Prognostic and predictive significance of circulating biomarkers in patients with advanced upper gastrointestinal cancer undergoing systemic chemotherapy.
The prognosis of patients with advanced cancers of the upper gastrointestinal (UGI) tract is poor. Systemic chemotherapy forms the basis for their treatment, with limited efficacy. Biomarkers have been introduced into clinical practice for cancer management. This study aimed to investigate the predictive and prognostic values of circulating biomarkers in patients with advanced esophageal and gastric cancers receiving chemotherapy. Overall, 92 patients with advanced esophageal squamous cell carcinoma (ESCC; n = 38) and gastric adenocarcinoma (GAC; n = 54) were enrolled. We analyzed the association of circulating lymphocyte subsets, inflammatory markers, and blood cell counts with treatment efficacy and patient survival. Significant differences were identified in peripheral blood parameters between the groups with different clinicopathological features. Hemoglobin (Hb, p = 0.014), eosinophil counts (p = 0.028), CD4+CD28+T/CD4+T percentage (p = 0.049), CD8+CD38+T/CD8+T percentage (p = 0.044), memory CD4+T (p = 0.007), and CD4+CD28+T (p = 0.007) were determined as predictors for achieving non-PD (progression disease) in the ESCC cohort. High levels of eosinophils (p = 0.030) and memory CD4+T cells (p = 0.026) and high eosinophil-to-lymphocyte ratio (ELR, p = 0.013) were predictors of non-PD in patients with GAC. The combined detection models exhibited good ability to distinguish between partial response (PR)/non-PR and PD/non-PD in patients with ESCC and GAC, respectively. Using the multivariate Cox model, the Eastern Cooperative Oncology Group (ECOG) score status (hazard ratio [HR]: 4.818, 95% confidence intervals [CI]: 2.076-11.184, p < 0.001) and eosinophil count (HR: 0.276, 95% CI: 0.120-0.636, p = 0.003) were independent prognostic factors of progression-free survival (PFS) in patients with ESCC. Metastatic sites (HR: 2.092, 95% CI: 1.307-3.351, p = 0.002) and eosinophil-to-lymphocyte ratio (ELR; HR: 0.379, 95% CI: 0.161-0.893, p = 0.027) were independent prognostic factors for overall survival (OS) in patients with ESCC. Differentiation (HR: 0.041, 95% CI: 0.200-0.803, p = 0.010), memory CD4+T (HR: 0.304, 95% CI: 0.137-0.675, p = 0.003), NK cells (HR: 2.302, 95% CI: 1.044-3.953, p = 0.037), and C-reactive protein-to-lymphocyte ratio (CLR; HR: 2.070, 95% CI: 1.024-4.186, p = 0.043) were independent prognostic factors for PFS in patients with GAC. Total lymphocyte counts (HR: 0.260, 95% CI: 0.086-0.783, p = 0.017), CD8+T (HR: 0.405, 95% CI: 0.165-0.997, p = 0.049), NK cells (HR: 3.395, 95% CI: 1.592-7.238, p = 0.002), and monocyte-to-lymphocyte ratio (MLR; HR: 3.076, 95% CI: 1.488-6.360, p = 0.002) were identified as independent prognostic factors associated with OS of GAC. Lymphocyte subsets, blood cell counts, and inflammatory parameters may predict the chemotherapeutic response and prognosis in ESCC and GAC. A combination of these markers can be used to stratify patients into risk groups, which could improve treatment strategies.


Nucleosome mobilization by ISW2 requires the concerted action of the ATPase and SLIDE domains.
The ISWI family of ATP-dependent chromatin remodelers represses transcription by changing nucleosome positions. ISWI regulates nucleosome positioning by requiring a minimal length of extranucleosomal DNA for moving nucleosomes. ISW2 from Saccharomyces cerevisiae, a member of the ISWI family, has a conserved domain called SLIDE (SANT-like ISWI domain) that binds to extranucleosomal DNA ~19 base pairs from the edge of nucleosomes. Loss of SLIDE binding does not perturb binding of the ATPase domain or the initial movement of DNA inside of nucleosomes. Not only is extranucleosomal DNA required to help recruit ISW2, but also the interactions of the SLIDE domain with extranucleosomal DNA are functionally required to move nucleosomes.


Food allergy in infants assessed in two German birth cohorts 10 years after the EuroPrevall Study.
The prevalence of food allergies (FA) in children increased rapidly at the turn of the century. The EuroPrevall study identified Germany as a country with very high prevalence of FA at that time. Using two large German birth cohorts, we provide an update of the status quo 10 years later. KUNO Kids and Ulm SPATZ Health studies are two ongoing prospective birth cohorts. Information on FA was obtained by questionnaires at birth and after 6, 12, and 24 months. Univariable and multivariable logistic regression analyses were performed to investigate risk factors during pregnancy, birth, and early childhood. In 1139 and 1006 children from KUNO Kids and SPATZ, the point prevalence of parent-reported FA symptoms at the ages of 1 and 2 years was 13.2% (95% CI: 11.2-15.2) and 13.9% (95% CI: 11.5-17.2) in KUNO Kids. Doctor's diagnosed FA at 1 and 2 years was 2.4% (95% CI: 1.6-3.4) and 2.7% (95% CI: 1.2-4.3) in KUNO Kids and 2.3% (95% CI: 1.3-3.6) and 3% (95% CI: 2.0-4.5) in SPATZ. Cow's milk and citrus fruits were most frequently suspected by parents to cause FA symptoms. Atopy in the child was associated with a higher frequency of FA at any time, whereas atopy in first-degree relatives was only associated with FA at year 1. Smoke exposure during pregnancy was a risk for FA at age 2. The prevalence of food allergy seems to have plateaued in the last 10 years in Germany. FA is often suspected by parents but only rarely diagnosed by oral food challenge. Risk factor analysis may help to establish personalized health approaches.


[Perspectives on treatment of hypertension in elderly patients].
Hypertension increases in prevalence with age. Population-based studies suggest that more than 50% of people over the age of 65 years may have chronic hypertension, defined as blood pressure (BP) > or = 140/90 mmHg. Hypertension, especially systolic hypertension, is the most common, powerful, however treatable risk factor for cardiovascular morbidity and mortality in the elderly. Large randomised trials have demonstrated that treating elderly and even very old persons (age > 80 years) is highly efficacious. A recent meta-analysis, comparing active treatment with placebo in isolated systolic hypertension demonstrated highly significant benefits: stroke was reduced by 30%, coronary heart disease events by 23%, all cardiovascular events by 26%, and cardiovascular deaths by 13%. The classic strategy of an initial thiazide or thiazide-like diuretic therapy has been verified by the most recent trials. Furthermore it is not appropriate to limit the choice of initial drugs for hypertensive older individuals to a single class of agents, since so many older people have other medical problems that affect this decision and reaching the target blood pressure is the determinant factor for cardiovascular risk reduction. Therefore single drug therapy with long-acting (dihydropyridine-type) calcium-antagonists, beta-blockers, angiotensin-converting-enzyme-inhibitors or angiotensin-receptor-blockers is justified with respect to individual efficacy and comorbidity. If a combination of antihypertensive drugs is needed to reach blood pressure goal, thiazides remain the cornerstone. First line therapy with an alpha-blocker is no longer recommended, even for men with hypertension and benign prostatic hypertrophy. Today, recommended blood pressure goals are the same for younger individuals and the elderly.


P53-mediated upregulation of DcR1 impairs oxaliplatin/TRAIL-induced synergistic anti-tumour potential in colon cancer cells.
Oxaliplatin has emerged as a major chemotherapeutic drug in the treatment of advanced colorectal cancer, yet like most conventional cancer therapeutics, its efficacy is often compromised due to p53 mutations. Unlike oxaliplatin, tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) triggers apoptosis in a p53-independent manner, and chemotherapy is known to overcome tumour resistance to TRAIL-induced cell death in most cancer cells. Using a panel of colon cancer cell lines, we assessed the ability of oxaliplatin to sensitize to TRAIL-induced apoptosis. We demonstrate that while both drugs additively or synergistically induced apoptosis in almost all cell lines tested, p53 wild-type colon cancer cells such as HCT116, LS513 or LS174T remained resistant. Impaired TRAIL-induced cell death resulted from a strong p53 dependent, oxaliplatin-mediated, DcR1 receptor expression increase. According to our finding, downregulation of DcR1 using siRNA, in p53 wild-type colon cancer cells, restored oxaliplatin/TRAIL synergistic apoptotic activity. On the contrary, exogenous DcR1 overexpression in SW480, a p53-mutated cell line, abolished the synergy between the two drugs. Altogether we demonstrate for the first time that p53 negatively regulates oxaliplatin-mediated TRAIL-induced apoptotic activity through DcR1 upregulation. Our findings could have important implications for future therapeutic strategies, and suggest that the association oxaliplatin/TRAIL should be restricted to patients harbouring a non-functional p53 protein.


Intravenous milrinone in cardiac surgery.
Phosphodiesterase inhibitors including milrinone produce positive inotropic effects by slowing the hydrolysis of cyclic adenosine monophosphate in the myocardium. With a loading dose of 50 microg/kg followed by an infusion of 0.5 microg x kg(-1) x min(-1), milrinone increases stroke volume index and left ventricular velocity of circumferential fiber shortening after weaning from cardiopulmonary bypass. Milrinone has potential for the treatment and prevention of internal mammary artery spasm because of its vasodilative effect, which is similar to that of papaverine, and is a potent pulmonary vasodilator for patients with right ventricular dysfunction and pulmonary vasoconstriction. Low-dose milrinone may have antiinflammatory properties and potentially can improve splanchnic perfusion.


Systemic lupus erythematosus in children: the complex problems of diagnosis and treatment encountered in 101 such patients at the Mayo Clinic.
Between 1945 and 1970, 101 children (86 girls and 15 boys) with systemic lupus erythematosus were evaluated at the Mayo Clinic. Only 9 children were less than 9 years old at the time of diagnosis. The most frequent presenting complaint was arthralgia; fever, fatigue, and a "butterfly" malar rash also were common. Renal involvement, found in more than 76 per cent of patients, was a prognostically poor sign. The overall survival of children with renal involvement is improved by the use of adequate steroid therapy.


Ectoine as a natural component of food: detection in red smear cheeses.
Ectoine is a compatible solute accumulated in halophilic bacteria in response to high salt concentrations and offers protection from osmotic stress. The occurrence of compatible solutes is widespread among bacteria, yet ectoine has never been detected in foods. The use of an ectoine producing microorganism (Brevibacterium linens) in the surface ripening of red smear cheeses led to the question whether ectoine can be found in cheese. Therefore we examined samples from a variety of cheese manufacturers and different types of red smear cheeses for the presence of ectoine using HPLC and HPLC/MS analysis. Ectoine solely appears in the rind and was detected up to 178 mg/200 g red smear cheese, depending on several factors like ripening status and conditions throughout the cheese production process (e.g. salt concentrations of used brine baths).


