[Urinary excretion rate of methylguanidine as a new maker of active oxygen in vivo: demonstration in hyperbaric oxygen therapy].
Methylguanidine (MG) which is known as a uremic toxin, is synthesized from creatinine (Cre). We have clarified that active oxygen plays an important role on MG synthesis in vitro and in rat hepatocytes. On the other hand, hyperoxia is very injurious in various tissues, and it has been reported that active oxygen produced in hyperoxia plays an important role on the tissue injury. This study was performed to investigate the effect of hyperoxia on MG synthesis in vivo. The subjects in this study were patients who were treated by hyperbaric oxygen therapy (HBO). Serum Cre, MG, and urinary Cre, MG before and after HBO were measured in these subjects. The subjects were classified into four groups. Group I-III were undergone HBO with condition of 100% O2, 2 atmosphere absolute (ATA), 1 hour, (I: Ccr less than 10 ml/min, II: 10 less than or equal to Ccr less than 50 ml/min, III: Ccr greater than or equal to 50 ml/min) and group IV (Ccr greater than or equal to 50 ml/min) with 100%O2, 3ATA, 1 hour. Urinary excretion rate of MG (urine MG/urine Cre) significantly increased after HBO therapy in every group. Urine MG/urine Cre/serum Cre ratio which was used as a index of MG synthesis rate also increased. In this study, it is clarified that MG excretion rate increases in hyperoxic condition. These results suggest that active oxygen plays an important role on MG synthesis in vivo, and that the urine MG/urine Cre/serum Cre ratio can be a useful maker of the active oxygen products in vivo.


Spontaneous and photosensitization-induced mutations in primary mouse cells transitioning through senescence and immortalization.
To investigate the role of oxidative stress-induced DNA damage and mutagenesis in cellular senescence and immortalization, here we profiled spontaneous and methylene blue plus light-induced mutations in the cII gene from λ phage in transgenic mouse embryonic fibroblasts during the transition from primary culture through senescence and immortalization. Consistent with detection of characteristic oxidized guanine lesions (8-oxodG) in the treated cells, we observed significantly increased relative cII mutant frequency in the treated pre-senescent cells which was augmented in their immortalized counterparts. The predominant mutation type in the treated pre-senescent cells was G:C→T:A transversion, whose frequency was intensified in the treated immortalized cells. Conversely, the prevailing mutation type in the treated immortalized cells was A:T→C:G transversion, with a unique sequence-context specificity, i.e. flanking purines at the 5' end of the mutated nucleotide. This mutation type was also enriched in the treated pre-senescent cells, although to a lower extent. The signature mutation of G:C→T:A transversions in the treated cells accorded with the well-established translesion synthesis bypass caused by 8-oxodG, and the hallmark A:T→C:G transversions conformed to the known replication errors because of oxidized guanine nucleosides (8-OHdGTPs). The distinctive features of photosensitization-induced mutagenesis in the immortalized cells, which were present at attenuated levels, in spontaneously immortalized cells provide insights into the role of oxidative stress in senescence bypass and immortalization. Our results have important implications for cancer biology because oxidized purines in the nucleoside pool can significantly contribute to genetic instability in DNA mismatch repair-defective human tumors.


Readmissions after stroke: linked data from the Australian Stroke Clinical Registry and hospital databases.
To assess the feasibility of linking a national clinical stroke registry with hospital admissions and emergency department data; and to determine factors associated with hospital readmission after stroke or transient ischaemic attack (TIA) in Australia. Data from the Australian Stroke Clinical Registry (AuSCR) at a single Victorian hospital were linked to coded, routinely collected hospital datasets for admissions (Victorian Admitted Episodes Dataset) and emergency presentations (Victorian Emergency Minimum Dataset) in Victoria from 15 June 2009 to 31 December 2010, using stepwise deterministic data linkage techniques. Association of patient characteristics, social circumstances, processes of care and discharge outcomes with all-cause readmissions within 1 year from time of hospital discharge after an index admission for stroke or TIA. Of 788 patients registered in the AuSCR, 46% (359/781) were female, 83% (658/788) had a stroke, and the median age was 76 years. Data were successfully linked for 782 of these patients (99%). Within 1 year of their index stroke or TIA event, 42% of patients (291/685) were readmitted, with 12% (35/286) readmitted due to a stroke or TIA. Factors significantly associated with 1-year hospital readmission were two or more presentations to an emergency department before the index event (adjusted odds ratio [aOR], 1.57; 95% CI, 1.02-2.43), higher Charlson comorbidity index score (aOR, 1.19; 95% CI, 1.07-1.32) and diagnosis of TIA on the index admission (aOR, 2.15; 95% CI, 1.30-3.56). Linking clinical registry data with routinely collected hospital data for stroke and TIA is feasible in Victoria. Using these linked data, we found that readmission to hospital is common in this patient group and is related to their comorbid conditions.


