Airways antiallergic effect and pharmacokinetics of alkaloids from Alstonia scholaris.
Alstonia scholaris (L.) R. Br. (Apocynaceae), an important herbal medicine, has been widely used to treat respiratory tract diseases, such as cough, asthma, phlegm, and chronic obstructive pulmonary disease. To evaluate pharmacological effect of alkaloids from A. scholaris on ovalbumin induced airways allergic inflammatory model, and explore whether the dosing frequency is related to pharmacokinetics. After oral administration of total alkaloids, the pharmacokinetic study of it was investigated. In addition, anti-allergic studies were carried out on ovalbumin-sensitized airways allergic inflammatory model of mice. The pharmacokinetics of total alkaloids (TA) was investigated in SD rat plasma by a fully-validated LC-MS/MS method. Then, an ovalbumin (OVA)-sensitized airways allergic inflammatory model was established, in which mice were intra-gastrically administrated by 3 times a day (8.3 and 16.7mg/kg) based on the pharmacokinetic behavior of TA) and single (25, 50mg/kg) treatment regimen. Dexamethasone was used as a positive control for corticosteroid drugs. Cellular infiltration was assessed in the broncho-alveolar lavage fluid (BALF). Expressions of interleukin-4 (IL-4) and interleukin-10 (IL-10) in the BALF were determined, levels of immunoglobulin E (IgE) and eotaxin in serum were measured, and superoxide dismutase (SOD) activities as well as malondialdehyde (MDA) content in the serum and BALF were examined. Finally, histopathological examination in the lung was assessed by H. E. staining. The time course of plasma concentration of 4 bioactive indole alkaloids fitted an open two-compartment model after oral administration of total alkaloids at doses of 10, 25, and 50mg/kg. The area under the curve and the maximum concentration values of four major alkaloids increased dose-dependently, and half-life suggested a short-lasting pharmacological effect. Then, an ovalbumin-provoked airways allergic inflammatory model indicated that the pharmacological effect of administration of total alkaloids 3 times a day was a little better than that of single dose daily. The percentage of eosinophils in BALF was reduced obviously and the pathological damage of lung was also attenuated. There was also a significant reduction in IL-4 and promotion in IL-10 in the BALF. Serum IgE and eotaxin expression also significantly decreased in treated animals. Furthermore, the activity of SOD elevated remarkably and lipid peroxidation product (MDA) decreased in the administrated mice. The pharmacological effects administrated for 3 times a day had precedence over single dose daily, which was related to the prolonged retention time and the maintained plasma concentration. Moreover, scholaricine and vallesamine might be responsible for the treatment of allergic asthma, mainly in total alkaloids.


Optimization of crude oil degradation by Dietzia cinnamea KA1, capable of biosurfactant production.
The aim of this study was isolation and characterization of a crude oil degrader and biosurfactant-producing bacterium, along with optimization of conditions for crude oil degradation. Among 11 isolates, 5 were able to emulsify crude oil in Minimal Salt Medium (MSM) among which one isolate, named KA1, showed the highest potency for growth rate and biodegradation. The isolate was identified as Dietzia cinnamea KA1 using morphological and biochemical characteristics and 16S rRNA gene sequencing. The optimal conditions were 510 mM NaCl, pH 9.0, 35 °C, and minimal requirement of 46.5 mM NH4 Cl and 2.10 mM NaH2 PO4 . Gravimetric test and Gas chromatography-Mass spectroscopy technique (GC-MS) showed that Dietzia cinnamea KA1 was able to utilize and degrade 95.7% of the crude oil after 5 days, under the optimal conditions. The isolate was able to grow and produce biosurfactant when cultured in MSM supplemented with crude oil, glycerol or whey as the sole carbon sources, but bacterial growth was occurred using molasses with no biosurfactant production. This is the first report of biosurfactant production by D. cinnamea using crude oil, glycerol and whey and the first study to report a species of Dietzia degrading a wide range of hydrocarbons in a short time.


Intrauterine Adhesions Following Miscarriage: Look and Learn.
To examine the incidence of intrauterine adhesions (IUA) following the management of miscarriage in women with previously documented normal uterine cavities. We conducted a retrospective cohort study from two fertility clinics with standard practice protocols for evaluating the uterine cavity prior to infertility treatment and following clinical pregnancy loss. A database query and manual chart review identified 144 women with normal uterine cavities who experienced a miscarriage between January 2010 and November 2012 and returned to the clinic for follow-up hysteroscopy. Following documentation of a non-viable clinical pregnancy using transvaginal ultrasound, patients chose expectant, medical, or surgical management according to standardized clinical protocols. The primary outcome was the detection of IUA. Secondary outcomes included the presence of retained products of conception and various risk factors associated with the development of IUA. The incidence of IUA following early pregnancy loss was 6.3%. There were no significant differences in patient characteristics between those with and without IUA. There was a significant association between IUA and increasing uterine size, particularly in the presence of multiple gestation (P = 0.039). Mechanical suction dilatation and curettage (D&C) was a risk factor for IUA, but manual vacuum aspiration was not a risk factor (P = 0.003). Retained products of conception were found in 13.9% of study participants, and the incidence did not differ among management options. This appears to be the first documentation of IUA that were entirely attributable to the index miscarriage or its management. There appears to be an increased risk of IUA following D&C with larger uteri and multiple pregnancies and following mechanical suction D&C.


Increased circulating and intrahepatic T-cell-specific chemokines in chronic hepatitis C: relationship with the type of virological response to peginterferon plus ribavirin combination therapy.
To determine the serum and intrahepatic levels of T-helper-1-associated chemokines in patients with chronic hepatitis C before, during and after peginterferon plus ribavirin combination therapy and to search for correlations with baseline characteristics of hepatitis C virus-related chronic liver disease and type of therapeutic response. Serum chemokine levels were determined by enzyme-linked immunosorbent assays and intrahepatic chemokine messenger RNA and protein levels were tested by ribonuclease protection assay and immunohistochemistry. Serum and intrahepatic chemokine levels were elevated in all patients with chronic hepatitis C and showed a marked decrease in patients who obtained a virological response vs. non-responders. Increased serum interferon-gamma-inducible protein-10 levels at baseline in genotype 1-infected patients were significantly associated with greater degrees of intrahepatic inflammation and fibrosis (P = 0.0046 and P = 0.02, respectively) and with virological non-response (P = 0.01). In patients with genotype 1, basal serum interferon-gamma-inducible protein-10 levels greater than 299 pg/mL identified 80% of non-responders and lower than 299 pg/mL identified 63% of responders. Circulating and intrahepatic T-helper-1-associated chemokines are abnormally elevated in patients with chronic hepatitis C. Increased baseline serum interferon-gamma-inducible protein-10 levels in genotype 1-infected patients are associated with virological non-response to peginterferon plus ribavirin combination therapy.


TORC1 controls G1-S cell cycle transition in yeast via Mpk1 and the greatwall kinase pathway.
The target of rapamycin complex 1 (TORC1) pathway couples nutrient, energy and hormonal signals with eukaryotic cell growth and division. In yeast, TORC1 coordinates growth with G1-S cell cycle progression, also coined as START, by favouring the expression of G1 cyclins that activate cyclin-dependent protein kinases (CDKs) and by destabilizing the CDK inhibitor Sic1. Following TORC1 downregulation by rapamycin treatment or nutrient limitation, clearance of G1 cyclins and C-terminal phosphorylation of Sic1 by unknown protein kinases are both required for Sic1 to escape ubiquitin-dependent proteolysis prompted by its flagging via the SCF(Cdc4) (Skp1/Cul1/F-box protein) ubiquitin ligase complex. Here we show that the stabilizing phosphorylation event within the C-terminus of Sic1 requires stimulation of the mitogen-activated protein kinase, Mpk1, and inhibition of the Cdc55 protein phosphatase 2A (PP2A(Cdc55)) by greatwall kinase-activated endosulfines. Thus, Mpk1 and the greatwall kinase pathway serve TORC1 to coordinate the phosphorylation status of Sic1 and consequently START with nutrient availability.


Daily rapid blood glucose variations may condition antithrombin III biologic activity but not its plasma concentration in insulin-dependent diabetes. A possible role for labile non-enzymatic glycation.
The effect of rapid daily variation of glycemia and labile HbA1 on both antithrombin III (ATIII) activity and plasma concentration in ten insulin dependent diabetics has been evaluated. The variations of both plasma glucose and labile HbA1 were inversely correlated to the alterations of ATIII activity (r = -0.71 and r = -0.73, respectively, p less than 0.001), while no change in ATIII plasma concentration was present. These data suggest a direct role of glucose in determining rapid alteration of ATIII biologic activity, in vivo, in diabetes, probably mediated by labile non-enzymatic glycation.


New Drugs for the Treatment of Tuberculosis.
Tuberculosis (TB) has now surpassed HIV as the leading infectious cause of death, and treatment success rates are declining. Multidrug-resistant TB, extensively drug-resistant TB, and even totally drug-resistant TB threaten to further destabilize disease control efforts. The second wave in TB drug development, which includes the diarylquinoline, bedaquiline, and the nitroimidazoles delamanid and pretomanid, may offer options for simpler, shorter, and potentially all-oral regimens to treat drug-resistant TB. The "third wave" of TB drug development includes numerous promising compounds, including less toxic versions of older drug classes and candidates with novel mechanisms of action.


Links between deprivation and risk of violence-related injury: a qualitative study to identify potential causal mechanisms.
Deprivation has been shown to have a greater effect on risk of violent injury among adolescent girls than boys, but the mechanisms underlying this association have not been identified. In this qualitative study designed to identify causal mechanisms, focus groups involving girls aged 14-16 years attending secondary schools in South Wales, UK, were convened. Schools were recruited based on a measure of area-level deprivation. Discussions were audio-recorded and transcripts analysed thematically. Girls from more deprived areas tended not to participate in organized activities, obtained alcohol from multiple sources, consumed alcoholic drinks of varying strengths in both supervised and unsupervised settings, and tended not to feel trusted by their parents; this led to poor adolescent-parent communication. Girls from less deprived areas tended to participate in organized activities, obtain alcohol from parents, consume low strength alcohol in supervised settings, and have a trusting and communicative relationship with their parents. Deprivation may increase risk of adolescent girls sustaining violence-related injury by increasing their time spent in unsupervised environments, with alcohol and without parental knowledge.


Uptake of 99Tcm-gluconate and early histologic findings in induced myocardial infarcts in dogs.
A new isotope, 99Tcm-gluconate, for detection of myocardial infarction has been investigated experimentally. Great differences in isotope uptake between infarcted myocardium and normal myocardium were found in dogs. A close correlation was found between the isotope uptake and the uptake of basic fuchsin (HBFP) at microscopy in early myocardial infarcts.


