Modified reverse aortoplasty versus extended anastomosis in patients with coarctation of the aorta and distal arch hypoplasia.
The aim of our prospective randomized study was to compare modified reverse aortoplasty (MRA) and extended end-to-end anastomosis (EEA). We have assessed the operative correction results in 54 infants with coarctation of the aorta and distal aortic arch hypoplasia who underwent primary repair in our institute between July 2013 and February 2014. All of the patients were <12 months old when they had the operation. Patients were randomly assigned to 2 arms: modified reverse subclavian flap angioplasty (MRA group; n = 27) or extended end-to-end anastomosis (EEA group; n = 27). We found a significant difference in residual arterial hypertension between the groups: 2 (7.7%) patients in the MRA group and 8 (30.8%) patients in the EEA group, respectively (P = 0.03). Risk factors for arterial hypertension were precoarctation area rigidity and endocardial fibroelastosis. During the last follow-up visit, recoarctation was observed in 1 (3.8%) patient in the MRA group and in 2 (7.7%) patients in the EEA group, respectively (P = 0.50). Cox regression analysis showed that the only risk factor for recoarctation was the patient's low weight before surgery [odds ratio (95% confidence interval) 0.016 (0.001-0.51), P = 0.047]. An aortic aneurysm developed in 2 (7.7%) patients in the MRA group; however, no significant difference was found between the groups (P = 0.15). The results of surgical repair of coarctation of the aorta using modified reverse subclavian flap angioplasty and extended anastomosis were comparable in mortality rates and early postoperative complications. Nevertheless, MRA may be beneficial in reducing residual arterial hypertension at the mid-term follow-up visit.


Prasugrel Versus Ticagrelor in Patients with Acute Coronary Syndrome Undergoing Percutaneous Coronary Intervention: a Systematic Review and Meta-analysis of Randomized Trials.
Dual antiplatelet therapy (DAPT) with aspirin and ticagrelor or prasugrel is the mainstay of treatment for patients with acute coronary syndrome (ACS) undergoing percutaneous coronary intervention (PCI). We aimed to systematically perform a head-to-head comparison of ticagrelor vs prasugrel in terms of efficacy and safety. We searched PubMed/Medline, EMBASE and Cochrane Central Register of Controlled Trials (CENTRAL) for relevant published randomized controlled trials (RCTs). The primary outcome was adverse cardiovascular events and secondary outcome was bleeding events. A random-effects meta-analysis was used to obtain the pooled estimate of each outcome. Nine RCTs with a total number of 6990 patients (3550 treated with prasugrel and 3481 treated with ticagrelor) were included. No significant difference between prasugrel and ticagrelor was observed in terms of mortality (OR 0.86, 95% CI 0.66 to 1.13, P = 0.28), major adverse cardiovascular events (MACEs) (OR 0.85, 95% CI 0.70 to 1.03, P = 0.10), non-fatal myocardial infarction (OR 0.78, 95% CI 0.57 to 1.06, P = 0.11), stroke (OR 1.02, 95% CI 0.60 to 1.72, P = 0.95), stent thrombosis (OR 0.76, 95% CI 0.47 to 1.21, P = 0.25), thrombolysis in myocardial infarction (TIMI) defined major (OR 0.94, 95% CI 0.19 to 4.67, P = 0.94), minor (OR 0.35, 95% CI 0.08 to 1.62, P = 0.18) and minimal (OR 0.48, 95% CI 0.19 to 1.18, P = 0.11) bleeding and Bleeding Academic Research Consortium (BARC) defined bleeding (OR 1.06, 95% CI 0.82 to 1.36, P = 0.68). In patients with ACS undergoing PCI, both prasugrel and ticagrelor were associated with similar cardiovascular outcomes and adverse bleeding events.


HECTD3 promotes gastric cancer progression by mediating the polyubiquitination of c-MYC.
The E3 ubiquitin ligase HECTD3 is homologous with the E6 related protein carboxyl terminus, which plays a vital role in biological modification, including immunoreactivity, drug resistance and apoptosis. Current research indicates that HECTD3 promotes the malignant proliferation of multiple tumors and increases drug tolerance. Our study primarily explored the important function and effects of HECTD3 in gastric cancer. Here, we discovered that HECTD3 is abnormally activated in gastric cancer, and the clinical prognosis database suggested that HECTD3 was strongly expressed in gastric cancer. Depletion of HECTD3 restrained the proliferative and clone abilities of cells and induced the apoptosis of gastric cancer cells. Mechanistically, our findings revealed that interaction between HECTD3 and c-MYC, and that the DOC domain of HECTD3 interacted with the CP and bHLHZ domains of c-MYC. Furthermore, we discovered that HECTD3 mediates K29-linked polyubiquitination of c-MYC. Then, our research indicated that cysteine mutation at amino acid 823 (ubiquitinase active site) of HECTD3 reduces the polyubiquitination of c-MYC. Our experimental results reveal that HECTD3 facilitates the malignant proliferation of gastric cancer by mediating K29 site-linked polyubiquitination of c-MYC. HECTD3 might become a curative marker.


Lowering the legal alcohol limit in Belgium: Potential effects on the number of traffic victims.
Since 1994, the legal limit of Blood Alcohol Concentration (BAC) is 0.5 g/L for the general drivers' population in Belgium. Since 2015, this limit has been lowered to 0.2 g/L for professional drivers. So far, no specific limitation has been adopted for novice drivers in Belgium. Recently, two bills were submitted to the House of Representatives: the first one proposed to impose a zero-limit for every driver, the second one proposed to restrict this zero-limit only to novice drivers. The present study evaluated the potential impact of the two bills that aim to reduce the legal BAC limit from 0.5 to zero, either for all drivers or for novice drivers only. We elaborated three scenarios related to the BAC categories for which lowering the legal BAC limit to zero would affect drink-driving behaviour: "Targeted BAC levels" scenario assumed that the new policy would impact only the specifically targeted BAC category, i.e., drivers in the category BAC below 0.5 g/L. "Extended impact" scenario corresponded to the "Targeted BAC levels" scenario to which we added a "halo effect" on drivers in the BAC category "0.5 g/L ≤ BAC < 0.8 g/L". "Broad range impact" scenario corresponded to the "Extended impact" scenario to where we extended the "halo effect" to drivers in the BAC category "0.8 g/L ≤ BAC < 1.2 g/L". The effect estimates were based on firstly, scientific literature on risks related to drink-driving at different BAC-levels, secondly data on crashes in Belgium and thirdly data on drink-driving behaviour in Belgium and in other European countries. In case of a zero limit for all drivers, an annual reduction can be expected of 10 to 17 fatalities (i.e., a decrease of 2.4% to 3.9%), 8 to 20 severe injuries (i.e., a decrease of 0.3% to 0.8%) and 135 to 315 slight injuries (i.e., a decrease of 0.4% to 0.8%). In case a zero limit is only applied to novice drivers, an annual decrease can be expected by 2 to 4 fatalities (i.e., a decline of 3.7% to 6.2%), 8 to 16 serious injuries and 135 to 262 slight injuries (i.e., a decline of 1.7% to 3.2% in both cases). There is no clear evidence on which of the three scenarios would be the most plausible. As the relative risk of a car crash increases strongly with the BAC level, the success of either measure will strongly depend on its ability to also affect drink driving at concentrations that are forbidden already. This also means that most of the casualties could be prevented if compliance with current rules increased.


Infiltration of Leu-3a+3b-positive lymphocytes and expression of human leukocyte antigen DR on tumor tissue induced by radiation therapy.
Tumor infiltrating lymphocytes induced by radiotherapy were analyzed by the biotin-avidin-horseradish peroxidase method. At the early dose of 4 Gy (400 rad) of irradiation (2 Gy X 2), remarkable infiltration of lymphocytes was observed surrounding cancer cells. This infiltration was mainly composed of Leu-3a+3b-positive lymphocytes, and human leukocyte antigen DR-positive tumor and interstitial cells were remarkably observed.


Mapping out hippocampal inhibition.
A synthesis of opto- and pharmacogenetics is beginning to reveal how various interneuron types direct incoming traffic in the hippocampus to help create a map of the environment.


Terbium chelates for fluorescence immunoassays.
The sensitization of terbium(III) ion luminescence in the presence of 1-ethyl-14-dihydro-6,7-methylenedioxy-4-oxoquinoline-3-carboxylic (oxolinic) acid was studied. The terbium label is bound to the antibody with diethylenetriamine-pentaacetic acid anhydride (DTPAA). Optimum luminescence intensity is observed at pH 7.5 and the luminescence significantly increases in the presence of cationic surfactant, cetyltrimethylammonium bromide. The sensitivity of Tb(III) detection is 5 x 10(14) mol l(-1). This luminescence system is proposed for time-resolved fluoroimmunoassay.


POD promoted oxidative gelation of water-extractable arabinoxylan through ferulic acid dimers. Evidence for its negative effect on malt filterability.
As a major component of non-starch polysaccharide in barley, arabinoxylan (AX) plays an important role in quality traits of malt and the final beer product. The Chinese barley malt has encountered filterability problems for a long time. The main reason caused by barley cultivar has been accepted in the malting and brewing industries. In our previous proteomic study, the peroxidase (POD) BP1 was found to be in quite high abundant in the filterability defect Chinese barley malt. Therefore, the present study tried to verify its negative effect on filterability, by surveying its activity in different malt samples and detecting effects of POD on AX gelation and filterability. The results showed that the activity of POD, as well as the content of AX bounded ferulic acid, were both negatively correlated with filterability, while the feruloyl esterase activity was positively correlated with it. In addition, AX gelation catalyzed by POD caused worse filterability, and the natural inhibitor of POD, vitamin C, could blocked the cross linking catalyzed by POD and thus improve the filterability. These results all suggested the great negative effect of POD on malt filterability.