Cytotoxicity of troglitazone through PPARγ-independent pathway and p38 MAPK pathway in renal cell carcinoma.
Agonists of peroxisome proliferator-activated receptor gamma (PPARγ) have been examined as chemopreventive and chemotherapeutic agents. The aim was to investigate the cytotoxicity of troglitazone (TGZ) and its mechanisms in terms of PPARγ dependency and the p38 mitogen-activated protein kinase (MAPK) pathway in three human renal cell carcinoma (RCC) cell lines, 786-O, Caki-2 and ACHN cells. TGZ induced apoptosis and exerted cytotoxicity in a PPARγ-independent manner. We demonstrated that TGZ activated the p38 MAPK pathway and was involved in the cytotoxicity of TGZ. It was also revealed that TGZ induced G(2)/M cell cycle arrest through activation of p38 MAPK.


Studies on the mechanism of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU)-induced hepatotoxicity. III. Ultrastructural characterization of bile duct injury.
The antineoplastic nitrosourea CCNU is a known hepatotoxin which has been shown to cause hyperbilirubinemia and reduction in bile flow. We studied morphological alterations in the common bile duct and interlobular bile ducts at 6, 12, and 24 h in male rats given a single oral dose (50 mg/kg) of CCNU. The portal vein was perfused with 1.0% glutaraldehyde fixative. Portal areas and the common bile duct were selectively dissected and processed using standard methods for light and transmission electron microscopy. The epithelial cells of larger common bile duct and interlobular bile ducts showed increased rough endoplasmic reticulum, markedly increased free ribosomes, and mitochondrial degeneration at 6 and 12 h after CCNU. There was also bile imbibition and loss of microvilli, which increased in severity at 12 and 24 h. The interstitium showed infiltration by acute inflammatory cells and dilated capillaries at 6 h. By 24 h, degeneration of epithelial cells was extensive; cells became necrotic and sloughed into the duct lumen. The smaller bile ductules showed no significant degenerative changes; adjacent hepatocytes were unremarkable. Early CCNU injury appears localized in the large bile ducts and reflects inflammatory edema, bile stasis, and degeneration of epithelial cells. Our studies suggest that this ductal injury may reflect metabolism of CCNU to reactive species within the bile ducts.


Incidence and neurological outcomes of aneurysm rupture during interventional neuroradiology procedures in a hybrid operating suite.
A hybrid operating suite, where craniotomy, brain CT scanning, fluoroscopy, and angiography can be performed on the same operating table, is becoming popular among neurosurgeons. However, whether use of a hybrid operating suite can improve neurological outcome has not been studied. We reviewed the incidence and outcomes of ruptures of cerebral vascular lesions during interventional neuroradiology procedures performed in our hybrid operating suite. Of 602 patients who underwent coil embolization under general anesthesia, five cases of intraprocedural rupture of the aneurysms were identified. Despite emergency neurosurgical intervention within 2 h of rupture of the aneurysms, two of these five patients died and one patient had permanent neurological deficit postsurgery. Whether use of a hybrid operating suite improves neurological outcome remains uncertain.


Cobalamin C Deficiency Induces a Typical Histopathological Pattern of Renal Arteriolar and Glomerular Thrombotic Microangiopathy.
Cobalamin C (cblC) deficiency is the most common inborn error of vitamin B12 metabolism. Renal failure attributed to thrombotic microangiopathy (TMA) has occasionally been described in the late-onset presentation of cblC deficiency, but kidney lesions associated with cblC deficiency remain poorly defined. This study aims to describe the characteristics of kidney disease in cblC deficiency, and to provide a comparative histological analysis with cblC-independent renal TMA. We performed a multicenter retrospective study including 7 patients with cblC deficiency and 16 matched controls with cblC-independent TMA. The patients included were aged 6 to 26 years at the time of the first manifestations. All patients presented with acute renal failure, proteinuria, and hemolysis; 5 patients required dialysis. The histological study revealed arteriolar and glomerular TMA in all patients. After comparison with the cblC-independent TMA control group, a vacuolated aspect of the glomerular basement membrane and the intensity of glomerular capillary wall IgM deposits were more present in cblC deficiency patients than in controls. Six patients were treated with hydroxycobalamin. All of them improved, with disappearance of hemolysis, and 3 of the 4 patients requiring renal replacement therapy were weaned off dialysis. This study provides a precise description of kidney pathology in cblC deficiency. Due to major therapeutic implications, we suggest that patients with renal TMA be screened for cblC deficiency regardless of age, particularly when the kidney biopsy provides evidence of long-lasting TMA, including a vacuolated aspect of the glomerular basement membrane and glomerular capillary wall IgM deposition.


[A 3H-thymidine paper strip method for stimulation of lymphocytes. I. Culture conditions for human or murine lymphocytes and reproducibility of the paper strip method].
A simple method for stimulation of human or murine lymphocytes is described. As a possible more rapid test than the standard estimation of [3H]-thymidine uptake after solubilisation of cell sediments, paper strip counting in a liquid scintillation counter was studied in 80 individuals and 20 mice. Counting efficiency approaches 1%, the reproducibility is good within the acceptable 16% range.


Biological evaluation and docking studies of natural isocoumarins as inhibitors for human kallikrein 5 and 7.
Human kallikrein 5 and 7 (KLK5 and KLK7) are trypsin-like and chymotrypsin-like serine proteases, respectively, and promising targets for the treatment of skin desquamation, inflammation and cancer. In an effort to develop new inhibitors for these enzymes, we carried out enzymatic inhibition assays and docking studies with three isocoumarin compounds. Some promising inhibitors were uncovered, with vioxanthin and 8,8'-paepalantine being the most potent competitive inhibitors of KLK5 (K(i)=22.9 μM) and KLK7 (K(i)=12.2 μM), respectively. Our docking studies showed a good correlation with the experimental results, and revealed a distinct binding mode for the inhibitors at the binding sites of KLK5 and KLK7. In addition, the docking results suggested that the formation of hydrogen bonds at the oxyanion hole is essential for a good inhibitor.


A new technique for localization of cellulase in situ using silver nitrate.
A new staining technique has been developed for the histochemical localization of cellulase in plant tissues by light microscopy. The products of cellulolysis are reducing sugars which can reduce the salts of heavy metals under appropriate conditions. The present technique relies on the deposition of black silver oxide due to reduction of alkaline silver nitrate to detect cellulase in tissues.


Alternation of inverse problem approach and deep learning for lens-free microscopy image reconstruction.
A lens-free microscope is a simple imaging device performing in-line holographic measurements. In the absence of focusing optics, a reconstruction algorithm is used to retrieve the sample image by solving the inverse problem. This is usually performed by optimization algorithms relying on gradient computation. However the presence of local minima leads to unsatisfactory convergence when phase wrapping errors occur. This is particularly the case in large optical thickness samples, for example cells in suspension and cells undergoing mitosis. To date, the occurrence of phase wrapping errors in the holographic reconstruction limits the application of lens-free microscopy in live cell imaging. To overcome this issue, we propose a novel approach in which the reconstruction alternates between two approaches, an inverse problem optimization and deep learning. The computation starts with a first reconstruction guess of the cell sample image. The result is then fed into a neural network, which is trained to correct phase wrapping errors. The neural network prediction is next used as the initialization of a second and last reconstruction step, which corrects to a certain extent the neural network prediction errors. We demonstrate the applicability of this approach in solving the phase wrapping problem occurring with cells in suspension at large densities. This is a challenging sample that typically cannot be reconstructed without phase wrapping errors, when using inverse problem optimization alone.


Modeling the behavior of an underwater acoustic relative positioning system based on complementary set of sequences.
The great variability usually found in underwater media makes modeling a challenging task, but helpful for better understanding or predicting the performance of future deployed systems. In this work, an underwater acoustic propagation model is presented. This model obtains the multipath structure by means of the ray tracing technique. Using this model, the behavior of a relative positioning system is presented. One of the main advantages of relative positioning systems is that only the distances between all the buoys are needed to obtain their positions. In order to obtain the distances, the propagation times of acoustic signals coded by Complementary Set of Sequences (CSS) are used. In this case, the arrival instants are obtained by means of correlation processes. The distances are then used to obtain the position of the buoys by means of the Multidimensional Scaling Technique (MDS). As an early example of an application using this relative positioning system, a tracking of the position of the buoys at different times is performed. With this tracking, the surface current of a particular region could be studied. The performance of the system is evaluated in terms of the distance from the real position to the estimated one.


Corrigendum: Ionizing Particle Radiation as a Modulator of Endogenous Bone Marrow Cell Reprogramming: Implications for Hematological Cancers.
[This corrects the article on p. 231 in vol. 5, PMID: 26528440.].


Hydroxycitric acid reverses tamoxifen resistance through inhibition of ATP citrate lyase.
Lipid metabolic reprogramming is involved in mediating tamoxifen (TAM) response in breast cancer cells. Published microarray data indicated that ATP citrate lyase (ACLY) is overexpressed in TAM-resistant BC cells. Hydroxycitric acid (HCA) is a powerful competitive inhibitor of the enzyme ACLY, which links carbohydrates and lipids metabolism. However, whether inhibition of ACLY could modulate TAM response in TAM-resistant BC cells remained unexplored. Thus the current study aimed to explore the effect of ACLY inhibition on TAM-resistant BC cells. The cytotoxicity of TAM and/or HCA on LCC2 and its TAM-sensitive counterpart MCF7 cells was evaluated. Also, the effect of TAM and/or HCA treatments on ACLY protein levels were investigated by western blotting. In addition, the effects of TAM and/or HCA on caspase-3, Bax, and Bcl2 levels were evaluated by ELISA.; besides, and flow cytometric analysis was performed for the detection of apoptosis. Moreover, cholesterol and triglyceride contents of LCC2 and MCF7 were quantified colorimetrically. Our results demonstrated that TAM/HCA co-treatment synergistically diminished LCC2 and MCF7 cell viability, with the effect being more significant on LCC2. Mechanistically, TAM/HCA co-treatment decreases the expression level of ACLY in LCC2 by 74 %, while in MCF7 by only 59 %. Moreover, apoptosis marker caspase-3 and Bax were increased, while the anti-apoptotic Bcl2 was decreased. Furthermore, the cholesterol and TG contents were increased in LCC2 than in MCF7. Our data revealed that ACLY plays a key role in TAM resistance and ACLY inhibition by HCA-mediated sensitization of BC-resistant cells to TAM.