Resistance of the shell membrane and mineral layer to diffusion of oxygen and water in flexible-shelled eggs of the snapping turtle (Chelydra serpentina).
At oviposition, flexible eggshells of many turtles have an outer mineral layer and an inner membrane layer of approximately equal thickness. We measured conductances of both layers to H2O and O2 at various levels of eggshell hydration. Both the mineral layer of the eggshell and the shell membrane offer significant resistance to diffusion of water vapor and oxygen in eggshells of the snapping turtle, Chelydra serpentina. Conductance to water vapor increases in both the membrane and mineral layer with increasing hydration of the eggshell, but conductance to oxygen decreases under similar conditions. Removal of the mineral layer increases conductance to oxygen in moist and dry eggshells, but decreases conductance at intermediate levels of dehydration. Removal of the mineral layer consistently increases conductance to water vapor. The eggshell membrane accounts for 24-76% of overall resistance to diffusion of water vapor. These results suggest that bulk flow of H2O or physical changes in the shell may interact with diffusion to limit gas exchange through the turtle eggshell.


Dummy molecular imprinted polymers coated with silver microspheres via surface enhanced Raman scattering for sensitive detection of benzimidazole.
Dummy molecular imprinted polymers (MIPs) with carbendazim as a dummy template coated with Ag microspheres were fabricated in N, N-dimethylformamide solution via a surface-enhanced Raman scattering (SERS) enhancement for detection of benzimidazole by using methylacrylamide and ethylene glycol dimethacrylate as the functional monomer and cross-linker, respectively. The scanning electron microscope, transmission electron microscope, energy dispersive spectrometer, particle size distribution analysis, and X-ray diffraction were used to perform the morphology, size, and crystal structure for prepared Ag@dummy MIPs. Under the optimal conditions, the SERS detection method presented good linearity with concentrations in the range of 1.0 × 10-3 ~ 1.0 × 10-8 mol L-1 for benzimidazole at 774 cm-1 and 1004 cm-1, respectively. And the minimum detection concentration of this method was as low as 1.0 × 10-8 mol L-1. Besides, Ag@dummy MIPs exhibited satisfactory sensitivity and selectivity to benzimidazole instead of carbendazim due to the dummy imprinting technology to eliminate the background noise interference. The reusability result of Ag@dummy MIPs was shown that the characteristic peaks of benzimidazole are still obvious after four times of repeated detection. This method provided an effective way to develop a qualitative and semi-quantitative analysis of benzimidazole in complex matrices.


Synthesis and turn-over of the replicative Cdc6 protein during the HeLa cell cycle.
The human replication protein Cdc6p is translocated from its chromatin sites to the cytoplasm during the replication phase (S phase) of the cell cycle. However, the amounts of Cdc6p on chromatin remain high during S phase implying either that displaced Cdc6p can rebind to chromatin, or that Cdc6p is synthesized de novo. We have performed metabolic labeling experiments and determined that [35S]methionine is incorporated into Cdc6p at similar rates during the G1 phase and the S phase of the cell cycle. Newly synthesized Cdc6p associates with chromatin. Pulse-chase experiments show that chromatin-bound newly synthesized Cdc6p has a half life of 2-4 h. The results indicate that, once bound to chromatin, pulse-labeled new Cdc6p behaves just as old Cdc6p: it dissociates and eventually disappears from the nucleus. The data suggest a surprisingly dynamic behaviour of Cdc6p in the HeLa cell cycle.


Preventive effect of taurine against acute paraquat intoxication in beagles.
The continuous infusion of taurine markedly protected against paraquat (PQ)-induced oliguria in beagles. Pharmacokinetic studies revealed that taurine infusion increased the blood concentration of PQ and reduced the net content in the kidney and, to a lesser extent, in the lung. The excretion of PQ into the urine was unaltered. The infusion of glycine did not have such effects.


Hepatitis B virus core protein inhibits TRAIL-induced apoptosis of hepatocytes by blocking DR5 expression.
Hepatitis B virus (HBV) causes chronic hepatitis in hundreds of millions of people worldwide, which can eventually lead to hepatocellular carcinoma (HCC). The molecular mechanisms underlying HBV persistence are not well understood. TRAIL, the TNF-related apoptosis-inducing ligand, has recently been implicated in hepatocyte death during HBV infection. We report here that the HBV core protein (HBc) is a potent inhibitor of TRAIL-induced apoptosis. Overexpressing HBc significantly decreased TRAIL-induced apoptosis of human hepatoma cells, whereas knocking-down HBc expression in hepatoma cells transfected with HBV genome enhanced it. When present in the same cell, HBc blocked the pro-apoptotic effect of the HBV X protein (HBx). The resistance of HBc-expressing cells to TRAIL-induced apoptosis was associated with a significant reduction in death receptor 5 (DR5) expression. Upon transfection, HBc significantly repressed the promoter activity of the human DR5 gene. Importantly, HBc gene transfer inhibited hepatocyte death in a mouse model of HBV-induced hepatitis; and in patients with chronic hepatitis, DR5 expression in the liver was significantly reduced. These results indicate that HBc may prevent hepatocytes from TRAIL-induced apoptosis by blocking DR5 expression, which in turn contributes to the development of chronic hepatitis and HCC. They also call into question the potential side effects of HBc-based vaccines.


Short jump vein graft for the treatment of symptomatic Riles type 1A occlusion of common carotid artery: case report and review of literature.
A 62-year-old man was presented with transient ischemic attack 1 day after percutaneous coronary intervention. Magnetic resonance imaging demonstrated fresh cerebral infarction in the left hemisphere. Digital subtraction angiography showed left Riles type 1A common carotid artery occlusion (CCAO). Blood flow in the internal carotid artery (ICA) was derived from the external carotid artery, which came through the anastomosis between the left occipital artery and a muscular branch of left vertebral artery. We performed short jump graft from CCA to ICA using saphenous vein, followed by ligation of CCA. The graft remained patent at the 1-year follow-up.


Femtosecond lasers for high-precision orthopedic surgery.
Laser micromachining with ultrashort pulses has shown great promise for clean, safe surgical treatment of bone tissue. However, comparisons of performance and development of "best practice" have been hampered by the difficulty of comparing results across a wide variety of experimental approaches and under surgically irrelevant conditions (e.g., dried, dead bone). Using a femtosecond (fs) pulsed laser system (τ = 140 fs, repetition rate = 1 kHz, λ = 800 nm), a comprehensive study of femtosecond laser microsurgery using the standard metrics of laser micromachining (ablation threshold, incubation effects, ablation rates, effect of focal point depth within the material and heat affected zone (HAZ)) was conducted on live, freshly harvested bovine and ovine cortical bone. Three important points of optimism for future implementation in the surgical theatre were identified: (1) the removal of material is relatively insensitive to the focal point depth within the material, removing the need for extreme depth precision for excellent performance; (2) femtosecond laser ablation of fresh bone demonstrates very little incubation effect, such that multiple passes of the laser over the same region of bone removes the same amount of material; and (3) the complete absence of collateral damage, heat- or shock-induced, on both the macro- and microscopic scales can be achieved readily, within a broad parameter range. Taken together, these results indicate a handheld or robotic deployed fiber laser platform for femtosecond laser microsurgery is a very viable prospect.


Assessing bladder capacity in children: is it feasible at all?
The aim of the study was to assess correlations between body parameters and bladder capacity in healthy schoolboys and schoolgirls. On the basis of those anthropometric data functional bladder capacity (FBC) in children was estimated. The questionnaires were sent to 1,055 parents of children aged from 6 to 14 years. Voided volumes were assessed during uroflowmetry, which was performed under typical conditions in separate rooms (Urodyn 1,000, Dantec, Skovlunde, Denmark). Residual volume was measured thereafter. Precise anthropometric parameters were obtained in standing the position. Children with dysfunctional problems, those who voided less than 100 ml and those with residual volumes > 5% of voided volume were excluded. Statistical analysis was done on the basis of the results gathered from 288 children. Pearson's correlation coefficient between body measurements and voided volume was calculated (P < 0.05). Regression modeling was used to propose the best formula for calculating FBC in children. Mean FBC is higher in girls than in boys (P < 0.05). The correlation between FBC and flow parameters was found (0.3 < "r" < 0.5). The value of the coefficient indicates loose relationship. We revealed correlation between weight, height, and length of the trunk, pelvic dimension and FBC. The best mathematical equation to estimate FBC in children based on the body measurements fits to 54% of population (FBC = -268.4 + 7.38 x flow time + 3.15 x suprasteral height + 1.25 x thickness of fat fold.


Formation of reactive oxygen species following bioactivation of gentamicin.
The present study investigated the ability of gentamicin to catalyze free radical reactions and probed the underlying mechanisms by hydroethidine imaging, oxygen consumption, and reduction of cytochrome c. In Epstein-Barr virus-transformed lymphoblastoid cells, a respiratory burst was induced by phorbol ester and detected by hydroethidine, a fluorescent indicator of superoxide radical. The addition of gentamicin increased the fluorescence two-fold while gentamicin did not produce fluorescence in the absence of phorbol ester. In membrane preparations, gentamicin did not enhance NADPH consumption ruling out a direct activation of NADPH oxidase. The formation of reactive oxygen species by gentamicin was additionally supported by experiments that showed gentamicin increased oxygen consumption two-fold in intact cells and a cell-free system. In addition, generation of superoxide was indicated by the gentamicin-stimulated reduction of cytochrome c. The stimulation by gentamicin depended upon the presence of iron (FeII/FeIII) and of arachidonic acid as an electron donor. These results support the hypothesis that an iron-gentamicin complex can increase reactive oxygen species in nonenzymatic and in biological systems. The requirement for a reductive activation in intact cells (e.g., by a respiratory burst) is interpreted as the conversion of an inactive FeIII-gentamicin to a redox-active FeII-gentamicin complex.


Dibenzophosphaborin: a hetero-pi-conjugated molecule with fluorescent properties based on intramolecular charge transfer between phosphorus and boron atoms.
[reaction: see text] The first dibenzo-1,4-phosphaborins were synthesized and characterized by X-ray crystallographic analysis. The phosphaborins exhibited UV/vis absorption derived from intramolecular charge transfer from the phosphorus atom to the boron atom. The phosphaborins showed different fluorescent properties depending on the substituents on the phosphorus atom.


