Gene silencing of TNF-alpha in a murine model of acute colitis using a modified cyclodextrin delivery system.
Inflammatory bowel disease (IBD) is a chronic relapsing inflammation of the gastrointestinal tract. The cytokine TNF-alpha (TNF-α) plays a pivotal role in mediating this inflammatory response. RNA interference (RNAi) holds great promise for the specific and selective silencing of aberrantly expressed genes, such as TNF-α in IBD. The aim of this study was to investigate the efficacy of an amphiphilic cationic cyclodextrin (CD) vector for effective TNF-α siRNA delivery to macrophage cells and to mice with induced acute-colitis. The stability of CD.siRNA was examined by gel electrophoresis in biorelevant media reflecting colonic fluids. RAW264.7 cells were transfected with CD.TNF-α siRNA, stimulated with lipopolysaccharide (LPS) and TNF-α and IL-6 responses were measured by PCR and ELISA. Female C57BL/6 mice were exposed to dextran sodium sulphate (DSS) and treated by intrarectal administration with either CD.siRNA TNF-α or a control solution. In vitro, siRNA in CD nanocomplexes remained intact and stable in both fed and fasted simulated colonic fluids. RAW264.7 cells transfected with CD.TNF-α siRNA and stimulated with LPS displayed a significant reduction in both gene and protein levels of TNF-α and IL-6. CD.TNF-α siRNA-treated mice revealed a mild amelioration in clinical signs of colitis, but significant reductions in total colon weight and colonic mRNA expression of TNF-α and IL-6 compared to DSS-control mice were detected. This data indicates the clinical potential of a local CD-based TNF-α siRNA delivery system for the treatment of IBD.


Lectins as probes for the assay of rhabdovirus infections in plants.
Thirteen different, biotinylated plant lectins were tested for their ability to recognize specifically the glycoproteins of the two different plant rhabdoviruses potato yellow dwarf virus and eggplant mottled dwarf virus. All viruses were propagated on the same plant host species, Nicotiana rustica L. The lectin-binding to the viral proteins was tested after electrophoretic separation and transfer to nitrocellulose membranes. Besides purified virus also partially pure virus preparations were used for the tests, in order to determine the specificity. The lectins had been selected for specificities to either one of the following monosaccharides: mannose, glucose, galactose, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine and fucose. In the test panel of thirteen lectins, seven were found to react with the viral glycoproteins. Among these, four (LCA, VFA, PSA, Con A) belonged to the mannosyl- or glycosyl-specific group. However, these four lectins reacted also with other host proteins when partially pure virus preparations were used as samples. The other three lectins (GSA2b, STA, WGA) were specific for N-acetyl-D-glucosamine and detected almost exclusively the viral glycoproteins. Two of these lectins, STA and WGA, were extremely suitable for virus-specific assays, since they did not react with glycoproteins in healthy controls that were identical or comparable in their electrophoretic mobility with the rhabdovirus glycoproteins. No binding to viral glycoproteins was observed with galactose-, N-acetyl-galactosamine- and fucose-specific lectins. The assay for rhabdovirus glycoproteins in plants with the lectins was approximately 8-16 times less sensitive than with virus-specific antibodies.


Role of the interferon regulatory factors (IRFs) in virus-mediated signaling and regulation of cell growth.
As a response to viral infection, cells express the early inflammatory genes that encode small proteins generally called cytokines or chemokines. These protein can activate immune responses to viral infection as well as to modulate directly the outcome of viral infection. The group of proteins with the direct antiviral effects have been called interferons. The stimulation of interferon synthesis in infected cells is regulated on a transcriptional level and two families of cellular transcriptional factors seem to play a critical role in the transcriptional activation of interferon genes. The first one are the proteins of NF-kappaB family and the second is the family of the interferon responsive factors. While both of the types of the transcriptional factors are important for the induction of interferon beta gene, the NF-kappaB factor do not seems to participate in the induction of interferon alpha genes. The present review is focused on the recently identified new members of cellular IRF family and their role in virus mediated response, responses and cell growth. In addition the HHV-8 encoded vIRFs are described.


Significance of meconium during labor.
Continuous fetal heart rate (FHR) monitoring and routine fetal scalp blood sampling was utilized in the evaluations of 366 fetuses during labor. One hundred and six patients had meconium in the amniotic fluid at some time during labor. A total of 26,110 uterine contractions were monitored during these 366 labors. The incidence of FHR patterns as a percentage of uterine contractions was calculated for the meconium and nonmeconium groups. Although there was a 3 1/2-fold increase in the incidence of low five-minute Apgar scores (less than 7) in the meconium group, signs of fetal distress were, with rare esception, not significantly different from those in the nonmeconium group. The presence of meconium in the amniotic fluid without signs of fetal asphyxia (late decelerations and acidosis) is not a sign of fetal distress and need not be an indication for active intervention. The combination of fetal asphyxia and meconium staining of the amniotic fluid, however, does enhance the potential for meconium aspiration and a poor neonatal outcome. Universal fetal heart rate monitoring and appropriate fetal acid-base evaluation is recommended for following patients with meconium in the amniotic fluid during labor.


Cohesive zone modeling of mode I tearing in thin soft materials.
The use of modeling and simulation is growing rapidly in applications such as surgery simulations, injury mechanics and tissue engineering. The aim of this study is to model and simulate tissue tearing and the resulting failure for use in such applications. In particular, our goal is to characterize the mechanics of mode I tearing in thin soft materials. We use the cohesive zone modeling approach to characterize the propagation of tears in a processed meat product (PMP). The bulk response of the PMP is modeled with a hyperelastic material model and the interface with a cohesive zone model. A multistep parameter estimation approach is developed to determine the bulk and the cohesive model parameters from uniaxial extension and tearing experiments. Results show that the proposed approach is able to capture both material and geometrical nonlinearities inherent to such problems, and accurately model the overall force-displacement response of thin soft materials during tearing at slow rates.


Synthesis and anticonvulsant activity of aromatic tetramethylcyclopropanecarboxamide derivatives.
As part of our ongoing research on potential new antiepileptic drugs (AEDs), a series of tetramethylcyclopropanecarboxamide derivatives containing benzene ring were designed, synthesized, and evaluated for anticonvulsant activities in the murine maximal electroshock (MES) and subcutaneous pentylenetetrazole (scMet) seizure tests. The most potent compound emerging from this study was N-(2,2,3,3-tetramethylcyclopropanecarboxamide)-p-phenyl-sulfonamide (21), possessing an ED(50) value of 26mg/kg in the rat-MES test and a remarkable PI (PI=TD(50)/ED(50)) value above 19. The better anticonvulsant potency of compound 21 and its wider safety margin compared to valproic acid (VPA) and zonisamide make it a potential candidate to become a new AED second-generation to VPA.


Fabrication of g-C3N4 Nanosheets Anchored With Controllable CdS Nanoparticles for Enhanced Visible-Light Photocatalytic Performance.
Herein, g-C3N4/CdS hybrids with controllable CdS nanoparticles anchoring on g-C3N4 nanosheets were constructed. The effects of CdS nanoparticles on photocatalytic H2 production and organic molecule degradation for g-C3N4/CdS hybrids were investigated. The maximum rate of H2 production for g-C3N4/CdS sample was 1,070.9 μmol g-1 h-1, which was about four times higher than that of the individual g-C3N4 nanosheet sample. The enhanced photocatalytic performance for prepared hybrids could be mainly attributed to the following causes: the formed heterojunctions can contribute to the light absorption and separation of photogenerated electrons and holes, the two-dimensional layered structure facilitates the transmission and transfer of electrons, and high specific surface area could provide more exposed active sites.


Preoperative gender differences in pulmonary gas exchange in morbidly obese subjects.
Morbidly obese men may have poorer pulmonary gas exchange compared to morbidly obese women (see Zavorsky et al., Chest 131:362-367, 2007). The purpose was to compare pulmonary gas exchange in morbidly obese men and women at rest and throughout exercise. Twenty-five women (age=38+/-10 years, 164+/-7 cm, body mass index or BMI = 51+/-7 kg/m(2), peak oxygen consumption or VO(2peak)=2.0+/-0.4 l/min) and 17 men (age=43+/-9 years, 178+/-7 cm, BMI=50+/-10 kg/m(2), VO(2peak)=2.6+/-0.8 l/min) were recruited to perform a graded exercise test on a cycle ergometer with temperature-corrected arterial blood-gas samples taken at rest and every minute of exercise, including peak exercise. At rest, women were 98% predicted for pulmonary diffusion compared to 88% predicted in men. At rest, women had better pulmonary gas exchange compared to the men which was related to women having a lower waist-to-hip ratio (WHR; p<0.01). Only 20% of the subjects had an excessive alveolar-to-arterial oxygen partial pressure difference (>or=25 mmHg) at peak exercise, but 75% of the subjects showed inadequate compensatory hyperventilation at peak exercise (arterial carbon dioxide pressure >35 mmHg), and both were not different between genders. At rest, morbidly obese men have poorer pulmonary gas exchange and pulmonary diffusion compared to morbidly obese women. The better gas exchange in women is related to the lower WHR in the women. During exercise, few subjects showed disturbances in pulmonary gas exchange despite demonstrating poor compensatory hyperventilation at peak exercise.


Preparation and utilization of fluorescent synthetic peptides.
In this study, several methods for controlled labelling of synthetic peptides by the use of fluorescent compounds (fluorescein isothiocyanate and dimethylaminonaphthalene sulfonyl chloride) were investigated. The first reagent yielded monofluoresceinated, active compounds only when the peptides lacked lysine residues. Monolabelling of peptides in solution with dimethylaminonaphthalenesulphonyl chloride was hindered by the broad reactivity of the reagent, but was achieved by reacting the fluorochrome on protected resin-bound peptides in solid-phase synthesis. The remarkable stability of the linkage allowed the cleavage of the peptide from the resin and deprotection of side-chain functions without hydrolysis of the labelled group. The binding of antipeptide antibodies to the labelled fragments was then estimated using different techniques.


Molecular Acceptors Based on a Triarylborane Core Unit for Organic Solar Cells.
Triarylboranes that exhibit p-π* conjugation serve as versatile building blocks to design n-type organic/polymer semiconductors. A series of new molecular acceptors based on triarylborane is reported here. These molecules are designed with a boron atom that bears a bulky 2,4,6-tri-tert-butylphenyl (Mes*) substituent at the core and strong electron-withdrawing 2-(3-oxo-2,3-dihydroinden-1-ylidene)malononitrile (IC) units as the end-capping groups that are linked to the core by bithiophene bridges. Butyl or butoxy groups are introduced to the bithiophene units to tune the optoelectronic properties. These molecules show nearly planar backbones with highly localized steric hindrance at the core, low LUMO/HOMO energy levels, and broad absorption bands spanning the visible region, which are all very desirable characteristics for use as electron acceptors in organic solar cell (OSC) applications. The attachment of butyl groups to the bithiophene bridges brings about a slightly twisted backbone, which in turn promotes good solubility and homogeneous donor/acceptor blend morphology, whereas the introduction of butoxy groups leads to improved planarity, favorable stacking in the film state, and a greatly reduced band gap. OSC devices based on these molecules exhibit encouraging photovoltaic performances with power conversion efficiencies reaching up to 4.07 %. These results further substantiate the strong potential of triarylboranes as the core unit of small molecule acceptors for OSC applications.