Epistasis regulates the developmental stability of the mouse craniofacial shape.
Fluctuating asymmetry is a classic concept linked to organismal development. It has traditionally been used as a measure of developmental instability, which is the inability of an organism to buffer environmental fluctuations during development. Developmental stability has a genetic component that influences the final phenotype of the organism and can lead to congenital disorders. According to alternative hypotheses, this genetic component might be either the result of additive genetic effects or a by-product of developmental gene networks. Here we present a genome-wide association study of the genetic architecture of fluctuating asymmetry of the skull shape in mice. Geometric morphometric methods were applied to quantify fluctuating asymmetry: we estimated fluctuating asymmetry as Mahalanobis distances to the mean asymmetry, correcting first for genetic directional asymmetry. We applied the marginal epistasis test to study epistasis among genomic regions. Results showed no evidence of additive effects but several interacting regions significantly associated with fluctuating asymmetry. Among the candidate genes overlapping these interacting regions we found an over-representation of genes involved in craniofacial development. A gene network is likely to be associated with skull developmental stability, and genes originally described as buffering genes (e.g., Hspa2) might occupy central positions within these networks, where regulatory elements may also play an important role. Our results constitute an important step in the exploration of the molecular roots of developmental stability and the first empirical evidence about its genetic architecture.


Palladium-catalyzed stereocontrolled vinylation of azoles and phenothiazine.
[reaction: see text] Vinylation of various azoles (pyrrole, indole, carbazole, and their derivatives) and phenothiazine with vinyl bromides catalyzed by palladium-phosphine complexes results in the respective N-vinylazoles in 30-99% yields. This reaction with cis- and trans-beta-bromostyrenes is stereospecific giving the respective products with full retention of configuration.


Assessment of waist-to-hip ratio attractiveness in women: an anthropometric analysis of digital silhouettes.
The low proportion of waist to hip size in females is a unique and adaptive human feature. In contemporary human populations, the waist-to-hip ratio (WHR) is negatively associated with women's health, fecundity, and cognitive ability. It is, therefore, hypothesized that men will prefer women with low WHR. Although this prediction is supported by many studies, considerable disagreement persists about which WHR values are the most attractive and the importance of WHR for attractiveness of the female body. Unfortunately, the methods applied thus far are flawed in several ways. In the present study, we investigated male preferences for female WHR using a high precision assessment procedure and digitally manufactured, high quality, anthropometrically informed stimuli which were disentangled from body mass covariation. Forty men were requested to choose the most attractive silhouette consecutively from six series (2 levels of realism × 3 levels of body mass), each consisting of 26 female images that varied in WHR (from .60 to .85 by .01). Substantial inter-individual variation in the choices made was observed. Nevertheless, low and average WHR values were chosen more frequently than above-average values or values below the normal variation of the trait. This preference pattern mirrors the relationship between WHR and mate value, suggesting that the preferences are adaptive.


Interactions of the AP-1 Golgi adaptor with the polymeric immunoglobulin receptor and their possible role in mediating brefeldin A-sensitive basolateral targeting from the trans-Golgi network.
We provide morphological, biochemical, and functional evidence suggesting that the AP-1 clathrin adaptor complex of the trans-Golgi network interacts with the polymeric immunoglobulin receptor in transfected Madin-Darby canine kidney cells. Our results indicate that immunofluorescently labeled gamma-adaptin subunit of the adaptor complex and the polymeric immunoglobulin receptor partially co-localize in polarized and semi-polarized cells. gamma-Adaptin is co-immunoisolated with membranes expressing the wild-type receptor. The entire AP-1 adaptor complex could be chemically cross-linked to the receptor in filter-grown cells. gamma-Adaptin could be co-immunoprecipitated with the wild-type receptor, with reduced efficiency with receptor mutant whose basolateral sorting motif has been deleted, and not with receptor lacking its cytoplasmic tail. Co-immunoprecipitation of gamma-adaptin was inhibited by brefeldin A. Mutation of cytoplasmic serine 726 inhibited receptor interactions with AP-1 but did not abrogate the fidelity of its basolateral targeting from the trans-Golgi network. However, the kinetics of receptor delivery to the basolateral cell surface were slowed by the mutation. Although surface delivery of the wild-type receptor was inhibited by brefeldin A, the delivery of the mutant receptor was insensitive to the drug. Our results are consistent with a working model in which phosphorylated cytoplasmic serine modulates the recruitment of the polymeric immunoglobulin receptor into AP-1/clathrin-coated areas in the trans-Golgi network. This process may regulate the efficiency of receptor targeting from the trans-Golgi network.


Trypsinogen activation and glutathione content are linked to pancreatic injury in models of biliary acute pancreatitis.
In models of biliary acute pancreatitis, which might resemble the situation in humans, premature activation of trypsinogen inside the pancreas ("autodigestion") occurs and is correlated with the extent of ductal and parenchymal injury. It is accompanied by a critical spending of protease inhibitors and glutathione, compromising important acinar cell defense and maintenance mechanisms. Premature activation of pancreatic digestive enzymes and profound changes of levels of certain biochemical compounds have been implicated in the pathophysiology of acute pancreatitis. Hitherto, little information on their role in biliary acute pancreatitis has been available. Three types of injury to the pancreaticobiliary duct system of various severity were induced in rats--ligation of the common bile-pancreatic duct, retrograde infusion of electrolyte, or retrograde infusion of taurocholate solution--and were compared to sham-operated animals. Trypsin, trypsin inhibitory capacity (TIC), reduced glutathione (GSH), and other compounds were measured in pancreatic tissue. Histopathology, as well as serum amylase, lipase, and gamma-glutamyl transferase (gamma GT) were assessed. Histopathology and elevated activity of gamma GT in the serum revealed increasing severity of pancreatic injury from sham operation through retrograde duct infusion with taurocholate. GSH was diminished even in macroscopically normal-appearing tissue, but significantly lower in altered (hemorrhagic)-looking sections. Conversely, tissue levels of trypsin were significantly increased. TIC was elevated only in the duct obstruction model, whereas it was reduced in the retrograde duct infusion models.


Biologic treatments in Takayasu's Arteritis: A comparative study of tumor necrosis factor inhibitors and tocilizumab.
To compare the treatment outcomes of TNF inhibitors and tocilizumab (TCZ) in patients with Takayasu arteritis. Takayasu arteritis patients who were refractory to conventional immunosuppressive (IS) drugs and received biologic treatment were included in this multicenter retrospective cohort study. Clinical, laboratory and imaging data during follow-up were recorded. Remission, glucocorticoid (GC) sparing effect, drug survival was compared between TNF inhibitor and TCZ treatments. Also, a subgroup matched comparison was performed between groups. One hundred and eleven (F/M: 98/13) patients were enrolled. A total of 173 biologic treatment courses (77 infliximab, 49 TCZ, 33 adalimumab, 9 certolizumab, 3 rituximab, 1 ustekinumab and 1 anakinra) were given. Tocilizumab was chosen in 23 patients and TNF inhibitors were chosen in 88 patients as first-line biologic agent. Complete/partial remission rates between TCZ and TNF inhibitors were similar at 3rd month and at the end of the follow-up. GC dose decrease (≤4 mg) or discontinuation of GCs was achieved in a similar rate in both groups (TNF inhibitors vs TCZ: 78% vs 59%, p = 0.125). Drug survival rate was 56% in TNF inhibitors and 57% in TCZ group (p = 0.22). The use of concomitant conventional ISs did not affect the drug survival (HR =0.78, 95% CI =0.42-1.43, p = 0.42). The match analysis showed similar results between groups in terms of relapse, decrease in GC dose, surgery need and mortality. The efficacy and safety outcomes and drug survival rates seem to be similar for TNF inhibitors and tocilizumab in patients with Takayasu arteritis.


A fast and highly efficient automated DNA extraction method from small quantities of bone powder from aged bone samples.
An efficient extraction method is important for obtaining high-quality DNA from degraded aged bone samples. An automated full-demineralization method using the EDTA and DNA Investigator Kit (Qiagen) combined with Qiagen's biorobots was optimized in our laboratory in the past to extract the DNA from 500 mg of aged bone samples. The purpose of this research was to further improve the method with the aim of reducing the required sample material, shortening the extraction time, and achieving higher throughput. To process extremely small samples, the amount of bone powder was reduced to 75 mg, EDTA was replaced with reagents from the Bone DNA Extraction Kit (Promega), and decalcification was shortened from overnight to 2.5 h. Instead of 50 ml tubes, 2 ml tubes were used, which allows higher throughput. The DNA Investigator Kit (Qiagen) and EZ1 Advanced XL biorobot (Qiagen) was used for DNA purification. A comparison between both extraction methods was made on 29 Second World War bones and 22 archaeological bone samples. The differences between both methods were explored by measuring nuclear DNA yield and STR typing success. After cleaning the samples, 500 mg of bone powder was processed using EDTA, and 75 mg of powder from the same bone was processed using the Bone DNA Extraction Kit (Promega). DNA content and DNA degradation were determined using PowerQuant (Promega), and the PowerPlex ESI 17 Fast System (Promega) was used for STR typing. The results showed that the full-demineralization protocol using 500 mg of bone was efficient for Second World War and archaeological samples, and the partial-demineralization protocol using 75 mg of bone powder was only efficient for the Second World War bones. The improved extraction method-for which significantly lower amounts of bone powder can be used, the extraction process is faster, and higher throughput of bone samples is possible-is applicable for genetic identification of relatively well-preserved aged bone samples in routine forensic analyses.


Chronic torsion of the wandering spleen.
Torsion of a wandering spleen can occasionally produce chronic disabling clinical findings that are mistaken for a variety of digestive disturbances. Since the clinical complaints are misleading, a correct and timely radiographic analysis can prevent an extensive but unnecessary pursuit of other more common conditions. Images of an ectopic, enlarged, disoriented spleen with a devitalized parenchyma and adherent pseudocapsule can quickly establish the diagnosis of chronic splenic torsion.


ABL allosteric inhibitors synergize with statins to enhance apoptosis of metastatic lung cancer cells.
Targeting mitochondrial metabolism has emerged as a treatment option for cancer patients. The ABL tyrosine kinases promote metastasis, and enhanced ABL signaling is associated with a poor prognosis in lung adenocarcinoma patients. Here we show that ABL kinase allosteric inhibitors impair mitochondrial integrity and decrease oxidative phosphorylation. To identify metabolic vulnerabilities that enhance this phenotype, we utilized a CRISPR/Cas9 loss-of-function screen and identified HMG-CoA reductase, the rate-limiting enzyme of the mevalonate pathway and target of statin therapies, as a top-scoring sensitizer to ABL inhibition. Combination treatment with ABL allosteric inhibitors and statins decreases metastatic lung cancer cell survival in vitro in a synergistic manner. Notably, combination therapy in mouse models of lung cancer brain metastasis and therapy resistance impairs metastatic colonization with a concomitant increase in animal survival. Thus, metabolic combination therapy might be effective to decrease metastatic outgrowth, leading to increased survival for lung cancer patients with advanced disease.