The Significance of Equivocal Exercise Treadmill ECG for Intermediate Risk Chest Pain Assessment - Insight From Coronary CT Angiography Data.
Exercise treadmill test (ETT) is commonly the first-line investigation in troponin-negative chest pain patients. Inconclusive results are common and often lead to repeated functional testings. Coronary computed tomographic angiography (CCTA) has excellent negative predictive value for coronary artery disease detection and may play an important role in their diagnostic workup. We aim to analyse ETT and CCTA findings to understand their modern roles in intermediate risk chest pain population with inconclusive ETT. Patients who underwent both ETT and CCTA at our institution between August 2011 and February 2013 were retrospectively investigated. The ETTs were blindly reviewed, with equivocal ETTs defined as any indeterminate results not strictly adhering to ACC/AHA guidelines for positive ETT. Baseline demographics, clinical characteristics and investigation results were collated. Follow-up outcome data for subsequent investigations, representations, major cardiac adverse events (MACE) and unexpected revascularisations were also analysed. 346 consecutive patients were identified (age 57±10years, 53% females, body mass index (BMI) 28±4, Dukes Clinical Score 48±30%, mean follow-up 2.1±0.4years). The ETT was positive in 31%, equivocal in 54% and negative in 15%. Obstructive coronary artery disease (CAD) prevalence was 25% (29% males, 13% females). Those with negative ETTs had obstructive CAD in 8%, with no adverse outcomes during follow-up. Obstructive CAD was seen in 20% with "equivocal" ETTs and 29% with "positive" ETTs. In females, obstructive CAD prevalence was low, and similar in those with equivocal or positive ETT (16% and 11% respectively). In males, obstructive CAD was almost 50% in those with positive ETT (p=0.005). Obstructive CAD was found in one in five "equivocal" ETTs, hence, not all should be considered negative. Obstructive CAD was only found in one in three "positive" ETTs, hence not all should proceed to invasive angiography. CCTA may be an important gatekeeper test in those with low-intermediate pre-test probability of obstructive CAD.


Measurement of non-methane hydrocarbons in Taipei city and their impact on ozone formation in relation to air quality.
Air pollutants data from semi-continuous measurements at multiple sampling sites in Taipei metropolitan area of Taiwan was obtained by collecting air samples in canisters. The hydrocarbon composition was determined by using GC/MS and GC/FID. The air samples were pre-concentrated onto glass beads prior to separation by PLOT and DB-1 columns of GC. The method showed detection limit of <1 ppb and relative standard deviation in the range of 5-30% for different compounds. Aromatic hydrocarbons (toluene, benzene, etc.) and aliphatic hydrocarbons (ethylene, acetylene, propane, etc.) were correlated primarily to determine the source of emission. The estimated hydrocarbons were ranked according to their abundance and photochemical reactivity. The criteria pollutants, ozone and NO2 were measured by UV-differential optical absorption spectroscopy (UV-DOAS), and were utilized to determine the relative importance of non-methane hydrocarbons (NMHC) and significant contribution of NO2 in limiting ozone formation. The obtained results suggest that ozone formation in Taipei city is probably limited by the supply of non-methane hydrocarbons. The concentration profile of targeted pollutants was compared to other metropolitan areas to determine air quality and the pollutant sources.


A comparison of the Fujita classification of awake and drug-induced sleep endoscopy patients.
Only a few studies have compared the outcomes of patients kept awake during endoscopic examination and subjects submitted to drug-induced sleep endoscopy. This study aimed to compare the endoscopic findings of patients submitted to outpatient endoscopy and endoscopic examination with sedation by propofol based on the Fujita Classification. This cross-sectional cohort study enrolled 34 patients. The subjects underwent ENT examination, nasal endoscopy with Müller's maneuver, and drug-induced sleep endoscopy with propofol. The Fujita Classification was used to compare the two modes of endoscopic examination. The examinations were correlated to patient clinical data such as BMI, age, and OSAS severity. There was no agreement between the two modes of endoscopic examination, whether for the group in general or for the analyzed subgroups. There was no agreement between the endoscopic findings of endoscopic examinations done with the patient awake or in drug-induced sleep.


Inhibition of T-type Ca(2+) currents in mouse spermatogenic cells by gossypol, an antifertility compound.
Gossypol, a male antifertility compound isolated from cotton, has been proved to inhibit capacitation and the acrosome reaction in human and mammalian sperm. Here, by using whole-cell recording, we observed the effects of gossypol on Ca(2+) and Cl(-) currents in mouse spermatogenic cells obtained by mechanical dissociation. The results showed that gossypol concentration-dependently and irreversibly inhibited T-type Ca(2+) currents in the cells. When the concentration of gossypol was > or =5 microM, the currents were blocked completely. The time to current block was progressively shortened as the gossypol concentration was increased from 5 to 80 microM. Moreover, the drug increased the time constant of inactivation in a concentration-dependent manner, while it did not affect the activation of the current. The inhibitory effect on the T-type Ca(2+) current did not correlate with signaling mediated by G proteins and tyrosine phosphorylation. No obvious effect of gossypol on Cl(-) currents was observed. These data suggest that the gossypol-induced inhibition of T-type Ca(2+) currents could be responsible for the antifertility activity of the compound, indicating a possibility to use gossypol as a local contraceptive drug.


Neuroprotective effects of carvedilol in diabetic rats: prevention of defective peripheral nerve perfusion and conduction velocity.
The effects of carvedilol (10 mg kg-1 day-1), a peripheral vasodilator and oxygen free radical scavenger, on nerve conduction and blood flow were examined in streptozotocin-diabetic rats. Rats were treated from induction of diabetes for 1 month in a preventive experiment; effects of treatment in non-diabetic rats were also examined. Diabetes caused a 20.0% (P < 0.001) reduction in sciatic motor conduction and a 14.7% (P < 0.001) deficit in saphenous sensory conduction. Diabetic rats given carvedilol treatment had motor and sensory conduction velocities which did not differ significantly from those of non-diabetic controls, but were greater than for untreated diabetes (P < 0.001). Carvedilol treatment did not significantly alter nerve conduction in non-diabetic rats. The nutritive (capillary) component of sciatic endoneurial blood flow, measured by microelectrode polarography and hydrogen clearance, was 45.8% (P < 0.001) reduced by diabetes. This was completely corrected (P < 0.001) by carvedilol treatment in diabetic rats, flow being in the upper half of the non-diabetic range. In non-diabetic rats, carvedilol caused a 30.6% (P < 0.05) elevation in blood flow. Hydrogen clearance data also revealed a shift away from non-nutritive (arterio-venous anastomotic) towards nutritive flow in treated diabetic rats (P < 0.05). We conclude that the data support a neurovascular hypothesis of diabetic neuropathy. Carvedilol, via vasodilator and anti-oxidant actions, prevents the nutritive blood flow changes that lead to nerve dysfunction.(ABSTRACT TRUNCATED AT 250 WORDS)


Three techniques for the determination of perindopril through derivatization with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole.
In addition to its pure form, three accurate, rapid, and simple methods have been established for determining perindopril (PRD) in its tablet form. At pH 9.0 using a borate buffer, developing the three designated methods was successful according to the reaction between PRD and 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) and the formation of a chromogen (with a yellow color) measurable at 460 nm using the spectrophotometric method (Method I). In addition, the produced chromogen was assessed using the spectrofluorimetric method (Method II) at 535 nm following excitation at 461 nm. Afterward, the same reaction product was separated and determined using the HPLC method with fluorescence detection (Method III). A Promosil C18 stainless steel column (Q7 5 mm particle size, 250-4.6 mm) has proven suitable for separation. The mobile phase adjustment was made at pH 3.0, with a 1.0 mL min -1 flow rate; its composition was methanol-sodium dihydrogen phosphate, 0.02 M (60: 40, v/v). Through concentration ranges of 5.0-60.0, 0.5-6.0, and 1.0-10.0 μg mL-1, the calibration curves were rectilinear for Methods I, II, and III, respectively, with limits of quantification (LOQ) of 1.08, 0.16 and 0.19 μg mL-1 as well as limits of detection (LOD) of 0.36, 0.05 and 0.06 μg mL-1. The developed methods were implemented to estimate PRD in tablets, and a comparison between the obtained outcomes utilizing the developed methods as well as obtained from the official method revealed that they were comparable. The official BP method was based on dissolving PRD in anhydrous acetic acid and titrating with 0.1 M perchloric acid, then the potentiometric determination of the end-point. The designated methods were also implemented in content uniformity testing with satisfying results. The reaction pathway proposal was speculated, and according to ICH Guidelines, the statistical evaluation of the data was performed. The three proposed methods were confirmed to be green, eco-friendly and safe to environment using Green Analytical procedure index (GAPI) method.


[Expression and characterization of subunit C of mouse lactate dehydrogenase in Escherichia coli].
To construct a prokaryotic recombinant vector for mouse lactate dehydrogenase-C and to detect its expression in BL21. The coding sequence of mouse lactate dehydrogenase subunit C was amplified from mouse testis RNA with specific primers, and cloned into pGEX-2T after the restriction digestion with BamH I and EcoR I. GST fusion protein was expressed after induction with IPTG. Sequencing and restriction digestion of the recombinant plasmid revealed the existence of coding sequence for mouse lactate dehydrogenase subunit C. A protein band of about 60,000 could be induced by IPTG in the recombinant plasmid. The coding sequence of mouse lactate dehydrogenase subunit C was introduced into the pGEX-2T plasmid and a GST-fused protein could be induced at a high level.


I. Antidepressants and sexual behavior: Weekly ketamine injections increase sexual behavior initially in female and male rats.
The present study characterized the effects of weekly ketamine injections on sexual behavior and anxiety in female and male rats, using a dosing protocol that mimics human therapeutic treatment for depression. In Experiment 1A, ketamine (10 mg/kg, i.p.) or saline was injected once per week for four consecutive weeks. The partner preference paradigm was used to measure sexual motivation 30 min after each weekly injection. Briefly, subjects were first given a 10-min test during which they could choose to spend time in the vicinity of a sexually receptive female stimulus or a sexually experienced male stimulus, however physical contact was restricted (no-contact). Immediately after, subjects were given unrestricted access to the stimulus animals (contact). After a washout period, subjects received four additional weekly injections of ketamine or saline, and then were tested for anxiety-like behavior on the elevated plus maze (EPM) after the last injection (Experiment 1B). For Experiment 2, similar procedures were used to test the effects of weekly ketamine injections on sexual motivation (Experiment 2A) and anxiety (Experiment 2B) in male subjects. In female subjects, ketamine increased sexual motivation as measured by greater time spent with the male stimulus, decreased likelihood of leaving after receiving mounts, and shorter return latencies after receiving intromissions, when compared to saline controls. In male subjects, ketamine shortened latency to first mount and first intromission, as well as increased time spent with the female stimulus. Very little anxiety was observed in either group (ketamine or saline) of female or male subjects when tested on the EPM. In conclusion, even after four weeks of ketamine exposure, sexual dysfunction did not emerge in either females or males. In contrast, ketamine increased sexual motivation in both females and males, with an initial robust response. However, as both groups gained sexual experience, the impact of ketamine diminished.