Clozapine use reduced psychiatric hospitalization and emergency room visits in patients with bipolar disorder independent of improved treatment regularity in a three-year follow-up period.
The efficacy of clozapine in bipolar disorder remains to be systemically examined. In the current study, we sought to disentangle the effect of clozapine from that of improved treatment regularity and to compare the effect of clozapine with the effect of regular treatment for bipolar disorder by exploring the complete 10-year clozapine prescription data from a Taiwanese total population health claims database. In the period between 2000 and 2009, 3,874 (3.3%) out of the 117,785 patients identified as having bipolar disorder in a Taiwanese total population health claims database were ever prescribed clozapine. Among them, 920 patients with bipolar disorder who had good pre-clozapine medication compliance and received at least two clozapine prescriptions were further categorized according to their clozapine medication possession ratio (MPR) as regular users (MPR ≥ 0.8; n = 476) and irregular users (MPR < 0.8; n = 444). Using a mirror-image design, we compared the numbers of emergency room (ER) visits, hospitalizations and hospital days, and the average durations of a single hospitalization during the pre- and post-clozapine mirror periods with a follow-up time of up to three years, controlling for time-variant course confounders. The patterns of change in outcome indices from the pre-clozapine period to the post-clozapine period differed significantly between the two clozapine-user groups. Clinical outcome indices improved only in regular users, while they deteriorated in irregular users. Over the three-year follow-up period, the irregular users consistently had a higher adjusted risk for increased numbers of ER visits [odds ratio (OR): 2.06-2.43], hospitalizations (OR: 2.52-3.22), and total hospital days (OR: 2.42-2.91) when compared to the regular users. Thus, effects of clozapine were consistently demonstrated in one- to three-year mirror comparison periods. Clozapine, when used with high treatment regularity (MPR > 0.8), was effective in reducing the numbers of ER visits, hospitalizations, and total hospital days in patients with bipolar disorder with previous frequent hospitalizations and ER visits despite regular pre-clozapine treatment for bipolar disorder. However, high early attrition and suboptimal treatment compliance need to be rectified in order to optimize the outcome of clozapine treatment in bipolar disorders.


Genome-wide analysis of DNA methylation and gene expression changes in two Arabidopsis ecotypes and their reciprocal hybrids.
Heterosis is a fundamental biological phenomenon characterized by the superior performance of a hybrid over its parents in many traits, but the underlying molecular basis remains elusive. To investigate whether DNA methylation plays a role in heterosis, we compared at single-base-pair resolution the DNA methylomes of Arabidopsis thaliana Landsberg erecta and C24 parental lines and their reciprocal F1 hybrids that exhibited heterosis. Both hybrids displayed increased DNA methylation across their entire genomes, especially in transposable elements. Interestingly, increased methylation of the hybrid genomes predominantly occurred in regions that were differentially methylated in the two parents and covered by small RNAs, implying that the RNA-directed DNA methylation (RdDM) pathway may direct DNA methylation in hybrids. In addition, we found that 77 genes sensitive to methylome remodeling were transcriptionally repressed in both reciprocal hybrids, including genes involved in flavonoid biosynthesis and two circadian oscillator genes circadian clock associated1 and late elongated hypocotyl. Moreover, growth vigor of F1 hybrids was compromised by treatment with an agent that demethylates DNA and by abolishing production of functional small RNAs due to mutations in Arabidopsis RNA methyltransferase HUA enhancer1. Together, our data suggest that genome-wide remodeling of DNA methylation directed by the RdDM pathway may play a role in heterosis.


Comparative Study of Serum Progesterone Levels at the Time of Human Chorionic Gonadotropin Trigger and Ovum PickUp in Predicting Outcome in Fresh in vitro Fertilization Cycles.
Studies have shown that premature rise of progesterone in controlled ovarian stimulation (COS) at the time of human chorionic gonadotropin (hCG) trigger is negatively associated with in vitro fertilization (IVF) outcome in fresh IVF cycles. Some authors have failed to demonstrate this. One large single centre retrospective cohort study has compared the pre and post hCG progesterone and observed that the ratio of the rise in progesterone could be a positive predictor. There is paucity of literature on this aspect. To compare the serum progesterone at hCG trigger and ovum pick-up (OPU) with IVF outcome by estimating the respective paired hormone levels. Serum progesterone levels at hCG trigger and OPU are compared retrospectively in 301 fresh IVF cycles with IVF outcome by long protocols with GnRH agonists for two years. Parametric and nonparametric testing of null hypothesis is performed. P value <0.05 is taken as significant. There is no predictive association of IVF outcome with either progesterone levels. The ratio of rise in progesterone is strongly positively associated with IVF outcome (P < 0.001). However, after adjusting for confounders and modifiers the retrieved number of oocytes are positively associated with IVF outcome (P = 0.044). The ratio of rise in progesterone is significantly associated with number of oocytes retrieved which in turn is associated with successful IVF outcome in fresh cycles. Ratio of rise in progesterone seems to be therefore an indirect parameter for predicting successful IVF outcome in fresh cycles.


Effectiveness of Reference Signal-Based Methods for Removal of EEG Artifacts Due to Subtle Movements During fMRI Scanning.
Subtle motion of an epileptic patient examined with co-registered EEG and functional MRI (EEG-fMRI) may often lead to spurious fMRI activation patterns when true epileptic spikes are contaminated with motion artefacts. In recent years, methods relying on reference signals for correcting these subtle movements in the EEG have emerged. In this study, the performance of two reference-based devices are compared to the template-based method with regard to their ability to remove movement-related artifacts in EEG measured during scanning. Measurements were performed with a novel double layer cap consisting of 29 EEG and 29 reference electrodes, and with a current loop cap consisting of 60 electrodes and three current loop wires attached to the cap. EEG was acquired inside the scanner during resting state, as well as when the subject was performing a cued movement task. For the double layer cap recordings, newly developed artifact removal algorithms are introduced and both reference signal-based methods are compared to a template-based correction method. The BCG artifacts occurring at resting state could be removed successfully by both the reference signal-based methods as well as by the template-based method. However, the reference signal-based methods were also capable of removing EEG artifacts induced by subtle movements, whereas the template-based method failed to remove these artifacts. Reference signal-based methods enable to correct for artifacts due to subtle movements, which are not removed by commonly used template-based removal algorithms. Sensitivity of EEG-fMRI analysis in patients with focal epilepsy is improved by avoiding erroneous detections of subtle movements as epileptic spikes in the EEG.