Effects of clinical mastitis and puerperal diseases on reproductive efficiency of dairy cows.
The objective of this study was to evaluate the effects of clinical mastitis (CM) occurring before or after the first AI postpartum, and puerperal diseases (PD) on the pregnancy per artificial insemination (P/AI), number of AI/conception, and days open (DO) of two different dairy herds (Girolando and Holstein). The CM, PD (retained placenta and metritis), and reproductive data were collected from two dairy farms throughout 1 year. Both farms were located in the southern region of Minas Gerais State, Brazil. One herd was composed of Girolando cows and the other of Holstein cows. Cows were inseminated after estrus detection or submitted to timed AI. Only CM cases (clots in milk accompanied or not by udder inflammation) that occurred before or after first AI postpartum (from calving until 35 days after the first AI) were considered. There were no effects of CM, PD, or both diseases on the reproductive efficiency of the Girolando herd. In the Holstein herd, a reduce P/AI and prolonged DO were verified for those affected by ≥ 2 CM cases. Holstein cows with CM also required more inseminations to become pregnant. A decrease in the P/AI and an increase in the number of AI/conception and DO were observed in cows of the Holstein herd that developed only CM, only PD, and for those diagnosed with both diseases. In summary, considering that some management differences exist between the two dairy farms, CM occurrence (before or after the first AI postpartum) and puerperal diseases negatively affected the reproductive efficiency of the Holstein herd. However, these diseases did not compromise the reproductive efficiency of the Girolando herd.


Safety and Efficacy of Tenapanor for Long-term Serum Phosphate Control in Maintenance Dialysis: A 52-Week Randomized Phase 3 Trial (PHREEDOM).
Treating hyperphosphatemia is a tenet of dialysis care. This trial assessed the safety and efficacy of tenapanor for the management of hyperphosphatemia. In this 52-week phase 3 study (NCT03427125), participants receiving maintenance dialysis with both hyperphosphatemia (serum phosphate 6.0-10.0 mg/dl) and a 1.5 mg/dl increase after phosphate binder washout were randomized (3:1) to tenapanor 30 mg twice daily for 26 weeks (randomized treatment period) or sevelamer carbonate (52-week safety control). Participants completing 26 weeks of treatment with tenapanor were rerandomized (1:1) to tenapanor or placebo for 12 weeks (randomized withdrawal period), and were eligible to enter the 14-week safety extension period. With input from the US Food and Drug Administration, the primary efficacy end point was the difference in the change in serum phosphate from the end of the randomized treatment period to the end of the randomized withdrawal period, among participants who achieved ≥1.2 mg/dl decrease in serum phosphate during the randomized treatment period (efficacy analysis set). Efficacy was also evaluated in the intention-to-treat (ITT) analysis set. Of 564 eligible participants randomized to receive tenapanor (n=423) or sevelamer carbonate (n=141) during the randomized treatment period, 255 (60%) in the tenapanor group subsequently were rerandomized to tenapanor (n=128) or placebo (n=127) during the randomized withdrawal period. In the efficacy analysis set (n=131), the difference in estimated mean change in serum phosphate level between tenapanor and placebo from the beginning to the end of the randomized withdrawal period was -1.4 mg/dl (P<0.0001); in the ITT analysis set (n=243), the estimated mean difference was -0.7 mg/dl (P=0.002). Loosened stools were the most frequently reported adverse event (53% during the randomized treatment period). Serious adverse events were reported more frequently for participants treated with sevelamer carbonate (16%-23% across the three study periods) compared with tenapanor (11%-17%). Tenapanor reduced serum phosphate concentrations and maintained control of serum phosphate in participants receiving maintenance dialysis, with an acceptable safety and tolerability profile.


Design and synthesis of novel quinazolinone-based fibrates as PPARα agonists with antihyperlipidemic activity.
Aiming to discover new antihyperlipidemic agents, a new set of quinazolinone-fibrate hybrids 9a-r bearing the essential features for peroxisome proliferator-activated receptor-α (PPARα) agonistic activity was synthesized and the structures were confirmed by different spectral data. All the target compounds were screened for their PPARα agonistic activity. Compounds 9o and 9q exhibited potent activity, with EC50 values better than that of fenofibrate by 8.7- and 27-fold, respectively. Molecular docking investigations were performed for all the newly synthesized compounds in the active site of the PPARα receptor to study their interactions and energies in the receptor. Moreover, the antihyperlipidemic and antioxidant activities of compounds 9o and 9q were determined using Triton WR-1339-induced hyperlipidemic rats. Compound 9q exhibited effective hypolipidemic activity in a dose-dependent manner, where it significantly reduced the serum levels of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and very-low-density lipoprotein cholesterol and increased the level of high-density lipoprotein cholesterol. Furthermore, it possesses a powerful antioxidant profile where it significantly elevated the levels of reduced glutathione as well as the total antioxidant capacity and significantly decreased the malondialdehyde level. The histopathological studies revealed that compound 9q improved the aortic architecture and hepatic steatosis. These findings support that compound 9q could be a promising lead compound for the development of new antihyperlipidemic agents.


p16INK4a is a beta-catenin target gene and indicates low survival in human colorectal tumors.
Human colorectal carcinomas display an infiltrative front of invasion where tumor cells undergo an epithelomesenchymal transition associated with low survival. Epithelomesenchymal transition is regulated by a nuclear beta-catenin accumulation, and subsequently, activation of beta-catenin/TCF4 target genes similar to CYCLIN D(1). Unexpectedly, these tumor cells are characterized by low proliferation, which correlates with the expression of the cell cycle inhibitor p16(INK4A). Therefore, we investigated the molecular mechanism of the transcriptional regulation of p16(INK4A) in colorectal cancer and its correlation with survival. Molecular biological techniques were used for investigating the transcriptional mechanisms of the p16(INK4A) gene regulation. Moreover, p16(INK4A) expression was correlated with the 10-year survival of patients with colorectal carcinomas. In colorectal carcinomas, expression of the p16(INK4A) gene is regulated by beta-catenin/TCF4 and correlates with low survival rates of patients with tumors displaying an infiltrative front of invasion. beta-catenin/TCF4 regulates cell cycle promoting (c-MYC, CYCLIN D(1)) and inhibiting genes (p16(INK4A)) at the same time in the mesenchymally differentiated tumor cells at the front of invasion. The function of p16(INK4A) seems to supersede in this context thus leading to low proliferation. Moreover, these tumor cells seem to govern the outcome of colorectal cancer independently of their proliferation.


Naloxone does not improve hemodynamics following graded hemorrhage in a canine model.
Two recent studies have demonstrated no improvement in hemodynamic parameters or survival following naloxone administration in hemorrhagic shock. This finding is in contrast to those of earlier studies, which consistently demonstrated a beneficial effect. The current study evaluated naloxone's ability to improve hemodynamics when administered following a fixed-volume hemorrhage, during a period of partial hemodynamic compensation. Thirteen conditioned beagles were anesthetized with pentobarbital (25 mg/kg intravenously), endotracheally intubated, and instrumented with a femoral arterial line and a pulmonary artery thermodilution catheter. Animals were then subjected to an estimated 50% graded hemorrhage (45 ml/kg) over one hour. Following hemorrhage, animals were observed for 90 minutes, then reinfused with shed blood over 30 minutes, and finally observed for an additional 60 minutes. Six animals received naloxone (2 mg/kg intravenously) 30 minutes after completion of hemorrhage and then 2 mg/kg/hr for the duration of the study. Seven control animals received an equivalent volume of saline. Heart rate, mean arterial pressure, central venous pressure, cardiac output, arterial and mixed venous blood gases, and serum lactate level were measured at regular intervals throughout the study. Cardiac index and systemic vascular resistance index were calculated at the same intervals. Overall, there were no statistically significant differences in the mean data values for mean arterial pressure, cardiac index, systemic vascular resistance index, or lactate, although lactate values were consistently higher in the naloxone group (two-tailed independent Student's t test). We conclude that naloxone does not significantly improve hemodynamics when administered after a fixed-volume hemorrhage.


Isolation of two hemorrhagic toxins from Crotalus basiliscus basiliscus (Mexican west coast rattlesnake) venom and their effect on blood clotting and complement.
1. Two hemorrhagic toxins of mol. wt 27,000 (B1) and 27,500 (B2) and pI 9.8 and 5.2 respectively were isolated from Crotalus basiliscus venom. 2. The two proteinases did not cross-react antigenically. 3. Both toxins caused hemorrhage in mice and each was capable of hydrolyzing hide power azure, casein, collagen and fibrin. 4. B1 hydrolyzed the A alpha, B beta and gamma chains of fibrinogen. B2 hydrolyzed the A alpha and B beta chains of fibrinogen, but not the gamma chain. 5. Both proteinases inactivated guinea pig complement.


Commitment of the mentally ill to outpatient treatment.
Outpatient commitment is examined in order to determine what types of civil commitment respondents are ordered to outpatient treatment. The sample consisted of 1226 adult civil commitment respondents having initial hearings in North Carolina. Respondents placed on outpatient commitment are compared with respondents who were released and respondents who were committed to the hospital. Respondents ordered to outpatient commitment are significantly more likely to be of the chronic, revolving door population of mental patients. Potential problems in evaluating the success of outpatient commitment are discussed.


Impact on Survival of Pretransplant Portal Vein Thrombosis.
Portal vein thrombosis (PVT) still remains a major challenge in managing liver transplant recipients. The aim of this study was to describe PVT prevalence in our health district and identify potential risk factors involved. Although further research is needed to make some general appointments, we identified anticoagulation therapy before transplant as a potential protective factor against PVT.