In vitro beta 2-microglobulin (beta 2m) secretion by normal and leukaemic B-cells: effects of recombinant cytokines and evidence for a differential response to the combined stimulus of phorbol ester and calcium ionophore.
Due to the increasing therapeutic use of immunoregulatory agents and the potential effects on cellular function, we examined the modulation of in vitro beta 2-microglobulin (beta 2m) production rates by 'normal' tonsil and leukaemic B-cells in response to a number of these agents. Tonsil B-cells responded to phorbol ester (TPA) by an increased beta 2m production rate, which was further enhanced by the combined stimuli of TPA plus the calcium ionophore A23187. In marked contrast, however, lymphocytes from a majority (8/11) of B-cell malignancies showed a suppression of the TPA-induced beta 2m production rate in response to the combined TPA/A23187 stimulus. These different responses of 'normal' and malignant B-cells were not apparent when IgM production rates were examined. The recombinant cytokines IL-1, IL-2, IFN-alpha, IFN-gamma and TNF also enhanced beta 2m production rates of both normal and leukaemic B-cells, but to a considerably lesser extent than did TPA. Bryostatin-1 increased beta 2m production to a level intermediate between that obtained by TPA and the cytokines. It is suggested that beta 2m production rates may correspond to the degree of B-cell differentiation, and/or to the degree of cellular 'activation'. The results further indicate that the in vitro measurement of beta 2m production provides a different index of the cellular response than that obtained by the conventional measurement of IgM production.


The species composition of Antarctic phytoplankton interpreted in terms of Tilman's competition theory.
An attempt was made, to test for the impact of resource competition on Antarctic marine phytoplankton. According to theory, species composition near competitive equilibrium should be determined by the ratios of limiting resources. Enrichment bioassays identified silicon and nitrogen as limiting nutrients for some of the most important phytoplankton species during early austral summer in the region near the Antarctic Peninsula. Together with the generally acknowledged limiting resource light, this gave three meaningful ratios of essential resources (Si:N, Si:light, N:light) and one ratio of substitutable resources (NO3:NH4). Phytoplankton species assemblages were found to be well separated by the ratios of the essential resources and by mixing depth. Nine out of 12 individual species were found to be separated along at least one of the gradients of resource ratios. Where comparison with competition experiments was available, predicted and realized distributions of species were compatible.


Interstitial cystitis: early diagnosis, pathology, and treatment.
In a retrospective review, 52 patients with interstitial cystitis have been studied. Patients with persistent lower tract irritative symptoms, repeatedly sterile urine, and negative urine cytology must be suspected of having interstitial cystitis, and a diagnosis of urethral syndrome in such patients is highly questionable until cystoscopy under anesthesia has been performed. We believe that the finding of multiple petechia-like hemorrhages (glomerulations) on the second distention of the bladder is the hallmark of interstitial cystitis, and that a reduced bladder capacity and a Hunner's ulcer represent a different (classic) stage of this disease. In all stages, the characteristic histologic finidng is submucosal edema and vasodilation. The presence of eosinophils and mast cells is variable, and even in the classic disease the muscularis often appears to be normal. Immuno fluorescent studies and laboratory tests, including the fluorescent antinuclear antibody test (FANA), have not helped us to diagnose (or investigate) interstitial cystitis. Bladder instillations with a 0.4 per cent solution of oxychlorosene sodium (Clorpactin WCS-90) have provided remarkable relief for many patients with this disease, particulary those with the classic form.


Transnasal endoscopic retrograde chalangiopancreatography using an ultrathin endoscope: a prospective comparison with a routine oral procedure.
To investigate if transnasal endoscopic retrograde cholangiopancreatography (n-ERCP) using an ultrathin forward-viewing scope may overcome the disadvantages of conventional oral ERCP (o-ERCP) related to the large-caliber side-viewing duodenoscope. The study involved 50 patients in whom 25 cases each were assigned to the o-ERCP and n-ERCP groups. We compared the requirements of esophagogastroduodenoscopy (EGD) prior to ERCP, rates and times required for successful cannulation into the pancreatobiliary ducts, incidence of post-procedure hyperamylasemia, cardiovascular parameters during the procedure, the dose of a sedative drug, and successful rates of endoscopic naso-biliary drainage (ENBD). Screening gastrointestinal observations were easily performed by the forward-viewing scope and thus no prior EGD was required in the n-ERCP group. There was no significant difference in the rates or times for cannulation, or incidence of hyperamylasemia between the groups. However, the cannulation was relatively difficult in n-ERCP when the scope appeared U-shape under fluoroscopy. Increments of blood pressure and the amount of a sedative drug were significantly lower in the n-ERCP group. ENBD was successfully performed succeeding to the n-ERCP in which mouth-to-nose transfer of the drainage tube was not required. n-ERCP is likely a well-tolerable method with less cardiovascular stress and no need of prior EGD or mouth-to-nose transfer of the ENBD tube. However, a deliberate application is needed since its performance is difficult in some cases and is not feasible for some endoscopic treatments such as stenting.


The nipple-sparing mastectomy: early results of a feasibility study of a new application of perioperative radiotherapy (ELIOT) in the treatment of breast cancer when mastectomy is indicated.
Breast cancer surgery has become less and less mutilating, however a mastectomy is required in the case of multicentric, large tumors or recurrences after conservative treatment. The removal of the nipple areola complex during the mastectomy dramatically increases the feeling of mutilation. To reduce this negative psychological impact, in cancers located outside of the central area of the breast, we propose a new type of nipple-sparing mastectomy associated with intraoperative electron beam radiotherapy (ELIOT) delivered on the region of the areola. The nipple-sparing mastectomy is performed leaving 5 mm of glandular tissue behind the nipple areola complex to preserve its blood supply. The reconstruction is immediately performed with a prosthesis or an autologous flap. Twenty-five patients were included in the study; two of them had a bilateral nipple-sparing mastectomy. Pathological examinations demonstrated the presence of 19 infiltrating carcinomas and 8 ductal carcinoma in situ. Two patients had a superficial skin areolar slough followed by spontaneous healing. One necrosis of the areola occurred due to extensive retroareolar dissection. In the early follow-up, the color of the areola was preserved. All patients except one expressed their satisfaction of having kept their areola. These preliminary results are encouraging but they require further studies to evaluate the long-term results, the local recurrence rate and the psychological impact.


Pretreatment with acetylaminofluorene enhances the repair of O6-methylguanine in DNA.
Epidemiological and experimental evidence suggests that the interplay of environmental factors may be particularly relevant to the induction of cancer in man. We have investigated the possible interrelationships, at the level of DNA repair, of two chemically different hepatocarcinogens administered to rats. Animals were pretreated for several weeks by inclusion of 2-acetylaminofluorene (AAF) in the diet and a study was made of the capacity of liver to repair lesions subsequently introduced into DNA by a pulse of dimethylnitrosamine (DMN). Of particular interest was the repair of adducts at the O6 position of guanine as this product has been implicated as a critical reaction site for carcinogenesis by the monofunctional alkylating agents. We show here that the capacity of liver to repair O6-methylguanine in DNA is enhanced by prolonged exposure of rats to the chemically unrelated agent, AAF.


Coexistence of mutualists and non-mutualists in a dual-lattice model.
Evolution and maintenance of mutualism have been one of the major questions in evolutionary ecology, because it is often susceptible of invasion of non-mutualistic strategy. Some previous studies using dual-lattice model suggest that spatial structures of habitat can prevent non-mutualism from prevailing over mutualism, while the detail of the dynamics is not fully revealed. Here we explore population dynamics of the two strategies (mutualism and non-mutualism) in two species engaged in Prisoner's Dilemma game on a dual-lattice space, especially focusing on whether mutualists and non-mutualists can coexist in long-term dynamics. The habitat consists of two layers, each of which a population of species inhabits, and interspecific interaction is restricted between two corresponding sites of the layers. Each individual of the both species is either a mutualist or a non-mutualist and only the former pay cost c for benefit of the partner b. The payoff of the game affects the individuals' fecundity, while the mortality is constant. Reproduction is restricted to neighboring vacant sites of the focal individuals. Our computer simulations of the model show that even if b/c ratio remains constant, mutualists become dominant in both species over wider ranges of basic reproduction rate (reproduction rate without interspecific interaction) as b and c increase. If basic reproduction rates are asymmetric between the species or basic reproduction rates were sufficiently large, mutualists and non-mutualists can coexist in one or both species, while their population sizes often fluctuate. Transition of the final state between mutualism and non-mutualism happens rather discontinuously, then total population sizes change drastically at the transition. Moreover, we also find paradoxical cases of unilateral exploitation, i.e. one species consists of mutualists and other species non-mutualists. Additional simulations reveal that accidental extinction of the non-mutualists of one species can result in extinction of mutualist of the other species.


Fate of N-nitrosodimethylamine in recycled water after recharge into anaerobic aquifer.
Laboratory and field experiments were undertaken to assess the fate of N-nitrosodimethylamine (NDMA) in aerobic recycled water that was recharged into a deep anaerobic pyritic aquifer, as part of a managed aquifer recharge (MAR) strategy. Laboratory studies demonstrated a high mobility of NDMA in the Leederville aquifer system with a retardation coefficient of 1.1. Anaerobic degradation column and (14)C-NDMA microcosm studies showed that anaerobic conditions of the aquifer provided a suitable environment for the biodegradation of NDMA with first-order kinetics. At microgram per litre concentrations, inhibition of biodegradation was observed with degradation half-lives (260±20 days) up to an order of magnitude greater than at nanogram per litre concentrations (25-150 days), which are more typical of environmental concentrations. No threshold effects were observed at the lower ng L(-1) concentrations with NDMA concentrations reduced from 560 ng L(-1) to <6 ng L(-1) over a 42 day 14C-NDMA aerobic microcosm experiment. Aerobic (14)C-NDMA microcosm studies were also undertaken to assess potential aerobic degradation, likely to occur close to the recharge bore. These microcosm experiments showed a faster degradation rate than anaerobic microcosms, with a degradation half-life of 8±2 days, after a lag period of approximately 10 days. Results from a MAR field trial recharging the Leederville aquifer with aerobic recycled water showed that NDMA concentrations reduced from 2.5±1.0 ng L(-1) to 1.3±0.4 ng L(-1) between the recharge bore and a monitoring location 20 m down gradient (an estimated aquifer residence time of 10 days), consistent with data from the aerobic microcosm experiment. Further down gradient, in the anaerobic zone of the aquifer, NDMA degradation could not be assessed, as NDMA concentrations were too close to their analytical detection limit (<1 ng L(-1)).