Surgical pathology of the lung in anti-basement membrane antibody-associated Goodpasture's syndrome.
We report the findings of lung biopsies from five patients with anti-basement membrane antibody-associated Goodpasture's syndrome (ABMA-GS). In four patients, pulmonary capillaritis with hemorrhage was found on lung biopsy, confirming that pulmonary capillaritis can be found in ABMA-GS. Although alveolar hemorrhage was the dominant pathologic finding in four of our five patients, all five patients had evidence of injury at the level of the alveolar wall, manifested by hyaline membranes and widening of alveolar walls by edematous connective tissue. In one of our patients, alveolar hemorrhage was only focal, and diffuse alveolar damage was the dominant pathologic finding. This pattern of lung disease has not been previously described in ABMA-GS.


Congenital visceral myopathy with a predominantly hypertrophic pattern treated by multivisceral transplantation.
Chronic intestinal pseudoobstruction due to visceral myopathy may cause intestinal failure. The anatomical pathology of visceral myopathy is dominated by an atrophic muscularis propria. We present a case that is overwhelmingly dominated by a hypertrophic muscularis propria. The hypertrophic pattern was documented from ages 2 to 28 years. Most descriptions of visceral myopathy focus on the atrophic pattern of injury to the muscularis propria, typified by vacuolar degeneration of myocytes, loss of muscle fibers, and frequently with a honeycomb pattern of fibrosis. Hypertrophic changes can be observed in visceral myopathy, but the hypertrophy typically represents only a minor feature, typically limited to the internal layer of the muscularis propria. In our patient, the observed muscular hypertrophy differed from most descriptions because it (a) was the dominant pattern of injury and (b) involved both layers of muscularis propria rather than just the inner circular layer. The patient had intestinal failure and the complications induced cirrhosis, eventually treated by multivisceral transplantation to correct the intestinal failure and cirrhosis.


The use of egocentric and exocentric location cues in saccadic programming.
Theoretically, the location of a visual target can be encoded with respect to the locations of other stimuli in the visual image (exocentric cues), or with respect to the observer (egocentric cues). Egocentric localization in the oculomotor system has been shown to rely on an internal representation of eye position that inaccurately encodes the time-course of saccadic eye movements, resulting in the mislocalization of visual targets presented near the time of a saccade. In the present investigation, subjects were instructed to localize perisaccadic stimuli in the presence or absence of a visual stimulus that could provide exocentric location information. Saccadic localization was more accurate in the presence of the exocentric cue, suggesting that localization is based on a combination of exocentric and egocentric cues. These findings indicate the need to reassess previously reported neurophysiological studies of spatial accuracy and current models of oculomotor control, which have focused almost exclusively on the egocentric localization abilities of the brain.


Hydroxymethylcytosine and demethylation of the γ-globin gene promoter during erythroid differentiation.
The mechanism responsible for developmental stage-specific regulation of γ-globin gene expression involves DNA methylation. Previous results have shown that the γ-globin promoter is nearly fully demethylated during fetal liver erythroid differentiation and partially demethylated during adult bone marrow erythroid differentiation. The hypothesis that 5-hydroxymethylcytosine (5 hmC), a known intermediate in DNA demethylation pathways, is involved in demethylation of the γ-globin gene promoter during erythroid differentiation was investigated by analyzing levels of 5-methylcytosine (5 mC) and 5 hmC at a CCGG site within the 5' γ-globin gene promoter region in FACS-purified cells from baboon bone marrow and fetal liver enriched for different stages of erythroid differentiation. Our results show that 5 mC and 5 hmC levels at the γ-globin promoter are dynamically modulated during erythroid differentiation with peak levels of 5 hmC preceding and/or coinciding with demethylation. The Tet2 and Tet3 dioxygenases that catalyze formation of 5 hmC are expressed during early stages of erythroid differentiation and Tet3 expression increases as differentiation proceeds. In baboon CD34+ bone marrow-derived erythroid progenitor cell cultures, γ-globin expression was positively correlated with 5 hmC and negatively correlated with 5 mC at the γ-globin promoter. Supplementation of culture media with Vitamin C, a cofactor of the Tet dioxygenases, reduced γ-globin promoter DNA methylation and increased γ-globin expression when added alone and in an additive manner in combination with either DNA methyltransferase or LSD1 inhibitors. These results strongly support the hypothesis that the Tet-mediated 5 hmC pathway is involved in developmental stage-specific regulation of γ-globin expression by mediating demethylation of the γ-globin promoter.


Combinations of Beauveria bassiana and spinetoram for the management of four important stored-product pests: laboratory and field trials.
The current study examines the efficacy of the semi-synthetic insecticide spinetoram and entomopathogenic fungi Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Hypocreales: Cordycipitaceae) as wheat protectants against the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), the granary weevil, Sitophilus granarius (L.) (Coleoptera: Curculionidae), and the khapra beetle, Trogoderma granarium Everts (Coleoptera: Dermestidae), under laboratory and field trials. One dose of B. bassiana, i.e., 1 × 107 conidia/kg wheat, two doses of spinetoram, i.e., spine1: 0.05 ppm (mg/kg wheat), spine2: 0.1 ppm, and their combinations (Bb + spine1, Bb + spine2) were evaluated at 20, 25, and 30 °C. All treatments provided significantly higher mortality at 30 °C compared with the other two temperatures. Maximum mortality levels were observed in the treatments where B. bassiana was combined with the higher dose of spinetoram (0.1 ppm). All treatments reduced progeny production in comparison with the control groups. Maximum progeny reduction was observed at 30 °C, on wheat treated with the Bb + spine2 combination. The combination Bb + spine2 also provided elevated mortality rates in both laboratory and field persistence trials, but at 180 days caused moderate mortality to all tested insect species. Concerning progeny, at laboratory persistence trials, the combination Bb + spine2 exhibited the lowest offspring emergence to all tested species compared to the other treatments and control. Overall, our study showed that R. dominica was the most susceptible species followed by S. granarius, T. castaneum, and T. granarium. Our findings revealed that the combination of B. bassiana and spinetoram may be a useful tool for efficient and advanced integrated pest management strategies for long storage periods under multiple temperatures.


Predicting drug-drug interactions through drug structural similarities and interaction networks incorporating pharmacokinetics and pharmacodynamics knowledge.
Drug-drug interactions (DDIs) may lead to adverse effects and potentially result in drug withdrawal from the market. Predicting DDIs during drug development would help reduce development costs and time by rigorous evaluation of drug candidates. The primary mechanisms of DDIs are based on pharmacokinetics (PK) and pharmacodynamics (PD). This study examines the effects of 2D structural similarities of drugs on DDI prediction through interaction networks including both PD and PK knowledge. Our assumption was that a query drug (Dq) and a drug to be examined (De) likely have DDI if the drugs in the interaction network of De are structurally similar to Dq. A network of De describes the associations between the drugs and the proteins relating to PK and PD for De. These include target proteins, proteins interacting with target proteins, enzymes, and transporters for De. We constructed logistic regression models for DDI prediction using only 2D structural similarities between each Dq and the drugs in the network of De. The results indicated that our models could effectively predict DDIs. It was found that integrating structural similarity scores of the drugs relating to both PK and PD of De was crucial for model performance. In particular, the combination of the target- and enzyme-related scores provided the largest increase of the predictive power.Graphical abstract.


Shelf life estimation of refrigerated vacuum packed beef accounting for uncertainty.
This study estimates the shelf life of vacuum packed beef meat (three muscles: striploin (longissimus thoracis et lumborum, LTL), tenderloin (psoas major, PM) and outside chuck (trapezius thoracis, TT)) at refrigeration temperatures (0 °C-10 °C) based on modelling the growth of two relevant groups of spoilage microorganisms: lactic acid bacteria (LAB) and Enterobacteriaceae. The growth models were developed combining a two-step and a one-step approach. The primary modelling was used to identify the parameters affecting the growth kinetics, guiding the definition of secondary growth models. For LAB, the secondary model included the effect of temperature and initial pH on the specific growth rate. On the other hand, the model for Enterobacteriaceae incorporated the effect of temperature on the specific growth rate and the lag phase; as well as the effect of the initial pH on the specific growth rate, the lag phase and the initial microbial count. We did not observe any significant effect of the type of muscle on the growth kinetics. Once the equations were defined, the models were fitted to the complete dataset using a one-step approach. Model validation was carried out by cross-validation, mitigating the impact of an arbitrary division between training and validation sets. The models were used to estimate the shelf life of the product, based on the maximum admissible microbial concentration (7 log CFU/g for LAB, 5 log CFU/g for Enterobacteriaceae). Although LAB was the dominant microbiota, in several cases, both LAB and Enterobacteriaceae reached the critical concentration practically at the same time. Furthermore, in some scenarios, the end of shelf life would be determined by Enterobacteriaceae, pointing at the potential importance of non-dominant microorganisms for product spoilage. These results can aid in the implementation of effective control measures in the meat processing industry.


Abnormal glial and glycoconjugate dispositions in the somatosensory cortical barrel field of the early postnatal reeler mutant mouse.
During early postnatal development in reeler mutant mice, lectin binding delineates prospective abnormal barrels as they will appear in the adult mutant somatosensory cortex. Glial fibers also may be more condensed within fascicles in developing reeler barrels. These fibers also appear to be misaligned, coursing predominantly in the tangential plane within the abnormal reeler barrel sides as opposed to having a radial orientation as seen in normal mouse barrels. The thalamic barreloid complex, however, reveals a disposition of glycoconjugates that is completely normal in reeler. Thus, there are anomalies in glia and associated glycoconjugates during mainly cortical development in the reeler mutant mouse that might be related to the primary action of the abnormal gene.


Post-translational oxidative modification of β2-glycoprotein I and its role in the pathophysiology of the antiphospholipid syndrome.
Vascular thrombosis and/or recurrent miscarriages are the main characteristics defining Antiphospholipid Syndrome (APS). Currently there is no well-defined clinical features and/or laboratory tests that predicts the risk of adverse prognostic outcomes in APS. In this short review, we report the importance of posttranslational modification of beta2 glycoprotein I, the major autoantigen in the APS beta2 glycoprotein I that may, in part, explain possible mechanisms for the generation of auto antibodies to beta2 glycoprotein I. A specific ELISA measuring the level of oxidised beta2 glycoprotein I could be used as a potential new laboratory test - along with other laboratory tests - to more accurately predict the risk of having a clinical event in patients with APS.