Human trophoblast-derived exosomes attenuate doxorubicin-induced cardiac injury by regulating miR-200b and downstream Zeb1.
Human trophoblast stem cells (TSCs) have been confirmed to play a cardioprotective role in heart failure. However, whether trophoblast stem cell-derived exosomes (TSC-Exos) can protect cardiomyocytes from doxorubicin (Dox)-induced injury remains unclear. In the present study, TSC-Exos were isolated from the supernatants of human trophoblasts using the ultracentrifugation method and characterized by transmission electron microscopy and western blotting. In vitro, primary cardiomyocytes were subjected to Dox and treated with TSC-Exos, miR-200b mimic or miR-200b inhibitor. Cellular apoptosis was observed by flow cytometry and immunoblotting. In vivo, mice were intraperitoneally injected into Dox to establish a heart failure model. Then, different groups of mice were administered either PBS, adeno-associated virus (AAV)-vector, AAV-miR-200b-inhibitor or TSC-Exos via tail vein injection. Then, the cardiac function, cardiac fibrosis and cardiomyocyte apoptosis in each group were evaluated, and the downstream molecular mechanism was explored. TSC-Exos and miR-200b inhibitor both decreased primary cardiomyocyte apoptosis. Similarly, mice receiving TSC-Exos and AAV-miR-200b inhibitor exhibited improved cardiac function, accompanied by reduced apoptosis and inflammation. The bioinformatic prediction and luciferase reporter results confirmed that Zeb1 was a downstream target of miR-200b and had an antiapoptotic effect. TSC-Exos attenuated doxorubicin-induced cardiac injury by playing antiapoptotic and anti-inflammatory roles. The underlying mechanism could be an increase in Zeb1 expression by the inhibition of miR-200b expression. In summary, this study sheds new light on the application of TSC-Exos as a potential therapeutic tool for heart failure.


Too much of a good thing: the Prl-3 in p53's oyster.
In this issue of Molecular Cell, Basak et al. (2008) identify Prl-3 as a p53-inducible gene acting in DNA damage-induced cell-cycle arrest. Both deletion and overexpression of Prl-3 can be cytostatic, adding to a growing list of genes whose expression must be carefully titrated for proper cell proliferation.


Genome sequences of four divergent multidrug-resistant Acinetobacter baumannii strains isolated from patients with sepsis or osteomyelitis.
Acinetobacter baumannii is a Gram-negative bacterium that causes nosocomial infections worldwide, with recent prevalence and higher frequency in wounded military personnel. Four A. baumannii strains from the Walter Reed Army Medical Center (WRAMC) isolated between 2008 and 2009 were sequenced, representing diverse, multidrug-resistant isolates from osteomyelitis or septic patients.


Encapsulation of sesamol and retinol using alginate and chitosan-coated W/O/W multiple emulsions containing Tween 80 and Span 80.
The conditions of production of multiple W/O/W nanoemulsions containing sesamol and retinol were optimized using response surface methodology (RSM). Span 80 (5, 10, and 15 % w/v), Tween 80 (1, 5.5, and 10 % w/v), and water in oil ratio (W/O) (20, 30, and 40 %) were considered as independent variables while encapsulation efficiency (EE%) and particle size were taken as dependent variables. Alginate (Alg) and chitosan (CS) were also applied to form a deposit layer. An optimum sample with an EE of 92.93 % and particle size of 381.94 nm was produced when Tween 80, Span 80, and W/O were 6.24 %, 10.84 %, and 37.70 %, respectively. Based on the Fourier transform infrared spectroscopy (FTIR), detection of hydrophobic band (2899 cm-1) approved the physical entrapment of biomolecules. Differential scanning calorimetry (DSC) indicated an endothermic peak at 236.48 °C associated with the ionic interactions of Alg-CS. Confocal laser scanning microscopy (CLSM) indicated Alg-CS complex deposit layer formed by electrostatic attraction surrounding the W/O/W multiple layers. The in vitro release of sesamol and retinol was 39 % of sesamol and 22 % of retinol in simulated gastric fluid (SGF) and 56 % and 22 % in simulated intestinal fluid (SIF), respectively.


Alteration of neutrophil (PMN) function by heparin, dexamethasone, and enalapril.
The aim of this study is to investigate the effect of a seemingly divergent class of pharmacologic agents, each having been reported to suppress intimal hyperplasia, on neutrophil (PMN) function. Human PMNs were isolated and exposed for 30 min to either saline or one of three different pharmacologic agents, each tested at three different concentrations: Group 1, saline (control, n = 14); Groups 2-4, heparin (5000 units, n = 8; 2500 units, n = 6; 1250 units, n = 6) respectively; Groups 5-7, dexamethasone (4 mg, n = 8; 2 mg, n = 6; 1 mg, n = 6), respectively; and Groups 8-10, enalapril (1.25 mg, n = 8; 0.62 mg, n = 6; 0.31 mg, n = 6). Superoxide anion production was measured by the reduction of cytochrome c in a spectrophotometric assay. Chemotaxis was evaluated by the number of PMNs migrating across a filter using a Neuro Probe chamber. Phagocytosis was determined by the ingestion of opsonized zymosan particles by PMNs. Serum obtained from each PMN donor was used both to opsonize the zymosan and as a chemoattractant in the chemotaxis assay. No agent, at any dose, significantly changed superoxide production when compared to control cells. All three agents significantly inhibited PMN chemotaxis at every dose tested (P less than 0.01). In the phagocytosis assay, both heparin (at high and intermediate doses) and enalapril (at all doses) significantly reduced phagocytic activity (P less than 0.01); however, dexamethasone (at high and intermediate doses) produced a marked stimulation (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Dietary restriction suppresses proteotoxicity and enhances longevity by an hsf-1-dependent mechanism in Caenorhabditis elegans.
Dietary restriction increases lifespan and slows the onset of age-associated disease in organisms from yeast to mammals. In humans, several age-related diseases are associated with aberrant protein folding or aggregation, including neurodegenerative disorders such as Alzheimer's, Parkinson's, and Huntington's diseases. We report here that dietary restriction dramatically suppresses age-associated paralysis in three nematode models of proteotoxicity. Similar to its longevity-enhancing properties, dietary restriction protects against proteotoxicity by a mechanism distinct from reduced insulin/IGF-1-like signaling. Instead, the heat shock transcription factor, hsf-1, is required for enhanced thermotolerance, suppression of proteotoxicity, and lifespan extension by dietary restriction. These findings demonstrate that dietary restriction confers a general protective effect against proteotoxicity and promotes longevity by a mechanism involving hsf-1.


Redefining ADHD Using an Adult Population: Should Inattention be Viewed as a Separate Dimension From Cognitive and Physiological Activity Level?
Objective: The purpose of this study was to reexamine the latent structure of ADHD and sluggish cognitive tempo (SCT) due to issues with construct validity. Two proposed changes to the construct include viewing hyperactivity and sluggishness (hypoactivity) as a single continuum of activity level, and viewing inattention as a separate dimension from activity level. Method: Data were collected from 1,398 adults using Amazon's MTurk. A new scale measuring activity level was developed, and scores of Inattention were regressed onto scores of Activity Level using curvilinear regression. Results: The Activity Level scale showed acceptable levels of internal consistency, normality, and unimodality. Curvilinear regression indicates that a quadratic (curvilinear) model accurately explains a small but significant portion of the variance in levels of inattention. Conclusion: Hyperactivity and hypoactivity may be viewed as a continuum, rather than separate disorders. Inattention may have a U-shaped relationship with activity level. Linear analyses may be insufficient and inaccurate for studying ADHD.


Non-contact ultra-widefield retinal imaging and fundus fluorescein angiography of an infant with incontinentia pigmenti without sedation in an ophthalmic office setting.
When fluorescein angioscopy or angiography is required in an infant, it is usually performed in the operating theater or neonatal unit. We report a case of an infant with incontinentia pigmenti in whom we were able to acquire angiographic information in an office setting by using an ultra-widefield non-contact system with oral fluorescein.


Perceptions of organizational culture among infection preventionists in Israel, the United States, and Thailand: Results from national infection prevention surveys.
In national surveys of infection preventionists in Israel (n = 15), the United States (n = 415), and Thailand (n = 100), we found that views of organizational culture track well with these countries' cultural dimension scores of power distance and individualism. Our findings highlight the importance of considering cultural dimensions when implementing infection prevention efforts.


catena-Poly[bis(1,3-benzo-thia-zol-3-ium) [[di-chlorido-anti-monate(III)]-di-μ-chlorido-μ-oxido-[chlorido-anti-monate(III)]-μ-chlorido]].
The title compound, {(C7H6NS)2[Sb2Cl6O]} n , contains two benzo-thia-zolidium cations and one tri-μ-chlorido-tri-chlorido-μ-oxido-di-anti-monate(III) anion. The structure of the inorganic cation may be described as as being built up from two polyhedra, i.e. a square-pyramidal SbCl4O and a distorted octa-hedral SbOCl5 unit, sharing a common face (comprising the O atom and two Cl atoms). The two benzo-thia-zole cations are quasi-planar and subtend a dihedral angle of 19.93 (5)°. The crystal packing can be described by alternating (100) layers and [001] chains of the organic cations and inorganic anions connected through an extensive three-dimensional network of N-H⋯Cl, C-H⋯O and C-H⋯Cl hydrogen bonds. This is consolidated by slipped π-π stacking, with centroid-to-centroid distances between the benzo-thia-zole rings of 3.7111 (18)-3.8452 (16) Å. These inter-actions link the mol-ecules within the layers and also link the layers together and reinforce the cohesion of the ionic structure.


Identification and spatial patterns of coastal water pollution sources based on GIS and chemometric approach.
Comprehensive and joint applications of GIS and chemometric approach were applied in identification and spatial patterns of coastal water pollution sources with a large data set (5 years (2000-2004), 17 parameters) obtained through coastal water monitoring of Southern Water Control Zone in Hong Kong. According to cluster analysis the pollution degree was significantly different between September-next May (the 1st period) and June-August (the 2nd period). Based on these results, four potential pollution sources, such as organic/eutrophication pollution, natural pollution, mineral/anthropic pollution and fecal pollution were identified by factor analysis/principal component analysis. Then the factor scores of each monitoring site were analyzed using inverse distance weighting method, and the results indicated degree of the influence by various potential pollution sources differed among the monitoring sites. This study indicated that hybrid approach was useful and effective for identification of coastal water pollution source and spatial patterns.


Cannabis use among patients with cutaneous lymphoma: A cross-sectional survey.
To evaluate patterns of cannabis use in patients with cutaneous lymphoma (CL), as well as the association between cannabis use and itch specifically. Cross-sectional survey created in partnership with the Cutaneous Lymphoma Foundation (CLF). The online survey was distributed electronically via email to the CLF listserv and links posted to social media over a 2-week period. Respondents were classified as current cannabis users, prior users, and never users. A visual analog scale (VAS) was used to assess itching severity, improvement of itch, and interest in learning about cannabis. A total of 119 patient responses (61% female, mean age 59 y) were included in analysis. The majority had mycosis fungoides or Sézary syndrome (74%; 88/119) and early stage (IA-IIA) disease (56%; 48/86). Mean VAS itch score was 3.2 + 2.8 for the cohort. Over half (55%; 60/110) reported ever having used cannabis, with 22% (24/110) endorsing current cannabis use. Common methods of cannabis use were smoking (54%) and vaporizing (46%). 25% (6/24) of current users reported using cannabis specifically to treat itch; these respondents noted that cannabis resulted in moderate improvement of itching (mean 6.6/10). There was strong interest in learning more about cannabis and cancer, and most desired this information from their CL doctor/nurse. Cannabis use is common among patients with CL, and patients report improvement of itching as a result of using cannabis. Further studies are needed to elucidate the risks and benefits of cannabis use in this patient population.