Lack of somatic ErbB2 tyrosine kinase domain mutations in hepatocellular carcinoma.
Small molecules targeting the epidermal growth factor receptor (EGFR) intracellular tyrosine kinase domain have shown promising activity in cancer therapeutics. Recent reports suggest activity of erlotinib, an ErbB1 inhibitor, and lapatinib, a dual inhibitor of ErbB1 and ErbB2, in hepatocellular carcinoma (HCC). Activating ErbB1 somatic mutations may predict treatment responses. We have previously reported ErbB1 tyrosine kinase domain mutations to be rare or absent in HCC, but data on the frequency of ErbB2 tyrosine kinase domain mutations in HCC is currently limited, apart from reports of a missense mutation identified in 11% of a small Caucasian sample. We studied exons 18-23 of the ErbB2 gene from tumor DNA of 100 Asian human HCC and found no exonic mutations of potential significance. Alternative mechanisms may be responsible for the observed therapeutic efficacy of ErbB1 and ErbB2 tyrosine kinase inhibitors.


Trends, Costs, and Complications of Anterior Cervical Discectomy and Fusion With and Without Bone Morphogenetic Protein in the United States Medicare Population.
Retrospective database review. After the Food and Drug Administration approved bone morphogenetic protein-2 (BMP) in 2002, BMP was used off-label in the cervical spine to increase bone growth and bony fusion. Since then, concerns have been raised regarding complication rates and safety. This study was conducted to examine the use of BMP in anterior cervical discectomy and fusion (ACDF) in the Medicare population and to determine risk of complications and associated costs within 90 days of surgery. Patients who underwent ACDF were identified using Current Procedural Terminology (CPT) and International Classification of Diseases, Ninth Revision Procedure codes (ICD9-P). Complications were identified using ICD9 diagnostic codes. Charges were calculated as amount billed, and reimbursements were calculated as amounts paid by Medicare. Data for these analyses came from a nationwide claims database. A total of 215 047 patients were identified who had ACDF from 2005 to 2011. For the majority of the procedures (89.0%), BMP was not used. BMP use rose from 11.84% in 2005 to a peak of 16.73% in 2007 before decreasing to 12.01% in 2011. BMP was used 16% more in women than men. BMP use was the highest in the West (13.6%) followed by Midwest (11.8%), South (10.6%), and Northeast (7.5%). There was a higher overall complication rate in the BMP group (2.1%) compared with the non-BMP group (1.9%) (odds ratio [OR] = 1.11, 95% CI = 1.01-1.22). The BMP group also had a higher rate of wound complications (0.98% vs 0.76%, OR = 1.29, 95% CI = 1.12-1.48). In this study population, there was no difference in dysphagia/hoarseness, neurologic, medical, or other complications. During the 90-day perioperative period, BMP surgeries were charged at 17.6% higher than non-BMP surgeries. The use of BMP in ACDF in the Medicare population has decreased since a peak in 2007. The rate of wound and overall complications for BMP use with ACDF was higher than without. Our results regarding dysphagia/hoarseness did not show a statistically meaningful difference, which is in contrast with many other studies. Charges associated with BMP use were higher during the 90-day perioperative period.


Undergraduate nursing students' perspectives on clinical assessment at transition to practice.
Assessment of clinical competence requires explicitly defined standards meeting the national standards of the nursing profession. This is a complex process because of the diverse nature of nursing practice. To explore the perceptions of final-year undergraduate nursing students regarding clinical assessment at transition to practice. An exploratory qualitative approach was adopted. Twenty-four students participated in three focus group discussions. Thematic analysis was conducted. Five themes emerged: the need for a valid and reliable clinical assessment tool, the need for a flexible style of reflection and specific feedback, the dynamic clinical learning environment, students' efforts in learning and assessment, and the unclear support system for preceptors. Workload, time, resource availability, adequate preparation of preceptors, and the provision of valid and reliable clinical assessment tools were deemed to influence the quality of students' clinical learning and assessment. Nursing leadership in hospitals and educational institutions has a joint responsibility in shaping the clinical learning environment and providing clinical assessments for the students.


Differences in Learning and Persistency Characterizing Behavior in Chronic Pain for the Iowa Gambling Task: Web-Based Laboratory-in-the-Field Study.
Chronic pain is a significant worldwide health problem. It has been reported that people with chronic pain experience decision-making impairments, but these findings have been based on conventional laboratory experiments to date. In such experiments, researchers have extensive control of conditions and can more precisely eliminate potential confounds. In contrast, there is much less known regarding how chronic pain affects decision-making captured via laboratory-in-the-field experiments. Although such settings can introduce more experimental uncertainty, collecting data in more ecologically valid contexts can better characterize the real-world impact of chronic pain. We aim to quantify decision-making differences between individuals with chronic pain and healthy controls in a laboratory-in-the-field environment by taking advantage of internet technologies and social media. A cross-sectional design with independent groups was used. A convenience sample of 45 participants was recruited through social media: 20 (44%) participants who self-reported living with chronic pain, and 25 (56%) people with no pain or who were living with pain for <6 months acting as controls. All participants completed a self-report questionnaire assessing their pain experiences and a neuropsychological task measuring their decision-making (ie, the Iowa Gambling Task) in their web browser at a time and location of their choice without supervision. Standard behavioral analysis revealed no differences in learning strategies between the 2 groups, although qualitative differences could be observed in the learning curves. However, computational modeling revealed that individuals with chronic pain were quicker to update their behavior than healthy controls, which reflected their increased learning rate (95% highest-posterior-density interval [HDI] 0.66-0.99) when fitted to the Values-Plus-Perseverance model. This result was further validated and extended on the Outcome-Representation Learning model as higher differences (95% HDI 0.16-0.47) between the reward and punishment learning rates were observed when fitted to this model, indicating that individuals with chronic pain were more sensitive to rewards. It was also found that they were less persistent in their choices during the Iowa Gambling Task compared with controls, a fact reflected by their decreased outcome perseverance (95% HDI -4.38 to -0.21) when fitted using the Outcome-Representation Learning model. Moreover, correlation analysis revealed that the estimated parameters had predictive value for the self-reported pain experiences, suggesting that the altered cognitive parameters could be potential candidates for inclusion in chronic pain assessments. We found that individuals with chronic pain were more driven by rewards and less consistent when making decisions in our laboratory-in-the-field experiment. In this case study, it was demonstrated that, compared with standard statistical summaries of behavioral performance, computational approaches offered superior ability to resolve, understand, and explain the differences in decision-making behavior in the context of chronic pain outside the laboratory.


Femoral revision using the Wagner stem: results at 2-9 years.
We followed 39 patients with 40 revision hip arthroplasties using the Wagner stem. Mean follow-up was 65 (29-108) months. There were 12 intraoperative proximal femoral fractures, which all united, five postoperative dislocations not related to subsidence, and two loose stems that were revised. Clinical improvement was best for pain. Once the stem was osteointegrated the clinical and radiological result remained stable for up to 9 years.


Identification of 10 Hub genes related to the progression of colorectal cancer by co-expression analysis.
Colorectal cancer (CRC) is the third most common cancer in the world. The present study is aimed at identifying hub genes associated with the progression of CRC. The data of the patients with CRC were obtained from the Gene Expression Omnibus (GEO) database and assessed by weighted gene co-expression network analysis (WGCNA), Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses performed in R by WGCNA, several hub genes that regulate the mechanism of tumorigenesis in CRC were identified. Differentially expressed genes in the data sets GSE28000 and GSE42284 were used to construct a co-expression network for WGCNA. The yellow, black and blue modules associated with CRC level were filtered. Combining the co-expression network and the PPI network, 15 candidate hub genes were screened. After validation using the TCGA-COAD dataset, a total of 10 hub genes (MT1X, MT1G, MT2A, CXCL8, IL1B, CXCL5, CXCL11, IL10RA, GZMB, KIT) closely related to the progression of CRC were identified. The expressions of MT1G, CXCL8, IL1B, CXCL5, CXCL11 and GZMB in CRC tissues were higher than normal tissues (p-value < 0.05). The expressions of MT1X, MT2A, IL10RA and KIT in CRC tissues were lower than normal tissues (p-value < 0.05). By combinating with a series of methods including GO enrichment analysis, KEGG pathway analysis, PPI network analysis and gene co-expression network analysis, we identified 10 hub genes that were associated with the progression of CRC.


Immunophenotypical characterization and influence on liver homeostasis of depleting and repopulating hepatic macrophages in rats injected with clodronate.
Hepatic macrophages (including Kupffer cells) play a crucial role in the homeostasis and act as mediators of inflammatory response in the liver. Hepatic macrophages were depleted in male F344 rats by a single intravenous injection of liposomal clodronate (CLD; 50mg/kg body weight), and immunophenotypical characteristics of depleting and repopulating macrophages were analyzed by different antibodies specific for macrophages. CD163(+) Kupffer cells were almost completely depleted on post-injection (PI) days 1-12. Macrophages reacting to CD68, Iba-1, and Gal-3 were drastically reduced in number on PI day 1 and then recovered gradually until PI day 12. MHC class II(+) and CD204(+) macrophages were moderately decreased during the observation period. Although hepatic macrophages detectable by different antibodies were reduced in varying degrees, Kupffer cells were the most susceptible to CLD. Liver situation influenced by depleted hepatic macrophages was also investigated. No marked histological changes were seen in the liver, but the proliferating activity of hepatocytes was significantly increased, supported by changes of gene profiles relating to cell proliferation on microarray analysis on PI day 1; the values of AST and ALT were significantly elevated; macrophage induction/activation factors (such as MCP-1, CSF-1, IL-6 and IL-4) were increased exclusively on PI day 1, whereas anti-inflammatory factors such as IL-10 and TGF-β1 remained significantly decreased after macrophage depletion. The present study confirmed importance of hepatic macrophages in liver homeostasis. The condition of hepatic macrophages should be taken into consideration when chemicals capable of inhibiting macrophage functions are evaluated.


Role of AF6 protein in cell-to-cell spread of Herpes simplex virus 1.
AF6 and its rat homologue afadin are multidomain proteins localized at cell junctions and involved in intercellular adhesion. AF6 interacts via its PDZ domain with nectin-1 at epithelial adherens junctions. Nectin-1 serves as a mediator of cell-to-cell spread for Herpes simplex virus 1 (HSV-1). We analyzed the role of AF6 protein in the viral spread and nectin-1 clustering at cell-cell contacts by knockdown of AF6 in epithelial cells. AF6 knockdown reduced efficiency of HSV-1 spreading, however, the clustering of nectin-1 at cell-cell contacts was not affected. Thus, AF6 protein is important for spreading of HSV-1 in epithelial cells, independently of nectin clustering, possibly by stabilization of the E-cadherin-dependent cell adhesion.