Helicobacter pylori: Helicobacter pylori gastritis--a novel distinct disease entity.
A global consensus report on Helicobacter pylori gastritis has been developed. Topics discussed include whether dyspepsia caused by H. pylori infection is separate from functional dyspepsia or not, the evaluation method for H. pylori-induced gastritis, eradication therapy for H. pylori gastritis to prevent gastric carcinogenesis and management after H. pylori eradication.


Fetal DNA quantitation in peripheral blood is not useful as a marker of disease severity in women with preeclampsia.
Trophoblast migration into the maternal circulation is increased in preeclampsia and can trigger the endothelial dysfunction that characterizes the clinical disease. We hypothesised that 'fetal trafficking' is increased in women with severe preeclampsia and that the quantity of trafficking is greater in women with more severe disease. To test the hypothesis, we used a technique that quantifies a genetic marker specific for the fetomaternal unit, that is, the SRY gene in a pregnant woman carrying a male fetus. Thirty two women with pre-eclampsia and 32 control women (women without preeclampsia) were recruited. Preeclampsia was defined according to the International Society for the Study of Hypertension in Pregnancy (ISSHP). All subjects with preeclampsia had evidence of the multisystemic nature of the disease. DNA was extracted from maternal peripheral blood and RTQ PCR analysis was performed to quantify the fetal DNA (SRY) and the total DNA (beta-actin) in each sample. The ratio of fetal to total DNA was calculated and compared between women with preeclampsia and controls. The women with preeclampsia and the control women did not differ in parity, blood pressure at booking, and gestational age at sampling. The groups differed significantly in age (29 +/- 5.7 vs 25 +/- 5.1 years; P =.007), diastolic blood pressure (DBP) at sampling (101 +/- 9.5 vs 70 +/- 5.5 mm Hg; P <.0001), gestational age at delivery (33 +/- 4.3 vs 39 +/- 1.8 weeks; P <.001), and fetal weight (1.98 +/- 1 vs 3.35 +/- 0.5 kgs; P <.0001). SRY was detected in 31 out of 32 women with preeclampsia and in 24 out of 32 control women (P <.001). The median SRY copy number per micro L was greater in women with preeclampsia (10.6, interquartile range 12.89) than in the control women (8.6, interquartile range 20.1) but these differences were not statistically significant at P =.75. The median ratio of fetal to total DNA was almost identical in both groups (0.06, interquartile range 0.13) in PET compared to (0.06, interquartile range 0.17) the control women. No correlation was found between the quantity of fetal DNA and disease severity. Fetal trafficking is more likely to be detected in women with preeclampsia compared to control women but the quantity does not appear to correlate with disease severity.


Inhibitory tendon-evoked reflex is attenuated in the torque-depressed isometric steady-state following active shortening.
Residual torque depression (rTD) is the reduction in steady-state isometric torque following an active shortening contraction when compared with an isometric contraction at the same muscle length and activation level. We have shown that spinal excitability increases in the rTD state, yet the mechanisms remains unknown. Percutaneous electrical tendon stimulation was used to induce tendon-evoked inhibitory reflexes. We demonstrated that in the rTD state, reduced torque contributes to a reduction in inhibitory afferent feedback, which indicates that the history-dependent properties of muscle can alter spinal excitability and the voluntary control of submaximal contractions through changes in peripheral afferent feedback. Novelty Residual force depression is a basic property of skeletal muscle, which can influence spinal and supraspinal excitability via inhibitory reflex activity. Residual force depression alters the voluntary control of force.


Soft X-ray beamline BL1N2 at Aichi Synchrotron Radiation Center and its industrial use.
BL1N2 is a soft X-ray XAFS (X-ray absorption fine structure) beamline that is well suited for industrial use. User service started in 2015. The beamline is a grazing optical system with a pre-mirror, an inlet slit, two mirrors for three gratings, an outlet slit and a post-mirror. Light of 150 eV to 2000 eV is available, and K-edge measurements of elements from B to Si are covered. The O K-edge is most often measured; transition metals such as Ni and Cu at their L-edges and lanthanoids at their M-edges are also often measured. Here, basic information about BL1N2, the effect of ageing by synchrotron radiation to remove mirror contamination, and a compatible sample handling system and transfer vessels to allow a one-stop service at three soft X-ray beamlines at AichiSR are described.


Influence of intestinal efflux pumps on the absorption and transport of furosemide.
Furosemide is a commonly used diuretic which is used in the treatment of edema, congestive heart failure, hypertension and renal failure. Its absorption exhibits inter- and intra-subject variability that can be attributed to many factors including the intestinal efflux pumps such as the P-glycoprotein (P-gp). This study was done due to the great disagreement between what is published in the literature regarding the influence of P-gp on furosemide and at the same time due to the importance of this drug in the treatment of different conditions as described above. In addition, an investigation of the effect of two of the commonly used pharmaceutical excipients (hydroxypropyl β-cyclodextrin [HPβCD] and Tween 80) and also a P-gp inhibitor (verapamil hydrochloride) on the intestinal absorption of this drug were also done. The study utilized the everted intestinal sacs technique to investigate both the effect of the efflux transporter (P-gp) on furosemide absorption and also the effect of the chosen excipients. The absorption of furosemide was significantly influenced by the P-gp as confirmed by the everted vis the non-everted sacs together with the verapamil study in which the transport of furosemide was inhibited by verapamil. In addition, Tween 80 was also shown to inhibit the P-gp pump whereas the HPβCD did not significantly influence the efflux of furosemide in this study. P-glycoprotein and some of the used excipients in the formulation play a very important role in the transport of furosemide and other drugs. Thus excipients that affect the activity of P-gp should be avoided when formulating drugs that are substrate for the P-gp or other efflux pumps.


A new alternative NF-κB pathway mediated the neuroprotection of GDNF on 6-OHDA-induced DA neurons neurotoxicity.
Glial cell line-derived neurotrophic factor (GDNF) is a potent protective factor for dopaminergic (DA) neurons, but the signaling mechanisms underlying the effect of GDNF on these neurons remain obscure. Here, both our in vivo and in vitro studies demonstrate that the majority of DA neurons express the NF-κB-inducing kinase (NIK), which is the essential kinase for mediating activation of the new alternative NF-κB signaling pathway. Additionally, we also show that GDNF induced the time/dose-dependent phosphorylation of IκB kinase α (IKKα) and p100, facilitated the processing of p100 to p52 and accelerated the translocation of NF-κB dimmers into the nuclei of DA neurons. We furtherly found that the dimmer which translocate into the nucleus was RelA/p52 not RelB/p52. Meanwhile, the attenuation of 6-OHDA-induced DA neuronal apoptosis due to GDNF was reversed subsequent to the inhibition of p100 expression by RNAi while the neuroprotective effect of GDNF on injured DA neurons was strengthened by the overexpression of p100. Our data, therefore, indicate that a new alternative NF-κB signaling pathway, which was not the classic pathway but different from the non-canonical pathway, exists in DA neurons and mediates the neuroprotective effect of GDNF on these neurons.


Comparative study on organic effluent degradation capabilities and electrical transport properties of polygonal ZnCo2O4 spinels fabricated using different green fuels.
The present work highlights biosynthesis of nano-sized heterometalic spinel ZnCo2O4 particles using different green extracts as capping agent. In this work we have fabricated polygonal ZnCo2O4 with Punica granatum peel extract, Camellia sinensis extract, Moringa oleifera leaf extract and green coffee beans extract in an effortless green pathway. Phase pure material synthesis was confirmed using XRD. Microstructural, morphological, compositional and optical characterisations has been carried out using TEM, FESEM, EDX, FTIR, photoluminescence and UV-Vis spectroscopy. Punica granatum peel extract assisted ZnCo2O4 sample shows superior catalytic efficiency of ~84.96% for Rhodamine B pollutant. ZnCo2O4 sample synthesized using pomegranate peel extract shows highest conductivity of ~8.074 × 10-5 Ω-1 cm-1 with activation energy of 2.099 eV at 503 K. Synthesized nanoparticles also show antibacterial activity for B. megaterium, B. subtilis and B. cereus. To the best of our knowledge, synthesis of ZnCo2O4 using these four green extracts and their comparative degradation capability, electrical properties and antibacterial study is explained for the first time in this work.


Synthetic musk fragrances in sediments from a subtropical river-lake system in eastern China: occurrences, profiles, and ecological risks.
Synthetic musk fragrances (SMFs) in aquatic environments have been of increasing concern because of their potential characteristic of persistent, bioaccumulated, and ecological harm. However, little is known about the distribution of SMFs in river-lake systems. In this study, the occurrence and risks of six SMFs measured in sediments from Lake Chaohu (eastern China) and the rivers flowing into it were investigated. The total sedimentary SMF concentrations ranged from 2.43 to 15.5 ng/g in Lake Chaohu (median = 5.17 ng/g), and 2.34-104 ng/g in the rivers (median = 10.6 ng/g). Overall, moderate levels of SMFs were found in comparison with previous results from other areas. Galaxolide and tonalide dominated in the rivers whereas cashmeran was dominant in Lake Chaohu. A source assessment indicated that the discharge from industries contributed importantly to the pollution of SMFs in the studied waters, in addition to the inputs from domestic sewage. Our estimates suggested that the current sedimentary SMF concentrations were likely to pose extremely low ecological risk to aquatic organisms. However, more studies are needed to focus on the spatial and temporal trends in distribution as well as the ecotoxicological implications of SMFs in the Lake Chaohu area because there is a general lack of relevant information.


The effect of hypercarbia on age-related changes in cerebral glucose transport and glucose-modulated agonal glycolytic rates.
This study examined the effect of hypercarbia on cerebral agonal glycolytic rates and brain lactate accumulation after complete ischemia induced by cardiac arrest. Before cardiac arrest, the blood plasma glucose concentration in seven newborn (113 d postconception; normal gestation, 115 d) and seven 1-mo-old (144 d postconception) piglets was adjusted to a specific value (range, 1 to 64 mM), and then inspired ventilation gases were changed to 10:50:40 CO2:O2:N2 for 20 min. The agonal glycolytic rate was measured by monitoring the rate of cerebral lactate formation in vivo using proton nuclear magnetic resonance spectroscopy, and postmortem brain lactate concentrations were measured biochemically in tissue extracts obtained 40 to 45 min after cardiac arrest. These data were compared with 21 normocarbic piglets of similar age, nine examined as part of the present study and 12 examined previously (Corbett RJT, Laptook AR, Ruley JI, Garcia D: Pediatr Res 30:579-586, 1991). There was a nonlinear relationship between the final postmortem brain lactate concentration and preischemia blood plasma glucose concentration that was most prominent in newborn piglets and previously had gone unnoticed. When analyzed using a steady-state model for glucose transport, this relationship revealed that normocarbic newborns had a lower preischemia affinity constant for the transport mechanism for glucose (2.8 +/- 1.5 mM) and lower cerebral glucose utilization rate relative to transport rate (0.12 +/- 0.04), compared with 1-mo-olds (4.5 +/- 1.4 mM and 0.30 +/- 0.03, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)


Inborn errors of immunity to infection: the rule rather than the exception.
The immune system's function is to protect against microorganisms, but infection is nonetheless the most frequent cause of death in human history. Until the last century, life expectancy was only approximately 25 years. Recent increases in human life span primarily reflect the development of hygiene, vaccines, and anti-infectious drugs, rather than the adjustment of our immune system to coevolving microbes by natural selection. We argue here that most individuals retain a natural vulnerability to infectious diseases, reflecting a great diversity of inborn errors of immunity.