The compensatory effect of exercise on physical activity and energy intake in young men with overweight: The EFECT randomised controlled trial.
The compensatory effect of exercise on total volume of physical activity and food intake has been described as a possible explanation for the limited body weight loss observed during exercise interventions. To investigate the effect of different exercise intensities on total volume of physical activity and energy intake amongst active men with overweight. Young men with overweight from a naval academy (n = 72; mean ± SD, age 21 ± 2 years, BMI 27.9 ± 2.13 kg/m2) were randomised to a control group (CG), moderate-intensity (MEG), or vigorous-intensity exercise group (VEG). MEG and VEG performed exercise sessions three times per week, for 60 min, during a 2-week period. Physical activity was assessed using triaxial accelerometers for 13 days. Energy intake was assessed at four time-points by 24-hour food recall. Intention-to-treat analyses were performed using linear mixed effect models. MEG and VEG presented a greater compensatory effect in the total volume of physical activity over time compared to CG, with a significant difference in the rate of change between VEG and CG (∆ = -250,503 counts vs. ∆ = -61,306 counts, respectively; p = 0.01), and MEG and CG (∆ = -253,336 counts vs. ∆ = -61,306 counts, respectively; p = 0.01). There was no difference between MEG and VEG (p = 0.97). Changes in energy intake were not different between groups (p = 0.18); however, MEG presented greater energy intake compared to CG (β=491 kcal/day; p = 0.01) and VEG (β=319 kcal/day; p = 0.07). VEG presented a greater reduction in body weight compared to MEG (-1.3 kg vs. -0.4 kg; p = 0.03) and CG (-1.3 kg vs. -0.6 kg; p = 0.07). Two weeks of exercise promoted a compensatory effect in total volume of physical activity in active men with overweight, regardless of exercise intensity. The compensatory effect was not observed for energy intake, although there was a trend for higher absolute energy intake in the MEG. Consequently, individuals in the VEG showed greater reduction in body weight over the intervention period.


Therapeutic effect of repeated natural killer T cell stimulation in mouse cholangitis complicated by colitis.
Primary sclerosing cholangitis is often complicated by ulcerative colitis. Recently, we reported on Th1-dominant cholangitis associated with experimental colitis, and natural killer T (NKT) cells might play an important role in this model. The aim of this study was to clarify the immunopathogenic role of NKT cells in this model using alpha-galactosylceramide. CD-1 mice were administered 2.0% dextran sulfate sodium for 29 days and injection of alpha-galactosylceramide was performed every 5 days, then inflammation was assessed. Mononuclear cells from the liver were analyzed with respect to cytokine production and the surface marker. alpha-Galactosylceramide improved survival rate, weight gain, and inflammation score. Also, interferon-gamma release from MNC, CD4/CD8 ratio, NKT cell population, and NK cell population were decreased by this treatment. These findings indicate that repeated stimulation of NKT cells modifies the Th1/Th2 balance to reduce Th1 dominance, and this may be a mechanism by which alpha-galactosylceramide has a therapeutic effect.


GM crops dancing to different tunes.
MEDIAWATCH: Parts of the British media are keeping up a campaign against the introduction of genetically modified crops ahead of the governemnet's assessment of the trial programme this autumn, often to the detriment of the arguments.


Evaluation of efforts to reduce sodium and ensure access to healthier beverages in four healthcare settings in Massachusetts, US 2016-2018.
Considerable science links diets lower in sodium and sugar-sweetened beverage consumption with better health outcomes. This study describes the evaluation process and outcomes of intervention strategies to reduce sodium in foods and sugar in beverages as part of a collaborative partnership between state public health, academic, community, and healthcare partners in Massachusetts, US. This quasi-experimental, pre-post study used nutrient data linked to observations of foods and beverages available in cafeterias and vending machines in four community healthcare settings to inform intervention strategies and evaluate changes. At post-assessment, beverages with no or very low sugar were significantly more prevalent in vending machines (OR = 1.93, p < 0.001) and cafeterias (OR = 1.83, p = 0.01) and low-sodium packaged foods were significantly more prevalent in cafeterias (OR = 2.45, p < 0.001), but not vending machines. These types of partnerships and tailored feedback and technical assistance strategies may support healthier food and beverage options within healthcare settings that serve patients, their families, and employees each day.


Phase I Study of High-Dose L-methylfolate in Combination with Temozolomide and Bevacizumab in Recurrent IDH wild-type High-Grade Glioma.
IDH mutations in low-grade gliomas (LGGs) results in improved survival and DNA hypermethylation compared to IDH wild-type LGGs. IDH-mutant LGGs become hypomethylated during progression. It's uncertain if methylation changes occur during IDH wild-type GBM progression and if the methylome can be reprogrammed. This phase I study evaluated the safety, tolerability, efficacy and methylome changes after L-methylfolate (LMF) treatment, in combination with temozolomide and bevacizumab in patients with recurrent high-grade glioma. Fourteen patients total, 13 with GBM, one with anaplastic astrocytoma, all IDH wild-type were enrolled in the study. All patients received LMF at either 15, 30, 60, or 90 mg daily plus temozolomide (75mg/m2 5 days per month) and bevacizumab (10mg/kg every two weeks). No MTD was identified. LMF treated had mOS of 9.5 months (95% CI, 9.1-35.4) comparable to bevacizumab historical control 8.6 months (95% CI, 6.8-10.8). Six patients treated with LMF survived more than 650 days. Across all treatment doses the most adverse events were diarrhea (7%, 1 patient, grade 2), reflux (7%, 1 patient, grade 2), and dysgeusia (7%, 1 patient, grade 2). In the six brains donated at death, there was a 25% increase in DNA methylated CpGs compared to the paired initial tumor. LMF in combination with temozolomide and bevacizumab was well tolerated in patients with recurrent IDH wild-type high-grade glioma. This small study did not establish a superior efficacy with addition of LMF compared to standard bevacizumab therapy, however, this study did show methylome reprogramming in high-grade glioma.


Effects of individual progressive single- and dual-task training on gait and cognition among older healthy adults: a randomized-controlled comparison study.
Dual-task training has beneficial effects on older individuals for gait and cognition. This study was aimed to make a comparison between the effects of individual progressive single- and dual-task training on gait and cognition among healthy older individuals. A total of 32 participants were divided randomly into two groups as the single-task group (n = 16, 64.6 ± 3.3 years, 7 males and 9 females) and dual-task group (n = 16, 65.6 ± 2.6 years, 8 males and 8 females). The 10-m walk test with the LEGSys device was used to assess spatio-temporal gait parameters. The cognitive parameters were evaluated using the Standardized Mini-Mental State Exam and Stroop Test. An individual progressive 60 min single- and dual-task training programs were applied twice per week for a period of 6 weeks. There were significant differences for both gait and cognition variables in the dual-task training group (p < 0.05), according to the comparison of pre- and post-treatment results. In the single-task training group, there were significant differences only in gait parameters with single-task conditions (p < 0.05). The comparisons of the delta values between the groups indicated that the dual-task training group was better compared to the single-task training group in gait speed, cadence, and many cognitive variables (p < 0.05). Individual progressive dual-task training is an effective and useful method that improves gait performance and cognitive skills among older individuals. NCT03777111, 12/13/2018.


Fatalities in Swedish skydiving.
Exact risk patterns in skydiving fatalities are not well known, but incomplete world injury data indicate that many are preventable. A comprehensive national material for Sweden of 37 skydiving fatalities 1964-2003 were reviewed to identify risk factors. In relation to jump volume, the period 1994-2003 had a fatality rate 11 times lower than 1964-1973. Student skydivers had the highest risk of fatal outcome, often caused by instability in freefall leading to unstable parachute activation with subsequent line entanglement, or parachute activation failure. Unintentional water landings also contributed to student fatality, with life jacket malfunctions, neglect to use life jackets, and automatic reserve parachute activation devices activated by water as aggravating factors. One-third of all fatalities had an inflated and operational parachute at some point prior to injury. A drastic worldwide increase in fatal landing incidents with fast wing parachutes during the 1990s did not occur in Sweden. Every fourth fatality caused by rapid deceleration against ground or water survived impact and died during transports or in hospitals. Rescue units and health care providers can improve management of skydiving incidents from knowledge about the incident and injury mechanisms we have described, and the skydiving community can target risk factors in preventive safety work.


Molecular design and synthesis of 1,4-disubstituted piperazines as α(1)-adrenergic receptor blockers.
A new series of 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-3-carboxylic acid amide and 3,5,6,8-tetrahydropyrido[4',3':4,5]thieno[2,3-d]pyrimidin-4-one derivatives were designed, synthesized, their binding and functional properties as α1-adrenoreceptors blockers were evaluated. A new validated α1-adrenoreceptor blocker pharmacophore model (hypothesis) was generated using Discovery Studio 2.5. The compare-fit study for the designed molecules with the generated hypothesis was fulfilled and several compounds showed significant high fit values. Compounds IVa-c, VIIa-d, VIIIa-c, Xa-c, XIa-d have shown blocking activity ranging from 46.73% up to 94.74% compared to 99.17% for prazosin.


Ecotoxicity of naproxen and its phototransformation products.
The occurrence of pharmaceuticals in the environment is of great concern and only few data are available about the adverse effects of such molecules and their derivatives on non-target aquatic organisms. This study was designed to assess the toxic potential of Naproxen, a nonsteroidal anti-inflammatory, Naproxen Na, its freely water soluble sodium salt and their photoproducts in the aquatic environment. Bioassays were performed on algae, rotifers and microcrustaceans to assess acute and chronic toxicity. Furthermore, possible genotoxic effects of photoderivatives were investigated using SOS chromotest and Ames fluctuation test. The results showed that photoproducts were more toxic than the parent compounds both for acute and chronic values, while genotoxic and mutagenic effects were not found. These findings suggested the opportunity to consider derivatives in ecotoxicology assessment of drugs.


Neural control of a cyclic postural behavior in the crayfish, Procambarus clarkii: the pattern-initiating interneurons.
As part of its repertoire of defensive behaviors, the crayfish, Procambarus clarkii, may respond to mildly threatening tactile or visual stimuli from the front of its body by walking backwards. During this behavior, the abdomen undergoes complex cyclical movements involving flexion and extension of the postural musculature which cause the tail to alternately contact and withdraw from the substrate. Intracellular neuropil recordings and dye injections were used to search for the interneurons responsible for initiating this postural motor pattern in the crayfish abdomen. Several diverse morphological types of interganglionic pattern-initiating (PI) interneurons were found. Each interneuron, when driven intracellularly, was capable of eliciting the same motor program, in its entirety, throughout the abdominal nerve cord. During pattern generation, PI interneurons exhibited a burst of spikes preceding the motor output. Silencing single PI interneurons with hyperpolarizing current during pattern generation failed to affect the motor program, indicating a redundancy of pattern-initiating function. The observations of extensive dye-coupling with other parallel axons, consistent dye-coupling with other identified cells in the pattern-initiating system, and the presence of multiple spike amplitudes in the bursts suggested electrotonic coupling among the PI interneurons. An additional group of interganglionic interneurons, the partial pattern-initiating (PPI) interneurons, were found to comprise a significant subset of the pattern-initiating system. As with the PI cells, the PPI interneurons exhibited a complex burst of spikes just preceding the patterned motor program. However, the PPI interneurons were only capable of eliciting an incomplete, though recognizable, postural motor pattern. Silencing any PPI interneuron during pattern generation caused a deficit in the motor pattern, indicating either an absence or lesser degree of functional redundancy within the PPI interneuron population compared to that occurring within the PI interneuron group. We conclude that a large number of PI interneurons are presynaptic to a relatively small group of PPI interneurons which, in turn, conduct pattern-initiating signals to the ganglionic oscillators. Our results indicate that pattern-initiation is accomplished through a command system involving multiple command elements organized in a coordinated interganglionic network.