Sonographic and Clinical Features of Typhlitis in Pediatric Cancer Patients on Chemotheaphy at Tikur Anbessa Specialized Hospital, Ethiopia, 2021.
Typhlitis, (neutropenic enterocolitis), is a necrotizing enteropathy of the right colon, and is characterized by the clinical triad of fever, abdominal pain, neutropenia and imaging findings of right-side colonic inflammation. It is seen in the setting of severe neutropenia in immune suppressed patients who undergo treatment for malignancies, in those who have organ transplant(s) or congenital or other acquired immunosuppression. We report the clinical and imaging findings of typhlitis in pediatric cancer patients who had received chemotherapy in the largest tertiary center in Addis Ababa, Ethiopia over a period of 20 months. The medical records of hospitalized cancer patients on treatment and with suspected typhlitis and with ultrasound reports were screened (November 2018- July 2020). Retrospective analysis of the clinical and sonographic data of those with typhlitis was done. Typhlitis was identified in 4.2% (12/286) of the patients on chemotherapy. 11 (91.7%) had hematologic malignancies (leukemia, lymphoma), one had a solid tumor (Head and neck embryonal RMS). Most (83.3%) had abdominal pain, diarrhea and neutropenia. Fever was identified in 67.7%. All had ultrasound evidence of typhlitis. and treated with IV antibiotics. Neither complications requiring surgical intervention nor death were seen. The magnitude of disease was comparable to what had previously been reported in other studies. While the presence of clinical a triad should prompt suspicion for the diagnosis, sonography can be used for confirmation and follow up obviating radiation, with good access in a resource limited setting.


How public can public goods be? Environmental context shapes the evolutionary ecology of partially private goods.
The production of costly public goods (as distinct from metabolic byproducts) has largely been understood through the realization that spatial structure can minimize losses to non-producing "cheaters" by allowing for the positive assortment of producers. In well-mixed systems, where positive assortment is not possible, the stable production of public goods has been proposed to depend on lineages that become indispensable as the sole producers of those goods while their neighbors lose production capacity through genome streamlining (the Black Queen Hypothesis). Here, we develop consumer-resource models motivated by nitrogen-fixing, siderophore-producing bacteria that consider the role of colimitation in shaping eco-evolutionary dynamics. Our models demonstrate that in well-mixed environments, single "public goods" can only be ecologically and evolutionarily stable if they are partially privatized (i.e., if producers reserve a portion of the product pool for private use). Colimitation introduces the possibility of subsidy: strains producing a fully public good can exclude non-producing strains so long as the producing strain derives sufficient benefit from the production of a second partially private good. We derive a lower bound for the degree of privatization necessary for production to be advantageous, which depends on external resource concentrations. Highly privatized, low-investment goods, in environments where the good is limiting, are especially likely to be stably produced. Coexistence emerges more rarely in our mechanistic model of the external environment than in past phenomenological approaches. Broadly, we show that the viability of production depends critically on the environmental context (i.e., external resource concentrations), with production of shared resources favored in environments where a partially-privatized resource is scarce.


Characterization of urinary metabolites as biomarkers of colistin-induced nephrotoxicity in rats by a liquid chromatography/mass spectrometry-based metabolomics approach.
Colistin is a polypeptide antibiotic that effectively treats infections caused by multidrug-resistant Gram-negative bacteria, but its clinical use is limited due to nephrotoxicity. The purpose of the present study was to identify biomarkers of colistin-induced nephrotoxicity and to further characterize the mechanisms underlying this process by analyzing urinary metabolites using untargeted metabolomic approach. Rats receiving intraperitoneal administration of colistin sodium methanesulfonate (CMS) (25 or 50mg/kg) exhibited histopathological changes in the kidney and increased blood urea nitrogen levels. Additionally, the levels of phenylalanine, tryptophan, and tyrosine in the urine of the CMS-treated group were significantly higher than those of the control group, suggesting that colistin caused proximal tubular damage. Urinary acetylcarnitine and butyrylcarnitine levels also increased after CMS treatment, but the levels of purine metabolites and metabolites related to the tricarboxylic acid cycle were reduced. The most significant increase in the CMS-treated groups was observed in creatine levels. CMS-induced selective nephrotoxicity may be attributed to relatively high tissue concentrations of colistin in the kidney. Taken together, our results indicate that high levels of colistin in the kidney caused perturbations in the tricarboxylic acid cycle, amino acid metabolism, creatine metabolism, and purine metabolism and ultimately led to kidney injury.


An extraordinary case: suicide with insulin in a grave dug by the victim himself.
The undressed body of a young man was discovered in a pit in a forest. The victim was a 27 year old depressed man, suffering from a severe diabetes. Besides personal objects, three empty insulin vials (400 I.U./vial) and empty ampules of distilled water, injection-needles and a syringe were also found in the pit. The investigation carried out at the scene, the questioning of his family and physicians, and the autopsy clarified this unusual case. In conclusion the hypothesis of "suicide" was maintained: the man administered himself a large dose of insulin; in hypoglycaemic coma he died from a bronchopneumonia in a "grave" dug by himself.


Ultra-High-Field Imaging of the Pediatric Brain and Spinal Cord.
Neuroimaging with ultra-high field magnets (≥7T) provides superior signal-to-noise, spatial resolution and tissue contrast; but also greater safety concerns, longer scanning times, and increased distortion and field inhomogeneity. Brain and spinal cord anatomic microstructure and function imaged in greater detail offers improved lesion detection, delineation, and characterization. The ongoing development of novel imaging contrasts and translation of cutting-edge sequences will aid more accurate, sensitive, and precise diagnosis, interventional planning, and follow-up for a variety of pathologic conditions.


Acute abdominal conditions in children and adolescents.
Evaluation and management of the acute abdomen in childhood require a thorough familiarity with the numerous disorders that may cause abdominal emergencies. Many of these entities are rarely seen in adults or have clinical features that are very different from those observed in adults. Diagnosis is often confounded by difficulties in obtaining an accurate history and physical examination. In infants and young children, complications of various congenital abnormalities are more common causes of abdominal pain, whereas acquired disorders are observed more frequently in older children and adolescents.


Comparison of geriatric nutritional risk index and creatinine index in short-term mortality prediction in maintenance hemodialysis patients.
The aim of this study is to analyze and compare the predictive values of the Geriatric Nutritional Risk Index (GNRI) and Creatinine Index (CI) in the short-term mortality of maintenance hemodialysis patients and to determine their best cut-offs. A total of 169 adult hemodialysis patients were included in this retrospective, cross-sectional, and single-center study. The demographic, clinical, and laboratory data of the month in which the patients were included in the study were obtained from their medical files and computer records. All-cause death was the primary outcome of the study during a 12-month follow-up after baseline GNRI and CI calculations. The mean age of the study population was 57 ± 16 years (49.7% were women, 15% were diabetic). During the one-year observation period, 19 (11.24%) of the cases died (8 CV deaths). The optimal cut-off value for GNRI was determined as 104.2 by ROC analysis [AUC = 0.682 ± 0.06, (95% CI, 0.549-0.815), p = 0.01]. The low GNRI group had a higher risk for all-cause and CV mortality compared to the higher GNRI group (p = 0.02 for both in log-rank test). The optimal sex-specific cut-off was 12.18 mg/kg/day for men [AUC = 0.723 ± 0.07, (95% CI, 0.574-0.875), p = 0.03] and was 12.08 mg/kg/day for females [AUC = 0.649 ± 0.13, (95% CI, 0.384- 0.914), p = 0.01]. Patients with lower sex-specific CI values had higher all-cause and CV mortality (p = 0.001 and p = 0.009 in log-rank test, respectively). In multivariate cox models, both GNRI [HR = 4.904 (% 95 CI, 1.77-13.56), p = 0.002] and sex-specific CI [HR = 5.1 (95% CI, 1.38-18.9), p = 0.01] predicted all-cause mortality. The association of GNRI with CV was lost [HR = 2.6 (CI 95%, 0.54-13.455), p = 0.22], but low CI had a very strong association with CV mortality [HR = 11.48 (CI 95%, 1.25 -104), p = 0.03]. In hemodialysis patients, GNRI and CI have similar powers in predicting all-cause short-term mortality. The association of CI with all-cause death depends on gender. On the other hand, sex-specific CI predicts CV mortality better than GNRI.


Cutting errors in total knee replacement: assessment by computer assisted surgery.
The observed errors in the position of the implanted prosthesis can be due to a number of potential causes. One of these is the potential error during execution of the bone cuts. However, there is only minimal information on this in the current literature. The amount of cutting errors in 40 consecutive total knee replacements was reported. All the operations were done by the same surgeon. The amount of cutting error was measured by the use of computer navigation system. It was hypothesized that there was no difference in the amount of error between bone cut through the cutting slot (slotted cutting) and bone cut done on the surface of the cutting guide (open cutting). It was found that the average absolute cutting error was 1 masculine in the coronal plane and 1.4 masculine in the sagittal plane. Significantly more outlier (more than 3 masculine) was observed in the errors in the sagittal plane (P = 0.014, chi square test). Open cutting resulted in less error in the sagittal plane of the tibial cut when compared with slotted cutting (P = 0.031, Mann-Whitney U Test). This was attributed by the use of a thicker saw blade with higher stiffness in the open cutting method.


Ah locus: genetic differences in susceptibility to cataracts induced by acetaminophen.
The Ahb/Ahb homozygous and the Ahb/Ahd heterozygous inbred mouse strains from the (C57BL/6)(DBA/2)F1 X DBA/2 backcross are genetically responsive to 3-methylcholanthrene. They both also develop, within 6 hours after a large intraperitoneal dose of acetaminophen, an irreversible opacity in the anterior portion of the lens. Such cataract formation does not occur in similarly treated nonresponsive inbred strains or nonresponsive Ahd/Ahd individuals from the same backcross. Differences in acetaminophen metabolism and toxicity are associated with the Ah locus in the mouse, and differences in heritability at the Ah locus exist in the human. Our ophthalmologic findings may be important clinically to certain patients receiving either a single large overdose of this drug or high doses over a long period.


Nurse managers' perceptions of care environment supporting older people's ability to function in nursing homes.
To describe nurse managers' perceptions of the care environment in nursing homes and how the residents' ability to function may be improved. The care environment, considered under physical, social and symbolic dimensions, can be used to maintain and enhance the residents' ability to function. An exploratory, descriptive qualitative research design based on focus groups (n = 4) was used. Fourteen nurse managers from six nursing homes in Southern Finland participated during 2014. Data were analysed using content analysis. The physical environment enables self-management, by facilitating a cosy, home-like and aesthetically pleasing environment and by the design of internal structures and assistive devices. The social environment was described as enabling by providing both private and communal spaces, encouraging personal privacy and self-determination whilst supporting communal interaction and involvement. The symbolic environment was described as supporting a personal culture, care culture development and connecting the components of care perspectives in the community including nursing principles, recognizing and accepting variation in approaches to care. Development in the care environment requires more innovative approaches especially in the social and symbolic dimensions. The results provide a framework for nurse managers for analysing and developing the care environment supporting residents' ability to function.


