesting situation of propagation of a faster wavefront with a slower waveback. For the case of a non-vanishing de-differentiation probability, the cell growth rate and wavefront propagation speed are both enhanced, and the wavefront speeds can be obtained analytically and confirmed by numerical solution of the reaction-diffusion equations.


Immunohistochemical detection of E-cadherin in differentiated thyroid carcinomas correlates with clinical outcome.
A retrospective immunohistochemical analysis of the adhesion molecule E-cadherin (E-CD) was performed in 112 differentiated thyroid carcinomas and 38 synchronous and 20 relapse metastases primarily from operations performed at the Medical School Hanover between 1982 and 1992. E-CD-specific antibody 5H9 was applied to paraffin-embedded tissues. All patients were clinically followed for a maximal period of 12 years. Lack of E-CD expression i.e., <5% of tumor cells positive) occurred in 18 of 112 (16.1%) cases, whereas the majority showed either low (24.1%), medium (35.7%), or high (24.1%) positivity. No difference was found between papillary (n = 88) and follicular (n = 24) carcinomas. Univariate statistical analysis for survival (Kaplan-Meier) showed that lack of E-CD expression (P < 0.024) is an adverse prognostic factor for differentiated thyroid carcinomas. The highest significance was seen among patients without lymph node involvement at first presentation (pN(0); P = 0.0068) and among females (P = 0.0033). Multivariate analysis (Cox model) indicated that E-CD staining is an independent prognostic factor (corrected risk factor, 3.7; P < 0.03) in addition to distant metastasis (pM1) and tumor size. A comparison of E-CD stainings between primary tumors and their metastatic lesions showed similar results in both synchronous and relapse metastases after therapy. In conclusion, E-CD immunostaining is an independent prognostic indicator for differentiated thyroid carcinomas. It may help to uncover the small group of patients with differentiated thyroid carcinomas carrying a high risk of suffering an unfavorable clinical outcome.


Heterogeneity in the Dorsal Subiculum of the Rat. Distinct Neuronal Zones Project to Different Cortical and Subcortical Targets.
The aim of the present study was to relate the distribution of efferents of the dorsal subiculum to their origin along the proximodistal axis of the subiculum. The distribution of subicular projections was studied in detail by means of the sensitive anterograde tracer Phaseolus vulgaris-leucoagglutinin (PHA-L), and the precise origin of these projections analysed with retrogradely transported fluorescent tracers, using double- and triple-labelling protocols. Injections of PHA-L in the proximal part of the dorsal subiculum, i.e. that part which borders field CA1, result in labelling of the infralimbic, entorhinal and perirhinal cortices, the nucleus accumbens and the lateral septal region, the interanteromedial nucleus of the thalamus, the core of the nucleus gelatinosus, and the mammillary nuclei, in particular in the rostral parts of the medial nucleus. In contrast, injections in the distal part of the dorsal subiculum, i.e. that part which borders the presubiculum, give rise to labelling in the retrosplenial and postrhinal cortices, the presubiculum, the anterior thalamic complex, the shell of the nucleus gelatinosus, and the mammillary nuclei, preferentially in the caudal part of the medial nucleus. The results of injections of different retrograde tracers, simultaneously placed in two or three targets of the subicular efferents, confirm the results of the anterograde tracing experiments. Moreover, they clearly demonstrate that the population of subicular neurons which, for example, projects to the nucleus accumbens and the interanteromedial nucleus of the thalamus is almost completely segregated from the population that projects to the retrosplenial cortex and the anterior complex of the thalamus. Thus within the dorsal subiculum, populations of neurons can be differentiated so that each population projects to a unique set of target structures. These cell populations are differentially positioned along the proximo-distal axis. In view of additional evidence indicating that some of the major afferents to the subiculum are organized along the same axis, we suggest that the heterogeneity of the dorsal subiculum along the proximo-distal axis reflects a general organizational characteristic of this hippocampal field.


Finding your unique path in science.
I always found it curious that in science, we value unique, creative thinkers, but we teach scientists to progress in a formulaic manner that rarely takes each person's individual strengths into account. Surprisingly, when we break the mold, we are often rewarded for it. This cycle of learning to survive using conventional wisdom but being rewarded for a unique path outside of it seems to be an unspoken key to success. I am honored to be awarded the 2020 Women in Cell Biology Junior Award for Excellence in Research and am thrilled to share some of the unconventional guiding principles that brought me to where I am in this rich scientific landscape. The game changers in the early phase of my career were informal mentors, open scientific communication, and persistence in pursuing difficult scientific questions.


Different durations of whole raw soybean supplementation during the prepartum period: Milk fatty acid profile and oocyte and embryo quality of early-lactating Holstein cows.
The objective of this study was to evaluate different durations of whole raw soybeans (WS) supplementation during the prepartum period on nutrient digestibility, milk yield and composition, energy balance, blood metabolites, and oocyte and embryo quality of transition cows. Thirty-one Holstein cows were used in a completely randomized design and assigned to 4 experimental groups (G): G90, G60, G30, and G0 (control), supplemented with a diet containing 12% of WS from 90, 60, 30, and 0 d relative to the calving date, respectively. Cows were dried off 60 d before the expected calving date. After parturition, all cows were fed a diet containing 12% of WS until 84 DIM. Blood samples were collected on d -49, -35, -21, -14, -7, 0, 7, 14, 21, 35, and 70 relative to partum. Ovum pick-ups were performed on d 21 ± 3, 42 ± 7, 63 ± 7, and 84 ± 7 of lactation. Different durations of WS supplementation did not affect DMI and apparent total-tract digestibility in either the pre- or postpartum periods. Duration of WS supplementation had no effect on milk yield and composition nor energy balance of cows. However, the duration of WS supplementation had several effects on milk fatty acid (FA) profile of cows, including a linear decrease in concentrations of cis-9 C18:1, unsaturated C18, total monounsaturated, and unsaturated FA. Further, the milk contents of cis-9,cis-12 C18:2 FA, cis-9,trans-11 C18:2 FA, and total polyunsaturated FA were increased when WS were fed to cows from 30 d but not from 60 or 90 d of the expected calving date. The length of WS supplementation in the prepartum period linearly increased blood cholesterol concentration of cows during the prepartum period, but it had no effect on blood glucose and nonesterified FA concentrations in the pre- and postpartum periods. Duration of WS supplementation during the prepartum period increased the average number of grade 2 oocytes, notably in G60, but it had no effect on embryo production and cleavage proportion of early-lactation cows. The duration of WS supplementation in the prepartum period had no effect on milk yield and energy balance of the subsequent lactation, but it altered milk FA profile in early lactation by decreasing unsaturated FA content, notably when starting to supplement WS at 90 and 60 d from the expected calving date. Our results also showed that the duration of WS supplementation during the prepartum period does not improve oocyte quality in the subsequent lactation of cows.