Multi-Centered Field Evaluation of a Salmonella spp. Point-of-Care PCR Assay Using Equine Feces and Environmental Samples.
The introduction of microfluidic card technology has opened the field for rapid point-of-care (POC) molecular assays, including fecal and environmental Salmonella spp. testing. The purpose of this study was to evaluate a novel POC PCR assay for the detection of Salmonella spp. in feces and environmental samples. A total of 143 fecal samples and 132 environmental samples were collected for POC PCR Salmonella spp. testing as well as qPCR testing. Each sample was inoculated into selenite broth and incubated for 18 to 24 hours. For the POC PCR assay, 14 μl of selenite broth were mixed with 126 μl of PCR reaction mix and pipetted into a microfluidic test card targeting the invA and ttrC gene of Salmonella enterica. For qPCR analysis, 200 µl of the selenite broth were processed for DNA purification and Salmonella spp. testing targeting the invA gene. The overall agreement between the POC PCR Salmonella spp. assay and qPCR assay was 88.1% for feces and 97.0% for environmental samples. Strong agreement and short turn-around-time make the POC device the first molecular diagnostic platform allowing detection of Salmonella spp. in a hospital setting without having to ship out samples to a veterinary diagnostic laboratory. The availability of an accurate POC PCR assay for the detection of Salmonella spp. will enhance the diagnostic capability of equine veterinarians to timely support a diagnosis of salmonellosis and also monitor the environment in order to reduce the risk of nosocomial infections.


Diabetes-induced changes of nitric oxide influence on the cardiovascular action of secretin.
The modulation by condition of the lack or the excess of nitric oxide (NO) on cardiovascular action of secretin in diabetic rats was investigated. In vitro the isolated heart function and in vivo, the systolic (SBP), diastolic (DSP) blood pressure and heart rate (HR) were measured. Secretin evoked inotropic positive effect and increased coronary outflow (CO), in vivo did not increase systemic pressure and the highest dose of the peptide increased the heart rate. NO synthase inhibitor, N(G) nitro-L-arginine methyl ester (L-NAME) deeply increased the systemic pressure and in vitro decreased coronary outflow. L-arginine and sodium nitroprusside (SNP) did not influence the isolated heart function and in vivo decreased the systemic pressure. L-NAME preserved the inotropic positive effect of secretin and the increase of the coronary outflow. In vivo co-administration of L-NAME+secretin evoked hypotensive effect and abolished the increase of the heart rate after the highest dose of the peptide. L-arginine abolished inotropic positive effect of the peptide and the increase of coronary outflow. In vivo co-administration of these substances caused hypotension and attenuated the increase of the heart rate after the highest dose of secretin. Co-injection of SNP and secretin preserved the inotropic effect of secretin and abolished the increase of the coronary outflow. In vivo infusion of SNP+secretin evoked hypotension and similarly to L-arginine, SNP abolished tachycardia induced by the highest dose of secretin. Both the lack and the excess of nitric oxide changed the cardiovascular action of secretin in diabetic rats.


Effect of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis on solid spinocellular carcinoma.
Phosphatidylinositol-specific phospholipase C (PIPLC) from Bacillus thuringiensis (0.11 units per ml) inhibited growth of IC-Sofia carcinoma cell line in culture by 64%. The growth of transplanted carcinoma in hamsters was also inhibited by 29% at a dose of 4.9 units of PIPLC/kg animal/day. In addition, degeneration processes and a significant fibrosis of the tumour occurred. The functional, clinical, biochemical and haematological parameters studied were consistent with the established antitumour effect in vivo.


Functional properties of an endothelial cell cofactor for thrombin-catalyzed activation of protein C.
Thrombin-catalyzed activation of Protein C is accelerated by a human endothelial cell surface cofactor. The cofactor occurs also on mouse hemangioma cells (a transformed endothelial cell line), but not on cultured human smooth muscle cells or fibroblasts. The cofactor remains bound to the cell surface during Protein C activation. The cofactor is saturable with respect to both Protein C (Km = 0.72 +/- 0.07 microM) and thrombin (Km = 0.48 +/- 0.05 nM). Diisopropylphosphoryl-thrombin is a competitive inhibitor of the cofactor-dependent reaction with Ki = 0.56 +/- 0.1 nM. Prothrombin Fragment 1, the peptide derived from prothrombin that retains phospholipid binding capacity, does not inhibit activation of Protein C when present in a 7:1 molar excess over Protein C. Platelet Factor 4 (20 microgram/ml) also fails to inhibit Protein C activation. It is concluded that the endothelial cell provides a surface on which Protein C can be activated under physiological conditions.


Ligand binding by murine IgM antibodies: intramolecular heterogeneity exists in certain, but not all, cases.
The ligand binding properties of eight hybridoma-derived murine anti-DNP IgM(kappa) antibodies were analysed by equilibrium dialysis. Four of these proteins exhibited the expected valances of approximately 10 and relatively low affinities (less than or equal to 2.2 x 10(5) M-1). The remaining four proteins exhibited valences of considerably less than 10 (less than or equal to 8) and relatively high affinities (greater than 10(6) M-1). When these proteins were subjected to two cycles of lyophilization, those of the former group were observed to still exhibit approximately 10 sites per molecule with homogeneous affinities similar to those of the respective untreated molecules. However, molecules in the latter group (valences of less than or equal to 8) were observed to exhibit only five to six binding sites subsequent to lyophilization with no changes in affinities. When the reductive subunits from each of the IgM(kappa) proteins were subjected to trypsinization, two different patterns were observed in terms of the yields of Fab mu fragments. Each of the proteins originally exhibiting approximately 10 binding sites yielded greater than 90% of the expected Fab mu fragments. In contrast each of the proteins exhibiting less than or equal to 8 binding sites yielded only approximately 50% of the expected Fab mu fragments. Collectively these results indicate the existence of at least two different forms of murine IgM molecules, those with approximately 10 homogeneous, relatively stable sites and those with only approx. five stable sites. It is suggested that these intramolecular functional differences may be attributable to intramolecular conformational differences.


Dendritic cells as accessory cells in antigen-specific murine T lymphocyte proliferation in vitro.
The role of dendritic cells in antigen-induced murine T lymphocyte activation was studied by addition of purified dendritic cells to purified lymph node T lymphocytes from ovalbumin-primed mice. In the presence of the priming antigen T cells generated an antigen-specific response. The response was at least 3-fold higher with the use of a modified IMDM culture medium. The complete requirement for accessory cells was demonstrated only when nylon wool-purified T lymphocytes were thoroughly depleted of Ia antigen-expressing cells. Dendritic cells as well as peritoneal exudate macrophages were equally effective as antigen-presenting cells.


Periodic potentials in hybrid van der Waals heterostructures formed by supramolecular lattices on graphene.
The rise of 2D materials made it possible to form heterostructures held together by weak interplanar van der Waals interactions. Within such van der Waals heterostructures, the occurrence of 2D periodic potentials significantly modifies the electronic structure of single sheets within the stack, therefore modulating the material properties. However, these periodic potentials are determined by the mechanical alignment of adjacent 2D materials, which is cumbersome and time-consuming. Here we show that programmable 1D periodic potentials extending over areas exceeding 104 nm2 and stable at ambient conditions arise when graphene is covered by a self-assembled supramolecular lattice. The amplitude and sign of the potential can be modified without altering its periodicity by employing photoreactive molecules or their reaction products. In this regard, the supramolecular lattice/graphene bilayer represents the hybrid analogue of fully inorganic van der Waals heterostructures, highlighting the rich prospects that molecular design offers to create ad hoc materials.


The effect of bleomycin on DNA in Escherichia coli K12 cells.
In Escherichia coli cells treated to reduce colony-forming ability to about 10%, bleomycin causes fewer than six randomly located DNA single-strand breaks or three double-strand breaks per genome. This is many fewer than produced by strand-breaking agents such as ionizing radiations in cells with similar loss of colony-forming ability. Bleomycin treatment to this level of colony-forming ability does affect the intracellular DNA, as shown by a change in the sedimentation rate of the chromosomal structure found in lysates made with sodium dodecyl sulfate. Bleomycin may act on only a limited part of the chromosome of such cells, perhaps the part associated with the outer cell membrane, or it may make strand breaks that are less repairable than those formed by ionizing radiations. Extensive DNA degradation in heavily treated cells (colony-forming ability 1% or less) could be from the action on DNA of bleomycin entering freely through membranes which are no longer intact, or from enzymatic degradation in heavily damaged cells.


Occurrence, use and potential toxic effects of metals and metal compounds.
Metals and metal compounds are constituents of our natural environment. Their distribution depends on the existence of natural sources (e.g. volcanoes or erosion) and their use in human's activity. They are transformed naturally (e.g. by bacterial activity) with formation of organic species that influence their mobility and accumulation in abiotic as well as biotic systems. Up to date metal species are released into the environment questioning their influence on human health. Due to their widespread use in human activities such as industry, agriculture and even as medicine (e.g. As, Se, Pt), numerous health risks may be associated with exposure to these substances. Different reports on metal intoxication are documented and studies especially on neurotoxicity, genotoxicity, or carcinogenicity, are previously published in numerous articles. This mini-review gives an overview on the use and the actions of selected metal species of actual scientific concern, with a focus on neuronal cells.


Time-to-event analysis with treatment arm selection at interim.
This paper discusses the application of an adaptive design for treatment arm selection in an oncology trial, with survival as the primary endpoint and disease progression as a key secondary endpoint. We carried out treatment arm selection at an interim analysis by using Bayesian predictive power combining evidence from the two endpoints. At the final analysis, we carried out a frequentist statistical test of efficacy on the survival endpoint. We investigated several approaches (Bonferroni approach, 'Dunnett-like' approach, a conditional error function approach and a combination p-value approach) with respect to their power and the precise conditions under which type I error control is attained.


The use of Fourier-transform infrared spectroscopy for the quantitative determination of glucose concentration in whole blood.
Fourier-transform infrared transmission spectroscopy has been used for the determination of glucose concentration in whole blood samples from 28 patients. A 4-vector partial least-squares calibration model, using the spectral range 950-1200 cm(-1), yielded a standard-error-of-prediction of 0.59 mM for an independent test set. For blood samples from a single patient, we found that the glucose concentration was proportional to the difference between the values of the second derivative spectrum at 1082 cm(-1) and 1093 cm(-1). This indicates that spectroscopy at these two specific wavenumbers alone could be used to determine the glucose concentration in blood plasma samples from a single patient, with a prediction error of 0.95 mM.