Microcirculatory assessment of vascular acrosyndrome in anorexia nervosa and analysis of manifestation factors.
Acrocyanosis (AC) is a common manifestation of starving syndrome in anorexia nervosa. We characterized microvascular changes associated with AC and determined discriminating factors between acrally symptomatic and nonsymptomatic patients. We examined 34 patients with anorexia nervosa (15 restrictive-anorectic type, 19 binge-eating/purging type, duration 1-25 years). Nineteen were symptomatic (SP) and 15 were nonsymptomatic (NSP). All underwent photo-pletysmography, sonography of the brachial artery, capillary microscopy and laboratory analysis. Disease characteristics and body mass index did not differ between SP and NSP. In SP more dilated efferent capillary loops and venoles were present (P<.001) and capillary flow velocities were reduced (0.21+/-0.12 ml/min vs. 0.34+/-0.15 ml/min; P=.015). Flow-mediated and nitroglycerin-induced dilatation showed no differences. Symptomatic patients had lower leukocyte counts (P=.008), lower eosinophils (P=.003) and lower LDL (P=.045) concentrations. A logistic regression model identified only leukocytes (P=.017) and eosinophils (P=.023) to be associated with AC. In acrally symptomatic patients the typical microvascular features of AC are present. AC is associated with lower leukocyte counts and lower eosinophils.


Formulation and Evaluation of Dermatological Product Containing Niacinamide.
In this study, niacinamide-based skin creams were formulated and evaluated. Niacinamide (also known as nicotinamide, 3-pyridinecarboxamide) is a physiologically active form of niacin or vitamin B3. Niacinamide, a derivative of niacin, has the ability to treat some skin conditions including aging skin. Quality of the product was assessed by using high-performance thin-layer chromatography (HPTLC) method to determine the content of niacinamide. No change in the physical properties was observed; the pH was in a proper range. The formulations showed good spreadability, no evidence of phase separation, and good consistency during this study period. From this study, it can be concluded that it is possible to develop creams that contain niacinamide having antiaging property and used as a barrier to protect skin.


Diagnostic utility of EUS-FNA in distinguishing hepatocellular carcinoma from gastric submucosal tumor: A case report.
Extrahepatically growing hepatocellular carcinoma (HCC) account for only 0.3 %-2.4 % of all hepatocellular carcinoma cases. We present the rare case of a patient in whom endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) was used to distinguish an extrahepatically growing HCC from a gastric submucosal tumor. The patient was a 44-year-old man who underwent an esophagogastroduodenoscopy (EGD) and was found to have a submucosal tumor (SMT). A gastrointestinal stromal tumor was suspected. Computed tomography (CT) showed an unclear connecting between the tumor and the liver. On immunohistochemical analysis, the tumor cells were positive for CK7, PIVKA-II, and glypican-3, and negative for C-kit. Based on these findings, HCC was suspected, and ethoxybenzyl diethylenetriamine-enhanced magnetic resonance imaging (EOB-MRI) was performed. EOB-MRI revealed the connecting of the tumor and the liver. Thus, the patient was diagnosed as having an extrahepatically protruding HCC, and laparoscopic S2 partial hepatectomy was performed. Pathologically, the tumor was identified as a moderately differentiated HCC. According to the 8th edition of the UICC TNM Classification, the tumor was classified as T1bN0M0 and stage IB. Owing to its rarity, the diagnosis of an extrahepatically protruding HCC is often difficult when the differentiation of a gastric SMT and HCC is unclear on CT. As in this case, if an HCC is also differentiated on hematoxylin and eosin staining after EUS-FNA, immunohistochemistry findings can help in the diagnosis. The findings show that EUS-FNA could be helpful for diagnosis when an HCC needs to be differentiated from a gastric SMT.


Optimisation of a 5-[3-phenyl-(2-cyclic-ether)-methyl-ether]-4-aminopyrrolopyrimidine series of IGF-1R inhibitors.
Taking the pyrrolopyrimidine derived IGF-1R inhibitor NVP-AEW541 as the starting point, the benzyl ether back-pocket binding moiety was replaced with a series of 2-cyclic ether methyl ethers leading to the identification of novel achiral [2.2.1]-bicyclic ether methyl ether containing analogues with improved IGF-1R activities and kinase selectivities. Further exploration of the series, including a fluorine scan of the 5-phenyl substituent, and optimisation of the sugar-pocket binding moiety identified compound 33 containing (S)-2-tetrahydrofuran methyl ether 6-fluorophenyl ether back-pocket, and cis-N-Ac-Pip sugar-pocket binding groups. Compound 33 showed improved selectivity and pharmacokinetics compared to NVP-AEW541, and produced comparable in vivo efficacy to linsitinib in inhibiting the growth of an IGF-1R dependent tumour xenograft model in the mouse.


Efficiency of the male effect with photostimulated bucks does not depend on their familiarity with goats.
In ewes, the ovulatory response of females exposed to familiar rams is lower than the response of those exposed to novel ones. In goats, males rendered sexually active by exposure to long days are more efficient to induce ovulation in seasonal anestrous females than untreated males. Two experiments were conducted to determine 1) whether male goats remain familiar to females after 45days of separation; and 2) whether photostimulated males are able to stimulate the sexual activity of females, independently of their familiarity with them. In Experiment 1, three groups of goats (n=10 goats per group) were put in contact with males (n=2 per group) during 10days in November (familiarization period). These males were called familiar males. After 15, 30 and 45days of separation from the males, females of each group were exposed to familiar or novel males during 10min. In each test, goats in contact with novel males displayed more distress bleats, escapes, head butts, and sniffing than those in contact with familiar males (P<0.05). In Experiment 2, we used sexually inactive (n=4 control males), and sexually active males (n=4 photostimulated males). In February, two groups of goats (n=50 each) were put in contact with control or photostimulated males (n=2 each) during 10days ("familiar" control or photostimulated male, respectively). After 45days of separation from the males, both groups of females were further divided into two groups (n=25 goats per group). In April, two groups were re-exposed to "familiar" control or "familiar" photostimulated males (n=2 per group), whereas the other two groups were exposed to "novel" control or "novel" photostimulated males (n=2 per group). The photostimulated males displayed a higher level of sexual behavior than the controls. The proportion of goats that ovulated and displayed estrus was higher when exposed to the photostimulated males than when exposed to control ones (≥80% vs. 0%; P<0.05). These proportions did not differ between groups exposed to familiar or novel photostimulated males (P>0.05). We concluded that after 45days of separation, males are still familiar to females. The photostimulated males are able to induce the sexual activity of seasonally anestrous goats independently of their familiarity with them.


Isolation of prostaglandin E2-like material from osteonecrosis induced by steroids and its prevention by kallikrein-inhibitor, aprotinin. An experimental study in rabbits.
Prostaglandin E2 (PGE2)-like activity was measured in osteonecrotic femoral heads caused by high doses of methyl prednisolone acetate in rabbits. Serum levels of cholesterol, total lipids, free fatty acids, alkaline phosphatase, SGOT and lipase were also determined. There were increases in local tissue PGE2-like activity, serum cholesterol, total lipid and free fatty acid levels in rabbits with steroid-induced osteonecrosis as compared to the controls. The rabbits given aprotinin together with the steroid showed significant decreases in serum cholesterol and lipid levels coupled with a marked increase in the local PGE2-like activity. Aprotinin prevented osteonecrosis induced by steroids. The results are discussed in the light of the relevant literature.


From the identification of biopsychosocial risk factors to an increase in pain-related self-efficacy (IDRIS) - The online-based conveyance of an explanatory model for chronic back pain: Study protocol of a cohort multiple randomized controlled trial.
Back pain has a high global prevalence and carries a substantial risk for chronification. Biopsychosocial factors are assumed to be critical in the transition from acute to chronic back pain. Digital interventions are a promising tool to educate patients about their complaints. Thus, providing patients with an explanatory model regarding their individual risk factors in the early stage of their complaints via the internet might thus be a valuable approach in treatment. The objectives of the present online study are to investigate the influence of a personalized psychoeducational intervention on self-efficacy and functioning and to examine biopsychosocial risk factors for symptom chronification. The intervention is based on a current model summarizing the empirical knowledge on relevant factors for persistent somatic symptoms, which has not been studied in back pain patients yet. An observational cohort of patients with acute and subacute back pain (N = 564) will be asked about biopsychosocial risk factors via online survey at baseline, 4-week, and 12-week follow-up. Within this cohort, a randomly selected group of 132 participants (intervention group) with psychological burden (MCS-12 score of the SF-12 ≤ 50) and relevant somatic symptom intensity and interference (mean sum score of two numeric rating scales ≥5) and no prior psychotherapeutic treatment will be offered a personalized explanatory model in the form of an animated psychoeducational video. The video will be personalized in terms of participants' individual symptom profile and will be made accessible to watch online for 7 days. Participants will be compared to a control group receiving no treatment regarding change in pain-specific self-efficacy after 1 month as primary outcome, and change in functioning after 1 and 3 months, respectively, as secondary outcomes. Acceptance and usefulness of the intervention will be evaluated using the number of video views and a numeric rating scale. This is the first investigation of a personalized, video-animated online psychoeducation based on patients' individual risk factors for the chronification of back pain and the first systematic evaluation of the risk factors included in a comprehensive aetiological model on persistent somatic symptoms in back pain patients. This way, this study contributes to the understanding of cross-disorder psychopathological factors and a stronger consideration of biopsychosocial factors in the treatment of persistent somatic symptoms. If proven effective, the internet-based intervention will make an important contribution to the early treatment of back pain. The study was approved by the Local Psychological Ethics Committee (LPEK) at the Center for Psychosocial Medicine of the University Medical Center Hamburg-Eppendorf. The study was registered at the German Clinical Trials Register in December 2021 (registration trial number: DRKS00025445).


Indoleamine 2,3 dioxygenase and regulation of T cell immunity.
Regulation of adaptive immune responses is critically important to allow the adaptive immune system to eradicate infections while causing minimal collateral damage to infected tissues, as well as preventing autoimmune disease mediated by self-reactive lymphocytes. Tumors and pathogens that cause persistent infections can subvert immunoregulatory processes to protect themselves from destruction by T cells, to the detriment of patients. A growing body of evidence supports the hypothesis that specialized subsets of dendritic cells expressing indoleamine 2,3 dioxygenase (IDO), which catalyzes oxidative catabolism of tryptophan, play critical roles in regulation of T cell-mediated immune responses. IDO-dependent T cell suppression by dendritic cells suggests that biochemical changes due to tryptophan catabolism have profound effects on T cell proliferation, differentiation, effector functions, and viability. This has critical implications for immunotherapeutic manipulations designed for patients with cancer and chronic infectious diseases. In this review, I focus on dendritic cells that can express IDO, and which acquire potent T cell regulatory functions as a consequence.