Case-mix adjusting performance measures in a veteran population: pharmacy- and diagnosis-based approaches.
To compare the rankings for health care utilization performance measures at the facility level in a Veterans Health Administration (VHA) health care delivery network using pharmacy- and diagnosis-based case-mix adjustment measures. The study included veterans who used inpatient or outpatient services in Veterans Integrated Service Network (VISN) 20 during fiscal year 1998 (October 1997 to September 1998; N = 126,076). Utilization and pharmacy data were extracted from VHA national databases and the VISN 20 data warehouse. We estimated concurrent regression models using pharmacy or diagnosis information in the base year (FY1998) to predict health service utilization in the same year. Utilization measures included bed days of care for inpatient care and provider visits for outpatient care. Rankings of predicted utilization measures across facilities vary by case-mix adjustment measure. There is greater consistency within the diagnosis-based models than between the diagnosis- and pharmacy-based models. The eight facilities were ranked differently by the diagnosis- and pharmacy-based models. Choice of case-mix adjustment measure affects rankings of facilities on performance measures, raising concerns about the validity of profiling practices. Differences in rankings may reflect differences in comparability of data capture across facilities between pharmacy and diagnosis data sources, and unstable estimates due to small numbers of patients in a facility.


Direct methods for detecting picorna-like virus from dead and alive triatomine insects.
In this work we report four different destructive and non-destructive methods for detecting picorna-like virus particles in triatomines. The methods are based on direct observation under transmission electron microscope and they consist of four ways to prepare samples of presumable infected material. The samples are prepared processing dead or alive insect parts, or even dry or fresh insect feces. The methods can be used as analytical or preparative techniques, for quantifying virus infection and checking virus integrity as well. In this work the four methods are applied in order to detect Triatoma virus (TrV) particles in T. infestans colonies.


Serum immunoglobulin free light chain correlates with tumor burden markers in Waldenstrom macroglobulinemia.
The serum IgM level has been utilised as a marker of tumor progression and to assess response to therapy in patients with Waldenstrom macroglobulinemia (WM). However, there are many limitations to the IgM protein level. The objective of this study was to evaluate the association of known tumor burden markers and prognostic factors with serum free light chain (sFLC) in 98 patients with WM. We demonstrated that sFLC measurement accurately differentiated IgM-MGUS compared with WM reflecting a measurement of tumor burden. In univariate and multivariate analysis, median sFLC at the cut-off at 60 mg/L was higher for WM patients with low hemoglobin and high beta2M, when we applied the WM-IPSS cut-offs, but showed no association with IgM level. This study demonstrates that sFLC is a new marker in WM disease. Further analysis is required to prospectively study the role of sFLC in monitoring response to therapy and as a prognostic marker in WM patients.


Characterisation of non-obese diabetic patients with marked insulin resistance identifies a novel familial partial lipodystrophy-associated PPARγ mutation (Y151C).
Familial partial lipodystrophy (FPLD) is a rare metabolic disorder with clinical features that may not be readily recognised. As FPLD patients require a specific therapeutic approach, early identification is warranted. In the present study we aimed to identify cases of FPLD among non-obese patients with type 2 diabetes mellitus and marked insulin resistance. We searched the databases of three diabetic outpatient clinics for patients with marked insulin resistance, arbitrarily defined as the use of ≥100 U insulin/day, and BMI ≤ 27 kg/m(2). In all patients, metabolic variables and anthropomorphic measurements were evaluated and DNA was sequenced for mutations in the genes encoding lamin A/C (LMNA), peroxisome proliferator-activated receptor γ (PPARγ) and cell death-inducing DFFA-like effector c (CIDEC). Out of 5,221 diabetic individuals, 24 patients fulfilled all criteria. Twelve patients were willing to participate, of whom five showed clinical features of lipodystrophy. In three of these patients the clinical diagnosis of FPLD was confirmed by the presence of mutations in LMNA or PPARG; one patient harboured a novel heterozygous mutation (Y151C) in PPARG. The Y151C mutant displayed impaired DNA-binding capacity and hence reduced transcriptional activity compared with wild-type PPARγ. Dominant-negative activity was absent. The combination of BMI ≤ 27 kg/m(2) and the use of >100 U insulin/day increases the chance of identifying lipodystrophy. Thus careful assessment of clinical features of FPLD should be considered in these patients, allowing earlier therapeutic interventions.


Aspects of our liver support systems using extracorporeal xenoperfusion of pig or baboon liver: review.
Artificial liver support systems using xenoperfusion of pig or baboon liver have metabolic activity and there is the possibility that they could substitute for total liver functions; however, several problems have yet to be solved. In our early clinical experience, a method of cross-hemodialysis with interposed cuprophane membrane was employed in order to avoid immunological reactions in patients. Sixteen patients with hepatic failure were treated by this method. Although the coma grade was ameliorated in 65% of the patients, the ultimate survival rate was 18.9%. In this clinical trial, the indication for liver support was clarified based on hepatic mitochondrial functions. This unsatisfactory result could also be attributed to insufficient effects of the device, due to the interposed membrane, and also to damage of the supporting livers due to hyperacute xenoperfusion injury. Recent investigations in the field of xenotransplantations have shown us possibilities for controlling xenogeneic hyperacute rejection. Suppression of complement activation enabled long-term xenoperfusion of supporting livers with high metabolic activity. The administration of prostaglandin E1 or soluble complement receptor type 1, and the use of transgenic pig livers expressing human decay-accelerating factor, may be promising methods to establish highly active artificial liver support systems using xenoperfusion.