Draft genome sequence of Lactobacillus plantarum strain DMR17 isolated from homemade cow dahi of Sikkim Himalayan region: an evaluation of lactate fermentation and secondary metabolism.
Lactobacillus plantarum DMR17 was isolated from homemade cow dahi of Sikkim Himalayan region of India. Here, we report the draft genome sequence of this strain. A total of 21,176,638 paired-end reads were obtained which were assembled into 65 contigs. The reference genome used was L. plantarum WCFS1. The genome size of DMR17 was 3,228,341 bp with G + C content of 46.25%. 3302 coding sequences were predicted including 68 tRNA and 67 rRNA genes. More than 88% of the total pre-processed reads from the sample were mapped to the reference genome. The identified coding proteins were classified into 27 functional categories based on COG classification. The genome was found to possess genes for lactate and mixed acid fermentation. The genome also showed the presence of genes for catabolism of aromatic compounds, phosphorous, and other metabolism. The genome information of L. plantarum DMR17 provides the basis for understanding the functional properties and to consider its use as a potential component of functional foods especially dahi.


Neuraxial Anesthesia in Parturients with Thrombocytopenia: A Multisite Retrospective Cohort Study.
The primary aim of this study was to estimate the risk of neuraxial hematoma associated with neuraxial anesthetic procedures in thrombocytopenic parturients. A multicenter retrospective cohort study design was used to estimate the risk for spinal-epidural hematoma in parturients with a platelet count of <100,000/mm receiving neuraxial anesthesia and the risk of complications in thrombocytopenic parturients who receive general anesthesia. No cases of spinal hematoma were observed in 102 thrombocytopenic parturients receiving epidural analgesia or 71 receiving spinal anesthesia. Including data from the previous published series (total n = 499), the exact binomial 95% confidence interval for the risk of spinal-epidural hematoma was 0% to 0.6%. Given the small number of patients at each specific platelet count, the theoretical risks at individual platelet count strata are presented. Overall aggregate serious morbidity rate in women who received general anesthesia secondary to thrombocytopenia was 6.5% (95% confidence interval, 2.1%-14.5%). Our work supports the relative maternal safety of neuraxial anesthesia in parturients with mild thrombocytopenia and estimates the maternal complication rate associated with the avoidance of neuraxial anesthesia. Remaining uncertainties at lower platelet counts make a national "low platelet" registry critical to a more accurate assessment of the risk of epidural hematoma and would aid in standardization of anesthesia practice.


[Our experience, with the tube manometry in chronic otitis media. I: Equipment and methodology (author's transl)].
A simple method for clinical routine to check tube function in perforated tympanic membranes using an additional set developed by us for an impedanceaudiometer is reported. Before the possibilities to check tube function by tympanometric methods in normal tympanic membranes are discussed.


Incidence of Post-Heart Transplant Chronic Thyroiditis and Its Association With Pretransplant Amiodarone Use.
Chronic thyroiditis (CT) is a common cause of thyroid dysfunction and could therefore adversely affect outcomes in patients undergoing heart transplant (HT). The incidence of post-HT CT and whether amiodarone, a commonly used anti-arrhythmic drug in patients with heart failure during pre-HT period, is associated with the development of post-HT CT are unknown. A retrospective review of HT recipients from February 2, 2010 to October 16, 2018 was performed. Patients who lacked relevant pre-/post-HT records, underwent thyroidectomy, had pre-HT thyroid dysfunction or thyroiditis within 15 days post-HT, and those on amiodarone during the post-HT period were excluded, yielding a final cohort of 75 patients. Patients had a mean age of 63.3 ± 1.4 years and were predominantly male (90.7%) and white (80%). The incidence of post-HT CT was 32% with the majority (83.3%) manifesting as hypothyroidism. Median time to diagnosis of CT after transplant was 10.2 months (interquartile range, 4-27.4). Additionally, the CT group had higher pre-HT use of amiodarone (non-CT vs CT: 21.6% vs 50%, P = .01), higher prevalence of atrial fibrillation (non-CT vs CT: 23.5% vs 45.8%; P = .05), and more stage IV/V chronic kidney disease (non-CT vs CT: 2% vs 16.7%, P = .02). On multivariate analysis, pre-HT amiodarone use was associated with the development of post-HT CT after adjustment for age, sex, and chronic kidney disease (odds ratio, 3.65; 95% CI, 1.17-11.44; P = .03). The incidence of post-HT CT is high and is strongly associated with pre-HT amiodarone use underpinning the importance of closely following the post-HT thyroid profile in these patients.


Reactions of Fluoroalkanes with Mg-Mg Bonds: Scope, sp3 C-F/sp2 C-F Coupling and Mechanism.
sp3 C-F Bonds of fluoroalkanes (7 examples; 1°, 2° and 3°) undergo addition to a low-valent Mg-Mg species generating reactive organomagnesium reagents. Further reactions with a series of electrophiles results in a net C-F to C-B, C-Si, C-Sn or C-C bond transformation (11 examples, diversity). The new reactivity has been exploited in an unprecedented one-pot magnesium-mediated coupling of sp3 C-F and sp2 C-F bonds. Calculations suggest that the sp3 C-F bond activation step occurs by frontside nucleophilic attack of the Mg-Mg reagent on the fluoroalkane.


Prognostic significance of arterial blood gas analysis in the early evaluation of paraquat poisoning patients.
To examine the utility of arterial blood gas analysis (ABG) in early evaluation of prognosis in paraquat poisoning. Our aim was to summarize the case data of 138 patients poisoned with oral paraquat treated in the Emergency Department of 307 Hospital of the Chinese People's Liberation Army from June 2009 to Sept. 2010, and analyze the correlations between various indices of arterial blood gas analysis (including pH, PO(2), PCO(2), base excess [BE], HCO(3)(-)) to prognosis and blood PQ concentration of patients presenting within 24 h after taking paraquat. PCO(2), HCO(3)(-) and BE values in deceased patients were significantly lower than those in surviving patients, p values 0.0003, <0.0001, <0.0001; PCO(2), HCO(3)(-) and BE values in patients who died in < 3 d were significantly lower than those in those who died in 3-7 d and 7 d after taking paraquat (p < 0.0001). The results of Cox Regression Analysis showed that there was correlation between paraquat amount, blood paraquat concentration and BE values and patients' survival time; the larger the absolute BE value was, the higher the death rate. Nevertheless, there were no correlations between early pH or PO(2) and prognosis in these patients. BE values may be a reliable index in early evaluation of prognosis in paraquat poisoning.


Effect of omeprazole and sucralfate on gastrointestinal injury in a fasting/NSAID model.
Equine gastric ulcer syndrome (EGUS) is a common and significant cause of morbidity in horses, with a range of clinical signs, including inappetence, colic and poor performance. Hospitalised horses are exposed to factors that may induce EGUS, including fasting and nonsteroidal anti-inflammatory drug (NSAID) administration, and may be at risk for development of squamous (ESGD) and glandular gastric disease (EGGD). Prophylactic anti-ulcer medication is often prescribed for these patients, but drug selection is complicated by different aetiology and response to treatment of ESGD and EGGD. To establish the efficacy of sucralfate or omeprazole used prophylactically in horses exposed to a combined feed-fast and NSAID administration EGUS induction protocol. We hypothesised that these drugs would be equally effective for prevention of gastric lesions in the experimental cohort. Randomised crossover experimental design. Horses (n = 14) received either omeprazole (1 mg/kg PO q24h) or sucralfate (20 mg/kg PO q8h) while undergoing the feed-fast/NSAID protocol, allowed an 8-week washout period, and then administered the alternate treatment. Serial gastroscopy, ultrasound and haematology documented treatment effects. ESGD and EGGD score increased over time under both treatments. There was a significant effect of treatment on EGGD scores (P < .001), with post-treatment EGGD scores higher for horses receiving sucralfate (median 3; IQR 2.25,3) than omeprazole (1; 1,1). The effect of treatment on ESGD scores just achieved significance (P = .05), with post-treatment ESGD scores higher for sucralfate (4; 3,4) than omeprazole (2; 2,3). This study was performed in healthy horses, and response to treatment may differ in horses with clinical illness. Additional investigation in a larger population may be required to detect significant differences in other clinical parameters. Omeprazole was superior to sucralfate for mitigating gastric lesion severity in healthy horses exposed to a feed-fast/NSAID model.


Relationships of ultrasound measures of intrinsic foot muscle cross-sectional area and muscle volume with maximum toe flexor muscle strength and physical performance in young adults.
[Purpose] To investigate the relationships between toe flexor muscle strength with (TFS-5-toes) and without (TFS-4-toes) the contribution of the great toe, anatomical and physiological muscle cross-sectional areas (CSA) of intrinsic toe flexor muscle and physical performance were measured. [Subjects] Seventeen men (82% sports-active) and 17 women (47% sports-active), aged 20 to 35 years, volunteered. [Methods] Anatomical CSA was measured in two intrinsic toe flexor muscles (flexor digitorum brevis [FDB] and abductor hallucis) by ultrasound. Muscle volume and muscle length of the FDB were also estimated, and physiological CSA was calculated. [Results] Both TFS-5-toes and TFS-4-toes correlated positively with walking speed in men (r=0.584 and r=0.553, respectively) and women (r=0.748 and r=0.533, respectively). Physiological CSA of the FDB was significantly correlated with TFS-5-toes (r=0.748) and TFS-4-toes (r=0.573) in women. In men, physiological CSA of the FDB correlated positively with TFS-4-toes (r=0.536), but not with TFS-5-toes (r=0.333). [Conclusion] Our results indicate that physiological CSA of the FDB is moderately associated with TFS-4-toes while toe flexor strength correlates with walking performance.


Automated dispersive liquid-liquid microextraction coupled to high performance liquid chromatography - cold vapour atomic fluorescence spectroscopy for the determination of mercury species in natural water samples.
An automated, home-constructed, and low cost dispersive liquid-liquid microextraction (DLLME) device that directly coupled to a high performance liquid chromatography (HPLC) - cold vapour atomic fluorescence spectroscopy (CVAFS) system was designed and developed for the determination of trace concentrations of methylmercury (MeHg+), ethylmercury (EtHg+) and inorganic mercury (Hg2+) in natural waters. With a simple, miniaturized and efficient automated DLLME system, nanogram amounts of these mercury species were extracted from natural water samples and injected into a hyphenated HPLC-CVAFS for quantification. The complete analytical procedure, including chelation, extraction, phase separation, collection and injection of the extracts, as well as HPLC-CVAFS quantification, was automated. Key parameters, such as the type and volume of the chelation, extraction and dispersive solvent, aspiration speed, sample pH, salt effect and matrix effect, were thoroughly investigated. Under the optimum conditions, linear range was 10-1200ngL-1 for EtHg+ and 5-450ngL-1 for MeHg+ and Hg2+. Limits of detection were 3.0ngL-1 for EtHg+ and 1.5ngL-1 for MeHg+ and Hg2+. Reproducibility and recoveries were assessed by spiking three natural water samples with different Hg concentrations, giving recoveries from 88.4-96.1%, and relative standard deviations <5.1%.