Prediction of quantitative calibration factors of some organic compounds in gas chromatography.
A quantitative structure-property relationship (QSPR) methodology that involves multilinear (Hansch-type) and nonlinear (radial basis function neural network (RBFNN)) approaches was performed to correlate the quantitative molar calibration factors (f(M)) of 140 organic compounds against structural factors. The statistical characteristics provided by the multiple linear model (R(2) = 0.963; RMS = 0.089; AARD = 3.86% for test set) indicated satisfactory stability and predictive ability, while the predictive ability of the RBFNN model is somewhat superior (R(2) = 0.983; RMS = 0.075; AARD = 3.19% for test set). The multilinear model provided some insight into the main structure factors that modulate the quantitative calibration factor of the investigated compounds.


A form of circulating ICAM-1 in human serum.
A circulating form of the usually membrane-bound intercellular adhesion molecule-1 (ICAM-1) was identified and characterized in normal human serum, and in sera from patients with leukocyte adhesion deficiency (LAD). The molecule, designated circulating ICAM-1 (cICAM-1) was detected and quantitated by sandwich ELISA. Levels of cICAM-1 in sera from normal individuals ranged from 100 to 200 ng/ml. Sera from LAD patients had elevated cICAM-1 levels ranging from 200 to 700 ng/ml. The elevated levels of cICAM-1 in LAD sera may be due to an inability to adsorb cICAM-1 to cell-bound LFA-1 or may be an indirect result of the pathology accompanying the syndrome. cICAM-1 bound to mAb specific for four distinct ICAM-1 epitopes localized in domains D1, D2, D4, and D5, and displayed similar molecular size properties as recombinant soluble ICAM-1 on FPLC size-exclusion chromatography. When immobilized via a domain D5-specific mAb, cICAM-1 mediated function (LFA-1)-dependent lymphocyte adhesion equivalent to sICAM-1. These data indicate that cICAM-1 contains most, if not all, of the five extracellular domains of membrane ICAM-1, as well as the ability to bind specifically to LFA-1. The cellular source of cICAM-1 appeared to be from mononuclear cells; only lymphoid cell lines or primary PBMC cultures had detectable levels of cICAM-1 in cell culture supernatants. Because cICAM-1 retains the ability to bind specifically to LFA-1, it may act to regulate cell adhesion by promoting de-adhesion. Alternatively, cICAM-1 may be the indirect consequence of inflammation or tissue damage. As such, the detection of cICAM-1 could be useful as a marker of inflammatory disease.


Methoxyamine-induced mutagenesis of nucleic acids. A proton NMR study of oligonucleotides containing N4-methoxycytosine paired with adenine or guanine.
We report the solution structure of two heptanucleotides each containing a central N4-methoxycytosine, in one case paired with adenine on the opposite strand and the other with guanine. For the N4-methoxycytosine adenine pair, only the imino form of the N4-methoxycytosine residue is observed and base pairing is in Watson-Crick geometry. However, rotation of the methoxy group about the N-OCH3 bond is not constrained to a particular orientation although it must be anti to the N3 of N4-methoxycytosine. The slow exchange on a proton NMR time scale between the single strand and double strand forms is attributed to the strong preference of the cis conformation of the OCH3 group in the single strand, which inhibits base pair formation. For the N4-methoxycytosine that is base paired with guanine, we observe an amino form in Watson-Crick geometry in slow exchange with a base paired imino form in wobble geometry. The amino form is predominant at low temperature whereas the imino form predominates above 313 K. We have measured the exchange rate between the two forms at 303 K and observed a value of approximately 1 S-1. The relative ratio of amino and imino forms of N4-methoxycytosine is influenced by both the base that is in front and the temperature. Our results explain the preferential replacement of dTTP by N4-methoxycytosine in primer elongation.


Ambulatory arterial stiffness index and 24-hour ambulatory pulse pressure as predictors of mortality in Ohasama, Japan.
Ambulatory arterial stiffness index (AASI) and pulse pressure (PP) are indexes of arterial stiffness and can be computed from 24-hour blood pressure recordings. We investigated the prognostic value of AASI and PP in relation to fatal outcomes. In 1542 Ohasama residents (baseline age, 40 to 93 years; 63.4% women), we applied Cox regression to relate mortality to AASI and PP while adjusting for sex, age, BMI, 24-hour MAP, smoking and drinking habits, diabetes mellitus, and a history of cardiovascular disease. During 13.3 years (median), 126 cardiovascular and 63 stroke deaths occurred. The sex- and age-standardized incidence rates of cardiovascular and stroke mortality across quartiles were U-shaped for AASI and J-shaped for PP. Across quartiles, the multivariate-adjusted hazard ratios for cardiovascular and stroke death significantly deviated from those in the whole population in a U-shaped fashion for AASI, whereas for PP, none of the HRs departed from the overall risk. The hazard ratios for cardiovascular mortality across ascending AASI quartiles were 1.40 (P=0.04), 0.82 (P=0.25), 0.64 (P=0.01), and 1.35 (P=0.03). Additional adjustment of AASI for PP and sensitivity analyses by sex, excluding patients on antihypertensive treatment or with a history of cardiovascular disease, or censoring deaths occurring within 2 years of enrollment, produced confirmatory results. In a Japanese population, AASI predicted cardiovascular and stroke mortality over and beyond PP and other risk factors, whereas in adjusted analyses, PP did not carry any prognostic information.


PRMT5-mediated H4R3sme2 Confers Cell Differentiation in Pediatric B-cell Precursor Acute Lymphoblastic Leukemia.
Little is known about the function of histone arginine methylation in acute lymphoblastic leukemia (ALL). The objective was to evaluate whether protein arginine methyltransferase 5 (PRMT5) plays a role in pediatric ALL and to determine the possible mechanism of epigenetic regulation. We used bone marrow samples from patients with pediatric ALL, the Nalm6 cell line, mature B-cell lines, and mouse xenograft models to evaluate the function of PRMT5 in ALL tumorigenesis. This study showed that PRMT5 and the symmetric dimethylation of H4R3 (H4R3sme2) were upregulated in most initially diagnosed (n = 15; 100%) and relapsed (n = 4; 75%) bone marrow leukemia cells from patients with pediatric B-cell precursor ALL (BCP-ALL) and were decreased when the disease was in remission (n = 15; 6.7%). Downregulation of H4R3sme2 by PRMT5 silencing induced BCP-ALL cell differentiation from the pre-B to immature B stage, whereas overexpressed PRMT5 with enhanced H4R3sme2 promoted human mature B cells to dedifferentiate back to the pre-B II/immature B stages in vitro. High PRMT5 expression enhanced the proportion of CD43+/B220+/sIgM- B leukocytes in recipient mice. CLC and CTSB were identified as potential target genes of PRMT5 in BCP-ALL cells and were inhibited by H4R3sme2 in gene promoters. We demonstrate that enhanced PRMT5 promotes BCP-ALL leukemogenesis partially by the dysregulation of B-cell lineage differentiation. H4R3sme2 and PRMT5 may serve as potential sensitive biomarkers of pediatric BCP-ALL. Suppression of the activation of PRMT5 may offer a promising therapeutic strategy against pediatric BCP-ALL.