Development and optimization of Fe3+-crosslinked sodium alginate-methylcellulose semi-interpenetrating polymer network beads for controlled release of ibuprofen.
In this study, ionically crosslinked beads of sodium alginate (NaAlg) and methylcellulose (MC) were prepared as semi-interpenetrating polymer networks (semi-IPN) in the size range of 1.97 ± 0.09-1.22 ± 0.13 mm by crosslinking with FeCl3 and loaded with ibuprofen (IBU), which is a nonsteroidal anti-inflammatory drug. The highest 93.33% entrapment efficiency of IBU was achieved, and the drug release behaviors, mean particle size, and entrapment efficiency of beads were investigated in terms of the polymer composition, a ratio of ibuprofen to polymer, exposure time to crosslinker, and concentration of the crosslinking agent. Semi-IPN formulations prepared were also characterized using Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), X-ray Diffraction (X-RD), and scanning electron microscopy (SEM). It was observed that IBU-loaded beads displayed better release performance with an increase amount of NaAlg in the structure. Finally, the optimum IBU release was obtained as 93.9% for beads containing 66.7% (w/w) NaAlg, 33.3% (w/w) MC, IBU/polymer ratio of 1/4, FeCl3 concentration of 0.1 M, and crosslinking time of 90 min. The in vitro release rate was fitted to five empirical equations, and the diffusion exponent n, which indicated that the release mechanism of IBU from beads followed the Anomalous and Case II transport mechanism.


Immunoelectron microscopic localisation of keratin and luminal epithelial antigens in normal and neoplastic urothelium.
Immunoelectron microscope cytochemistry was carried out on 2% paraformaldehyde fixed, 50 mu sections of normal urothelium and bladder carcinoma cells in culture using antisera raised in rabbits to human 40-63 000 MW epidermal "broad spectrum" keratin and calf urothelial "luminal epithelial antigen" (aLEA) Both the unconjugated and indirect immunoperoxidase-DAB techniques were used before routine embedding. The localisation of both keratin and luminal epithelial antigen (LEA) was similar in normal and neoplastic cells and reaction product was associated not only with tonofilaments but also lining membrane vesicles and on fine filaments in the cytoplasmic ground substance.


Desensitization kinetics of a chloride acetylcholine response in Aplysia.
The kinetics of desensitization of acetylcholine-evoked Cl conductance increased response of Aplysia RC neurons of the abdominal ganglion were studied under voltage-clamp conditions for comparison with results of similar studies on acetylcholine Na and K responses. The response evoked by acetylcholine on RC neurons was an outward current at resting potential (about -45 mV) that reversed at about -65 mV and was blocked by D-tubocurarine and strychnine but not hexamethonium and was not activated by arecoline. From the current-voltage relation this response can be ascribed to a pure conductance increase to Cl. The apparent KD was 40.6 microM. Upon prolonged exposure to acetylcholine the response peaked within 200-400 ms, and then decayed to a plateau current in the continued presence of the agonist. The peak and plateau currents reversed at the same potential, indicating that there had not been significant redistribution of Cl. The current decay in every cell was best fit by a double exponential function plus a constant, and the average time constants were tau fast = 1.8 +/- 0.2 s and tau slow = 16.2 +/- 1.0 s. Both components were slowed by cooling. While tau fast did not change with dose, tau slow increased with dose. Both components accelerated with hyperpolarization and upon application of trifluoperazine (2 microM). These results are consistent with the interpretation that desensitization of the acetylcholine Cl response is composed of two independent processes. This conclusion is the same as that derived from studies of the acetylcholine Na and K responses, and is in general consistent with desensitization being a property of a common acetylcholine receptor, and independent of the ionic selectivity of the associated channel. There are, however, significant differences in voltage, temperature and trifluoperazine dependence of the two components of the three ionic responses which may reflect influence of the different ion channels and/or transduction mechanisms.


Results of a pilot study of hyperfractionated radiation therapy for children with brain stem gliomas.
The majority of children with brain stem gliomas develop progressive disease within 18 months of diagnosis and treatment. Radiotherapy (RT) is of transient benefit in most patients and higher total doses of RT have been related to improved survival. The amount of RT which can be given is limited by the tolerance of the surrounding brain. Hyperfractionated RT theoretically allows higher doses of RT to be tolerated by the brain. Sixteen children with brain stem gliomas were treated on a hyperfractionated RT schedule, receiving 120 cGy of RT twice daily, to a total dose of 6480 cGy. All patients tolerated treatment well. Eleven of 15 (73%) evaluable patients had a response to treatment and two (13%) others had stable disease. One patient developed progressive disease during treatment. All patients were tapered off steroids by the completion of treatment. Thirteen of 16 (81%) patients developed progressive disease at a median of 7 months after diagnosis and three remain in remission 8, 12, and 15 months following diagnosis. These results were similar to those of historical controls. Two patients were surgically explored at time of relapse and 5 have had an autopsy. No acute or subacute neurologic toxicity was seen; but long-term detrimental effects on brain could not be assessed. The implications of this study are that escalations of the dose of hyperfractionated RT can be entertained for children with brain stem gliomas.


A conductive polymer based electronic nose for early detection of Penicillium digitatum in post-harvest oranges.
We describe the construction of an electronic nose, comprising four chemiresistive sensors formed by the deposition of thin conductive polymer films onto interdigitated electrodes, attached to a personal computer via a data acquisition board. This e-nose was used to detect biodeterioration of oranges colonized by Penicillium digitatum. Significant responses were obtained after only 24 h of incubation i.e. at an early stage of biodeterioration, enabling remedial measures to be taken in storage facilities and efficiently distinguishing between good and poor quality fruits. The instrument has a very low analysis time of 40 s.


A single subcutaneous bolus of erythropoietin normalizes cerebral blood flow autoregulation after subarachnoid haemorrhage in rats.
Systemic administration of recombinant erythropoietin (EPO) has been demonstrated to mediate neuroprotection. This effect of EPO may in part rely on a beneficial effect on cerebrovascular dysfunction leading to ischaemic neuronal damage. We investigated the in vivo effects of subcutaneously administered recombinant EPO on impaired cerebral blood flow (CBF) autoregulation after experimental subarachnoid haemorrhage (SAH). Four groups of male Sprague-Dawley rats were studied: group A, sham operation plus vehicle; group B, sham operation plus EPO; group C, SAH plus vehicle; group D, SAH plus EPO. SAH was induced by injection of 0.07 ml of autologous blood into the cisterna magna. EPO (400 iu kg(-1) s.c.) or vehicle was given immediately after the subarachnoid injection of blood or saline. Forty-eight hours after the induction of SAH, CBF autoregulatory function was evaluated using the intracarotid (133)Xe method. CBF autoregulation was preserved in both sham-operated groups (lower limits of mean arterial blood pressure: 91+/-3 and 98+/-3 mmHg in groups A and B, respectively). In the vehicle treated SAH-group, autoregulation was abolished and the relationship between CBF and blood pressure was best described by a single linear regression line. A subcutaneous injection of EPO given immediately after the induction of SAH normalized autoregulation of CBF (lower limit in group D: 93+/-4 mmHg, NS compared with groups A and B). Early activation of endothelial EPO receptors may represent a potential therapeutic strategy in the treatment of cerebrovascular perturbations after SAH.


Prediction of Feed Efficiency and Performance-Based Traits in Fish via Integration of Multiple Omics and Clinical Covariates.
Fish aquaculture is a rapidly expanding global industry, set to support growing demands for sources of marine protein. Enhancing feed efficiency (FE) in farmed fish is required to reduce production costs and improve sector sustainability. Recognising that organisms are complex systems whose emerging phenotypes are the product of multiple interacting molecular processes, systems-based approaches are expected to deliver new biological insights into FE and growth performance. Here, we establish 14 diverse layers of multi-omics and clinical covariates to assess their capacities to predict FE and associated performance traits in a fish model (Oncorhynchus tshawytscha) and uncover the influential variables. Inter-omic relatedness between the different layers revealed several significant concordances, particularly between datasets originating from similar material/tissue and between blood indicators and some of the proteomic (liver), metabolomic (liver), and microbiomic layers. Single- and multi-layer random forest (RF) regression models showed that integration of all data layers provide greater FE prediction power than any single-layer model alone. Although FE was among the most challenging of the traits we attempted to predict, the mean accuracy of 40 different FE models in terms of root-mean square errors normalized to percentage was 30.4%, supporting RF as a feature selection tool and approach for complex trait prediction. Major contributions to the integrated FE models were derived from layers of proteomic and metabolomic data, with substantial influence also provided by the lipid composition layer. A correlation matrix of the top 27 variables in the models highlighted FE trait-associations with faecal bacteria (Serratia spp.), palmitic and nervonic acid moieties in whole body lipids, levels of free glycerol in muscle, and N-acetylglutamic acid content in liver. In summary, we identified subsets of molecular characteristics for the assessment of commercially relevant performance-based metrics in farmed Chinook salmon.


An Open-Source, Programmable Pneumatic Setup for Operation and Automated Control of Single- and Multi-Layer Microfluidic Devices.
Microfluidic technologies have been used across diverse disciplines (e.g. high-throughput biological measurement, fluid physics, laboratory fluid manipulation) but widespread adoption has been limited in part due to the lack of openly disseminated resources that enable non-specialist labs to make and operate their own devices. Here, we report the open-source build of a pneumatic setup capable of operating both single and multilayer (Quake-style) microfluidic devices with programmable scripting automation. This setup can operate both simple and complex devices with 48 device valve control inputs and 18 sample inputs, with modular design for easy expansion, at a fraction of the cost of similar commercial solutions. We present a detailed step-by-step guide to building the pneumatic instrumentation, as well as instructions for custom device operation using our software, Geppetto, through an easy-to-use GUI for live on-chip valve actuation and a scripting system for experiment automation. We show robust valve actuation with near real-time software feedback and demonstrate use of the setup for high-throughput biochemical measurements on-chip. This open-source setup will enable specialists and novices alike to run microfluidic devices easily in their own laboratories.