Mycelial growth and exo-biopolymer production by submerged culture of various edible mushrooms under different media.
The effect of synthetic media on the submerged mycelial growth and exo-biopolymer production in various edible mushrooms was investigated in shake flask culture. Among 19 mushrooms examined, the relatively high yield in mycelial biomass and exo-biopolymer production was achieved in potato malt peptone (PMP) medium. In particular, Ganoderma lucidum NO. 1 and Phellinus linteus KCTC 6190 showed favourable growth in PMP medium with exo-biopolymer concentration of 1170 and 1520 mg l(-1), respectively. Enhanced exo-biopolymer production was achieved from Ganoderma lucidum NO. 1 and Phellinus linteus KCTC 6190 in a 5L batch fermentor, indicating approximately 5000 and 2410 mg l(-1), respectively. The exo-biopolymer production and mycelial growth from various mushrooms were found to be strongly controlled by different complex media.


Postauricular incision versus conventional transcervical incision in submandibular gland excision: A systematic review and meta-analysis.
Surgical removal of the submandibular gland is indicated for different conditions affecting the gland, such as neoplasm, sialadenitis, and sialolithiasis, and different types of surgical approaches have been reported. The purpose of this meta-analysis was to evaluate the differences between the postauricular approach and conventional transcervical approach in submandibular gland excision. A systematic review was performed using PubMed, Embase and the Cochrane Library to identify studies comparing outcomes of submandibular gland surgery via the postauricular approach and conventional transcervical approach. The data of interest were analyzed with Comprehensive Meta-Analysis software (version 3; Biostat, Englewood, NJ). Dichotomous data and continuous data were analyzed by calculating the risk difference (RD) and the mean difference (MD) with the 95% confidence interval (CI), respectively. The results show that the postauricular incision is a feasible approach to access the submandibular gland, and compared with the conventional transcervical approach, it requires a longer operative duration but has a similar complication rate and yields better cosmetic outcomes.


Clinical anatomy of the retrocalcaneal bursa.
The goal of the study was to perform a detailed anatomical description of the retrocalcaneal bursa (RB). Its morphological arrangement was studied on 10 fresh and 30 embalmed lower extremities by microdissection and light microscopy. The RB was present constantly and in all the cases contained 1-2 cm long synovial fold, beginning on the upper wall of RB and distally interposed between the anterior surface of the Achilles tendon and the posterior surface of the calcaneal tuberosity. The volume of RB was 1-1.5 ml. The histological analysis confirmed that the inner surface of the superior and posterior wall of RB have been covered by unilayered synovial membrane, projecting into synovial villi of different shapes and sizes. In the ceiling of RB, delicate fascicle of skeletal muscle fibers was discovered, radiating distally into the regularly present synovial fold. The whole bottom of RB has been covered by 200-500 microm layer of fibrous cartilage into which the calcaneal tendon attached. The cartilagineous layer continued anteroproximally to cover the whole bursal surface of the calcaneal tuberosity, where the thickness of the cortical bone was reduced on mere 50 microm. The obtained results can be used in the improvement of the differential diagnostics and therapy of diagnostics and therapy of the retrocalcaneal bursitis as well as of other kinds of achillar enthesopathies and heel pain.


A case of abdominal CSF pseudocyst associated with silicone allergy.
The authors present a case of a patient with an abdominal CSF pseudocyst that resulted from an allergic reaction to silicone. The patient underwent repair surgery of the meningomyelocele associated with the Chiari II malformation, and the V-P shunt was instituted at 6 months of age. A formation of the abdominal CSF pseudocyst and the consequent shunt malfunction were observed 40 days after the V-P shunt. An increase in the number of the peripheral eosinophils and serum immunoglobulin E (IgE), and an infiltration of eosinophils in the specimen harvested from the pseudocyst wall suggested an allergic reaction as the cause of the pseudocyst. A sixth operation to revise the V-P shunt was performed using the shunt system made of "extracted silicone", which was produced extracting the allergic substances. The serum IgE was normalized after surgery and the abdominal CSF pseudocyst has not recurred for 22 months.


Alterations to N-linked oligosaccharides which affect intracellular transport rates and regulated secretion but not sorting of lysosomal acid phosphatase in Dictyostelium discoideum.
The importance of N-linked oligosaccharides and their associated modifications in the transport, sorting, and secretion of lysosomal acid phosphatase was investigated using three mutant Dictyostelium cell lines. These mutants synthesize altered N-linked oligosaccharides with the following properties: (i) in strain HL244 carbohydrate side chains lack mannose 6-sulfate residues, (ii) in strain M31 the side chains retain the two alpha-1,3-linked glucose residues resulting in less sulfate and methylphosphate modifications, and (iii) in strain HL243 the nonglucosylated branches are missing three of the outer mannose sugars and the oligosaccharides contain fewer sulfate and phosphate modifications. Lysosomal enzymes in both HL243 and HL244 are also missing a shared epitope termed common antigen-1 (CA-1), which consists in part of mannose 6-sulfate moieties. No increases were observed in the secretion of radiolabeled acid phosphatase or acid phosphatase activity during growth in any of the mutant cell lines, suggesting that the enzyme was correctly sorted to lysosomes. In support of this, Percoll gradient fractionations and indirect immunofluorescence microscopy indicated that acid phosphatase was transported to lysosomes in all cell lines. However, radiolabel pulse chase protocols indicated that newly synthesized acid phosphatase was transported out of the endoplasmic reticulum (ER) and into lysosomes at a two- to threefold slower rate in HL243 and at a sixfold slower rate in M31. The rate of transport of acid phosphatase from the ER to the Golgi was reduced only twofold in M31 as determined by digestion of newly synthesized enzyme with endoglycosidose H. This suggests that certain alterations in carbohydrate structure may only slightly affect transport of the enzyme from the ER to the Golgi but these alterations may greatly delay transport from the Golgi or post-Golgi compartments to lysosomes. Finally all three mutants secreted acid phosphatase at significantly lower rates than the wild-type strain when growing cells were placed in a buffered salt solution (conditions which stimulate the secretion of mature lysosomally localized enzymes). In contrast, alpha-mannosidase was secreted with similar kinetics from the mutant and wild-type strains. Together, these results suggest that the mechanism(s) operating to sort acid phosphatase in Dictyostelium can tolerate a wide range of changes in N-linked oligosaccharides including a reduction in phosphate and the absence of CA-1 and sulfate, while in contrast, these same alterations can profoundly influence the rate of transport of acid phosphatase from the ER and post-ER compartments to lysosomes as well as the secr


Cardiovascular remodeling as a result of fibroblast growth factor-23 (FGF-23)/Klotho imbalance in patients with CKD.
In chronic kidney disease (CKD) cardiovascular remodeling (CVR) is very frequent compared with general population and, as suppose, may be associated with «new» renal risk factors. The aim of study was to estimate association of new serum biomarkers (FGF-23, Klotho) and traditional biomarker of cardiac damage-serum Troponin I (sTr-I) with signs of CVR. One hundred thirty CKD G1-5D patients without cardiovascular disease (CVD) clinical manifestation were included. We measured serum FGF-23, Klotho and sTr-I. The instrumental methods were: echocardiography, SphygmoCor test [Pulse Wave Velocity (PWV), Central (aortic) Blood Pressure (CBP), Subendocardial Blood Supply (SBS)]. FGF-23 level correlated with: sTr-I (r = 0.512; p < 0.01), eccentric left ventricular hypertrophy, LVH (r = 0.543; p < 0.01), SBS (r = - 0.499; p < 0.05). There were no differences of FGF-23 level in patients with normal and high CBP. Klotho correlated with concentric LVH (r = - 0.451; p < 0.01), PWV (r = - 0.667; p < 0.001), Cardiac Calcification Score, CCS (r = - 0.581; p < 0.01). Multivariate analysis revealed positive independent association of FGF-23 with eccentric LVH (OR = 1.036, 95% CI (1.004-1.068); p = 0.038). Klotho was a negative determinant for concentric LVH (OR = 0.990, 95% CI 0.987-0.994; p < 0.001), increased PWV (OR = 0.984, 95% CI (0.977-0.991); p < 0.001) and CCS (OR = 0.991, 95% CI (0.988-0.995); p < 0.001). In addition, multivariate analysis revealed a relationship between serum Klotho (OR = 0.980, 95% CI (0.964-0.996); p = 0.016), FGF-23 (OR = 3.145, 95% CI (1.020-9.695); p = 0.046) and troponin I level. In CKD patients without CVD clinical manifestation increased serum FGF-23 level and decreased Klotho are associated with CVR: FGF-23 with eccentric LVH (independently of CBP), Klotho determinate concentric LVH, PWV and CCS. Moderately elevated sTr-I levels may be a manifestation of FGF-23/Klotho imbalance in CKD.


Microalbuminuria in non-diabetic patients with unstable angina/non ST-segment elevation myocardial infarction.
Microalbuminuria (MAU) is defined as an urinary albumin excretion rate between 20-200 mg/l or 30-300 mg/day. It is a surrogate marker for endothelial dysfunction and is independently associated with atherosclerotis in diabetic and in non-diabetic patients. We assessed the prevalence of MAU in non-diabetic patients who presented with UA/NSTEMI and the relation of MAU to the severity of coronary artery disease in patients at a cardiac care center in Iraq. Seventy non-diabetic patients referred to the Iraqi Center for Heart Disease, Baghdad, between November 1st 2010 and June 1st 2011 with the diagnosis of UA/NSTEMI were included in this study. Physical examination, ECG and echocardiography were performed on all patients. TIMI ("Thrombolysis in Myocardial Infarction") risk score was obtained. Urine samples were collected and sent for quantitative determination of MAU. All patients underwent diagnostic coronary angiography. Data are give as mean (quantitative and percent) ± SD. Fifty-three men (76%) and 17 (24%) women (mean age 56 ± 12 years) were investigated. Overall 37 (53%) individuals presented with arterial hypertension and 41 (59%) with a history of smoking. 58 patients (83%) had ischemic ECG changes (defined as ST segment depression more than 1 mm from baseline, and/or T wave inversion), 52 (74%) had echocardiographic findings indicative of ischemia (defined as segmental wall motion abnormalities). Twenty-one (30%) patients tested positive for MAU. There was a significant correlation of echocardiographic signs of ischemia and MAU, (n = 20 (38%), p < 0.01). There was a clear relationship between MAU and TIMI risk score. Additionally, MAU was more common in patients with multivessel coronary artery disease (CAD) (p < 0.001). There was no statistically significant correlation between MAU and mean age, sex, smoking, and blood pressure. In this analysis of patients with UA/NSTEMI we found a strong correlation of microalbuminuria with echocardiographic changes and findings in coronary angiography.