[A social cultural model of the "regular period" in the sex life of men].
In the experimental-testing examination of 957 patients, statistically significant results of summing up the information relevant to aspects of regular sexual intercourse in male subjects were obtained, which give good grounds for believing that the illicit/secret sexual relationship model prevailing in modern society is indeed that very social and psychological base on which further rise in venereal case rate is recordable among the population of Ukraine. And if we wish some kind of effective measures to be instituted if only to stabilize the level of venereal morbidity in the population we will certainly have to work out a comprehensive long-term state programme aimed to change motivation patterns of the social and culturological model of caring on extramarital sexual life.


[The trypsin-antitrypsin system and the content of blood middle-molecular peptides in experimental traumatic shock].
It was established that the activity of trypsin and the content of its inhibitors--alpha 1-inhibitor of proteinases and alpha 2-macroglobulin--undergo phase changes of different meaning and direction with the gradual development of traumatic shock. The character of these changes indicates that trypsinemia is an important pathogenetic link of traumatic shock and corresponds to its severity. The compensatory activation of the trypsin inhibitors is directed to increasing the resistance of the body to trypsinemia. Deficiency of the inhibition system leads to activation of organic proteolysis and growth of the content of middle-molecular peptides in the blood. Such a character of homeostasis disorders substantiates the need for the search for effective measures of correction of the trypsin-antitrypsin system and freeing the organism of the excess of middle-molecular molecules.


Meschiidae, a new family of Lygaeoidea (Hemiptera: Heteroptera) from India and Australia, with descriptions of a new genus and two new species .
Meschiidae fam. nov. is erected for the genera Meschia Distant, 1910, containing four species: M. pugnax Distant, 1910 and M. quadrimaculata Distant, 1910 from India, M. woodwardi Scudder, 1957 from Queensland, Australia, and M. barrowensis sp. nov. from Barrow Island, Western Australia, and the related Neomeschia gen. nov. with type species N. queenslandensis sp. nov. from Queensland, Australia. A key to all Australian species and genera is provided. Redescriptions are provided for Meschia, its type species M. pugnax, and M. woodwardi. Relationships, and reasons for the new family placement of Meschia, are discussed.


[German publications in leading English language journals].
The per capita publication output of German medical research is low compared not only to the English-speaking nations but also to Austria and Switzerland. We assessed the publications from German universities in the leading English language general medical journals: New England Journal of Medicine (NEJM), Lancet, Journal of the American Medical Association (JAMA), and British Medical Journal (BMJ). Medline (1988-5/2000) was searched for journal-articles with German origin published in the above journals and manually classified as to institution and speciality. 372 publications from Germany were found. 216 had been published in Lancet, 87 in NEJM, 52 in BMJ, and 17 in JAMA. 58 were excluded (2 from British military hospitals in Germany, 2 from pharmaceutical companies, 2 from physician associations, 18 from non-university basic research institutions or government agencies, 3 from non-medical university departments, 30 from non-university hospitals, 1 from a transfusion service of the German Red Cross). The remaining 314 publications were from 34 universities. The most speciality was internal Medicine with 140 university publications, the most represented university was the Ludwig-Maximilians-University in Munich with 43 publications. 5 universities had no publications in the analysed journals. Large differences were found between the German universities with respect to publications in the leading English Language general medical journals.


The Potential for Reducing Lynch Syndrome Cancer Risk with Nutritional Nrf2 Activators.
Lynch syndrome (LS), is an autosomal dominant disorder predisposing patients to multiple cancers, predominantly colorectal (CRC) and endometrial, and is implicated in 2-4% of all CRC cases. LS is characterized by mutations of four mismatch repair (MMR) genes which code for proteins responsible for recognizing and repairing DNA lesions occurring through multiple mechanisms including oxidative stress (OS). Increased OS can cause DNA mutations and is considered carcinogenic. Due to reduced MMR activity, LS patients have an increased risk of cancer as a result of a decreased ability to recognize and repair DNA lesions caused by OS. Due to its carcinogenic properties, reducing the level of OS may reduce the risk of cancer. Nutritional Nrf2 activators have been shown to reduce the risk of carcinogenesis in the general population through activation of the endogenous antioxidant system. Common nutritional Nrf2 activators include sulforaphane, curcumin, DATS, quercetin, resveratrol, and EGCG. Since LS patients are more susceptible to carcinogenesis caused by OS, it is hypothesized that nutritional Nrf2 activators may have the potential to reduce the risk of cancer in those with LS by modulating OS and inflammation. The purpose of this paper is to review the available evidence in support of this statement.


Is a Drain Tip Culture Required After Spinal Surgery?
The efficacy of use of a drain tip culture for early detection of surgical-site infection (SSI) was investigated in 329 patients after spinal surgery. To examine the efficacy of a wound drain tip culture for detection of SSI in spinal surgery. A complication of SSI after spinal surgery has high associated morbidity and mortality, and is often difficult to treat. The subjects were patients who underwent spinal surgery at our institution between January 2010 and March 2013. All subjects were treated with antimicrobial prophylaxis based on evidence-based guidelines and were followed for at least 6 months after surgery. Data from culture studies using the distal tip of the wound drain were used for analysis. Drain tip cultures were positive in 34 cases and there were 19 SSIs. Ten of the 34-tip culture-positive wounds developed SSI. Drain tip cultures had a sensitivity of 52%, specificity of 92%, positive predictive value (PPV) of 29%, and negative predictive value of 97% for predicting a wound infection. The association between a positive suction tip culture and wound infection was significant (P<0.05). The PPV for SSI was 60% in cases in which methicillin-resistant bacteria were detected in a drain tip, and the SSI rate in these cases differed significantly compared with those with non-methicillin-resistant bacteria (P=0.01). A drain tip culture is useful for early detection of SSI caused by methicillin-resistant bacteria.


Supporting research and development in ambulance services: research for better health care in prehospital settings.
This paper discusses recent developments in research support for ambulance trusts in England and Wales and how this could be designed to lead to better implementation, collaboration in and initiation of high-quality research to support a truly evidence-based service. The National Ambulance Research Steering Group was set up in 2007 to establish the strategic direction for involvement of regional ambulance services in developing relevant and well-designed research for improving the quality of services to patients. Ambulance services have been working together and with academic partners to implement research and to participate, collaborate and lead the design of research that is relevant for patients and ambulance services. New structures to support the strategic development of ambulance and prehospital research will help address gaps in the evidence for health interventions and service delivery in prehospital and ambulance care and ensure that ambulance services can increase their capacity and capability for high-quality research.