Fetal heart foramen ovale area by three-dimensional ultrasound using stic in the rendering mode: reference range and applicability in congenital heart diseases.
To determine reference values for the foramen ovale (FO) area of fetal hearts by three-dimensional (3D) ultrasound using spatio-temporal image correlation (STIC) method in the rendering mode, as well as applicability in fetuses with congenital heart disease (CHD). A retrospective and prospective study was performed of 242 normal fetuses and 36 fetuses with CHD between 20 and 33 + 6 weeks of gestation. The FO area was determined in the four-chamber view with manual delineation. To determine the reference curve of the FO area as a function of gestational age (GA), a linear regression model was utilized with an adjusted coefficient of determination (R2). For intra- and interobserver reproducibility calculations, the concordance correlation coefficient (CCC) was used. The mean ± standard deviation (SD) of the FO measurement area was 21.2 ± 1.8 and 48.1 ± 1.9 mm2 at 20 and 33 weeks' gestation, respectively. A linear correlation was observed between the FO area and GA (1.924*GA - 17.95; R2 = 0.91). Good intra- (CCC = 0.97) and interobserver (CCC = 0.94) agreement was observed for the FO area measurement. The mean difference in FO area between normal and CHD fetuses was - 14.4 mm2 (p < 0.001). Reference values for the FO area of fetal hearts were determined by 3D ultrasound using STIC in the rendering mode. This method showed good intra- and interobserver reproducibility and could be used to assess different CHD types.


Genetic deletion of S6k1 does not rescue the phenotypic deficits observed in the R6/2 mouse model of Huntington's disease.
Huntington's disease (HD) is a fatal inherited autosomal dominant neurodegenerative disorder caused by an expansion in the number of CAG trinucleotide repeats in the huntingtin gene. The disease is characterized by motor, behavioural and cognitive symptoms for which at present there are no disease altering treatments. It has been shown that manipulating the mTOR (mammalian target of rapamycin) pathway using rapamycin or its analogue CCI-779 can improve the cellular and behavioural phenotypes of HD models. Ribosomal protein S6 kinase 1 (S6K1) is a major downstream signalling molecule of mTOR, and its activity is reduced by rapamycin suggesting that deregulation of S6K1 activity may be beneficial in HD. Furthermore, S6k1 knockout mice have increased lifespan and improvement in age-related phenotypes. To evalute the potential benefit of S6k1 loss on HD-related phenotypes, we crossed the R6/2 HD model with the long-lived S6k1 knockout mouse line. We found that S6k1 knockout does not ameliorate behavioural or physiological phenotypes in the R6/2 mouse model. Additionally, no improvements were seen in brain mass reduction or mutant huntingtin protein aggregate levels. Therefore, these results suggest that while a reduction in S6K1 signalling has beneficial effects on ageing it is unlikely to be a therapeutic strategy for HD patients.


Unrelated donor bone marrow transplantation for children with juvenile myelomonocytic leukaemia.
Children with juvenile myelomonocytic leukaemia (JMML) have a poor outcome, with survival in a minority of patients. The major limitation on success of sibling donor bone marrow transplantation for JMML has been reported to be relapse. A total of 46 children with a diagnosis of JMML underwent unrelated donor marrow (URD) transplantation facilitated by the National Marrow Donor Program. Forty-three of 46 patients had neutrophil engraftment at a median of 20 d post transplant, with platelet recovery in 28 of 40 evaluable patients at a median of 34.5 d. Thirty-two of 44 evaluable patients developed acute graft-versus-host-disease (GVHD) (Grades 2-4) and chronic GVHD developed in 14 of 35 evaluable patients. At a median follow-up of 2.0 years, probabilities of survival and disease-free survival were 42% and 24% respectively. The probability of relapse was 58% at 2 years and represents the major cause of treatment failure. Multivariate analysis revealed that chronic GVHD was associated with reduced relapse [risk ratio 0.20 (95% CI 0.04-1.02, P=0.05)] improved survival [risk ratio 0.13 (95% CI 0.03-0.68, P=0.02)] and event-free survival [risk ratio 0.23 (95% CI 0.06-0.94, P=0.04)]. This study demonstrates that relapse is the major cause of treatment failure in patients with JMML undergoing URD transplantation. With lower relapse observed in patients with chronic GVHD, new treatment strategies that focus on enhancing the graft-versus-leukaemia effect may improve survival.


Zooplankton community structure driven by vertebrate and invertebrate predators.
A zooplankton community was established in outdoor experimental ponds, into which a vertebrate predator (topmouth gudgeon: Pseudorasbora parva) and/or an invertebrate predator (phantom midge larva: Chaoborus flavicans) were introduced and their predation effects on the zooplankton community structure were evaluated. In the ponds which had Chaoborus but not fish, small- and medium-sized cladocerans and calanoid copepods were eliminated while rotifers became abundant. A large-sized cladoceran Daphnia longispina, whose juveniles had high helmets and long tailspines as anti-predator devices, escaped from Chaoborus predation and increased. In the ponds which had fish but not Chaoborus, the large-sized Daphnia was selectively predated by the fish while small-and medium-sized cladocerans and calanoid copepods predominated. In the ponds containing both Chaoborus and fish, the fish reduced the late instar larvae (III and IV) of Chaoborus but increased the early instar larvae (I and II). Small- and large-sized cladocerans were scarcely found. The former might have been eliminated by predation of the early instar larvae of Chaoborus, while the latter was probably predated by fish. Consequently, the medium-sized cladocerans, which may have succeeded in escaping from both types of predator, appeared abundantly. The results suggest that various combinations of vertebrate and invertebrate predators are able to drive various kinds of zooplankton community structure.


Molecular epidemiology and antifungal susceptibility of Cryptococcus neoformans isolates from Ugandan AIDS patients.
Little is known of the antifungal susceptibility patterns and molecular epidemiology of Cryptococcus neoformans from tropical regions. We studied 164 clinical isolates of C. neofomans from 120 Ugandan AIDS patients with cryptococcal meningitis by analyzing their electrophoretic karyotypes and antifungal susceptibility profiles. Computer-assisted analysis of karyotype patterns was performed to generate dendrograms. MICs of fluconazole and flucytosine were determined by reference methods. A total of 43 distinguishable DNA types were identified among the 164 isolates. Only 30 patients (25%) were infected with their own unique strain of c. neoformans, whereas 75% of the patients shared their infecting strain with at least one other patient. Among 17 patients with more than one CSF isolate of C. neoformans, sequential isolates were identical or highly related in 12 (71%) and were different in five patients (29%). The isolates were susceptible to both fluconazole and flucytosine and there were no instances in which a stepwise increase in either fluconazole or flucytosine MICs was observed among serial isolates. These findings suggest that the epidemiology of cryptococcal disease in AIDS patients from tropical regions may be somewhat different from that observed in more temperate climates.


Topical negative pressure wound therapy enhances the local tissue perfusion - A pilot study.
Topical negative pressure wound therapy (TNPWT) is a regularly used method in modern wound treatment with a growing and diverse potential for clinical use. So far positive effects on microcirculation have been observed and examined, although precise statements on the underlying mechanism appear unsatisfying. The aim of our study was to extend the understanding of the effect of TNPWT on tissue perfusion and determine the time frame and the extent to which the tissue perfusion changes due to TNPWT. TNPWT was applied to the anterior thighs of 40 healthy individuals for 30 min, respectively. Before and up to 90 min after the application, measurements of the amount of regional haemoglobin (rHb), capillary venous oxygen saturation (sO2), blood flow (flow) and velocity were conducted with spectrophotometry (combining white light spectrometry and laser Doppler spectroscopy) within two different depths/skin layers. A superficial measuring probe for depths up to 3 mm and a deep measuring probe for up to 7 mm were used. All parameters show significant changes after the intervention compared to baseline measurements. The greater effect was seen superficially. The superficially measured rHb, sO2 and flow showed a significant increase and stayed above the baseline at the end of the protocol. Whereas deeply measured, the rHb initially showed a decrease. The flow and sO2 showed a significant increase up to 60 min after the intervention. The application of TNPWT on healthy tissue shows an increase in capillary-venous oxygen saturation and haemoglobin concentration of at least 90 min after intervention. A possible use in clinical practice for preconditioning to enhance wound healing for high-risk patients to develop wound healing disorder, requires further studies to investigate the actual duration of the effect.


Efficiency of a multi-barrier household system for surface water treatment combining a household slow sand filter to a Mesita Azul® ultraviolet disinfection device.
Low-cost household technologies for water treatment are crucial to improving drinking water quality and preventing health, social and economic impacts, mostly in middle- and low-income regions. This work assessed the removal efficiency of physical-chemical and bacteriological parameters from river water by a multi-barrier household water treatment system for 113 consecutive days. This system combines a pre-treatment step through a non-woven synthetic blanket, filtration by an intermittent household slow sand filter (HSSF) and a Mesita Azul® ultraviolet disinfection device. In general, the water quality was improved by the evaluated system. Turbidity was removed by an average of 73% (ranging from 33 to 94%), total coliforms (TC) of 3.88 log10 (ranging from 2.22 to 5.16 log10) and E. coli of 2.49 log10 (ranging from 1.81 to 3.30 log10). Filtration improvement was mostly correlated to HSSF biofilm development and influent water quality. Characterisation of HSSF schmutzdecke demonstrated a predominance of organic content, and a higher presence of carbohydrates than proteins on the sand and the blanket. Ultraviolet disinfection with Mesita Azul® inactivated most of the remaining bacteria after filtration and no regrowth was observed after 15 days of disinfection. In conclusion, the multi-barrier household water treatment system was efficient in treating river water, reducing risks of microbial contamination to achieve safe drinking water.


The neuroendocrine genesis of polycystic ovary syndrome: A role for arcuate nucleus GABA neurons.
Polycystic ovary syndrome (PCOS) is a prevalent and distressing endocrine disorder lacking a clearly identified aetiology. Despite its name, PCOS may result from impaired neuronal circuits in the brain that regulate steroid hormone feedback to the hypothalamo-pituitary-gonadal axis. Ovarian function in all mammals is controlled by the gonadotropin-releasing hormone (GnRH) neurons, a small group of neurons that reside in the pre-optic area of the hypothalamus. GnRH neurons drive the secretion of the gonadotropins from the pituitary gland that subsequently control ovarian function, including the production of gonadal steroid hormones. These hormones, in turn, provide important feedback signals to GnRH neurons via a hormone sensitive neuronal network in the brain. In many women with PCOS this feedback pathway is impaired, resulting in the downstream consequences of the syndrome. This review will explore what is currently known from clinical and animal studies about the identity, relative contribution and significance of the individual neuronal components within the GnRH neuronal network that contribute to the pathophysiology of PCOS. We review evidence for the specific neuronal pathways hypothesised to mediate progesterone negative feedback to GnRH neurons, and discuss the potential mechanisms by which androgens may evoke disruptions in these circuits at different developmental time points. Finally, this review discusses data providing compelling support for disordered progesterone-sensitive GABAergic input to GnRH neurons, originating specifically within the arcuate nucleus in prenatal androgen induced forms of PCOS.