Geometry and scale in species-area relationships.
Arising from F. He & S. P. Hubbell 473, 368-371 (2011). He and Hubbell developed a sampling theory for the species-area relationship (SAR) and the endemics-area relationship (EAR). They argued that the number of extinctions after habitat loss is described by the EAR and that extinction rates in previous studies are overestimates because the EAR is always lower than the SAR. Here we show that their conclusion is not general and depends on the geometry of habitat destruction and the scale of the SAR. We also question their critique of the Millennium Ecosystem Assessment estimates, as those estimates are not dependent on the SAR only, although important uncertainties remain due to other methodological issues.


[Health insurance scheme: main contributor to inequalities in COVID-19 mortality in ColombiaTipo de cobertura de assistência à saúde: o principal fator contribuinte para as desigualdades em mortalidade por COVID-19 na Colômbia].
To quantify socioeconomic inequalities in COVID-19 mortality in Colombia and to assess the extent to which type of health insurance, comorbidity burden, area of residence, and ethnicity account for such inequalities. We analyzed data from a retrospective cohort of COVID-19 cases. We estimated the relative and slope indices of inequality (RII and SII) using survival models for all participants and stratified them by age and gender. We calculated the percentage reduction in RII and SII after adjustment for potentially relevant factors. We identified significant inequalities for the whole cohort and by subgroups (age and gender). Inequalities were higher among younger adults and gradually decreased with age, going from RII of 5.65 (95% confidence interval [CI] = 3.25, 9.82) in participants younger than 25 years to RII of 1.49 (95% CI = 1.41, 1.58) in those aged 65 years and older. Type of health insurance was the most important factor, accounting for 20% and 59% of the relative and absolute inequalities, respectively. Significant socioeconomic inequalities exist in COVID-19 mortality in Colombia. Health insurance appears to be the main contributor to those inequalities, posing challenges for the design of public health strategies.


The risks and benefits of thrombolytic therapy in acute aortic and mitral prosthetic valve dysfunction: report of a case and review of the literature.
A patient with acute thrombosis of a St. Jude mitral prosthesis was treated with streptokinase with initial success. Subsequent recurrent thrombosis and embolism prompted operative revision. Because the use of thrombolytic agents in this setting is somewhat controversial, we searched the literature for all reports of aortic or mitral prosthetic valve dysfunction treated this way. The cases of 58 patients treated 62 times were reviewed for efficacy of therapy and morbidity. Thrombolytic therapy may be useful in patients with prosthetic valve thrombosis causing critical hemodynamic compromise. It is frequently the only treatment needed. Further, it may help reduce operative risk for those patients in whom complete resolution is not possible. The incidence of systemic embolism is 18%, however, neurological events are usually limited and transient.


The N-terminal domain of Escherichia coli ClpB enhances chaperone function.
ClpB/Hsp104 collaborates with the Hsp70 system to promote the solubilization and reactivation of proteins that misfold and aggregate following heat shock. In Escherichia coli and other eubacteria, two ClpB isoforms (ClpB95 and ClpB80) that differ by the presence or absence of a highly mobile 149-residues long N-terminus domain are synthesized from the same transcript. Whether and how the N-domain contributes to ClpB chaperone activity remains controversial. Here, we show that, whereas fusion of a 20-residues long hexahistidine extension to the N-terminus of ClpB95 interferes with its in vivo and in vitro activity, the same tag has no detectable effect on ClpB80 function. In addition, ClpB95 is more effective than ClpB80 at restoring the folding of the model protein preS2-beta-galactosidase as stress severity increases, and is superior to ClpB80 in improving the high temperature growth and low temperature recovery of dnaK756 DeltaclpB cells. Our results are consistent with a model in which the N-domain of ClpB95 maximizes substrate processing under conditions where the cellular supply of free DnaK-DnaJ is limiting.


Metabolic interaction of hydrogen peroxide and hypoxia in zebrafish fibroblasts.
Cells require oxygen for aerobic metabolism, which may also result in the production of reactive oxygen species (ROS) as a by-product. Under low oxygen conditions, ROS formation has been reported to either increase or decrease. We addressed this physiological response for the first time in zebrafish embryonic fibroblasts (Z3) and used a hydrogen peroxide (H2O2)-specific fluorescent protein (roGFP2-Orp1) either targeted to the mitochondria or expressed in the cytosol. Microfluidic live-cell imaging measurements showed that oxygen deprivation in Z3 cells results in decreased or stable H2O2 levels within the mitochondria or the cytosol, respectively, and that the reductive shift recorded in the mitochondrial matrix is directly dependent on oxygen concentration. The response was accompanied by a transient increase in extracellular acidification rate (ECAR) and a lower cellular reducing potential as assessed by the viability stain alamarBlue. Complex I and III inhibition with Rotenone and Antimycin A led to H2O2 production under normoxia but these inhibitors were not able to avert the reductive shift under hypoxia. Only by system-wide inhibition of flavin-containing oxidases with Diphenyleneiodonium (DPI) were we able to decrease the reductive shift, while selective inhibition of NADPH oxidases with the inhibitor Apocynin had no effect on the hypoxia response. Since DPI also led to a strong increase in ECAR we found that, in order to keep the cytosolic H2O2 levels stable, glycolytic metabolism was of fundamental importance. According to our experiments with the glucose-6-phosphate dehydrogenase inhibitor 6-Aminonicotinamide, this was attributable to the pentose phosphate pathway producing reducing equivalents required for ROS degradation.


Generation of reentry in anisotropic myocardium.
We investigated numerically the effects of the rotation of fiber axis orientation through the myocardial wall on wave propagation. We show that because of this rotation and inherent discrete properties of myocardium, a premature stimulus can create unidirectional conduction block leading to reentry. The dynamics of the subsequent reentrant patterns are complicated by the presence of rotational anisotropy, as the center of reentry drifts, and the reentry terminates in finite time when it collides with the domain boundary.


Target-cell-specific left-right asymmetry of NMDA receptor content in schaffer collateral synapses in epsilon1/NR2A knock-out mice.
Input-dependent left-right asymmetry of NMDA receptor epsilon2 (NR2B) subunit allocation was discovered in hippocampal Schaffer collateral (Sch) and commissural fiber pyramidal cell synapses (Kawakami et al., 2003). To investigate whether this asymmetrical epsilon2 allocation is also related to the types of the postsynaptic cells, we compared postembedding immunogold labeling for epsilon2 in left and right Sch synapses on pyramidal cells and interneurons. To facilitate the detection of epsilon2 density difference, we used epsilon1 (NR2A) knock-out (KO) mice, which have a simplified NMDA receptor subunit composition. The labeling density for epsilon2 but not zeta1 (NR1) and subtype 2/3 glutamate receptor (GluR2/3) in Sch-CA1 pyramidal cell synapses was significantly different between the left and right hippocampus with opposite directions in strata oriens and radiatum; the left to right ratio of epsilon2 labeling density was 1:1.50 in stratum oriens and 1.44:1 in stratum radiatum. No significant difference, however, was detected in CA1 stratum radiatum between the left and right Sch-GluR4-positive (mostly parvalbumin-positive) and Sch-GluR4-negative interneuron synapses. Consistent with the anatomical asymmetry, the amplitude ratio of NMDA EPSCs to non-NMDA EPSCs in pyramidal cells was approximately two times larger in right than left stratum radiatum and vice versa in stratum oriens of epsilon1 KO mice. Moreover, the amplitude of long-term potentiation in the Sch-CA1 synapses of left stratum radiatum was significantly larger than that in the right corresponding synapses. These results indicate that the asymmetry of epsilon2 distribution is target cell specific, resulting in the left-right difference in NMDA receptor content and plasticity in Sch-CA1 pyramidal cell synapses in epsilon1 KO mice.


Iodothyronine deiodinase activity in methionine-deficient rats fed selenium-deficient or selenium-sufficient diets.
We examined the effect of methionine deficiency on iodothyronine 5'-deiodinase activity in selenium-deficient rats or selenium-sufficient rats fed sodium selenate or selenomethionine. Forty-two weanling male Wistar rats were divided into six groups and pair fed the respective purified L-amino acid-based diets for 4 wk. L-methionine concentrations in the diet were 8.0 g/kg for sufficient rats, and 2.0 g/kg for deficient rats. Selenium concentrations in the diet were 0.5 mg/kg (as sodium selenate or selenomethionine) for selenium-sufficient rats and less than 0.005 mg/kg for selenium-deficient rats. Type I 5'-deiodinase activities were significantly lower in liver and higher in kidney of methionine-deficient rats than in those of methionine-sufficient rats fed either the selenium-sufficient or the selenium-deficient diets. The type I 5'-deiodinase activity in brain was significantly lower in the methionine-deficient rats than in the methionine-sufficient rats fed the selenium-deficient diet. Type II 5'-deiodinase activity in brain was significantly higher in the methionine-deficient rats than in the methionine-sufficient rats fed selenium-sufficient diet as sodium selenate. Both thyroxine and 3,3',5-triiodothyronine concentrations in plasma were significantly higher in the methionine-deficient rats than in the methionine-sufficient rats. It is suggested that the methionine deficiency affects the 5'-deiodinase activity and thyroid hormones level in the rats.


Advance in the pharmacological and comorbidities management of heart failure with preserved ejection fraction: evidence from clinical trials.
The prevalence of heart failure with preserved ejection fraction (HFpEF) accounts for approximately 50% of the total heart failure population, and with the aging of the population and the increasing prevalence of hypertension, obesity, and type 2 diabetes (T2DM), the incidence of HFpEF continues to rise and has become the most common subtype of heart failure. Compared with heart failure with reduced ejection fraction, HFpEF has a more complex pathophysiology and is more often associated with hypertension, T2DM, obesity, atrial fibrillation, renal insufficiency, pulmonary hypertension, obstructive sleep apnea, and other comorbidities. HFpEF has generally been considered a syndrome with high phenotypic heterogeneity, and no effective treatments have been shown to reduce mortality to date. Diuretics and comorbidity management are traditional treatments for HFpEF; however, they are mostly empirical due to a lack of clinical evidence in the setting of HFpEF. With the EMPEROR-Preserved and DELIVER results, sodium-glucose cotransporter 2 inhibitors become the first evidence-based therapies to reduce rehospitalization for heart failure. Subgroup analyses of the PARAGON-HF, TOPCAT, and CHARM-Preserved trials suggest that angiotensin receptor-neprilysin inhibitors, spironolactone, and angiotensin II receptor blockers may be beneficial in patients at the lower end of the ejection fraction spectrum. Other potential pharmacotherapies represented by non-steroidal mineralocorticoid receptor antagonists finerenone and antifibrotic agent pirfenidone also hold promise for the treatment of HFpEF. This article intends to review the clinical evidence on current pharmacotherapies of HFpEF, as well as the comorbidities management of atrial fibrillation, hypertension, T2DM, obesity, pulmonary hypertension, renal insufficiency, obstructive sleep apnea, and iron deficiency, to optimize the clinical management of HFpEF.