[Assessing nurses' knowledge of insulin administration and the impact of the introduction of concentrated insulins.]
Insulin is a high-risk medication, and even slight changes in blood levels can lead to serious side effects or can even result in death. Error in administering drugs is one of the main causes of over- or under-dosing, and the recent introduction of concentrated insulins (CI) has increased this risk. We assessed nurses' knowledge of these CI, their beliefs about the "insulin unit" (IU), and the impact that this knowledge had on the risk of making medication errors. A direct interview survey was conducted in eight departments of medicine and surgery in a university hospital. Sixty-eight nurses and midwives were interviewed. Twenty-six percent of them had already encountered a CI prescription and only 51.5 percent correctly defined the notion of IU. Only 18 percent responded correctly to a practical case of a CI prescription, whilst 35 percent multiplied the dose by two and 24 percent divided it by two. Sixty percent indicated that they regularly use a U-100 graduated insulin syringe to withdraw insulin from the pen. Insulin administration errors related to this misuse, which are very well documented in the literature, are linked to nurses' lack of knowledge about the true definition of IU. These administration errors have increased with the introduction of concentrated insulins.


A multifaceted investigation into molecular associations of chronic thromboembolic pulmonary hypertension pathogenesis.
Chronic thromboembolic pulmonary hypertension is characterized by incomplete thrombus resolution following acute pulmonary embolism, leading to pulmonary hypertension and right ventricular dysfunction. Conditions such as thrombophilias, dysfibrinogenemias, and inflammatory states have been associated with chronic thromboembolic pulmonary hypertension, but molecular mechanisms underlying this disease are poorly understood. We sought to characterize the molecular and functional features associated with chronic thromboembolic pulmonary hypertension using a multifaceted approach. We utilized functional assays to compare clot lysis times between chronic thromboembolic pulmonary hypertension patients and multiple controls. We then performed immunohistochemical characterization of tissue from chronic thromboembolic pulmonary hypertension, pulmonary arterial hypertension, and healthy controls, and examined RNA expression patterns of cultured lymphocytes and pulmonary arterial specimens. We then confirmed RNA expression changes using immunohistochemistry, immunofluorescence, and Western blotting in pulmonary arterial tissue. Clot lysis times in chronic thromboembolic pulmonary hypertension patients are similar to multiple controls. Chronic thromboembolic pulmonary hypertension endarterectomized tissue has reduced expression of both smooth muscle and endothelial cell markers. RNA expression profiles in pulmonary arteries and peripheral blood lymphocytes identified differences in RNA transcript levels related to inflammation and growth factor signaling, which we confirmed using immunohistochemistry. Gene expression data also suggested significant alterations in metabolic pathways, and immunofluorescence and Western blot experiments confirmed that unglycosylated CD36 and adiponectin expression were increased in chronic thromboembolic pulmonary hypertension versus controls. Our data do not support impaired clot lysis underlying chronic thromboembolic pulmonary hypertension, but did demonstrate distinct molecular patterns present both in peripheral blood and in pathologic specimens of chronic thromboembolic pulmonary hypertension patients suggesting that altered metabolism may play a role in chronic thromboembolic pulmonary hypertension pathogenesis.


Role of DNA gyrase and topoisomerase IV mutations in fluoroquinolone resistance of Capnocytophaga spp. clinical isolates and laboratory mutants.
Capnocytophaga spp. are often reported to cause bacteraemia and extra-oral infections and are characterized by their significant contribution to resistance to β-lactam and macrolide-lincosamide-streptogramin antibiotics in the human oral microbiota. The implication of mutations in the quinolone resistance-determining region (QRDR) of DNA gyrase A and B ( gyrA and gyrB ) and topoisomerase IV ( parC and parE ) of fluoroquinolone (FQ)-resistant Capnocytophaga spp., hitherto unknown, was explored in this study. Two reference strains ( Capnocytophaga gingivalis ATCC 33624 and Capnocytophaga sputigena ATCC 33612) and four Capnocytophaga spp. isolated from clinical samples were studied. Nine in vitro FQ-resistant mutants, derived from two reference strains and one FQ-susceptible clinical isolate, were selected by successive inoculations onto medium containing levofloxacin. MICs of ofloxacin, norfloxacin, ciprofloxacin, levofloxacin and moxifloxacin were determined. The presumed QRDRs of GyrA, GyrB, ParC and ParE from Capnocytophaga spp. were determined by sequence homology to Bacteroides fragilis and Escherichia coli . PCR primers were designed to amplify the presumed QRDR genetic region of Capnocytophaga spp. and sequence analyses were performed using the BLAST program at the National Center for Biotechnology Information. gyrA mutations leading to a substitution from amino acid position 80 to 86 were systematically detected in Capnocytophaga spp. with ciprofloxacin MIC >1 mg/L and considered as the primary target of FQs. No mutational alteration in the QRDR of gyrB was detected. Other mutations in parC and parE led to spontaneous amino acid substitutions of DNA topoisomerase IV subunit B with no alteration in FQ susceptibility.


Intracellular green fluorescent protein-polyalanine aggregates are associated with cell death.
Eight diseases, exemplified by Huntington's disease and spinocerebellar ataxia type 1, are caused by CAG-repeat expansion mutations. The CAG repeats are translated into expanded polyglutamine tracts, which are associated with deleterious novel functions. While these diseases are characterized by intraneuronal aggregate formation, it is unclear whether the aggregates cause disease. We have addressed this debate by generating intracellular aggregates with green fluorescent protein (GFP) fused to 19-37 alanines. No aggregates were seen in cells expressing native GFP or GFP fused to seven alanines. Aggregate-containing cells expressing GFP fused to 19-37 polyalanines show high rates of nuclear fragmentation compared with cells expressing the same constructs without aggregates, or cells expressing GFP fused to seven alanines. This suggests an association between aggregate formation and cell death.


[Effect of pentagastrin on the myoelectric activity of the pyloric sphincter, body of the stomach and the duodenum].
In chronic experiments on rabbits, i.v. administration of the pentagastrin (0.01-1.0 micrograms/kg) exerted inhibitory effects on the myoelectrical activity of the pylorus and stomach, whereas the effect of pentagastrin on the duodenum was rather activating. The threshold dose of pentagastrin for pylorus was 5 times smaller than for the duodenum and 10 times smaller than for the stomach. Possible mechanisms of the pentagastrin effect on the contractile activity of the pylorus, stomach and duodenum smooth muscles, are discussed.


Comparison of markers of bone formation and resorption in prostate cancer patients to predict bone metastasis.
We investigated the usefulness of two biochemical markers of bone formation (PICP, the carboxy-terminal propeptide of type I procollagen, and bone ALP, bone-derived alkaline phosphatase) and a marker of bone resorption (ICTP, the carboxy-terminal telopeptide of type I collagen), to determine whether the presence of bone metastasis in prostate cancer could be evaluated and the extent of bone metastasis could be stratified by the serum levels of these markers, compared to total alkaline phosphatase (T-ALP) and prostate-specific antigen (PSA). The serum levels of PICP, bone ALP, ICTP, T-ALP and PSA were significantly higher in patients with both prostate cancer and bone metastasis (n=49) than in patients with benign prostatic hyperplasia (n=35) and patients with prostate cancer without bone metastasis (n=70). The superiority of a marker in the rate of detection of bone metastasis was evaluated with receiver operating characteristic curves. The serum marker levels were compared as a function of metastatic burden in bone (i.e., the extent of disease, EOD grade). We found that bone ALP is the most suitable marker for evaluating bone metastasis, especially for stratifying the degree of bone metastasis. Both PICP and ICTP were useful in this respect, but rather inferior to bone ALP. T-ALP had the lowest ability for detecting bone metastasis, but its correlation with the EOD grade was excellent, second to that of bone ALP. PSA showed limited reliability for stratifying the extent of bone metastasis.