Absorption kinetics of regular and isophane (NPH) insulin in the normal dog.
Absorption kinetics of regular and isophane (NPH) insulins were evaluated in seven normal fasted dogs by measuring serial serum concentrations of insulin and glucose following the subcutaneous administration of regular and NPH insulins. These results were compared to serum insulin values determined after injecting similar doses of regular insulin intravenously. Regular insulin was better absorbed than NPH insulin (mean bioavailability index 64.6% vs. 41.1%, P less than .01) resulting in a significantly greater maximal increase in mean circulating insulin concentrations above baseline values (362.2 microU/ml vs. 147.8 microU/ml, P less than .05). The time interval between insulin injection and return of serum insulin values to basal concentrations was also significantly shorter for regular than for NPH insulin (4.9 hr vs. 8.6 hr, P less than .05). However, there were no significant differences between regular and NPH insulins in time to reach peak serum insulin concentrations, maximal reduction in serum glucose concentrations, or time of lowest circulating glucose levels. The results of this study support previously accepted values for time-action characteristics of regular insulin, but suggest that NPH insulin may have an earlier peak and shorter duration of action than has previously been proposed in the dog.


Helicobacter bilis infection accelerates and H. hepaticus infection delays the development of colitis in multiple drug resistance-deficient (mdr1a-/-) mice.
mdr1a-deficient mice lack P-glycoprotein and spontaneously develop colitis with age. Helicobacter spp. are gram-negative organisms that have been associated with colitis in certain mouse strains, but Helicobacter spp. have been excluded as contributing to the spontaneous colitis that develops in mdr1a-/- mice. We wished to determine whether infection with either H. bilis or H. hepaticus would accelerate the development of inflammatory bowel disease (IBD) in mdr1a-/- mice. We found that H. bilis infection induced diarrhea, weight loss, and IBD in mdr1a-/- mice within 6 to 17 weeks post-inoculation and before the expected onset of spontaneous IBD. Histopathology of H. bilis-induced IBD included crypt hyperplasia, inflammatory cell infiltrates, crypt abscesses, and obliteration of normal gut architecture. Reverse transcription-polymerase chain reaction and Taqman analysis from colonic tissue showed increased transcripts for interferon-gamma and interleukin-10 from H. bilis-infected colitic mdr1a-/- mice. Additionally, mesenteric lymph nodes had increased cellularity with expansion of CD4+ and CD8+ T cells and B cells and increased proliferation to soluble H. bilis antigens with elaboration of interferon-gamma, tumor necrosis factor-alpha and interleukin-10. In contrast, H. hepaticus infection of mdr1a-/- mice did not accelerate disease but rather delayed the onset of spontaneous colitis which was milder in severity. mdr1a-/- mice infected with Helicobacter spp. may provide a useful tool to explore the pathogenesis of microbial-induced IBD in a model with a presumed epithelial cell "barrier" defect.


Bilateral upper and lower eyelid margin swelling and madarosis due to lymphoma.
Over a 2-year period, swellings of all 4 eyelid margins developed in a 32-year-old woman and was accompanied by complete loss of eyelashes. An inflammatory dermatologic condition was considered the most likely cause. A full-thickness right lower eyelid biopsy revealed a multinodular lymphoid tumor at the eyelid margin which immunophenotypically and genetically was diagnosed as an extranodal marginal zone lymphoma. The mode of presentation of the disease was considered to be most unusual, as was its B cell lineage, since the majority of primary cutaneous lymphomas are of T-cell origin. Systemic workup demonstrated bilateral involvement of the external auditory canals.


A critical evaluation of copper metabolism in Indian Wilson's disease children with special reference to their phenotypes and relatives.
Wilson's disease is an autosomal recessive disorder of copper accumulation in various organs, with most common clinical manifestations such as hepatic, neurological, and renal dysfunctions. Serum copper and ceruloplasmin in Wilson's disease were significantly lower as compared to normals, controls, and relatives of Wilson's disease patients, whereas marked hypercupriuria (145+/-7 microg/24 h) was observed in Wilson's children only. A good correlation (r=0.92) was found between non-ceruloplasmin-bound copper and 24-h urinary copper excretion in Wilson's disease patients. Further, copper studies among the different phenotypes of Wilson's disease revealed substantially low serum ceruloplasmin and a marked hypercupriuria in Wilson's disease children associated with renal tubular acidosis as compared to the patients with either hepatological or neurological manifestations. Serum ceruloplasmin levels in 14 patients of Wilson's disease were between 14 and 20 mg/dL. These patients of Wilson's disease were confirmed by measuring liver biopsy copper, which was about nine times higher than normal hepatic copper content. During the family screening by copper studies, four asymptomatic siblings were diagnosed for Wilson's disease. These subjects were then started on D-penicillamine therapy because presymptomatic treatment prevents progression of the disease complications.


Predictors for persistent urodynamic stress incontinence following extensive pelvic reconstructive surgery with and without midurethral sling.
The objective of this study was to identify the predictors for persistent urodynamic stress incontinence (P-USI) in women following extensive pelvic reconstructive surgery (PRS) with and without midurethral sling (MUS). A total of 1,017 women who underwent pelvic organ prolapse (POP) surgery from January 2005 to December 2013 in our institutions were analyzed. We included 349 USI women who had extensive PRS for POP stage III or more of whom 209 underwent concomitant MUS. Of the women who underwent extensive PRS without MUS, 64.3 % (90/140) developed P-USI compared to only 10.5 % (22/209) of those who had concomitant MUS. Those with concomitant MUS and PRS alone were at higher risk of developing P-USI if they had overt USI [odds ratio (OR) 2.2, 95 % confidence interval (CI) 1.3-4.0, p = 0.014 and OR 4.7, 95 % CI 2.0-11.3, p < 0.001, respectively], maximum urethral closure pressure (MUCP) of < 60 cm H2O (OR 5.0, 95 % CI 3.0-8.1, p < 0.001 and OR 5.3, 95 % CI 2.7-10.4, p < 0.001, respectively), and functional urethral length (FUL) of < 2 cm (OR 5.4, 95 % CI 2.7-8.8, p < 0.001 and OR 3.9, 95 % CI 2.4-6.9, p < 0.001, respectively). Parity ≥ 6 (OR 3.9, 95 % CI 1.7-5.2, p < 0.001) and Prolift T (OR 3.1, 95 % CI 1.9-4, p < 0.001) posed a higher risk of P-USI in those with concomitant surgery. Perigee and Avaulta A seemed to be protective against P-USI in those without MUS. Overt USI with advanced POP together with low MUCP and FUL values have a higher risk of developing P-USI. Therefore, counseling these women is worthwhile while considering the type of mesh used.


Scalp hair and urine mercury content of children in the Northeast United States: the New England Children's Amalgam Trial.
Children may be at particular risk from toxic effects of mercury (Hg). Previous studies of hair (organic) and urine (inorganic) Hg levels in US children were unable to assess Hg levels while accounting for exposure to amalgam dental restorations. This analysis describes, over a 5-year period, levels and correlates/predictors of scalp hair (H-Hg) and urinary (U-Hg) mercury in 534 New England Children's Amalgam Trial (NECAT) participants, aged 6-10 years and without exposure to dental amalgam at baseline. Mean H-Hg levels were between 0.3 and 0.4 microg/g over 5 years. 17-29% of children had H-Hg levels > or = 0.5 microg/g, and 5.0 to 8.5% of children had levels > or = 1 microg/g, in any given study year. In adjusted models, fish consumption frequency was the most robust predictor of high H-Hg. U-Hg mean levels were between 0.7 and 0.9 microg/g creatinine over two years. The percentage of those with U-Hg > or 2.3 microg/g creatinine ranged from 4% to 6%. Number of amalgam restorations had a significant dose-response relationship with U-Hg level. Daily gum chewing in the presence of amalgam was associated with high U-Hg.


ALKBH5-mediated m6A modification of lincRNA LINC02551 enhances the stability of DDX24 to promote hepatocellular carcinoma growth and metastasis.
As the most important RNA epigenetic regulation in eukaryotic cells, N6-metheyladenosine (m6A) modification has been demonstrated to play significant roles in cancer progression. However, this modification in long intergenic non-coding RNAs (lincRNAs) and the corresponding functions remain elusive. Here, we showed a lincRNA LINC02551 was downregulated by AlkB Homolog 5 (ALKBH5) overexpression in a m6A-dependent manner in hepatocellular carcinoma (HCC). Functionally, LINC02551 was required for the growth and metastasis of HCC. Mechanistically, LINC02551, a bona fide m6A target of ALKBH5, acted as a molecular adaptor that blocked the combination between DDX24 and a E3 ligase TRIM27 to decrease the ubiquitination and subsequent degradation of DDX24, ultimately facilitating HCC growth and metastasis. Thus, ALKBH5-mediated LINC02551 m6A methylation was required for HCC growth and metastasis.


Analysis and comparison of the bacterial community in fermented grains during the fermentation for two different styles of Chinese liquor.
Bacterial populations in fermented grains during fermentation may play important roles in Chinese liquor flavor. PCR-based denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene library analysis were performed to analyze the bacterial community structure of two styles of liquor. The results of DGGE profiles showed that bacterial diversity decreased with the fermentation process and Lactobacillus acetotolerans became the predominant species at the end of the fermentation. But the obvious differences of bacterial community appeared in the middle stage of two styles of liquor fermentation, in which the different upstream production techniques were used. Moreover, 16S rRNA gene libraries of two styles were constructed. A total of 125 and 107 clones, chosen from two libraries, were grouped into 46 and 49 operational taxonomic units (OTUs) by amplified ribosomal DNA restriction analysis. According to sequencing results of clones, the predominant bacteria in strong aroma style fermented grains were those from the class Bacilli, Bacteroidetes, and Clostridia, whereas the predominant bacteria in fermented grains of roasted sesame aroma style belonged to Bacilli, Flavobacteria, and Gammaproteobacteria. Molecular analysis of the bacterial diversity of the liquor fermentation will benefit the analysis of important microorganisms playing key roles in the formation of liquor flavor components.