Minimally invasive dynamic hip screw for fixation of hip fractures.
We compared a minimally invasive surgical technique to the conventional (open approach) surgical technique used in fixation of hip fractures with the dynamic hip screw (DHS) device. Using a case-control design (44 cases and 44 controls), we tested the null hypothesis that there is no difference between the two techniques in the following outcome measures: duration of surgery, time to mobilisation and weight bearing postoperatively, length of hospital stay, mean difference of pre- and postoperative haemoglobin levels, position of the lag screw of the DHS device in the femoral head, and the tip-apex distance. The minimally invasive DHS technique had significantly shorter duration of surgery and length of hospital stay. There was also less blood loss in the minimally invasive DHS technique. The minimally invasive DHS technique produces better outcome measures in the operating time, length of hospital stay, and blood loss compared to the conventional approach while maintaining equal fixation stability.


Physical activity improves outcomes of combined lenvatinib plus anti-PD-1 therapy in unresectable hepatocellular carcinoma: a retrospective study and mouse model.
Physical activity is known to have anti-cancer effects, including immunomodulatory actions. This study investigated the hypothesis that physical activity synergizes with combined lenvatinib plus anti-PD-1 therapy to enhance efficacy in patients with unresectable HCC. The physical activity levels of patients with unresectable HCC receiving combined lenvatinib plus anti-PD-1 therapy were recorded by questionnaire. Patients were categorized according to physical activity levels (active vs. sedentary). The primary outcome was overall survival (OS). Secondary outcomes included objective response rate (ORR) and progression-free survival (PFS). A subcutaneous syngeneic HCC model was generated in C57BL/6 mice. Mice were randomized to receive placebo, combined lenvatinib plus anti-PD-1 antibodies or combination therapy plus physical activity. Tumors were measured every 3 days and harvested for immunohistochemistry analysis at 20 mm maximum diameter. Fifty-nine patients with unresectable HCC were categorized to active (n = 28) or sedentary (n = 31) groups. The active group had higher albumin and des-γ-carboxy prothrombin levels and lower hepatitis B virus load at baseline; other clinical and oncologic characteristics were comparable between the two groups. Patients in the active group had significantly longer OS (HR = 0.220, 95% CI 0.060-0.799) and PFS (HR = 0.158, 95% CI 0.044-0.562) and higher ORR (OR = 4.571, 95% CI 1.482-14.102) than patients in the sedentary group. Regular physical activity was independently associated with OS, PFS and ORR. The mouse model showed that physical activity significantly suppressed tumor growth and prolonged survival of tumor-bearing mice. Furthermore, physical activity inhibited Treg cell infiltration and immune checkpoint expression (including CTLA4, TIGIT and TIM3) induced by long-term combined lenvatinib plus anti-PD-1 therapy, improving efficacy. Regular physical activity was associated with improved outcomes in unresectable HCC receiving combined lenvatinib plus anti-PD-1 therapy. Physical activity may improve therapeutic efficacy by reprograming the tumor microenvironment from an immunosuppressive to immunostimulatory phenotype.


Metabolic bone disease in primary biliary cirrhosis at presentation.
Metabolic bone disease, particularly osteoporosis, is a complication of advanced primary biliary cirrhosis, but the extent of the problem is unclear. We present 33 patients who were investigated for bone disease at the time of diagnosis of their liver disease and who had received no prior treatment likely to influence their bones. Iliac crest bone biopsy showed no patient with osteoporosis, and mild osteomalacic changes in 1 patient. Slight elevations in appositional rate, osteoid volume, and resorption surface were compatible with a state of high bone turnover. Photon absorptiometry revealed a low forearm bone mineral content in 3 of 25 patients, calcium absorption was below normal in 14 of 24 patients, and there was evidence of fat malabsorption in 11 of 25 patients. Five patients also had low serum levels of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D. Thus, little evidence of significant metabolic bone disease was found in this group by these methods, but abnormalities were seen, such as poor calcium absorption, that may predispose to its later development.


Common Diarrheal Illnesses in the Elderly.
Diarrhea is a fairly common problem among the elderly that has a higher morbidity and mortality compared with the general population. There are multiple reasons for diarrhea in the elderly that can be stratified by different mechanisms: infectious, osmotic, secretory, inflammatory, and malabsorptive. Oral hydration and dietary management are the basic management principles for all forms of diarrhea but specific treatment should address the root cause of diarrhea in order to improve outcomes.


Biosynthesis of poly-beta-hydroxyalkanoates by Sphingopyxis chilensis S37 and Wautersia sp. PZK cultured in cellulose pulp mill effluents containing 2,4,6-trichlorophenol.
Poly-beta-hydroxyalkanoates (PHA) polymer is synthesized by different bacterial species. There has been considerable interest in the development and production of biodegradable polymers; however, the high cost of PHA production has restricted its applications. Kraft cellulose industry effluents containing 2,4,6-trichlorophenol (10 or 20 microg ml(-1)) were used by the bacteria Sphingopyxis chilensis S37 and Wautersia sp. PZK to synthesize PHA. In this condition, S. chilensis S37 was able to grow and degrade 2,4,6-trichlorophenol (ca. 60%) and 80% of these cells accumulated PHA. Wautersia PZK completely degraded 2,4,6-TCP and more than 90% of the cells accumulated PHA in 72 h. The PHA detection was performed by flow cytometry and polyester composition was characterized by gas chromatography-mass spectroscopy (GC-MS), indicating that these polymers are made by 3-hydroxybutyric acid and 3-hydroxyhexadecanoic acid for S37 and PZK strains, respectively. Results demonstrated that strains' growth and PHA production and composition are not modified in cellulose effluents with or without 2,4,6-TCP (10-20 microg ml(-1)). Therefore, our results indicate that S. chilensis S37 and Wautersia sp. PZK are able to degrade a toxic compound such as a 2,4,6-TCP and simultaneously produce a valuable biopolymer using low-value substrates.


Silent infarction in acute stroke patients. Prevalence, localization, risk factors, and clinical significance: the Copenhagen Stroke Study.
Our objective was to study age-specific prevalence, computed tomographic (CT) characteristics, risk factors, and the prognostic influence on stroke outcome of silent infarction in acute stroke patients. The study was prospective and community-based and included 801 acute stroke patients, of whom 587 had first-ever stroke. A CT scan was performed in 500 (85%) of the 587 patients with first-ever stroke. CT was reviewed blindly, and infarcts were classified according to patient history as silent or symptomatic. Patients were evaluated initially with the Mini-Mental State Examination (MMSE) and weekly with both the Scandinavian Stroke Scale (SSS) and the Barthel Index (BI) from the onset of stroke to completion of rehabilitation. CT characteristics, risk factors, and stroke outcome were compared in stroke patients with and without silent infarction. The prevalence of silent infarction in patients with first-ever stroke and recurrent strokes was similar, at 29% (group aged 0 to 54 years, 16%; 55 to 64 years, 22%; 65 to 74 years, 30%; 75 years or older, 33%). Silent infarcts were small and subcortical. Independent risk factors were increasing age (odds ratio [OR], 1.95 per 25 years; confidence interval [CI], 1.19 to 3.15), hypertension (OR, 1.75; CI, 1.13 to 2.70), claudication (OR, 1.74; CI, 1.01 to 3.00), and male sex (OR, 1.72; CI, 1.12 to 2.64); other stroke risk factors such as atrial fibrillation and former transient ischemic attack were not independent risk factors. Patients with and without silent infarction did not differ in frequency of prestroke home care (P = .2). MMSE (P = .56), initial BI (P = .62) and SSS score (P = .08), BI (P = .85) and SSS score (P = .75) after completion of rehabilitation, or in the speed of recovery (P = .85). Length of hospital stay, mortality rate, and discharge rate to nursing home also did not differ between the two groups. This community-based study shows that silent infarction in stroke patients is more related to certain stroke risk factors than others and that silent infarction does not seem to influence the prognosis of stroke.


[Therapeutical and inflammatory response in rats with secondary peritonitis and topical use of ampicilin/sulbactam].
The acute peritonitis is an important cause of sepsis and death on intensive care units and surgery. The treatment must include: systemical use of antibiotics, drainage of abscess and restauration of gastrointestinal integrity. The topical use of antibiotics in the peritoneal cavity is controversial. The aim of this study was to evaluate the use of topical use of ampicilin/ sulbactam in the treatment of peritonitis. We measured the plasmatic levels of nitric oxide, count of eosinophils, lymphocytes, monocytes, and neutrophils in blood and peritoneal cavity, using a model of peritonitis in rats (transfixation and ligature of cecum). Twenty four Wistar rats were divided in 4 groups (n = 6 each). group A: induction of peritonitis with ligature of cecum and topical treatment with saline; group B: induction of peritonitis with ligature of cecum and topical treatment with ampicilin/sulbactam; group C: transfixation of cecum; group D: laparotomy and peritoneal exsudate + blood sample. The transfixation-ligture of cecum remained for 24 hs before treatment. A relaparotomy was performed in 18 rats and peritoneal exsudate/blood were collected. Dosage of Nitric oxide, count of eosinophil, lymphocytes, monocytes, and neutrophils in blood and peritoneal exsudte were done. The difference was not significant in the levels of nitric oxide, eosinophil, lymphocytes, monocytes, and neutrophils in blood and peritoneal exsudate (p > 0,05) among the studied groups. The use of ampicilin associated to sulbactam via intraperitoneal in rats with fecal peritonitis did not change survival.; the levels of plama nitric oxide, count of eosinophil, lymphocytes, monocytes, and neutrophils in blood and peritoneal exsudate were not affected.


[New portfolio, a users' manual].
The portfolio, a tool for monitoring nursing students throughout their internship period, has now been modified. The new 2015 version can be used to monitor and trace the student's career as well as progress made. One of the major new points is the integration of an intermediate internship assessment.