[Comparative study of energy metabolism in the liver and skeletal muscles of rat and rabbit. Effect of starvation].
The steady state concentration of carbohydrate and adenosine phosphate metabolites in rat and rabbit liver and in rabbit skeletal muscle and oxidative phosphorylation parameters of rat and rabbit liver mitochondria were compared. The effects of 24 hr starvation on the energy metabolism of liver and skeletal muscle of the animals were investigated. The steady state concentrations of glycogen and phosphoenolpyruvate in normal rabbit liver were found to be much lower than in the rat and other mammalian livers (45.7 mumoles of glucose equivalents and 38 nmoles of PEP per 1 g of liver wet mass, respectively). On the contrast, the concentrations of glucose 6-phosphate, pyruvate and Pi in rabbit skeletal muscle were unusually high (up to 3, 1 and 15 mumoles per 1 g, respectively). In terms of glucose, pyruvate, lactate, Pi, adenine nucleotide contents and cytosolic NAD+/NADH ratio in the liver, and glycogen, glucose, lactate, creatine and adenosine phosphates in skeletal muscle and oxidative and phosphorylated properties of isolated liver mitochondria, no significant differences between rat and rabbit were found. During 24 hr starvation gluconeogenesis in rabbit liver occurred earlier and was more intensive than in rat liver. This is indicative of the existence of interspecies differences in the control mechanisms of carbohydrate and phosphorus metabolism.


Enhanced detection of live bacteria using a dendrimer thin film in an optical biosensor.
Here we describe the detection of live Pseudomonas aeruginosa using a sensing film containing a fourth-generation hydroxy-terminated polyamidoamine (PAMAM) dendrimer (i.e., G4-OH) and SYTOX Green fluorescent nucleic acid stain. The films are configured on simple, disposable plastic coupons or optical fibers and are interrogated using a miniature fiber-optic spectrometer. SYTOX Green is generally considered a dead cell stain because it is not able to cross the membranes of live cells. In the presence of PAMAM-OH (G4-OH) in water, the bacterial cell becomes permeable to the SYTOX dye and the fluorescence is significantly enhanced. The fluorescence increases with the bacteria concentration, and the intensity at 5.4 x 10(7) cells mL(-1) is 350% higher than the liquid controls without PAMAM-OH. We also demonstrate that dendrimers stabilize the sensing film. After drying and desiccation, the SYTOX Green/PAMAM-OH films are still able to quantitatively detect P. aeruginosa in water. Incorporation of glucose into the SYTOX Green/ PAMAM-OH film may improve the homogeneity of the film and enhances the fluorescence signal an additional 11-25%.


Nuclear crystalloids in sieve elements of Boraginaceae: a protein digestion study.
Nuclear crystalloids have been found in sieve elements of several Boraginaceae. Nuclei of differentiating sieve elements of Echium and other genera except Amsinckia contain one or more crystalloids composed of thin rods densely packed in parallel arrangement. After the nuclei disintegrate in the maturing sieve element the crystalloids are released into the cell lumen where they persist intact. In Amsinckia the crystalloid consists of two components: a dense component, similar to the crystalloid in the other genera and a loosely arranged paracrystalline component. The proteinaceous nature of the nuclear crystalloids and their possible similarity to P-protein was investigated by enzyme digestion techniques. Three proteolytic enzymes were employed in this study: protease, pepsin and trypsin. Successful digestion of the dense crystalloid in both Echium and Amsinckia was obtained with each enzyme tested. P-protein plugging the sieve plate pores was also digested. The loose component in Amsinckia and the aggregated and dispersed P-protein were not affected by the enzyme digestion procedures. These results seemed to indicate that the density or compactness of the proteinaceous inclusions may play a role in the differential response.


Kinins and angiotensins. Angiotensin I converting enzyme (kininase II) in endothelial cells cultured from human pulmonary arteries and veins.
Human pulmonary endothelial cells were used to study the metabolism of angiotensin I and bradykinin by angiotensin I converting enzyme (kininase II; ACE). The endothelial cells cultured from pulmonary arteries and veins differed with respect to their enzyme activity and their surface structure, as viewed with scanning electron microscopy. Cells from adult pulmonary arteries had greater ACE activity than cells from either adult pulmonary veins or fetal pulmonary arteries. In addition, cells from adult pulmonary arteries exhibited more prominent surface projections than cells from veins or fetal pulmonary arteries. A brief exposure to trypsin decreased the ACE activity in cells from arteries but not in cells from veins. Possible the differences in ACE activity between these cells are related to their surface structures.