Multicenter study of the evolution of different types of avulsion over the 12 months after delivery.
To perform a multicenter study of muscle recovery in levator ani muscle (LAM) avulsion during the first 12 months postpartum according to the type of LAM avulsion suffered. This was a multicenter prospective observational study including 242 primiparas. Transperineal ultrasound was performed at 6 months and 12 months after delivery. Type I LAM avulsion was present when most of the lateral fibers of the pubovisceral muscle were observed at the muscle's insertion at the pubic level. Type II LAM avulsion was defined as complete detachment of the pubovisceral muscle from its insertion at the pubic level. Among the 56 patients who completed the study (with ultrasound at 6 and 12 months after delivery), 76 avulsions (10 cases of bilateral avulsion) were identified at 6 months after delivery, and the total number of avulsions had decreased to 58 at 12 months after delivery (P < 0.001; 95% confidence interval [CI] 13.9%-33.5%). This decrease was due to the disappearance of 69.2% of the cases of Type I LAM avulsions (P < 0.001; 95% CI: 50.2%-88.2%). However, the number of Type II LAM avulsions remained constant at 6 months and 12 months after delivery. The spontaneous resolution of LAM avulsion during the first 12 months postpartum occurs in cases of Type I LAM avulsion but is not observed in Type II LAM avulsion.


Adenosine A1 receptors contribute to immune regulation after neonatal hypoxic ischemic brain injury.
Neonatal brain hypoxic ischemia (HI) often results in long-term motor and cognitive impairments. Post-ischemic inflammation greatly effects outcome and adenosine receptor signaling modulates both HI and immune cell function. Here, we investigated the influence of adenosine A1 receptor deficiency (A1R(-/-)) on key immune cell populations in a neonatal brain HI model. Ten-day-old mice were subjected to HI. Functional outcome was assessed by open locomotion and beam walking test and infarction size evaluated. Flow cytometry was performed on brain-infiltrating cells, and semi-automated analysis of flow cytometric data was applied. A1R(-/-) mice displayed larger infarctions (+33%, p < 0.05) and performed worse in beam walking tests (44% more mistakes, p < 0.05) than wild-type (WT) mice. Myeloid cell activation after injury was enhanced in A1R(-/-) versus WT brains. Activated B lymphocytes expressing IL-10 infiltrated the brain after HI in WT, but were less activated and did not increase in relative frequency in A1R(-/-). Also, A1R(-/-) B lymphocytes expressed less IL-10 than their WT counterparts, the A1R antagonist DPCPX decreased IL-10 expression whereas the A1R agonist CPA increased it. CD4(+) T lymphocytes including FoxP3(+) T regulatory cells, were unaffected by genotype, whereas CD8(+) T lymphocyte responses were smaller in A1R(-/-) mice. Using PCA to characterize the immune profile, we could discriminate the A1R(-/-) and WT genotypes as well as sham operated from HI-subjected animals. We conclude that A1R signaling modulates IL-10 expression by immune cells, influences the activation of these cells in vivo, and affects outcome after HI.


Perceptions of Type 2 Diabetes Patients Towards Diabetes Management Visits at Public Primary Health Care Centers with Diverse Opinions Towards Nurse-led Clinics in Muscat, Oman: A Pilot Qualitative Study.
The literature has described several positive outcomes related to diabetes management via nurse-led clinics. This is especially true where a shortage of physicians is recorded within a team-based approach. We sought to explore the perceptions of patients with type 2 diabetes towards the current diabetes management visits at public primary health care centers in Muscat, Oman and their opinions towards nurse-led diabetes management clinics. This pilot qualitative study included seven semi-structured interviews with type 2 diabetes patients from four purposely selected public primary health care centers in Muscat. Qualitative thematic analysis was applied. Patients with type 2 diabetes expressed their satisfaction with the present diabetes management visits at public primary health care. Their opinions towards nurse-led clinic were diverse and divided patients into three categories: those who totally refused the nurse-led clinics, those who accepted the clinics but with reservations, and patients that totally accepted the nurse-led clinics. The patients' main concern was the nurses' abilities to handle and understand the disease and its management. Our pilot study revealed type 2 diabetes patients' satisfaction with the current diabetes management clinics. However, transformation to nurse-led clinics within team-based approach requires further studies with a bigger sample size. Further studies on requirements related to the Omani health care system and a better understanding of patients' worries and their readiness to accept the concept of nurse-led clinics and their outcomes are also recommended.


Thyroid peroxidase activity in euthyroid and mild hypothyroid patients with Hashimoto's thyroiditis.
Thyroid peroxidase (TPO) activity was measured spectrophotometrically according to Hosoya's guaiacol method. The mean TPO activity in ten patients with Hashimoto's thyroiditis was 19.8 +/- 7.6 (mean +/- SE) in an arbitrary unit, which was not significantly different from the normal value in seven normal thyroid tissues (33.7 +/- 5.4). The ten patients were divided into two groups, euthyroid and mild hypothyroid, on the basis of their basal serum TSH. In the euthyroid group, TPO activity (8.17 +/- 1.3) was significantly less than the normal tissue (p less than 0.01). In the hypothyroid group, TPO activity (27.64 +/- 13.8) was almost similar to the normal tissue. A positive correlation was obtained between TPO activity and serum TSH in ten patients with Hashimoto's thyroiditis (r = 0.85, p less than 0.01). It was concluded that TPO activity is significantly decreased in Hashimoto's thyroiditis even when the thyroid function was still within normal range, but the activity might be restored to normal in hypothyroid patients by the stimulation of elevated TSH.


Relative effects of mammal herbivory and plant spacing on seedling recruitment following fire and mining.
There is much debate concerning which ecological constraints are the most limiting factors to seedling recruitment in disturbed communities. We provide the first comparison between selective herbivory and plant competition effects among two post-mined forest ecosystems (primary succession) and one post-fire woodland ecosystem (secondary succession). Animal exclosure assessments of nine common species across eight sites were performed for comparison within three locations separated by up to 200 km. Additionally, we asked whether pre-browsed plants differed in nutrient content between or within species in the separate systems. Among the nine common species, seven of these were affected by mammal herbivory while five shared a similar vulnerability to predation regardless of system. One species was limited by competition (planting density). There was a strong linear correlation between herbivore selectivity (% browsed) and impact (biomass loss) on the fertilized minesites, but not post-fire sites. Phosphorus and potassium were higher for most species in the post-mined system. Principal components analyses revealed that nutrients in shortest supply may be the most likely components of selection within each system. Among all locations, species with highest levels of phosphorus, ADF and leaf water content were often favoured, while high tannins and nitrogen content were generally selected against. Herbivory, rather than seedling competition, was the limiting factor for plant performance among post-fire and post-mined reclamation areas. The post-fire seedlings were smaller and more water and nutrient limited, nevertheless browsing prevalence was equivalent at all locations with nearly all seedlings predated. Kangaroo density in the post-fire community declined from the beginning of the experiment, while numbers in the post-mined revegetation increased fourfold within one year. Differences in water and nutrient availability may explain why herbivores are more likely to be attracted to post-mined communities.


Feasibility of mFOLFOX6 as the adjuvant treatment after curative resection of metastases from colorectal cancer in Japanese patients.
An oxaliplatin-based regimen as the adjuvant treatment for stage III colon cancer demonstrated a survival advantage over fluorouracil (FU) and leucovorin (LV) in the MOSAIC and NSABP C-07 trials. For adjuvant treatment after the resection of metastases from colorectal cancer), active chemotherapy regimens such as FOLFOX are recommended. However, the safety data of FOLFOX are insufficient for its use after metastasectomy of colorectal cancer in Japanese patients. The aim of this study was to evaluate the safety of mFOLFOX6 for adjuvant treatment after the resection of metastases from colorectal cancer. Among 67 consecutive patients who received mFOLFOX6 as the adjuvant treatment after resection of metastases from colorectal cancer between September 2002 and March 2009 in our institution, 51 patients who had not received preoperative chemotherapy were reviewed. The mFOLFOX6 treatment comprised oxaliplatin 85 mg/m(2) and l-leucovorin 200 mg/m(2) given intravenously over a 2-h period on day 1, followed by a 5-FU bolus of 400 mg/m(2) and a 46-h infusion of 5-FU 2400 mg/m(2), every 2 weeks for up to 12 cycles. National Cancer Institute Common Terminology Criteria for Adverse Events version 3.0 (NCI-CTC) grade 3-4 toxicities per patient were: peripheral neuropathy 8%, allergic reaction 4%, aspartate transaminase (AST) 4%, febrile neutropenia 4%, nausea 2%, anorexia 2%, fatigue 2%, alanine transaminase (ALT) 2%, bilirubin 2%, neutrophils 49%, leukocytes 6%, and hemoglobin 2%; 71% of the patients completed the scheduled 12 cycles. Adjuvant therapy with mFOLFOX6 after resection of metastases from colorectal cancer is feasible for Japanese patients.


Assisted Reproductive Techniques and Risk of Congenital Heart Diseases in Children: a Systematic Review and Meta-analysis.
Infertility is a growing phenomenon and leads to an increased use of assisted reproductive techniques (ARTs). In recent years, concerns about the safety of these procedures emerged and ARTs were hypothesized to be a risk factor for developing congenital heart diseases (CHDs) in offspring. Our aim is to investigate the association between ART and CHD, specifying results according to various subtypes of defects. We performed a systematic review and random-effects meta-analysis following the PRISMA guidelines. MEDLINE and Google Scholar were searched from January 2011 to May 2022. Data about incidence of CHD in ART were tabulated and extracted from all the studies included. Twenty-four studies were included. Pooled incidence of CHDs after IVF pregnancies was 3% (95% CI 0.3-0.4; I2 = 99%), decreasing to 1% (95% CI 0.00-0.01; I2 = 93%) for major CHDs only. An increased risk of CHDs, especially minor (i.e., not requiring surgical correction), seems to occur in ART compared with non-ART pregnancies [RR 1.71 (95% CI 1.25-2.34; I2 = 99%)]. For major CHDs, not enough evidence is available to assess the real risk. Moreover, some confounding factors (i.e., maternal age and male infertility) seem to play a critical role to determine an increased risk of CHDs. Conflicting results emerged among the studies, setting the need for further research to validate the actual evidence and state the real risk of CHD following ART pregnancies.