Evaluating Glaucoma Treatment Effect on Intraocular Pressure Reduction Using Propensity Score Weighted Regression.
Observational studies in glaucoma patients can provide important evidence on treatment effects, especially for combination therapies which are often used in reality. But the success relies on the reduction of selection bias through methods such as propensity score (PS) weighting. The objective of this study was to assess the effects of five glaucoma treatments (medication, laser, non-laser surgery (NLS), laser + medication, and NLS + medication) on 1-year intraocular pressure (IOP) change. Data were collected from 90 glaucoma subjects who underwent a single laser, or NLS intervention, and/or took the same medication for at least 6 months, and had IOP measures before the treatment and 12-months after. Baseline IOP was significantly different across groups (p = 0.007) and this unbalance was successfully corrected by the PS weighting (p = 0.81). All groups showed statistically significant PS-weighted IOP reductions, with the largest reduction in NLS group (-6.78 mmHg). Baseline IOP significantly interacted with treatments (p = 0.03), and at high baseline IOP medication was less effective than other treatments. Our findings showed that the 1-year IOP reduction differed across treatment groups and was dependent on baseline IOP. The use of PS-weighted methods reduced treatment selection bias at baseline and allowed valid assessment of the treatment effect in an observational study.


Endoscopic foregut surgery and interventions: The future is now. The state-of-the-art and my personal journey.
In this paper, I reviewed the emerging field of endoscopic surgery and present data supporting the contention that endoscopy can now be used to treat many foregut diseases that have been traditionally treated surgically. Within each topic, the content will progress as follows: "lessons learned", "technical considerations" and "future opportunities". Lessons learned will provide a brief background and update on the most current literature. Technical considerations will include my personal experience, including tips and tricks that I have learned over the years. Finally, future opportunities will address current unmet needs and potential new areas of development. The foregut is defined as "the upper part of the embryonic alimentary canal from which the pharynx, esophagus, lung, stomach, liver, pancreas, and part of the duodenum develop". Foregut surgery is well established in treating conditions such as gastroesophageal reflux disease (GERD), achalasia, esophageal diverticula, Barrett's esophagus (BE) and esophageal cancer, stomach cancer, gastric-outlet obstruction, and obesity. Over the past decade, remarkable progress in interventional endoscopy has culminated in the conceptualization and practice of endoscopic foregut surgery for various clinical conditions summarized in this paper. Regarding GERD, there are now several technologies available to effectively treat it and potentially eliminate symptoms, and the need for long-term treatment with proton pump inhibitors. For the first time, fundoplication can be performed without the need for open or laparoscopic surgery. Long-term data going out 5-10 years are now emerging showing extended durability. In respect to achalasia, per-oral endoscopic myotomy (POEM) which was developed in Japan, has become an alternative to the traditional Heller's myotomy. Recent meta-analysis show that POEM may have better results than Heller, but the issue of post-POEM GERD still needs to be addressed. There is now a resurgence of endoscopic treatment of Zenker's diverticula with improved technique (Z-POEM) and equipment; thus, patients are choosing flexible endoscopic treatment as opposed to open or rigid endoscopy options. In regard to BE, endoscopic submucosal dissection (ESD) which is well established in Asia, is now becoming more mainstream in the West for the treatment of BE with high grade dysplasia, as well as early esophageal cancer. In combination with all the ablation technologies (radiofrequency ablation, cryotherapy, hybrid argon plasma coagulation), the entire spectrum of Barrett's and related dysplasia and early cancer can be managed predominantly by endoscopy. Importantly, in regard to early gastric cancer and submucosal tumors (SMTs) of the stomach, ESD and full thickness resection (FTR) can excise these lesions en-bloc and endoscopic suturing is now used to close large defects and perforations. For treatment of patients with malignant gastric outlet obstruction (GOO), endoscopic gastro-jejunostomy is now showing better results than enteral stenting. G-POEM is also emerging as a treatment option for patients with gastroparesis. Obesity has become an epidemic in many western countries and is becoming also prevalent in Asia. Endoscopic sleeve gastroplasty (ESG) is now becoming an established treatment option, especially for obese patients with body mass index between 30 and 35. Data show an average weight loss of 16 kg after ESG with long-term data confirming sustainability. Finally, in respect to endo-hepatology, there are many new endoscopic interventions that have been developed for patients with liver disease. Endoscopic ultrasound (EUS)-guided liver biopsy and EUS-guided portal pressure measurement are exciting new frontiers for the endo-hepatologists.


Effect of norepinephrine on isolated coronary artery of normal and myocardial infarcted pigs.
The effect of norepinephrine, the adrenergic neurotransmitter, on isolated coronary arterial strips of normal and myocardial infarcted pigs was investigated with the view to characterize the nature of adrenoceptors in this tissue. In small, as well as large, isolated coronary arterial strips of normal pigs norepinephrine produced relaxation which was found to be mediated through activation of beta 1-adrenoceptors. While alpha adrenoceptors were also found to be present in large coronary arteries, they were absent in the small coronary arteries. In isolated coronary arterial strips dissected out from pigs in which myocardial infarction was induced, norepinephrine produced contraction, its alpha-adrenergic effect. This could be due to arterial hypoxia and/or sensitization of alpha-receptors present in the coronary artery.


Printed High-Adhesion Flexible Electrodes Based on an Interlocking Structure for Self-Powered Intelligent Movement Monitoring.
Two-dimensional transition metal carbide nitrides (MXenes) have been extensively explored in diverse areas, such as electrochemical energy storage and flexible electronics. Although the solution-processed MXene-based device has made significant achievements, it is still a challenge to develop large-scale and high-resolution printing methods for flexible printed electronics. In this work, we reported a novel strategy of a porous interlocking structure to obtain flexible MXene/laser-induced graphene (LMX) composite electrodes with enhanced adhesion and high printing resolution. In comparison to traditional printed MXene electrodes, the LMX electrode with an interlocking interface possesses enhanced mechanical properties (adhesive strength of 2.17 MPa) and comparable electrical properties (0.68 S/mm). Furthermore, owing to the outstanding stability and flexibility, the LMX-based triboelectric nanogenerator (TENG) can be used as a self-powered sensor to monitor finger-bending movement. A support vector machine (SVM)-assisted self-powered motion sensor can distinguish the bending angle with high recognition accuracy and can effectively identify different angles. The successful experience of directly printing flexible electrodes with excellent mechanical and electrical properties can be promoted to other solution-processed two-dimensional materials. Our strategy opens up a promising perspective to develop flexible and printed electronics.