Incidence of postoperative hyponatremia after endoscopic endonasal pituitary transposition for skull base pathologies.
Pituitary transposition is a novel surgical approach to access the retroinfundibular space and interpeduncular cistern. Few studies have evaluated post-surgical outcomes, including incidence of hyponatremia, following pituitary transposition. This is a retrospective study including 72 patients who underwent endoscopic endonasal surgery involving pituitary transposition for non-pituitary derived tumors over a decade at the University of Pittsburgh Medical Center. Anterior pituitary deficiencies and replacement therapy, tumor pathology and pre-operative serum sodium (Na) were recorded. Na was assessed at postoperative day 1, 3, 5, 7, and 10. Anatomical/surgical parameters included sellar height, sellar access angle to approach the tumor, and cranial extension of the tumor above the sellar floor (B) compared to the height of the gland (A) (B/A). T-test (normally distributed variables) and Wilcoxon rank-sum test (not-normally distributed) were applied for mean comparison. Logistic regression analyzed correlations between anatomical/surgical parameters and postoperative hyponatremia. 55.6% of patients developed post-operative transient hyponatremia. Two patients (5%) developed severe hyponatremia (sodium level < 120 mmol/L). Eleven (15.3%) patients required desmopressin replacement immediately post-operatively, and 2 other patients needed desmopressin after discharge and after sodium nadir developed. Hyponatremia was inversely associated with sellar access angle (p = 0.02) and the tumor cranial extension above the sellar floor showing a trend towards significance (p = 0.09). More than half of patients who had pituitary transposition developed transient hyponatremia. Hyponatremia was more common in those with narrower sellar access angle and smaller cranial extension of the tumor above the sellar floor. Anatomical/surgical parameters may allow risk-stratification for post-operative hyponatremia following pituitary transposition.


Patterns of E. coli leucine tRNA isoacceptors following bacteriophage MS2 infection.
The RNA extracted from MS2 phage particles can accept radioactive leucine and serine in the presence of tRNA activating enzymes. Leucine acceptance is due to the presence of E. coli leucine tRNA that binds very tightly to the virus particle. RPC-5 column chromatography shows that the pattern of virus associated leucyl-tRNA isoacceptors is different from that of normal E. coli leucyl-tRNA. It is also different from the pattern of host leucyl-tRNA isoacceptors found in E. coli lysate following MS2 phage infection. The RPC-5 pattern of the latter tRNA shows several new peaks of leucine tRNA isoacceptors. The possibility that these tRNAs are some modified forms of normal leucine tRNA isoacceptors is suggested.


Characterization of pediatric golf cart injuries to guide injury prevention efforts.
Golf cart injuries represent an increasing source of morbidity and mortality in the United States. Characterization of the circumstances of these injuries can inform injury prevention efforts. This study retrospectively reviews a prospective trauma registry at a level-one pediatric trauma center for golf cart-related injuries in patients under 18years of age admitted to the hospital between 2008 and 2016. The 40 identified crashes were associated with 82 hospital days, 17 ICU days, and more than $1 million in hospital charges over the study period. The median hospital stay was 1.5days, and the median hospital charge was $20,489. Severe injuries with an Injury Severity Score of >15 were identified in 25% of patients, and moderate injuries with scores between nine and 15 were identified in an additional 30%. The most common injures were head and neck (60%) and external injuries to the body surface (52.5%). Only a single child was wearing a seatbelt, and the vast majority was not using any safety equipment. Children as young as nine years old were driving golf carts, and child drivers were associated with the cart overturning (p=0.007). Golf cart crashes were a source of substantial morbidity at a level-one trauma center. Increased safety measures, such as higher hip restraints, seatbelts, and front-wheel breaks could substantially increase the safety of golf carts. Increased regulation of driving age as well as driver education may also reduce these injuries.


Evaluation of neuronal protective effects of xanthine oxidoreductase inhibitors on severe whole-brain ischemia in mouse model and analysis of xanthine oxidoreductase activity in the mouse brain.
Global cerebral ischemia and reperfusion (I/R) often result in high mortality. Free radicals play an important role in global cerebral I/R. Xanthine oxidoreductase (XOR) inhibitors, such as allopurinol, have been reported to protect tissues from damage caused by reactive oxygen species (ROS) by inhibiting its production through XOR inhibition. The recently introduced XOR inhibitor febuxostat, which is a more potent inhibitor than allopurinol, is expected to decrease free radical production more effectively. Here, we analyzed the effects of allopurinol and febuxostat in decreasing global severe cerebral I/R damage in mice. Mice were divided into three groups: a placebo group, an allopurinol group, and a febuxostat group. Pathological examinations, which were performed in each group in the CA1 and CA2 regions of the hippocampus 4 days after I/R surgery, revealed that there was a decrease in the number of neuronal cells in the 14-min occlusion model in both regions and that drugs that were administered to prevent this damage were not effective. The enzymatic activity was extremely low in the mouse brain, and XOR could not be detected in the nonischemic and ischemic mice brains with western blot analyses. Thus, one of the reasons for the decreased effectiveness of XOR inhibitors in controlling severe whole-brain ischemia in a mouse model was the low levels of expression of XOR in the mouse brain.


Nitrogenaseless mutants of Azotobacter vinelandii.
Mutants of Azotobacter which grow normally on excess ammonia under a variety of conditions and which grow slowly or not at all on atmospheric nitrogen have been isolated. Extracts of these strains have low or no detectable nitrogenase activity. There are three classes of mutants. Cell-free preparations of members of the first class possess an enhancement factor (EF+) which stimulates wild-type nitrogenase in vitro. Homogenates of members of the second class possess an enhanceable factor (EF-) which complements in vitro with extracts of the first class of mutants to give substantial nitrogenase activity. Preparations of members of the third class contain neither EF+ nor EF- activity. EF+ and EF- are repressed by the same conditions that repress nitrogenase. Molybdenum-deficient cells of the second class of mutants do not appear to contain EF- activity, but molybdenum deficient cells of the first class of mutants contain EF+. Because of these observations, EF+ is tentatively equated to azoferredoxin and EF- to molybdoferredoxin.