Dispersive liquid‒liquid microextraction combined with enzyme-linked immunosorbent assay for the analysis of chlorpyrifos in cereal samples.
In this paper, an analysis method for chlorpyrifos (CPF) in cereal samples was proposed using dispersive liquid‒liquid microextraction combined with an enzyme-linked immunosorbent assay. In the dispersive liquid‒liquid microextraction, deep eutectic solvents and fatty acids were used as solvents to extract, purify, and concentrate CPF in cereals. In the enzyme-linked immunosorbent assay, gold nanoparticles were utilized to enrich and conjugate more antibodies and horseradish peroxidase, while magnetic beads were used as solid supports to amplify the signal and shorten the detection time of CPF. The linearity range was 0.002-1 μg kg-1, and the limit of detection was 0.0006 μg kg-1. The extraction recoveries were 86.7-99.9% with a relative standard deviation of less than 7.0%. The proposed method was successfully used to analyze CPF in cereal samples (rice, wheat, maize, and millet) and has prospects for the pretreatment and detection of CPF residues in other food samples.


Preoperative Decision to Perform Portal Vein Resection Improves Survival in Patients With Resectable Pancreatic Head Cancer Adjacent to Portal Vein.
We hypothesized that preoperatively planned portal vein resection (PVR), which prevents from approaching tumors, improves survival in patients with resectable pancreatic head cancer adjacent to the portal vein (PhC-PV). The decision to perform PVR is difficult in patients with resectable PhC-PV. This is a retrospective, bi-institutional study of patients undergoing pancreatoduodenectomy (PD) for resectable PhC-PV from 2009 to 2018. We compared clinical data of patients who underwent PD with preoperatively planned PVR (planned PVR group) and those who underwent conventional PD (cPD) in which decision to perform PVR was made intraoperatively (cPD group). Among the study population of 176 patients, 53 patients (30.1%) underwent PD with planned PVR. The remaining 123 patients (69.9%) underwent cPD. Tumor characteristics were similar between the 2 groups. Operation time and major complication rates did not differ between the 2 groups. The local recurrence rate of patients in the planned PVR group (28.3%) was lower than that of the cPD group (44.7%; P = 0.041). Median overall survival (OS) was longer in the planned PVR group than in the cPD group (32 vs 27 months; P = 0.011). Multivariate analysis revealed that having undergone planned PVR was an independent factor for favorable OS (hazard ratio = 1.65; 95% confidence interval = 1.08-2.61; P = 0.021). The preoperative decision to perform PVR improves survival by enhancing local control of resectable PhC-PV.


Histone deacetylase-mediated morphological transition in Candida albicans.
Candida albicans is the most common opportunistic fungal pathogen, which switches its morphology from single-cell yeast to filament through the various signaling pathways responding to diverse environmental cues. Various transcriptional factors such as Nrg1, Efg1, Brg1, Ssn6, and Tup1 are the key components of these signaling pathways. Since C. albicans can regulate its transcriptional gene expressions using common eukaryotic regulatory systems, its morphological transition by these signaling pathways could be linked to the epigenetic regulation by chromatin structure modifiers. Histone proteins, which are critical components of eukaryotic chromatin structure, can regulate the eukaryotic chromatin structure through their own modifications such as acetylation, methylation, phosphorylation and ubiquitylation. Recent studies revealed that various histone modifications, especially histone acetylation and deacetylation, participate in morphological transition of C. albicans collaborating with well-known transcription factors in the signaling pathways. Here, we review recent studies about chromatin-mediated morphological transition of C. albicans focusing on the interaction between transcription factors in the signaling pathways and histone deacetylases.


Giant cell tumor-like cholangiocarcinoma associated with systemic cholelithiasis.
A cholangiocarcinoma of the hepatic hilus with an element of giant cell tumor that occurred in a 59-year-old man is reported. His medical history included systemic cholelithiasis and repeated operations on the biliary passages. Four years after the last operation, which was a hepatic segmentectomy, he was readmitted because of persistent fever. A computed tomography scan showed a low-density area and stones in the hepatic hilus. He died of hepatic failure approximately 1 month later. At autopsy, a fist-sized tumor and gallstones were found at the hepatic hilus. Histologically, the tumor mainly showed sarcomatoid features and some tubular adenocarcinoma. An element of giant cell tumor consisting of many osteoclast-type giant cells also was noted. The results of immunohistochemical studies showed a positive reaction to cytokeratin and vimentin in some of the spindle-shaped sarcomatoid cells. Sarcomatoid bile duct carcinomas are rare, as are those with osteoclast-type giant cells. The authors also discuss the histogenesis of these giant cells.


Immunopathogenicity and oncogenicity of murine leukaemia virus. IV. Antinuclear antibody response and tumour induction in B10.A recombinant mice.
The levels of murine leukaemia virus (MuLV) proteins p30 and gp70, antinuclear antibody (ANA), anti-soluble nuclear protein, anti-single-stranded DNA, anti-double-stranded DNA and anti-histone antibodies were measured in B10.A and B10.A recombinant mice neonatally infected with MuLV-Scripps (Lerner et al., 1972). The incidence and latency of lymphoma and the incidence of glomerulonephritis were also determined. The mice studied could be divided into high-responder and low-responder groups. Most of the high ANA antibody could be attributed to anti-histone antibody. High response was associated with the H-2b haplotype and recombinant strains on the B10 background which were identical at the I-A subregion derived from the H-2b parental stock. In contrast, low ANA response was associated with the I-A subregion derived from the H-2k haplotype. In all groups of virus-inoculated animals, most animals developed serum elevations of p30 and gp70 and at least 72% of the inoculated animals developed lymphomas. High serum p30 levels correlated inversely with latency and directly with gp70 values. From 4 to 28% of the animals in each virus-inoculated group had histological evidence of glomerulonephritis, although no clear genetic basis could be ascribed to the incidence of glomerulonephritis, serum p30 or gp70 levels, or latency of lymphoma development.


Simultaneous determination of a hydrophobic drug candidate and its metabolite in human plasma with salting-out assisted liquid/liquid extraction using a mass spectrometry friendly salt.
In previously reported applications of salting-out assisted liquid/liquid extraction (SALLE) with acetonitrile, only inorganic salts were evaluated and implemented as the salting-out reagents. A potential concern of the method for the subsequent LC-MS analysis of biological samples was that a portion of the added salt (typically of high concentration) might be extracted and affect the chromatography separation and ionization of chromatography effluents in a mass spectrometer. Here we report, for the first time, the use of a mass spectrometry friendly organic salt, ammonium acetate, as a salting-out reagent in SALLE with acetonitrile for the simultaneous quantitation of an Abbott investigational new drug ABT-869 and its hydrophilic metabolite in human plasma. The performance of SALLE with ammonium acetate was compared with that of a previously reported method with a conventional liquid/liquid extraction technique using a set of pooled incurred samples. The % differences of the measured concentrations for 24 samples from these two methods were found to be within acceptance criteria, demonstrating SALLE with ammonium acetate as a reliable sample preparation technique. The SALLE method is simple, fast (25 min/plate), easy for automation, free of drying down step, and environmentally friendly. SALLE with mass spectrometry friendly salts has been applied to regulated sample analysis of both hydrophilic and hydrophobic compounds. It is envisioned that SALLE with acetonitrile and ammonium acetate be a universal method for high throughput automated sample preparation for bioanalytical chemistry.


Platelet-Monocyte Aggregates and C-Reactive Protein are Associated with VTE in Older Surgical Patients.
Emerging evidence implicates platelets as key mediators of venous thromboembolism (VTE). Nevertheless, the pathways by which platelets and circulating procoagulant proteins synergistically orchestrate VTE remain incompletely understood. We prospectively determined whether activated platelets and systemic procoagulant factors were associated with VTE in 32 older orthopedic surgery patients. Circulating platelet-monocyte aggregates (PMAs), p-selectin expression (P-SEL), and integrin αIIbβ3 activation (PAC-1 binding) were assessed pre-operatively and 24 hours post-operatively. The proinflammatory and procoagulant molecule C-reactive protein (CRP), which induces PMA formation in vitro, along with plasma d-dimer and fibrinogen levels were also measured. The primary outcome was VTE occurring within 30 days post-operatively. Overall, 40.6% of patients developed VTE. Patients with VTE had a significant increase in circulating PMAs and CRP post-operatively, compared to those without VTE. Changes in PMA and CRP in VTE patients were significantly correlated (r(2) = 0.536, p = 0.004). In contrast, P-SEL expression and PAC-1 binding, fibrinogen levels, and d-dimers were not associated with VTE. This is the first study to identify that increased circulating PMAs and CRP levels are early markers associated with post-surgical VTE. Our findings also provide new clinical evidence supporting the interplay between PMAs and CRP in patients with VTE.


Therapeutic efficacy of salvage cystectomy. Do results reflect natural history of bladder cancer?
In a retrospective review of patients who underwent salvage cystectomy, patients with muscle-invasive cancers who had initially presented with tumors that were pathologically TA or T1 appeared to have a better prognosis than those who had muscle-invasive (T2 or T3) cancers on initial clinical presentation. Prognosis in each group appeared to correlate with the stage of tumor at initial clinical presentation rather than with stage of tumor at recurrence after radiation therapy. Prognosis also appeared to correlate with architectural configuration of the presenting tumor as well as the type of invasion. Thus, papillary lesions with muscle invasion by a broad front of histologically cohesive blocks of cells appeared to have a better prognosis than did solid or nodular lesions that appeared to invade the muscle wall in a tentacular fashion with fingerlike tumor cell extensions that seemed to percolate through the bladder wall. Taken together, the results of treatment in these patients may have represented the intrinsic nature of their particular tumors.


Influence of storage conditions on normal plasma amino-acid concentrations.
Conflicting information in the literature is given concerning the optimal preparation and storage conditions of plasma samples for amino-acid analysis. To assess the optimal pre-storage treatment, we compared several methods and studied their influence on plasma amino-acid levels of rats and humans, stored at different temperatures. In rat plasma, the frequently reported degradation of glutamine was not measurable at a storage temperature of -70 degrees C. However, storage of native, not deproteinised plasma at this temperature, resulted in a 32% decrease of arginine and a 30% increase in ornithine after 24 weeks. Deproteinisation prohibited this arginine decay. At -20 degrees C, arginine decay was even more pronounced, whereas glutamine decreased by 14% in untreated plasma, by 10% in sulfosalicylic acid deproteinised plasma and by 3% if the deproteinisation was followed by removal of the protein pellet and subsequent neutralisation. To confirm these unexpected results in humans, we repeated this experiment with plasma of 6 volunteers. In contrast to rat plasma, we did not observe any changes in arginine and ornithine concentrations in human plasma stored at -70 degrees C. At -20 degrees C the reduction in glutamine was only 4-5%. These results suggest that interspecies differences in enxymatic activity exist in plasma. Finally, having assessed the optimal treatment and storage conditions (deproteinisation followed by storage at -70 degrees C), samples were obtained from a total of 112 human volunteers, stratified for age and sex, and amino-acids were measured. In the female group, we found a tendency to a gradual increase in most amino-acid concentrations with advancing age, which however only reached significance for histidine, citrulline, alanine and leucine. These observations demonstrate that plasma samples for amino-acid analysis should be deproteinised and stored at -70 degrees C. Also important interspecies differences appear to exist in plasma enzymatic activity. Finally, control samples should be taken from an age and sex matched control group.