Tumor-related epilepsy in children.
A 10-year retrospective review of 15 children with cerebral tumors and seizures was conducted to study the factors responsible for delay in the diagnosis of tumors and to assess outcome following surgery. Mean duration of seizures prior to surgery was 37 months. Ninety-three percent had no focal neurologic deficits. Head computed tomography was abnormal in 64%, whereas magnetic resonance imaging was abnormal in all patients. Electroencephalography showed focal abnormalities ipsilateral to the tumor in 73%. There was no surgical mortality. Eighty percent were seizure free or had rare seizures following surgery. Factors contributing to a delayed diagnosis of the brain tumor included a nonfocal neurologic examination and delay in obtaining an appropriate neuroimaging study. We believe that head magnetic resonance imaging should be the investigation of choice in partial epilepsies.


Protective Effects of Astragaloside IV Combined with Budesonide in Bronchitis in Rats by Regulation of Nrf2/Keap1 Pathway.
BACKGROUND This study was conducted to evaluate the effects of astragaloside IV and budesonide on bronchitis in rats and to explore the mechanism involved. MATERIAL AND METHODS Eighty Sprague-Dawley (SD) rats were randomly divided into 5 groups, including a Bronchitis model group (BM), a Budesonide group (BG), an Astragaloside IV group (AG), an Astragaloside IV combined with Budesonide group (CG), and a blank control group (BC). Lung tissue was stained with hematoxylin and eosin (H&E). The activity of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) were detected by enzyme-linked immunosorbent assay (ELISA). The nuclear factor erythroid 2 [NF-E2]-related factor 2 (Nrf2), Kelch-like erythroid cell-derived protein with CNC homology [ECH]-associated protein 1 (Keap1), BTB and CNC homology 1 (Bach1), B-cell lymphoma-2(Bcl-2), and BCl-2-associated X protein (Bax) mRNA and protein were examined by RT-PCR and Western blot, respectively. RESULTS Compared with the Bronchitis model group, the lung tissue lesions in the Budesonide group, Astragaloside IV group, and Astragaloside IV combined with Budesonide group were effectively ameliorated and the airway resistance was significantly decreased. The activities of SOD, GSH-Px, and CAT were increased after treatment with drugs, while the content of MDA was decreased. The levels of Nrf2, Keap1, and Bcl-2 proteins were increased and the levels of Bach1 and Bax were decreased after treatment with Budesonide and Astragaloside IV. CONCLUSIONS Astragaloside IV combined with budesonide can ameliorate the lesions caused by bronchitis in rats through activating the Nrf2/Keap1 pathway, which plays a protective role on anti-oxidative stress injury.


Design and Evaluation of Transdermal Patches of Timolol Maleate.
Continuous intravenous infusion of controlled drug delivery has certain risks. This could be diligently duplicated devoid of its hassles by using the skin as the port of drug entry. Transdermal drug delivery system is the main route with discrete, self-contained dosage forms when placed on the skin, transporting the medicament through the skin into the systemic circulation in a wellcontrolled manner. The rationale of the current work was to formulate and evaluate a transdermal patch of an antihypertensive drug by using different grades of polymers with a view to circumvent the hepatic first pass metabolism and also to escalate its bioavailability. Solvent-casting method was used to prepare transdermal patches of timolol maleate using Eudragit RL100, Eudragit RS100, ethyl cellulose as polymers, and dibutyl phthalate as the plasticizer. The formulated patches were evaluated for their physiochemical parameters such as folding endurance, percentage moisture content, thickness, and water vapour transmission. The formulated patches were subjected to in-vitro permeation studies by using a Franz diffusion cell with a dialysis sac. The optimized formulation chosen on the basis of physiochemical characteristics and in-vitro studies was subjected to in-vivo studies on methyl prednisolone acetate-induced hypertensive rats. The data from release kinetics disclosed that the Korsmayer-Peppas could be the best fitting model. The results obtained from in-vitro studies disclosed that formulation with high proportion of Eudragit grade RL100 in acetone solvent system exhibited better drug release compared to rest of the formulations. The output obtained from in-vivo studies performed on rats revealed that the optimized formulation showed decrease in blood pressure from 158.53 ± 0.39 to 128.91 ± 0.50 mmHg. It was concluded that timolol maleate patch could be formulated into a matrix-type transdermal patch for the management of hypertension.


Adrenergic mechanisms in canine intralobar pulmonary arteries and veins.
The responses of canine intralobar pulmonary arteries (IPA) and veins (IPV) to transmural nerve stimulation (TNS), and exogenously administered norepinephrine (NE) were studied to evaluate the alpha-adrenergic neuroeffector system in the canine pulmonary vasculature. IPA and IPV elicited contractions in response to both NE (10(-10) to 10(-5) M) and TNS (0.5-32 Hz, 2 ms duration and delay). The equilibrium (steady state) contractile responses of IPA and IPV to TNS were abolished with the adrenergic neuronal blocking agents guanethidine and bretylium and the depolarization blocking agent tetrodotoxin in concentrations which did not affect the responses to NE or KCl. The contractile responses of IPA and IPV to TNS and NE were reduced in a concentration-dependent way, by the alpha-adrenergic receptor-blocking agents phentolamine, tolazoline and clonidine. The contractile responses of IPA and IPV to TNS and NE were enhanced after inhibition of neuronal reuptake of NE (uptake1) with cocaine, as well as after blockade of extraneuronal reuptake of NE (uptake2) with hydrocortisone. Analysis of the equilibrium responses of the IPA and IPV to TNS and NE with an Arunlakshana-Schild plot to define the concentration of alpha 1-receptor antagonists necessary to double the ED50 for TNS and NE (defined as the pA2), demonstrated that the postsynaptic alpha-receptors of IPV differed from that of IPA. These data support the conclusions that IPA and IPV 1) contain a functional adrenergic innervation and neuroeffector system, 2) contract in response to both TNS and NE, 3) demonstrate the presence of mechanisms for both uptake1 and uptake2 of NE, and 4) IPV contain postsynaptic alpha-receptors that may differ from each other.


[The effect of a low dose of piracetam on the activity of the dopaminergic system in the rat striatum].
The low-dose effect (100 mg/kg, intraperitoneally) of the nootropic drug pyracetam on some DA-ergic neurochemical parameters of the rat striatum, as well as on the locomotion activity of rats were studied using the "open-field" test. It was shown that pyracetam (l mM) in vitro increases the K(+)-stimulated (28 mM) DA release from the perfused isolated striatum to 148 +/- 14 pmole/mg tissue compared to the control animals: 101 +/- 10 pmole /mg (p < 0.05, Student's t-test). Pyracetam in a dose of 100 mg/kg increased the DA level and decreased the 5-HT level in the striatum homogenates: DA- to 121% and 5-HT-to 81% (p < 0.05), respectively. The content of DOPAC, HVA and 5-HIAA in the tissue remained the same. In addition to the mentioned effects pyracetam promoted the locomotion activity of rats in the "open field" -putative behavioral marker of the striatum DA-ergic function. Thus pyracetam is capable of modifying the DA-ergic activity of the rat striatum, thus stimulating the neuromediator release.