"Bubble-Diode" Breathable Electrodes for Fast Gas Transport.
Bubbles arising from wild gas evolution commonly exist in electrochemical systems, particularly in water electrolysis and rechargeable aqueous batteries (e.g., Zn-air batteries). Substantial energy dissipation occurs due to the obstruction of active sites and ion-conducting pathways by evolving bubbles. Efforts are made to elucidate effective strategies for fast gas transport, most of which focus on minimizing bubble size and facilitating their timely detachment through complex techniques such as constructing super-hydrophilic nano-structure electrodes, flowing electrolytes, and ultrasonic oscillation. Recently, an innovative, facile, and highly efficient method utilizing a breathable electrode design to promote gaseous molecules to the external environment emerges as a promising approach since it avoids remarkable bubble accumulation while remaining free of additional accessories. This concept highlights the origin and evolution of this promising design. Starting with introducing the basic concept of traditional breathable electrodes based on hydrophobic polymer networks and discussing the current progress made in underlying mechanisms, a detailed description of the advanced design inspired by a "bubble-diode" concept with superior breathability follows. This Concept aims to contribute to a deeper understanding of this technology and paves the way for further advancements in this renewable energy era.


The natural history of resected pancreatic cancer without adjuvant chemotherapy.
Diagnostic imaging, surgical care, and perioperative morbidity and mortality have significantly improved for patients undergoing resections for pancreatic adenocarcinoma. This study was undertaken to define the natural history and patterns of recurrence of resected pancreatic cancer without neoadjuvant or adjuvant therapies using current standards of care. Sixty-one patients underwent pancreatectomy without neoadjuvant or adjuvant therapy. Tumors were staged according to the American Joint Committee on Cancer (AJCC) classification system. CT scans were obtained every 3 months and recurrence categorized as: liver only, local, distant, multiple sites, or clinical. Median survival after pancreatectomy was 12 months. Cancer recurred in 51 (84%) patients. The radiographic site of initial recurrence did not generally impact survival; patients initially recurring at multiple sites had significantly abbreviated median survival of 5.6 months. AJCC stage was found to correlate with disease-free and overall survival, although tumor size alone did not. The presence of lymphatic metastasis correlated with disease-free but not overall survival. Overall survival after pancreaticoduodenectomy remains poor in the absence of neoadjuvant or adjuvant therapy. AJCC stage is the best predictor of disease-free and overall survival; tumor size, lymph node status, and site of recurrence alone do not impact survival in a meaningful way.


Quantitative determination of acemetacin and its metabolite indometacin in blood and plasma by column liquid chromatography.
A column liquid chromatographic (LC) method using UV detection for the determination of acemetacin and its metabolite indometacin in blood is described. The lower detection limit for both compounds is ca. 25 micrograms/l, the precision (coefficient of variation) is 6% for acemetacin and 10% for indometacin. The method is also suited for determination of both compounds in plasma, precisions in this case are even better than for blood, i.e. around 3% for both acemetacin and indometacin. Blood samples of three volunteers who had received 90 mg of acemetacin orally were analysed using the new method and very good agreement with results from a thin-layer chromatographic/fluorescence method was found.


Thrombotic Events in COVID-19 Are Associated With a Lower Use of Prophylactic Anticoagulation Before Hospitalization and Followed by Decreases in Platelet Reactivity.
Background: Coronavirus disease of 2019 (COVID-19) is associated with a prothrombotic state and a high incidence of thrombotic event(s) (TE). Objectives: To study platelet reactivity in hospitalized COVID-19 patients and determine a possible association with the clinical outcomes thrombosis and all-cause mortality. Methods: Seventy nine hospitalized COVID-19 patients were enrolled in this retrospective cohort study and provided blood samples in which platelet reactivity in response to stimulation with ADP and TRAP-6 was determined using flow cytometry. Clinical outcomes included thrombotic events, and all-cause mortality. Results: The incidence of TE in this study was 28% and all-cause mortality 16%. Patients that developed a TE were younger than patients that did not develop a TE [median age of 55 vs. 70 years; adjusted odds ratio (AOR) = 0.96 per 1 year of age, 95% confidence interval (CI) 0.92-1.00; p = 0.041]. Furthermore, patients using preexisting thromboprophylaxis were less likely to develop a thrombotic complication than patients that were not (18 vs. 54%; AOR = 0.19, 95% CI 0.04-0.84; p = 0.029). Conversely, having asthma strongly increased the risk on TE development (AOR = 6.2, 95% CI 1.15-33.7; p = 0.034). No significant differences in baseline P-selectin expression or platelet reactivity were observed between the COVID-19 positive patients (n = 79) and COVID-19 negative hospitalized control patients (n = 21), nor between COVID-19 positive survivors or non-survivors. However, patients showed decreased platelet reactivity in response to TRAP-6 following TE development. Conclusion: We observed an association between the use of preexisting thromboprophylaxis and a decreased risk of TE during COVID-19. This suggests that these therapies are beneficial for coping with COVID-19 associated hypercoagulability. This highlights the importance of patient therapy adherence. We observed lowered platelet reactivity after the development of TE, which might be attributed to platelet desensitization during thromboinflammation.