Clinical significance of spleen stiffness in patients with acute decompensated heart failure.
Congestive splenomegaly is a classic sign of organ congestion in acute decompensated heart failure (ADHF). Shear wave elastography (SWE) allows the measurement of spleen stiffness (SS). We hypothesized that SS could quantify the severity of splenic congestion and predict adverse events in ADHF. This study included two cohorts: a haemodynamic cohort (62 HF patients) and an outcome cohort (115 ADHF patients). SS was measured by two-dimensional SWE on the same day of right heart catheterization in the haemodynamic cohort. Right atrial pressure (RAP) independently correlated with SS (β = 0.32, P = 0.002). SS was measured in the outcome cohort before discharge. The 115 patients were divided into three groups on the basis of the tertile value of SS. The third tertile SS group had a higher prevalence of severe tricuspid regurgitation, higher N-terminal B-type natriuretic peptide (NT pro-BNP), and larger right ventricular diastolic diameter, than had the first tertile group and the second tertile group. During a median follow-up period of 105 (77-135) days, adverse events occurred in 25 patients (one death and 24 rehospitalizations for HF). The third tertile SS group had a significantly higher rate of adverse events (P < 0.001). A higher SS was independently associated with adverse events after adjusting for conventional validated risk score, liver function test, liver stiffness, and estimated RAP. The degree of SS at discharge can be used as a marker of residual splenic congestion, which is predictive of adverse events in patients with ADHF.


Primary headache and epilepsy: a multicenter cross-sectional study.
The prevalence and characteristics of interictal headache, epilepsy and headache/epilepsy comorbidity were assessed in 858 women and 309 men aged 18-81 years from headache and epilepsy centers in Italy. The research hypothesis was that comorbidity among patients with either disorder would be expected to be higher than in the general population. Interictal headache was diagnosed in 675 cases (migraine 482; tension-type headache 168; other types 25), epilepsy in 336 (partial 171; generalized 165) and comorbidity in 156 (1.6% from headache centers; 30.0% from epilepsy centers). Patients with epilepsy, headache and comorbidity differed in a number of demographic and clinical aspects. However, for both headache and epilepsy, a family history of the same clinical condition was equally prevalent in patients with and without comorbidity. These findings do not support the purported association between headache and epilepsy.


The inclusion of cognitive behavioural therapeutic components in the undergraduate nursing curriculum. A systematic integrative review of the evidence.
Despite the increasing evidence base for the use of cognitive behavioural interventions in nursing, the extent to which therapeutic interventions and their components are included in the undergraduate curriculum in Australia remains unclear. To explore if and how cognitive behavioural components are being taught to undergraduate nursing students to deliver outcome benefits to students and patients. An integrative literature review reporting links between CBT components and methods taught in pre-registration nurse education. Based on an analysis of current literature, it is suggested that evidence-based cognitive behavioural components are being taught to undergraduate nursing students. The study reports that teaching cognitive behavioural components can bring clinical benefits to positive patient outcome, to student wellbeing and study performance. However, the teaching of cognitive behavioural conceptual models as a therapeutic skill was found to be limited. Recommendations to incorporate a simple, evidence based cognitive behavioural conceptual tool, highlights how nurse educators can address the requirement to teach from the current evidence base.


Starmerella vitis f.a., sp. nov., a yeast species isolated from flowers and grapes.
A novel yeast species of Starmerella vitis f.a. sp. nov. is proposed to accommodate five strains isolated from flowers, grapes and an insect in the Azores, Canada, Hungary, Palau and Taiwan. As the strains were genetically distinct, we used parsimony network analysis based on ITS-D1/D2 sequences to delineate the species in a statistically objective manner. According to sequence comparisons and phylogenetic analysis, the novel species is most closely related to Starmerella lactis-condensi. The two species cannot be distinguished by conventional physiological tests. The type strain of Starmerella vitis f.a., sp. nov. is CBS 16418T; Mycobank number MB 835251.


Magnetic-field-induced band-structure change in CeBiPt.
We report on a field-induced change of the electronic band structure of CeBiPt as evidenced by electrical-transport measurements in pulsed magnetic fields. Above approximately 25 T, the charge-carrier concentration increases nearly 30% with a concomitant disappearance of the Shubnikov-de Haas signal. These features are intimately related to the Ce 4f electrons since for the non-4f compound LaBiPt the Fermi surface remains unaffected. Electronic band-structure calculations point to a 4f-polarization-induced change of the Fermi-surface topology.


Corynebacterium diphtheriae 67-72p hemagglutinin, characterized as the protein DIP0733, contributes to invasion and induction of apoptosis in HEp-2 cells.
Although Corynebacterium diphtheriae has been classically described as an exclusively extracellular pathogen, there is growing evidence that it may be internalized by epithelial cells. The aim of the present report was to investigate the nature and involvement of the surface-exposed non-fimbrial 67-72 kDa proteins (67-72p), previously characterized as adhesin/hemagglutinin, in C. diphtheriae internalization by HEp-2 cells. Transmission electron microscopy and bacterial internalization inhibition assays indicated the role of 67-72p as invasin for strains of varied sources. Cytoskeletal changes with accumulation of polymerized actin in HEp-2 cells beneath adherent 67-72p-adsorbed microspheres were observed by the Fluorescent actin staining test. Trypan blue staining method and Methylthiazole tetrazolium reduction assay showed a significant decrease in viability of HEp-2 cells treated with 67-72p. Morphological changes in HEp-2 cells observed after treatment with 67-72p included vacuolization, nuclear fragmentation and the formation of apoptotic bodies. Flow cytometry revealed an apoptotic volume decrease in HEp-2 cells treated with 67-72p. Moreover, a double-staining assay using Propidium Iodide/Annexin V gave information about the numbers of vital vs. early apoptotic cells and late apoptotic or secondary necrotic cells. The comparative analysis of MALDI-TOF MS experiments with the probes provided for 67-72p CDC-E8392 with an in silico proteome deduced from the complete genome sequence of C. diphtheriae identified with significant scores 67-72p as the protein DIP0733. In conclusion, DIP0733 (67-72p) may be directly implicated in bacterial invasion and apoptosis of epithelial cells in the early stages of diphtheria and C. diphtheriae invasive infection.


Aza-Ortho-Quinone Methides as Reactive Intermediates: Generation and Utility in Contemporary Asymmetric Synthesis.
The aza-ortho-quinone methide (aza-o-QM) chemistry has overwhelmingly progressed in the past few decades. This review aims to integrate various transition metal-catalyzed and organocatalytic strategies in taming aza-o-QM intermediates, including the aza-ortho-vinylidene quinone methide (aza-o-VQM), aza-ortho-alkynyl quinone methide (aza-o-AQM), aza-para-quinone methide (aza-p-QM), and indole-based aza-o-QM analog. These transient species are often utilized for the direct and enantioselective synthesis of complex (hetero)polycyclic or fused-ring molecular scaffolds such as tetrahydroquinoline and indoline, among others, which are abundant in many natural products, bioactive compounds, and pharmaceuticals.


Berberine and monacolin effects on the cardiovascular risk profile of women with oestroprogestin-induced hypercholesterolemia.
One of the most frequent side effect of oral contraceptives use is a stable alteration of the lipid profile. This could be even more relevant in women affected by polycystic ovary syndrome (PCOS). Considering the importance of a balanced lipid profile in cardiovascular prevention and that the exposure to the drugs could be many years long, our aim was to investigate the possible beneficial effect of a largely tested low-dosed combined lipid-lowering nutraceutical on dyslipidemias induced by oestroprogestins prescribed to young women for different indications. We prospectively enrolled 84 patients in primary cardiovascular disease prevention, with low estimated cardiovascular disease risk (<5 % according to the ESC/EAS guidelines), and LDL-C increased above normal value (LDL-C >130 mg/dL) after the use of at least two different oral oestroprogestins treatments. Forty-four women were prescribed oral oestroprogestins for PCOS, while 40 for pure contraception. The tested nutraceutical contained berberine 500 mg/tab and monacolins 3 mg/tab was prescribed to all enrolled patients, associated the previously prescribed standard lipid-lowering diet. After 3 months of nutraceutical treatment, we observed a significant improvement in BMI (-1.5 ± 0.8 %, p < 0.001), FPG (-6.9 ± 5.8 %, p < 0.001), HOMA index (-3.5 ± 5.6 %, p < 0.001), TC (-20.1 ± 6.6 %, p < 0.001), LDL-C (-25.3 ± 8.9 %, p < 0.001), HDL-C (+14.1 ± 2.2 %, p < 0.001), TG (-29.9 ± 25.2 %, p < 0.001) and hsCRP (-2.5 ± 2.4 %, p = 0.019). Similar results have been obtained even repeating the analysis by subgroups, beyond hsCRP that significantly improved in PCOS patients compared to both the baseline and the non-PCOS group. It appears that the tested combined lipid-lowering nutraceutical is able to equally improve lipid metabolism in oral contraceptive induced hypercholesterolemia in women affected or not by PCOS.


Minute virus of mice (MVM) mRNAs predominantly polyadenylate at a single site.
The polyadenylation sites for MVM(p) and MVM(i) mRNAs were determined by a quantitative hybridization-S1 protection assay. mRNAs produced by MVM(p) both early and late in infection of mouse A9 fibroblasts, and by MVM(p) and MVM(i) late in infection of human NB324K cells, polyadenylate predominantly at a single site, at nucleotide 4908 +/- 2 for MVM(p) and 4843 +/- 2 for MVM(i), shortly downstream of the final AATAAA in each viral genome. These results demonstrate that although the right-hand end of MVM has multiple AATAAA signals, and MVM(p) and MVM(i) vary significantly within this region, 3' end processing of viral mRNAs is not a prevalent mechanism for the regulation of MVM gene expression.


Cerebral blood volume and water extraction from cerebral parenchyma by hyperosmolar contrast media.
The attenuation coefficients of cerebral parenchyma and systemic blood were simultaneously measured at varying times after injection of two contrast agents with differing osmolarities. Cerebral parenchyma was in fact denser than the theory would predict. The osmolarity of the iodinated contrast agents seems to play only a very minor part. However, liposolubility could perhaps explain the leakage of iodine across the BBB and thus the increase in the attenuation coefficient of cerebral parenchyma.


Autoantibodies to Tamm-Horsfall protein associated with urinary tract infections in girls.
Girls with various forms of urinary tract infections and a reference material were analyzed for autoantibodies in serum to the Tamm-Horsfall glycoprotein. Such antibodies could be detected in all sera analyzed. In the control subjects cord blood contained very low IgA and IgM anti-TH, which increased significantly up to the age of 8 months. The IgG anti-TH levels in cord blood correlated with maternal levels. After the age of 2 months the IgG anti-TH followed the anti-TH levels of the other immunoglobulin classes. Among the infants aged 2 to 7 months with acute UTI, no anti-TH increases were found. In girls more than one year of age with acute nonobstructive UTI, IgG and IgA anti-TH levels were significantly higher in those with acute pyelonephritis and reflux, with or without parenchymal reduction, than in those with acute pyelonephritis and normal radiologic findings. The latter group had significantly higher levels of IgG and IgA but not IgM anti-TH than did those with acute cystitits. In contrast, girls with renal parenchymal reduction but no signs of infection at the time of testing had significantly depressed anti-TH levels compared to control values.