Group foraging increases foraging efficiency in a piscivorous diver, the African penguin.
Marine piscivores have evolved a variety of morphological and behavioural adaptations, including group foraging, to optimize foraging efficiency when targeting shoaling fish. For penguins that are known to associate at sea and feed on these prey resources, there is nonetheless a lack of empirical evidence to support improved foraging efficiency when foraging with conspecifics. We examined the hunting strategies and foraging performance of breeding African penguins equipped with animal-borne video recorders. Individuals pursued both solitary as well as schooling pelagic fish, and demonstrated independent as well as group foraging behaviour. The most profitable foraging involved herding of fish schools upwards during the ascent phase of a dive where most catches constituted depolarized fish. Catch-per-unit-effort was significantly improved when targeting fish schools as opposed to single fish, especially when foraging in groups. In contrast to more generalist penguin species, African penguins appear to have evolved specialist hunting strategies closely linked to their primary reliance on schooling pelagic fish. The specialist nature of the observed hunting strategies further limits the survival potential of this species if Allee effects reduce group size-related foraging efficiency. This is likely to be exacerbated by diminishing fish stocks due to resource competition and environmental change.


Making every Australian count: challenges for the National Disability Insurance Scheme (NDIS) and the equal inclusion of homeless Aboriginal and Torres Strait Islander Peoples with neurocognitive disability.
This article highlights the dearth of accurate evidence available to inform the National Disability Insurance Scheme (NDIS) regarding the extent and nature of neurocognitive disability amongst homeless Aboriginal and Torres Strait Islander people. Without accurate prevalence rates of neurocognitive disability, homeless Aboriginal and Torres Strait Islander people are in danger of not being counted by the NDIS and not receiving supports to which they are entitled. Addressing this knowledge gap is challenged by a range of factors, including: (1) the long-term effect of profound intergenerational disenfranchisement of Aboriginal and Torres Strait Islander people; (2) Aboriginal and Torres Strait Islander cultural perspectives around disability; (3) the generally unrecognised and poorly understood nature of neurocognitive disability; (4) the use of research methods that are not culturally safe; (5) research logistics; and (6) the absence of culturally appropriate assessment tools to identify prevalence. It is argued that an accurate evidence base that is informed by culturally safe research methods and assessment tools is needed to accurately guide the Commonwealth government and the National Disability Insurance Agency about the expected level of need for the NDIS. Research within this framework will contribute to the realisation of a truly inclusive NDIS.


Adrenal epinephrine and norepinephrine release to hypoglycemia measured by microdialysis in conscious rats.
Experiments were conducted in conscious male rats to determine whether hypoglycemia induced by insulin administration preferentially stimulated epinephrine (Epi) or norepinephrine (NE) adrenal medullary chromaffin cells. The release of Epi and NE from the adrenal medulla was continuously monitored using a microdialysis probe of novel design that had been inserted in the adrenal medulla approximately 16 h before the administration of insulin. Following insulin, 3 U/kg i.v., blood glucose declined and dialysate Epi levels rose. No measurable increment in dialysate NE was obtained. Similarly, plasma Epi increased with no detectable change in NE. Patterns of dialysate and plasma catecholamine changes were similar in two groups of animals that had been fed or fasted overnight before insulin treatment. However, the magnitude of the Epi increase was greater in the fasted animals. After recovery of the blood glucose concentration to preinsulin levels, dialysate and plasma catecholamine concentrations returned to control values. These experiments clearly demonstrate that adrenal medullary chromaffin cells that produce Epi are preferentially stimulated in response to insulin-induced hypoglycemia.


Donor P-gp polymorphisms strongly influence renal function and graft loss in a cohort of renal transplant recipients on cyclosporine therapy in a long-term follow-up.
Cyclosporin A (CsA) is a substrate for cytochrome P450 3A and the efflux transporter P-glycoprotein (P-gp; ABCB1), both abundantly expressed in the kidney. In a long-term follow-up of a cohort of patients who had received kidney transplants between the years 1990 and 2005, we retrospectively investigated the effect of CYP3A4, CYP3A5, and ABCB1 polymorphisms in kidney graft donors on recipients' renal function and risk of subsequent graft loss. DNA samples from 227 donors and clinical data from the 259 respective recipients were analyzed. Graft loss was significantly associated with the presence of the ABCB1 variant haplotype 1236T/2677T/3435T in the donor (1236T/2677T/3435T vs. other haplotypes: hazard ratio = 9.346; 95% confidence interval (CI) (2.278-38.461); P = 0.0019) and with previous episodes of acute organ rejection (hazard ratio = 3.077; 95% CI (1.213-7.812); P = 0.0178). The variant haplotype was also associated with a greater decrease in renal function (homozygotes for TTT -3.047 mlxmin(-1)/year; heterozygotes for TTT -4.435 mlxmin(-1)/year; others -2.186 mlxmin(-1)/year; P = 0.0240). The study showed that the presence of ABCB1 polymorphisms in donors influences long-term graft outcome adversely with decrease in renal function and graft loss in transplant recipients receiving CsA.


The experience of limits and the broadening of the horizons of consciousness.
The author traces the gradual evolution of the clinical and diagnostic category of the borderline in order to demonstrate how, in the course of a few decades, twentieth century psychiatric culture, with its own particular vision of the world, passed from an experience of limit to a concept of limit. In other words, as psychiatric culture became aware of a certain difficulty in defining an object under observation, it was gradually forced to modify its own descriptive categories and elaborate a new concept that contained within itself that primitive experience of indeterminacy. The author suggests that a similar phenomenon can also develop in other fields of human life, but here it takes place over such a long period of time that the relationship between the primitive experience and its subsequent conceptualization is difficult to trace. Through the borderline experience or the experience of limits, consciousness modifies its horizons, not in the sense of a simple integration of a new content but in the sense of a profound modification of the entire system of values in which consciousness identified itself up to that moment. It is in this key that the author attempts a psychological reading of Aeschylus' Prometheus Bound and Shakespeare's Hamlet, a reading that has also implications for the understanding of the themes emerging at present on the international scene.