Inducible production of interleukin-6 by human polymorphonuclear neutrophils: role of granulocyte-macrophage colony-stimulating factor and tumor necrosis factor-alpha.
The recent demonstration of the ability of human polymorphonuclear neutrophils (PMN) to secrete various cytokines in response to the granulocyte activator granulocyte-macrophage colony-stimulating factor (GM-CSF) but not to other cytokines, has led to the identification of PMN as biosynthetically active cells. In this study we have investigated the ability of PMN to secrete interleukin-6 (IL-6), a molecule known to be involved in inflammatory reactions. Using RNA blotting analysis and bioassays, we show that PMN could be induced to synthesize transcripts specific for IL-6, indistinguishable in size from IL-6 mRNA produced by activated human macrophages. Consequently, PMN released IL-6-like activity into their culture supernatants that could be neutralized by monospecific anti-IL-6 antibody. Interleukin-6 secretion by PMN, however, required previous stimulation with GM-CSF or tumor necrosis factor-alpha (TNF-alpha), whereas other cytokines, including interleukin-3 (IL-3), granulocyte-CSF (G-CSF), macrophage-CSF (M-CSF), interferon gamma (IFN-gamma), and lymphotoxin (LT), failed to induce IL-6 mRNA accumulation and protein secretion by PMN. Similar to GM-CSF and TNF-alpha, other compounds, including the inhibitor of protein synthesis cyclohexemide (CHX), endotoxin (Escherichia coli-derived lipopolysaccharide), and phorbol myristate acetate (PMA) (but not the chemoattractant N-formyl-methionyl-leucyl-phenylalanine [FMLP]), induced detectable levels of IL-6 transcripts in PMN.


Epigenetic repression of E-cadherin by human papillomavirus 16 E7 protein.
A common feature shared between several human cancer-associated viruses, such as Epstein-Barr virus, Hepatitis B virus and Hepatitis C virus, and Human papillomavirus (HPV) is the ability to reduce the expression of cellular E-cadherin. Since E-cadherin is used by Langerhans cells to move through the stratified epithelium, its reduction may affect the efficiency by which the immune system responds to HPV infection and the length of persistent HPV infections. We observed that the E7 protein of this virus (HPV16) is most efficient at reducing E-cadherin levels. This E7 activity is independent of retinoblastoma protein or AP-2alpha degradation. Instead it is associated with augmentation of cellular DNA methyltransferase I (Dnmt1) activity. Significantly, inhibition of Dnmt activity re-established E-cadherin levels of the cells, presenting the possibility that similar epigenetic intervention clinically may be a way to re-establish the influx of Langerhans cells into infected epithelium to counteract HPV persistence.


Oocyte cytoplasmic maturation: a key mediator of oocyte and embryo developmental competence.
Efforts have intensified to successfully mature and inseminate oocytes in vitro and then culture ensuing embryos to transferable stages from a large number of mammalian species. Success varies, but generally even for the most successful species it is only possible to obtain a maximum of a 40 to 50% development of zygotes to the blastocyst stage. Reduced oocyte developmental competence is suggested as a primary reason for the reduced potential of in vitro-produced embryos. The vast majority of in vitro-matured oocytes are meiotically competent; however, many do not attain an optimal oocyte diameter before insemination. Variations in oocyte in vitro maturation media can influence embryo development, blastocyst cell number, and apoptosis. In addition, studies have indicated that cytoplasmic donation from so-called competent to incompetent oocytes can improve developmental outcomes. Oocyte cytoplasmic maturation includes those events that instill upon the oocyte a capacity to complete nuclear maturation, insemination, early embryogenesis and thus provide a foundation for implantation, initiation of pregnancy, and normal fetal development. Although we can define oocyte cytoplasmic maturation, we are only now beginning to understand the molecular steps that underlie this process. In general terms, oocyte cytoplasmic maturation involves the accumulation of mRNA, proteins, substrates, and nutrients that are required to achieve the oocyte developmental competence that fosters embryonic developmental competence. Collectively we are beginning to specify oocyte cytoplasmic maturation, and eventually a coherent understanding of this critical event in oocyte biology will emerge.


Message design strategies to raise public awareness of social determinants of health and population health disparities.
Raising public awareness of the importance of social determinants of health (SDH) and health disparities presents formidable communication challenges. This article reviews three message strategies that could be used to raise awareness of SDH and health disparities: message framing, narratives, and visual imagery. Although few studies have directly tested message strategies for raising awareness of SDH and health disparities, the accumulated evidence from other domains suggests that population health advocates should frame messages to acknowledge a role for individual decisions about behavior but emphasize SDH. These messages might use narratives to provide examples of individuals facing structural barriers (unsafe working conditions, neighborhood safety concerns, lack of civic opportunities) in efforts to avoid poverty, unemployment, racial discrimination, and other social determinants. Evocative visual images that invite generalizations, suggest causal interpretations, highlight contrasts, and create analogies could accompany these narratives. These narratives and images should not distract attention from SDH and population health disparities, activate negative stereotypes, or provoke counterproductive emotional responses directed at the source of the message. The field of communication science offers valuable insights into ways that population health advocates and researchers might develop better messages to shape public opinion and debate about the social conditions that shape the health and well-being of populations. The time has arrived to begin thinking systematically about issues in communicating about SDH and health disparities. This article offers a broad framework for these efforts and concludes with an agenda for future research to refine message strategies to raise awareness of SDH and health disparities.