Ferric cacodylate efficiently stimulates growth of rat renal glomerular epithelial cells in vitro.
Rat renal glomerular epithelial cells (SGE1 cell line) can be maintained and grown continuously in serum-free medium supplemented with insulin, iron-saturated transferrin (Tr), selenium, bovine serum albumin (BSA), linoleic acid, and epidermal growth factor (EGF). Of the growth supplements used, Tr is essential for proliferation of the cells. In the present study, we describe the use of a unique iron-chelate complex, ferric cacodylate (Fe-Cac), positively charged molecules in neutral buffer, that could almost replace Tr in serum-free culture. It even stimulated the growth of SGE1 cells more efficiently than ferric chloride (FeCl(3)) and other iron-chelate complexes, such as ferric nitrilotriacetate (Fe-NTA) and ferric citrate (Fe-Cit). The growth-stimulatory activity of Fe-Cac was exerted at iron concentrations of more than 0.01 μg/ml, whereas a 10-fold excess of iron concentration was required with FeCl(3), Fe-NTA and Fe-Cit. We observed that SGE1 cells grew until confluent, then formed hemicysts (domes) in serum-free medium containing Fe-Cac, suggesting that Fe-Cac did not merely permit cell growth but also supported polarization and organization of the cells into a functional epithelial architecture. Moreover, since the stimulatory activity of Fe-Cac was completely abolished by desferrioxamine, a strong iron chelator, it is suggested that iron is crucial for growth of SGE1 cells. When the cells were treated with suramin, an inhibitor of cellular pinocytosis and endocytosis of a large spectrum of ligands including receptor-bound growth factors, growth-stimulatory activity of Tr was inhibited, whereas the activity of Fe-Cac was not affected. These results, taken together, strongly suggest that the growth-stimulatory activity of Fe-Cac is associated with iron delivery into the cells through the cell membrane by diffusion, which is different from Tr receptor-mediated endocytosis. The use of Fe-Cac for investigating iron-regulated cell proliferation is suggested.


Single-molecule imaging reveals how mavacamten and PKA modulate ATP turnover in skeletal muscle myofibrils.
Muscle contraction is controlled at two levels: the thin and the thick filaments. The latter level of control involves three states of myosin heads: active, disordered relaxed (DRX), and super-relaxed (SRX), the distribution of which controls the number of myosins available to interact with actin. How these are controlled is still uncertain. Using fluorescently labeled ATP, we were able to spatially assign the activity of individual myosins within the sarcomere. We observed that SRX comprises 53% of all heads in the C-zone compared with 35% and 44% in the P- and D-zones, respectively. The recently FDA-approved hypertrophic cardiomyopathy drug, mavacamten (mava), significantly decreased DRX, favoring SRX in both the C- and D-zones at 60% and 63%, respectively. Since thick filament regulation is in part regulated by the myosin-binding protein-C (MyBP-C), we also studied PKA phosphorylation. This had the opposite effect as mava, specifically in the C-zone where it decreased SRX to 34%, favoring DRX. These results directly show that excess concentrations of mava do increase SRX, but the effect is limited across the sarcomere, suggesting mava is less effective on skeletal muscle. In addition, we show that PKA directly affects the contractile machinery of skeletal muscle leading to the liberation of repressed heads. Since the effect is focused on the C-zone, this suggests it is likely through MyBP-C phosphorylation, although our data suggest that a further reserve of myosins remain that are not accessible to PKA treatment.


Fear of hypoglycaemia in adults with Type 1 diabetes.
The aim of this study was to examine the fear of hypoglycaemia and its association with demographic and disease-specific variables in a large and unselective population of adult patients with Type 1 diabetes. Questionnaires were sent by post to all patients with Type 1 diabetes who were identified in the local diabetes registries of two hospitals in Stockholm, Sweden (n=1387). Fear of hypoglycaemia was measured using the Swedish Hypoglycaemia Fear Survey, the Worry subscale and the Aloneness subscale. Demographic variables and disease-specific factors were collected from patients' self reports and medical records. Univariate analysis and multiple stepwise linear regression analysis were used in the statistical analyses of the data. Seven hundred and sixty-four (55%) patients participated in the study (mean age 43.3 years and mean HbA(1c) 7.0%, normal <5.0%). The Hypoglycaemia Fear Survey - Worry subscale was significantly associated with frequency of severe hypoglycaemia, number of symptoms during mild hypoglycaemia, gender, hypoglycaemic symptoms during hyperglycaemia and hypoglycaemic unawareness. The Hypoglycaemia Fear Survey - Aloneness subscale was significantly associated with frequency of severe hypoglycaemia, number of symptoms during mild hypoglycaemia, gender, frequency of mild hypoglycaemia, HbA(1c) , hypoglycaemic unawareness and visits to the emergency room because of severe hypoglycaemia. Fear of hypoglycaemia proved to be more prevalent in females and indicated a different pattern between genders in relation to factors associated with fear of hypoglycaemia. This study identifies the frequency of severe hypoglycaemia as the most important factor associated with fear of hypoglycaemia. Moreover, for the first time, we document gender differences in fear of hypoglycaemia, suggesting that females are more affected by fear of hypoglycaemia than men.


Storage of platelets in additive solutions: a pilot in vitro study of the effects of potassium and magnesium.
Platelet additive solutions (PAS) have been shown to be suitable for extended platelet storage but have required the carryover of substantial (30%) amounts of plasma for success. Improving platelet quality by optimizing the composition of PAS may allow a reduction to be made in the amount of plasma carried over. Reducing the proportion of plasma carried over would facilitate some methods of viral inactivation and make available greater amounts of plasma for other needs. Platelets from six pools of 25 buffy coat platelet units and five apheresis platelet units were aliquoted for storage in plasma, or converted to PAS units in either a specific additive solution (PAS-III), with 30% or 20% plasma, or a modification of PAS-III containing 5.0 mm potassium and 1.5 mm magnesium (PAS-IIIM), with 30% or 20% plasma. Units were stored at room temperature with agitation for 7 days with in vitro testing for biochemical, haematological and functional parameters. Storage of platelets in PAS-IIIM resulted in a reduced rate of glycolysis and better retention of pH, morphology score and ATP levels. Platelets initially showed less evidence of activation when stored in PAS-IIIM, with reduced P-selectin expression. Storage in PAS-IIIM with 20% (rather than the standard 30%) plasma appeared to result in the retention of in vitro properties, similarly to storage in standard PAS-III with 30% plasma. Storing platelets in an additive solution containing magnesium and potassium improves the functionality of the platelets, as measured by in vitro testing, and may allow a reduction of the amount of plasma required to be carried over to the final unit.


Molecular design of new aggrecanases-2 inhibitors.
Aggrecanases-2 is a very important potential drug target for the treatment of osteoarthritis. In this study, a series of known aggrecanases-2 inhibitors was analyzed by the technologies of three-dimensional quantitative structure-activity relationships (3D-QSAR) and molecular docking. Two 3D-QSAR models, which based on comparative molecular field analysis (CoMFA) and comparative molecular similarity analysis (CoMSIA) methods, were established. Molecular docking was employed to explore the details of the interaction between inhibitors and aggrecanases-2 protein. According to the analyses for these models, several new potential inhibitors with higher activity predicted were designed, and were supported by the simulation of molecular docking. This work propose the fast and effective approach to design and prediction for new potential inhibitors, and the study of the interaction mechanism provide a better understanding for the inhibitors binding into the target protein, which will be useful for the structure-based drug design and modifications.


Effect of age on kainate-induced seizure severity and cell death.
While the onset and extent of epilepsy increases in the aged population, the reasons for this increased incidence remain unexplored. The present study used two inbred strains of mice (C57BL/6J and FVB/NJ) to address the genetic control of age-dependent neurodegeneration by building upon previous experiments that have identified phenotypic differences in susceptibility to hippocampal seizure-induced cell death. We determined if seizure induction and seizure-induced cell death are affected differentially in young adult, mature, and aged male C57BL/6J and FVB/NJ mice administered the excitotoxin, kainic acid. Dose response testing was performed in three to four groups of male mice from each strain. Following kainate injections, mice were scored for seizure activity and brains from mice in each age group were processed for light microscopic histopathologic evaluation 7 days following kainate administration to evaluate the severity of seizure-induced brain damage. Irrespective of the dose of kainate administered or the age group examined, resistant strains of mice (C57BL/6J) continued to be resistant to seizure-induced cell death. In contrast, aged animals of the FVB/NJ strain were more vulnerable to the induction of behavioral seizures and associated neuropathology after systemic injection of kainic acid than young or middle-aged mice. Results from these studies suggest that the age-related increased susceptibility to the neurotoxic effects of seizure induction and seizure-induced injury is regulated in a strain-dependent manner, similar to previous observations in young adult mice.


[Response of an undifferentiated and nullipotential cell line derived from an A/Heston mouse teratocarcinoma to infection with type C ecotropic murine viruses].
The PCC6 cell line, derived from an A/Heston Mouse Teratocarcinoma, and composed of nullipotential embryonic carcinoma cells (ECC), is completely resistant to infection with type C murine ecotropic viruses. It reacts as other cell lines previously studied which are derived from a 129 Mouse teratocarcinoma and composed of multipotential ECC.


Importance of oxidative stress in the pathogenesis, diagnosis, and monitoring of patients with neuropsychiatric disorders, a review.
Oxidative stress is defined as the persistent imbalance between the activity of toxic reactive forms of both oxygen and nitrogen and the antioxidant defense. In low concentrations, they are essential for the proper functioning of the body. Still, their excessive amount contributes to the damage of the biomolecules, consequently leading to various pathologies of the organism. Due to the lipid-rich brain structure, enormous oxygen consumption, and the lack of a sufficient antioxidant barrier make it highly susceptible to oxidative imbalance. Hence, oxidative stress has been linked to various psychiatric disorders. These diseases include all behavioral, emotional, and cognitive abnormalities associated with a significant impediment to social life. Each of the diseases in question: Alzheimer's disease, schizophrenia, depression, and bipolar disorder, is characterized by excessive oxidative stress. Considerable damages to DNA, RNA, proteins, lipids, and mitochondrial dysfunction, are observed. All conditions show increased lipid peroxidation, which appears to be typical of psychiatric disorders because the brain contains large amounts of these types of molecules. In addition, numerous abnormalities in the antioxidant defense are noted, but the results of studies on the activity of antioxidant enzymes differ significantly. The most promising biomarkers seem to be GSH in Alzheimer's disease as an early-stage marker of the disease and thioredoxin in schizophrenia as a marker for therapy monitoring. Data from the literature are consistent with the decrease in antioxidants such as vitamin C, E, uric acid, albumin, etc. Despite these numerous inconsistencies, it seems that oxidative stress is present in the course of psychiatric diseases. Still, it cannot be conclusively determined whether it is the direct cause of development, a consequence of other abnormalities at the biochemical or molecular level, or the result of the disease itself.