Effects of the diuretic furosemide on the sensitivity of glycolysis and glycogen synthesis to insulin in the soleus muscle of the rat.
The diuretic furosemide (0.5 mmol/l) impaired glucose uptake in the soleus muscle of the rat by decreasing the sensitivity of glycolysis to insulin. In addition, at higher concentrations (1.0 mmol/l) the drug inhibited the basal rate of glycolysis. It did not, however, inhibit the rate of glycogen synthesis except at a concentration of 6.0 mmol/l. Since furosemide has some structural similarities to adenosine, the above effects on insulin sensitivity may be due to its ability to act as an adenosine receptor agonist in muscle. These effects of furosemide in skeletal muscle may contribute to the glucose intolerance following therapy with this and similar agents in man.


Changes in Type 2 Diabetes Trends in Children and Adolescents During the COVID-19 Pandemic.
There is concern that the growing incidence of pediatric type 2 diabetes (T2D) may have been further exacerbated by the COVID-19 pandemic. To examine whether trends in new-onset pediatric T2D-inclusive of patients requiring hospitalization and patients managed as outpatients-were impacted during the COVID-19 pandemic, and to compare patient characteristics prior to and during COVID-19. A retrospective single-center medical record review was conducted in a hospital which cares for 90% of Alabama's pediatric T2D patients. Patients with new-onset T2D referred from March 2017 to March 2021 were included. Counts of patients presenting per month ("monthly rates") were computed. Linear regression models were estimated for the full sample and stratified by Medicaid and non-Medicaid insurance status. Patient characteristics prior to vs during COVID-19 were compared. A total of 642 patients presented with new-onset T2D over this period. Monthly rates were 11.1 ± 3.8 prior to COVID-19 and 19.3 ± 7.8 during COVID-19 (P = .004). Monthly rates for Medicaid patients differed prior to and during COVID-19 (7.9 ± 3.4 vs 15.3 ± 6.6, P = .003) but not for non-Medicaid patients (3.3 ± 1.7 vs 4.0 ± 2.4, P = .33). Regression results showed significant increases in monthly rates during COVID-19 for the full sample (β= 5.93, P < .05) and for Medicaid enrollees (β= 5.42, P < .05) Hospitalization rate, severity of obesity, and hemoglobin A1c remained similar prior to and during COVID-19, though the proportion of male patients increased from 36.8% to 46.1% (P = .021). A rise in new-onset T2D was observed among Alabama's youth during the COVID-19 pandemic, a burden that disproportionately affected Medicaid enrollees and males. Future research should explore the pathways through which the pandemic impacted pediatric T2D.


Electrical properties of embryonic rat brainstem motoneurones in organotypic slice culture.
Organotypic co-cultures of embryonic E18-19 rat brainstem slices and tongue muscle maintained in vitro for more than 16 days were used to study the electrogenic properties of developing embryonic brainstem motoneurones derived from oculomotor, facial or hypoglossal nuclei. This preparation offers a unique opportunity to study the development of prenatal mammalian embryonic motoneurones. Our results show that embryonic rat brainstem motoneurones grown in organotypic culture with tongue muscle develop electrogenic membrane properties that can be compared with those described in newborn and adult animals in slice preparation. These motoneurones displayed a variety of sodium and calcium conductances. Two types of sodium conductances were present in all the recorded motoneurones. An Hodgkin-Huxley TTX-sensitive sodium conductance was involved in the spike potential, whereas a TTX-insensitive high threshold sodium conductance was uncovered when potassium and calcium currents were suppressed. Prominent calcium potentials contributed to the large delayed depolarizations following the spike potentials. High threshold calcium potentials were triggered with a slightly lower threshold than the sodium spike and contributed to the control of the pattern of discharge of the cell in response to slight shifts of membrane potentials. Low threshold calcium potentials were seen in 16% of the recorded motoneurones in hyperpolarized conditions. These calcium currents underlie the triggering of doublet spikes and rebound responses. Brainstem motoneurones in culture did not display differences that could be correlated with the origin of the motoneurone pool explanted and retained undifferentiated features suggesting that development of electrophysiological properties specific of each motoneurone pool are determined by presynaptic networks and target properties.


Ultrastructure of spermiogenesis and the spermatozoon in the genus Joyeuxiella Fuhrmann, 1935 (Cestoda, Cyclophyllidea, Dipylidiidae): comparative analysis of J. echinorhynchoides (Sonsino, 1889) and J. pasqualei (Diamare, 1893).
This paper describes the ultrastructure of spermiogenesis and the mature spermatozoon of two Dipylidiidae cestodes, Joyeuxiella echinorhynchoides and J. pasqualei. In both species, spermiogenesis follows the type III described by Bâ and Marchand for the cyclophyllideans. Nevertheless, it is interesting to note the presence of striated roots associated with the centrioles. The spermatozoon presents the same ultrastructural features in J. echinorhynchoides and J. pasqualei. The apical cone in the anterior extremity of the sperm is more than 2.0 microm long in both J. echinorhynchoides and J. pasqualei. Both species present a single crest-like body, 150 nm thick in J. echinorhynchoides and 75 nm thick in J. pasqualei. The cortical microtubules are spiralled at an angle of 40-45 degrees to the spermatozoon axis in both Joyeuxiella species. A periaxonemal sheath and glycogen granules are also described in the mature sperm. We also describe, for the first time, the disposition of glycogen granules in two opposed and spiralled cords in cestodes and the formation of the periaxonemal sheath in the final stage of spermiogenesis.


Pressed Lateral Flow Assay Strips for Flow Delay-Induced Signal Enhancement in Lateral Flow Assay Strips.
This paper proposes that the signal intensity of a lateral flow assay (LFA) strip can be increased by pressing the top of the strip, effectively reducing its flow rate. The reduced flow rate allows more time for antigen-antibody interactions to occur, resulting in increased signal intensity and an improved detection limit. To assess the potential of the pressed LFA (pLFA) strip, C-reactive protein (CRP) diluted in phosphate-buffered saline (PBS) and serum is detected, affording signal enhancement and a lowered limit of detection. Additionally, to show that the signal enhancement by pressure-induced flow delay applies to existing LFA products, commercially available COVID-19 antigen test strips are pressed, and signal enhancement is observed. Lastly, we show that the signal intensity of COVID-19 LFA kits can be increased by approximately two-fold at maximum by applying pressure on top of the manufactured product. This study suggests that pressed LFA strips can be used to reduce the chances of determining ambiguous signals as false-negative results and can potentially improve the detection sensitivity. The online version contains supplementary material available at 10.1007/s13206-022-00085-w.


Ecological traits do not predict the uptake of microplastics by fishes in a Neotropical River.
Pollution by synthetic polymers is even more problematic to the environment when this material is fragmented into small portions, forming microplastics (MPs). We analyzed the contamination of ichthyofauna by MPs in an important river of the Atlantic Rainforest in regard to abundance, diversity of morphotypes, polymers, colors, and sizes of the synthetic particles in 20 species of fish. Fish were collected in November 2019 and in March 2020 in five sites along the Pomba River. Of the 101 fish analyzed, 49 (49%) presented MPs in at least one organ. Of the 20 species of fish collected 13 included individuals with at least one MP in their analyzed organs. The organs, trophic categories and feeding areas did not affect the general abundance of MPs types. Blue MPs were predominant, followed by the colors black, red, and white. MP fibers represented 91% of total MPs. Most MPs were between 2 and 3 mm in size. Polyethylene terephthalate (PET), polypropylene (PP), polyamide (PA), polyvinylidene chloride "Nylon" (PVDC), and high-density polyethylene (HDPE) were detected in the fishes. The exposure of the fish species to MPs was associated mainly with individual size and species-specific aspects, regardless of ecological traits. Considering that 55% of the fish species studied are consumed by humans, it is necessary to study the potential impact of MP ingestion on human health and to understand to what extent we may be consuming both plastic particles and contaminants that are adsorbed to MPs.


Surgical modifications, additions, and alternatives to Kasai hepato-portoenterostomy to improve the outcome in biliary atresia.
Kasai hepato-portoenterostomy (HPE) is the most widely used surgical technique to restore bile flow in biliary atresia (BA). We aimed to review literature on HPE substitutes and additions to Kasai especially in advanced BA (ABA). A PubMed search was done for surgical procedures apart from or along with Kasai HPE for BA. Additional procedures to prevent cholangitis were also reviewed. Procedures and outcome were analysed. Alternative procedures done by the authors have also been described briefly. Results have been compiled in this review article. In ABA, with portal hypertension and liver cirrhosis, Kasai HPE is associated with poor outcome, increased morbidity, and even mortality. Most require liver transplant (LT). Some alternatives to HPE include exploration at porta hepatis to assess the bile flow yet avoid the major surgery (HPE) as a bridge to LT. Conduit diversion may help to combat cholangitis resistant to steroid therapy. Stoma formation is not preferred in cases listed for LT due to high risk of bleeding. Hepatocyte infusion, stem cell therapy, and synthetic liver are the future options to meet the challenges in BA. Various alternative procedures may become handy in the future especially in ABA.


Extended Follow-up of an Educational Intervention Encouraging Appropriate Use of Blood Transfusions.
A restrictive transfusion strategy of packed red blood cells (PRBCs) has been associated with at least non-inferior patient outcomes in a variety of clinical settings. In December 2014, we conducted an educational intervention which consisted of an oral presentation and computerized notifications at a single tertiary medical center. The aim of this study was to examine the long-term effects of a simple and low-cost educational intervention aimed to promote awareness to transfusion guidelines. We retrospectively analyzed all PRBC transfusions ordered between 2014 and 2017. The primary end point was defined as the percentage of PRBC transfused to patients with hemoglobin (Hb) ≥8 g/dL. Between 2014 and 2017, a total of 27,475 PRBCs were transfused in our medical center. There was a continuous reduction in the percentage of PRBCs transfused at a Hb level ≥8 g/dL between 2014 and 2017, with a matching increase in the PRBC percentage trans-fused at Hb <7 g/dL (OR reduction of 42%, 95% CI 0.54-0.62 and OR increase of 68% [95% CI 1.56-1.81], respec-tively). A simple educational intervention likely contributed to sustained improvement in the appropriateness of PRBC transfusions.