Factors influencing calcium-induced terminal differentiation in cultured mouse epidermal cells.
Mouse epidermal cells can be grown as a proliferating monolayer in medium containing 0.02-0.1 mM calcium. Terminal differentiation of these cells with formation of cornified cells and cell death is induced by elevating calcium in the medium to greater than 0.1 mM. A variety of agents were studied as potential modifiers of this calcium-induced terminal differentiation. Other than calcium, no cation tested was active in inducing or preventing epidermal maturation. Modifiers of calcium or sodium fluxes, local anesthetics and protease inhibitors were also without effect. Modulators or analogues of cyclic nucleotides did not influence epidermal differentiation, and cyclic nucleotide levels did not change significantly in the first 10 min after increasing calcium. Effective inhibition of calcium-induced differentiation, as estimated by morphology, ultrastructure and cornified envelope formation, was seen with the divalent cation ionophore A23187 and the Na+K+ATPase inhibitor ouabain. The well-known effects of ouabain on intracellular sodium and potassium suggested the possible involvement of these ions in the program of calcium-induced epidermal maturation. The increase in medium calcium produced an elevation of both intracellular sodium and potassium within 12-24 hours. The calcium-induced increase in intracellular potassium appears to be the more relevant of these changes since the increase was blocked by both ouabain and A23187. Other inhibitors of calcium-induced differentiation, including harmaline, 8(diethylamino)octyl 3,4,5-trimethoxybenzoate hydrochloride (TMB-8) and low potassium medium, also blocked the rise of intracellular potassium. The five inhibitors had no consistent effect on intracellular sodium. Thus, elevated intracellular potassium may be necessary for the later stages of epidermal differentiation. However, neither ouabain nor A23187 affected the assembly of desmosomes, the earliest ultrastructural change noted after increasing medium calcium. This rapid change in cell-cell contact, beginning within minutes after calcium elevation, appears to be independent of changes in sodium and potassium, but may instead be modulated by increased calcium at the cell surface.


Inhaled lipid nanocarriers for pulmonary delivery of glucocorticoids: Previous strategies, recent advances and key factors description.
In view of the strong anti-inflammatory activity of glucocorticoids (GC) they are used in the treatment of almost all inflammatory lung diseases. In particular, inhaled GC (IGC) allow high drug concentrations to be deposited in the lung and may reduce the incidence of adverse effects associated with systemic administration. However, rapid absorption through the highly absorbent surface of the lung epithelium may limit the success of localized therapy. Therefore, inhalation of GC incorporated into nanocarriers is a possible approach to overcome this drawback. In particular, lipid nanocarriers, which showed high pulmonary biocompatibility and are well known in the pharmaceutical industry, have the best prospects for pulmonary delivery of GC by inhalation. This review provides an overview of the pre-clinical applications of inhaled GC-lipid nanocarriers based on several key factors that will determine the efficiency of local pulmonary GC delivery: 1) stability to nebulization, 2) deposition profile in the lungs, 3) mucociliary clearance, 4) selective accumulation in target cells, 5) residence time in the lung and systemic absorption and 6) biocompatibility. Finally, novel preclinical pulmonary models for inflammatory lung diseases are also discussed.


Elevated levels of endothelin-1 in hepatic venous blood are associated with intrapulmonary vasodilatation in humans.
Hepatopulmonary syndrome is a pulmonary vascular complication of cirrhosis in which intrapulmonary vasodilatation (IPV) results in hypoxemia. Endothelin-1 (ET-1), produced by proliferating cholangiocytes, has been identified as a mediator of IPV in an animal model of HPS, but the pathophysiology of IPV in humans has not been defined. The purpose of this study was to assess whether cirrhosis with IPV, which often leads to HPS, is associated with increased hepatic venous ET-1 blood levels. We performed a prospective cohort pilot study of 40 patients with liver disease undergoing transjugular liver biopsy from November 1, 2008 to September 1, 2009. Patients were categorized according to absence (-) or presence (+) of IPV as determined by bubble-contrasted echocardiography. Hepatic venous blood was assayed for ET-1 by ELISA. The percent volume of cholangiocytes in the liver biopsy specimen was determined by morphometric analysis, as a measure of bile duct proliferation. Nine subjects were excluded, due to absence of cirrhosis (6) and patent foramen ovale (3). Of the remaining 31 subjects, IPV was present in 18 (58%). Median hepatic venous ET-1 was higher with IPV+ than IPV- at levels of 9.1 pg/mL (range 7.5-11.7) versus 2.1 pg/mL (1.3-5.6), respectively (P = 0.004). ET-1 levels correlated positively with cholangiocyte percent volume (r = 0.72, P < 0.001) but not with measures of liver dysfunction (bilirubin, INR, MELD score, or hepatic venous pressure gradient). In human cirrhosis, increased hepatic venous ET-1 is associated with IPV and increased hepatic cholangiocyte volume.


Mortality outcomes associated with invasive aspergillosis among acute exacerbation of chronic obstructive pulmonary disease patient population.
Literature regarding trends of mortality, and complications of aspergillosis infection among patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) is limited. Data from the National Readmissions Database (NRD) that constitutes 49.1% of the stratified sample of all hospitals in the United States (US), representing more than 95% of the national population were analyzed for hospitalizations with aspergillosis among AECOPD. Predictors and trends related to aspergillosis in AECOPD were evaluated. A Linear p-trend was used to assess the trends. Out of the total 7,282,644 index hospitalizations for AECOPD (mean age 69.17 ± 12.04years, 55.3% females), 8209 (11.2/10,000) with primary diagnosis of invasive aspergillosis were recorded in the NRD for 2013-2018. Invasive aspergillosis was strongly associated with mortality (OR 4.47, 95%CI 4.02-4.97, p < 0.001) among AECOPD patients. Malignancy and organ transplant status were predominant predictors of developing aspergillosis among AECOPD patients. The IA-AECOPD group had higher rates of multi-organ manifestations including ACS (3.7% vs 0.44%; p-value0.001), AF (20% vs 18.4%; p-value0.001), PE (4.79% vs1.87%; p-value0.001), AKI (22.3% vs17.5%; p-value0.001), ICU admission (16.5% vs11.9%; p-value0.001), and MV (22.3% vs7.31%; p-value0.001) than the AECOPD group. The absolute yearly trend for mortality of aspergillosis was steady (linear p-trend 0.22) while the yearly rate of IA-AECOPD had decreased from 15/10,000 in 2013 to 9/10,000 in 2018 (linear p-trend 0.02). Aspergillosis was related with high mortality among AECOD hospitalizations. There has been a significant improvement in the yearly rates of aspergillosis while the mortality trend was steady among aspergillosis subgroups. Improved risk factor management through goal-directed approach may improve clinical outcomes.


Evaluation for the clinical diagnosis of Pythium insidiosum using a single-tube nested PCR.
Pythiosis is a rare infectious disease caused by Pythium insidiosum, which typically occurs in tropical and subtropical regions. The high mortality rate may be in consequence of the lack of diagnosis. The objective of this study was to evaluate reliability of a new single-tube nested PCR for detection of P. insidiosum DNA. A total of 78 clinical isolates of various fungi and bacteria, 106 clinical specimens and 80 simulated positive blood samples were tested. The developed primer pairs CPL6-CPR8 and YTL1-YTR1 are located on 18S subunit of the rRNA gene of P. insidiosum. The specificity, negative and positive predictive values were 100, 100 and 87.5 %, respectively, as compared with direct microscopy and cultivation. The detection limit of the single-tube nested PCR was 21 zoospores corresponding to 2.7 pg of the DNA. The results demonstrate that the new single-tube nested PCR offers a highly sensitive, specific and rapid genetic method for detecting P. insidiosum.


A new proposed rodent model of chemically induced prostate carcinogenesis: distinct time-course prostate cancer progression in the dorsolateral and ventral lobes.
Characterization of novel rodent models for prostate cancer studies requires evaluation of either spontaneous and carcinogen-induced tumors as well as tumor incidence in different prostatic lobes. We propose a new short-term rodent model of chemically induced prostate carcinogenesis in which prostate cancer progression occurs differentially in the dorsolateral and ventral lobes. Adult gerbils were treated with MNU alone or associated with testosterone for 3 or 6 months of treatment. Tumor incidence, latency, localization, and immunohistochemistry (AR, PCNA, smooth muscle α-actin, p63, MGMT, and E-cadherin) were studied in both lobes. Comparisons between both lobes revealed that lesions developed first in the DL while the VL presented longer tumor latency. However, after 6 months, there was a dramatic increase in tumor multiplicity in the VL, mainly in MNU-treated groups. Lesions clearly progressed from a premalignant to a malignant phenotype over time and tumor latency was decreased by MNU + testosterone administration. Three-dimensional reconstruction of the prostatic complex showed that the DL developed tumors exclusively in the periurethral area and showed intense AR, PCNA, and MGMT immunostaining. Moreover, VL lesions emerged throughout the entire lobe. MNU-induced lesions presented markers indicative of an aggressive phenotype: lack of basal cells, rupture of the smooth muscle cell layer, loss of E-cadherin, and high MGMT staining. There are distinct pathways involved in tumor progression in gerbil prostate lobes. This animal provides a good model for prostate cancer since it allows the investigation of advanced steps of carcinogenesis with shorter latency periods in both lobes.


Activation of CD4+ T lymphocytes improves wound healing and survival after experimental myocardial infarction in mice.
The role of adaptive immunity, especially CD4(+) T-helper cells, has not yet been systematically investigated in wound healing and remodeling after myocardial infarction (MI). Therefore, we studied whether CD4(+) T cells become activated and influence wound healing after experimental MI in mice. When we compared sham versus MI in wild-type (WT) mice, T-cell receptor-dependent activation of both conventional Foxp3(-) and regulatory Foxp3(+) CD4(+) T cells could be demonstrated in heart-draining lymph nodes within the first week after MI. Concomitantly, we found infiltration of CD4(+) T cells in infarcted myocardium. To study the role of CD4(+) T cells in wound healing and remodeling, CD4(+) T-cell-deficient mice (CD4 knockout [KO], MHCII(Δ/Δ)) and T-cell receptor-transgenic OT-II mice recognizing an irrelevant ovalbumin-derived peptide were studied. Serial echocardiography up to day 56 after MI revealed increased left ventricular dilation in CD4 KO compared with WT mice. Within the infarcted myocardium, CD4 KO mice displayed higher total numbers of leukocytes and proinflammatory monocytes (18.3±3.0 10(4)/mg WT versus 75.7±17.0 10(4)/mg CD4 KO, P<0.05). MHCII(Δ/Δ) and OT-II mice displayed significantly greater mortality (21% WT versus 48% OT-II, P<0.05, and WT 22% versus 52% MHCII(Δ/Δ), P<0.05) and myocardial rupture rates than WT mice. Collagen matrix formation in the infarct zone was severely disturbed in CD4 KO and MHCII(Δ/Δ) mice, as well as in OT-II mice. The present study provides the first evidence that CD4(+) T cells become activated after MI, presumably driven by recognition of cardiac autoantigens, and facilitate wound healing of the myocardium.