The Penrose drain: a safe, atraumatic colostomy bridge.
Because of the problems associated with the large, bulky bridges presently used for construction of loop ostomies, particularly complications of leaks and skin excoriation, we have studied the use of the Penrose drain as an alternative. This method has been time-tested on 45 patients and has been found to be safe, reliable, and inexpensive and has gained popularity among patients and ostomy nurses. The bulky colostomy bridge should no longer be accepted as a standard of care.


Measurement of 33P in faeces by suspension counting of faecal ash.
The estimation of 33P in faecal samples by direct suspension counting of finely ground faecal ash particles is shown to be a convenient technique having good accuracy and reproducibility. The sensitivity of the method is such that 75 pCi 33P can be measured with a standard error of 2% in a counting period of 1 h. Using 50 mg of ash for counting, this activity requires a 33P concentration of about 40 pCi/g of faecal homogenate.


The effect of carbon dioxide on the diameter of brain capillaries.
Cerebral capillaries in cats subjected to variations in carbon dioxide tensions were studied using carbon black perfusion fixation. Five animals each were grouped into hypocapnia, normocapnia, and hypercapnia and 500 cortical, 250 white matter and 250 caudate nucleus capillaries per group were analyzed at 400 x magnification. Capillary diameters were found to change significantly in the cortex (6.1-10.0 microns), white matter (6.5-9.6 microns) and caudate nucleus (6.3-8.8 microns) from hypocapnia to hypercapnia, respectively. These findings suggest that capillary vasomotion occurs and that capillaries are not rigid tubes as previously portrayed.


Beta-hydroxysteroid dehydrogenase: activity in microemulsion and extraction from Pseudomonas testosteroni cells with microemulsion.
The stability of purified beta-hydroxysteroid dehydrogenase activity measured as a function of time was good in buffered cationic and non-ionic microemulsions. The use of 1-pentanol and 1-hexanol in place of 1-butanol as cosurfactant gave increased activity and stability. The NAD+ Michaelis constant was 0.22 mM in buffer and 3.5 mM in waterpool concentration in microemulsion. Proteins, among them beta-hydroxysteroid dehydrogenase, were extracted from Pseudomonas testosteroni with cationic microemulsion, thus indicating that microemulsions may be utilized in protein release from cells.


Near-infrared fluorescence imaging of mouse myocardial microvascular endothelium using Cy5.5-lectin conjugate.
Cy5.5-lectin, a non-toxic conjugate, combines the benefits of near-infrared (NIR) imaging, such as significant reduction of background fluorescence and increased tissue depth penetration, with its affinity for vascular endothelial cells. When compared to endothelial staining methods using FITC-lectin and ICAM2 antibodies, Cy5.5-lectin was confirmed to specifically bind endothelial cells and produce a fluorescence signal both in real-time and post-infusion. Ex-vivo experiments with isolated hearts demonstrated that binding was limited to perfused areas of the myocardium. With mouse in-vivo tail-vein injections, other organs such as the liver, spleen, and kidney were also stained and yielded similar quality images of the heart.


Regorafenib combined with PD1 blockade increases CD8 T-cell infiltration by inducing CXCL10 expression in hepatocellular carcinoma.
Combining inhibitors of vascular endothelial growth factor and the programmed cell death protein 1 (PD1) pathway has shown efficacy in multiple cancers, but the disease-specific and agent-specific mechanisms of benefit remain unclear. We examined the efficacy and defined the mechanisms of benefit when combining regorafenib (a multikinase antivascular endothelial growth factor receptor inhibitor) with PD1 blockade in murine hepatocellular carcinoma (HCC) models. We used orthotopic models of HCC in mice with liver damage to test the effects of regorafenib-dosed orally at 5, 10 or 20 mg/kg daily-combined with anti-PD1 antibodies (10 mg/kg intraperitoneally thrice weekly). We evaluated the effects of therapy on tumor vasculature and immune microenvironment using immunofluorescence, flow cytometry, RNA-sequencing, ELISA and pharmacokinetic/pharmacodynamic studies in mice and in tissue and blood samples from patients with cancer. Regorafenib/anti-PD1 combination therapy increased survival compared with regofarenib or anti-PD1 alone in a regorafenib dose-dependent manner. Combination therapy increased regorafenib uptake into the tumor tissues by normalizing the HCC vasculature and increasing CD8 T-cell infiltration and activation at an intermediate regorafenib dose. The efficacy of regorafenib/anti-PD1 therapy was compromised in mice lacking functional T cells (Rag1-deficient mice). Regorafenib treatment increased the transcription and protein expression of CXCL10-a ligand for CXCR3 expressed on tumor-infiltrating lymphocytes-in murine HCC and in blood of patients with HCC. Using Cxcr3-deficient mice, we demonstrate that CXCR3 mediated the increased intratumoral CD8 T-cell infiltration and the added survival benefit when regorafenib was combined with anti-PD1 therapy. Judicious regorafenib/anti-PD1 combination therapy can inhibit tumor growth and increase survival by normalizing tumor vasculature and increasing intratumoral CXCR3+CD8 T-cell infiltration through elevated CXCL10 expression in HCC cells.


Application of HACCP principles to control visitor health threats on dairy farms open to the general public.
An increasing number of Dutch dairy farmers have diversified their activities, often opening their farm up to visitors (tourist accommodation, farm shop, contact with livestock, etc). It is essential to prevent these visitors from having accidents or becoming ill, which could result in financial claims and might harm the reputation of the agricultural sector. This article describes how the hazard analysis critical control points concept and principles (HACCP) can be applied to these activities and integrated with on-farm operational herd health and production management programmes.


Will focal therapy become a standard of care for men with localized prostate cancer?
The current treatment choice for men with localized prostate cancer lies between active surveillance and radical therapy. The difference between these two extremes of care is 5% in terms of cancer-related absolute mortality at 8 years. It is generally accepted that this small difference will decrease for men diagnosed in the prostate-specific-antigen era. Radical therapy is associated with considerable adverse effects (e.g. incontinence, impotence, rectal problems) because it treats the whole gland, and damages surrounding structures in up to half of men. Men are being diagnosed at a younger age with lower-risk disease, and many have unifocal or unilateral disease. We propose a new concept whereby only the tumor focus and a margin of normal tissue are treated. This paradigm might decrease adverse effects whilst, at the same time, retaining effective cancer control. The arguments for and against active surveillance and radical therapy are reviewed in this article, with focal therapy presented as a means for bridging these two approaches.


Supernovae, neutron stars and biomolecular chirality.
Recent theoretical and experimental investigations of the origin of biomolecular chirality are reviewed briefly. Biotic and abiotic theories are evaluated critically with the conclusion that asymmetric photochemical processes with circulary polarized light (CPL), particularly asymmetric photolyses, constitute the most viable mechanisms. Solar CPL sources appear too weak and random to be effective. We suggest an alternative CPL source, namely, the synchrotron radiation from the neutron star remnants of supernova explosions. This could asymmetrically process racemic compounds in the organic mantles of the dust grains in interstellar clouds, and the resulting chiral molecules could be transferred to Earth by cold accretion as the solar system periodically traverses these interstellar clouds.


A detailed atlas of chick heart development in vivo.
Various model organisms such as mouse, xenopus, or zebrafish embryos have been studied in the past to gain insight into the complex processes driving normal and abnormal development of the vertebrate heart. Despite the fact that the chicken embryo has been a favored classic model system used by embryologists and cardiovascular scientists for centuries to illustrate the principles of basic vertebrate embryology and cardiovascular development, so far, no one has provided a thorough documentation of heart development in this model from early visual stages to the stage of a completely formed heart with (a) images and (b) video recordings of beating hearts. However, in vivo documentation of heart development stages is indispensable because the initially tubular embryonic heart not only undergoes dramatic morphological changes, but also intriguing functional changes during cardiogenesis, which, only if they follow and remain within the normal developmental pathway, lead to the establishment of the normal four-chambered heart. In this work we present the first reference catalogue of cardiac development in vivo with (1) 25 plates of high resolution colour images in different views from Hamburger-Hamilton (HH)-stage 12 (day 2, relatively straight heart tube, early myocardial contractions) through HH-stage 35 (day 9, four-chambered heart) in end-diastole and end-systole, including a plate with an overview of all these stages; (2) collection of 82 video recordings of beating hearts in different views corresponding to the stages shown in the plates.


Neonatal management and safe discharge of late and moderate preterm infants.
Late and moderate preterm infants form the majority of admissions for prematurity to special care neonatal nurseries. Although at risk for acute disorders of prematurity, they do not suffer the serious long term risks and chronic illnesses of the extremely premature. The special challenges addressed here are of transition and of thermal adaptation, nutritional compensation for postnatal growth restriction, the establishment of early feeding, and the avoidance of post-discharge jaundice or apnea. These 'healthy' premature infants provide challenges for discharge planning, in that opportunities may be available for discharge well before the expected date of delivery, which should be pursued. Barriers to early discharge are rigid conservative protocols and unwarranted investigations; facilitators of discharge are individualized care by nurses expert in cue-based feeding, early management of the thermal environment, support of family preferences and encouragement of mother-baby interactions. Safe discharge depends on recognizing these opportunities and applying strategies to address them.


Current topics in childhood lupus nephritis.
Lupus nephritis is a major predictor of the prognosis of systemic lupus erythematosus (SLE). The present paper discusses lupus nephritis from clinical and immunopathological points of view. Although recent advances in diagnosis and treatment improve the prognosis of children with SLE, there remain many unsolved clinical problems. One of the current topics in the treatment for SLE is intermittent intravenous cyclophosphamide therapy which is effective even for the steroid-resistant patients with severe lupus nephritis, at least for short-term observation. Immunopathologically, the following issues are discussed: (i) The C5b-9 terminal complement complex plays an important role in the pathogenesis of lupus nephritis. The possible interaction of vitronectin and SP-40,40 is also mentioned; (ii) A semi-quantitative analysis of the charge barrier of the glomerular basement membrane reveals that the charge barrier dysfunction plays an important role in the pathogenesis of proteinuria in lupus nephritis. This study also demonstrates that the charge of immune deposits is important for the initiation of glomerular injury in lupus nephritis; (iii) It is demonstrated that the histopathological diversity of lupus nephritis is based on biological properties of nephritogenic auto-antibodies in murine lupus models.