A Phase I trial of the farnesyltransferase inhibitor L-778,123 and radiotherapy for locally advanced lung and head and neck cancer.
Preclinical data have demonstrated that farnesyltransferaseinhibitors (FTIs) are radiation sensitizers in selected cell lines. The objective of this Phase I trial was to determine the maximally tolerated dose of the FTI L-778,123 in combination with radiotherapy in non-small cell lung cancer (NSCLC) and head and neck cancer (HNC). L-778,123 was given by continuous i.v. infusion and dose escalated in conjunction with standard radiotherapy. The presence of a ras mutation was not required for study entry. Nine patients (six NSCLC patients and three HNC patients) were enrolled on two dose levels of FTI. No dose-limiting toxicities were observed at the first dose level of 280 mg/m2/day during weeks 1, 2, 4, and 5 of radiotherapy. One episode of dose-limiting toxicity, grade IV neutropenia, was observed in one of three patients treated at 560 mg/m2/day during weeks 1, 2, 4, 5, and 7. No episodes of dose-limiting mucositis, esophagitis, or pneumonitis were observed. Of the four patients with NSCLC with evaluable disease, three patients had a complete response to treatment and one patient had a partial response. A complete clinical response to treatment was observed in two patients with HNC. In vitro studies in tumor cells obtained from a NSCLC patient on this trial showed radiosensitization with FTI and that tumor cells accumulated in G2-M after L-778,123 treatment. The combination of L-778,123 and radiotherapy at dose level 1 is associated with acceptable toxicity. Local responses have been observed in four NSCLC patients without a clear increase in radiotherapy-associated toxicities.


Deep Learning-Assisted Diagnosis of Cerebral Aneurysms Using the HeadXNet Model.
Deep learning has the potential to augment clinician performance in medical imaging interpretation and reduce time to diagnosis through automated segmentation. Few studies to date have explored this topic. To develop and apply a neural network segmentation model (the HeadXNet model) capable of generating precise voxel-by-voxel predictions of intracranial aneurysms on head computed tomographic angiography (CTA) imaging to augment clinicians' intracranial aneurysm diagnostic performance. In this diagnostic study, a 3-dimensional convolutional neural network architecture was developed using a training set of 611 head CTA examinations to generate aneurysm segmentations. Segmentation outputs from this support model on a test set of 115 examinations were provided to clinicians. Between August 13, 2018, and October 4, 2018, 8 clinicians diagnosed the presence of aneurysm on the test set, both with and without model augmentation, in a crossover design using randomized order and a 14-day washout period. Head and neck examinations performed between January 3, 2003, and May 31, 2017, at a single academic medical center were used to train, validate, and test the model. Examinations positive for aneurysm had at least 1 clinically significant, nonruptured intracranial aneurysm. Examinations with hemorrhage, ruptured aneurysm, posttraumatic or infectious pseudoaneurysm, arteriovenous malformation, surgical clips, coils, catheters, or other surgical hardware were excluded. All other CTA examinations were considered controls. Sensitivity, specificity, accuracy, time, and interrater agreement were measured. Metrics for clinician performance with and without model augmentation were compared. The data set contained 818 examinations from 662 unique patients with 328 CTA examinations (40.1%) containing at least 1 intracranial aneurysm and 490 examinations (59.9%) without intracranial aneurysms. The 8 clinicians reading the test set ranged in experience from 2 to 12 years. Augmenting clinicians with artificial intelligence-produced segmentation predictions resulted in clinicians achieving statistically significant improvements in sensitivity, accuracy, and interrater agreement when compared with no augmentation. The clinicians' mean sensitivity increased by 0.059 (95% CI, 0.028-0.091; adjusted P = .01), mean accuracy increased by 0.038 (95% CI, 0.014-0.062; adjusted P = .02), and mean interrater agreement (Fleiss κ) increased by 0.060, from 0.799 to 0.859 (adjusted P = .05). There was no statistically significant change in mean specificity (0.016; 95% CI, -0.010 to 0.041; adjusted P = .16) and time to diagnosis (5.71 seconds; 95% CI, 7.22-18.63 seconds; adjusted P = .19). The deep learning model developed successfully detected clinically significant intracranial aneurysms on CTA. This suggests that integration of an artificial intelligence-assisted diagnostic model may augment clinician performance with dependable and accurate predictions and thereby optimize patient care.


A new partially hydrolyzed whey-based follow-on formula with age-adapted protein content supports healthy growth during the first year of life.
Standard infant formulae often have higher protein content than breastmilk in order to compensate for potentially lower digestibility; excess protein intake may promote adverse effects later in life. A new partially hydrolyzed whey-based (pHF-W) follow-on formula (FoF) with age-adapted protein content was evaluated for growth and gastrointestinal (GI) tolerance in healthy infants. Formula-fed (FF) infants (n = 108) received standard pHF-W formula (1.9 g protein/100 kcal) from enrollment (age ≤ 30 days) until age 120 days followed by new pHF-W FoF (1.6 g protein/100 kcal) until 360 days. Weight gain velocity (WGV) (mean daily WG from enrollment to age 180 days) was compared to WHO growth standards and a breastfed (BF) reference group (n = 86) (non-inferiority margin -3 g/day). GI tolerance was assessed using a validated questionnaire (scale range 13-65). WGV in FF infants (mean ± SD 24.0 ± 4.4 g/day) was non-inferior to BF (23.7 ± 3.9 g/day) and WHO standards (all p ≤ 0.013). Weight-for-age, length-for-age, weight-for-length, and head circumference-for-age z-scores of FF infants were not significantly different from BF at any timepoint. Symptoms of GI intolerance were low (≤23) at all timepoints and similar between groups. A new pHF-W FoF with age-adapted protein content fed sequentially after standard pHF-W infant formula is safe, well-tolerated, and promotes a healthy growth pattern consistent with BF infants and WHO standards during the first year of life. [https://clinicaltrials.gov/], identifier [NCT03276663].


Maternal depression is associated with DNA methylation changes in cord blood T lymphocytes and adult hippocampi.
Depression affects 10-15% of pregnant women and has been associated with preterm delivery and later developmental, behavioural and learning disabilities. We tested the hypothesis that maternal depression is associated with DNA methylation alterations in maternal T lymphocytes, neonatal cord blood T lymphocytes and adult offspring hippocampi. Genome-wide DNA methylation of CD3+ T lymphocytes isolated from 38 antepartum maternal and 44 neonatal cord blood samples were analyzed using Illumina Methylation 450 K microarrays. Previously obtained methylation data sets using methylated DNA immunoprecipitation and array-hybridization of 62 postmortem hippocampal samples of adult males were re-analyzed to test associations with history of maternal depression. We found 145 (false discovery rate (FDR) q<0.05) and 2520 (FDR q<0.1) differentially methylated CG-sites in cord blood T lymphocytes of neonates from the maternal depression group as compared with the control group. However, no significant DNA methylation differences were detected in the antepartum maternal T lymphocytes of our preliminary data set. We also detected 294 differentially methylated probes (FDR q<0.1) in hippocampal samples associated with history of maternal depression. We observed a significant overlap (P=0.002) of 33 genes with changes in DNA methylation in T lymphocytes of neonates and brains of adult offspring. Many of these genes are involved in immune system functions. Our results show that DNA methylation changes in offspring associated with maternal depression are detectable at birth in the immune system and persist to adulthood in the brain. This is consistent with the hypothesis that system-wide epigenetic changes are involved in life-long responses to maternal depression in the offspring.