Farm income and production impacts from the use of genetically modified (GM) crop technology 1996-2020.
This paper updates previous estimates for the global value of using genetically modified (GM) crop technology in agriculture at the farm level. It examined impacts on yields, important variable costs of production, including the cost of the technology, direct farm (gross) income, and impacts on the production base of the main crops where the technology is used (soybeans, corn, cotton, and canola). Over the period 1996 to 2020, the economic benefits have been significant with farm incomes for those using the technology having increased by $261.3 billion US dollars. This equates to an average farm income gain across all GM crops grown in this period of about $112/hectare. In 2020, the farm income gains were $18.8 billion (average of $103/ha). The cumulative farm income gains have been divided 52% to farmers in developing countries and 48% to farmers in developed countries. Seventy-two percentage of the gains have derived from yield and production gains with the remaining 28% coming from cost savings. These yield and production gains have made important contributions to increasing global production levels of the four main crops, having, for example, added 330 million tonnes and 595 million tonnes respectively, to the global production of soybeans and maize since the introduction of the technology in the mid-1990s. In 2020, the extra global production of the four main crops in which GM technology is widely used (85 million tonnes), would have, if conventional production systems been used, required an additional 23.4 million ha of land to be planted to these crops. In terms of investment, for each extra dollar invested in GM crop seeds (relative to the cost of conventional seed), farmers gained an average US $3.76 in extra income. In developing countries, the average return was $5.22 for each extra dollar invested in GM crop seed and in developed countries the average return was $3.00.


Phylogenetic characterization of 'Candidatus Helicobacter bovis', a new gastric helicobacter in cattle.
Recently helicobacter-like organisms have been reported in the pyloric part of the abomasum of calves and adult cattle. Cultivation of these spiral bacteria has not been successful to date. In the present study, comparative 16S rDNA sequence analysis was used to determine the taxonomic position of these bacteria. Seven abomasal biopsies of adult cattle were sampled from different Belgian and Dutch farms. In all samples the presence of helicobacter-like organisms was demonstrated by biochemical, immunohistochemical and electron microscopical data. Bacterial 16S rDNA was amplified by PCR and sequences were determined either by direct or indirect sequence analysis. Pairwise comparisons revealed all sequences to be more than 99% homologous. Phylogenetic analysis placed the organism, corresponding to the reference sequence R2XA, within the genus Helicobacter. A diagnostic PCR assay was designed, differentiating all of the bovine 16S rDNA sequences from Helicobacter and Wolinella species. The low similarity level towards Helicobacter bilis (92.8%), its closest validly named neighbour, indicates that this novel taxon is indeed a novel Helicobacter species. An in situ hybridization procedure associated the bovine sequences to the helicobacter-like organisms in the abomasum. The name 'Candidatus Helicobacter bovis' is proposed for this new abomasal helicobacter from cattle.


Advanced cytologic approaches for the diagnosis of pancreatobiliary cancer.
The review will provide information on the use of advanced cytologic techniques such as fluorescence in-situ hybridization (FISH) and digital image analysis (DIA) for the detection of pancreatobiliary malignancy. FISH and DIA are techniques that can be used to assess cells for chromosomal alterations. Since tumor cells are frequently aneuploid, FISH and DIA can be used to detect malignancy. Recent studies have shown that the application of FISH and DIA to pancreatobiliary brushing specimens increases the sensitivity of detection of malignancy over conventional cytology. The diagnosis of malignancy in the pancreatobiliary tract confers a bleak prognosis due to the aggressive nature of these tumors and ineffective treatment options. Early detection of tumor is more likely to provide the patient with the option of surgical resection, which offers the best chance for cure. Current pathologic testing techniques, namely biopsy and routine cytology, are limited in their ability to detect tumor. FISH and DIA increase the sensitivity of pancreatobiliary malignancy compared to routine cytology.


The Harvard Trauma Questionnaire. Validating a cross-cultural instrument for measuring torture, trauma, and posttraumatic stress disorder in Indochinese refugees.
There are no valid and reliable cross-cultural instruments capable of measuring torture, trauma, and trauma-related symptoms associated with the DSM-III-R diagnosis of posttraumatic stress disorder (PTSD). Generating such standardized instruments for patients from non-Western cultures involves particular methodological challenges. This study describes the development and validation of three Indochinese versions of the Harvard Trauma Questionnaire (HTQ), a simple and reliable screening instrument that is well received by refugee patients and bicultural staff. It identifies for the first time trauma symptoms related to the Indochinese refugee experience that are associated with PTSD criteria. The HTQ's cultural sensitivity may make it useful for assessing other highly traumatized non-Western populations.


Management and investigation of neonatal encephalopathy: 2017 update.
This review discusses an approach to determining the cause of neonatal encephalopathy, as well as current evidence on resuscitation and subsequent management of hypoxic-ischaemic encephalopathy (HIE). Encephalopathy in neonates can be due to varied aetiologies in addition to hypoxic-ischaemia. A combination of careful history, examination and the judicious use of investigations can help determine the cause. Over the last 7 years, infants with moderate to severe HIE have benefited from the introduction of routine therapeutic hypothermia; the number needed to treat for an additional beneficial outcome is 7 (95% CI 5 to 10). More recent research has focused on optimal resuscitation practices for babies with cardiorespiratory depression, such as delayed cord clamping after establishment of ventilation and resuscitation in air. Around a quarter of infants with asystole at 10 min after birth who are subsequently cooled have normal outcomes, suggesting that individualised decision making on stopping resuscitation is needed, based on access to intensive treatment unit and early cooling. The full benefit of cooling appears to have been exploited in our current treatment protocols of 72 hours at 33.5°C; deeper and longer cooling showed adverse outcome. The challenge over the next 5-10 years will be to assess which adjunct therapies are safe and optimise hypothermic brain protection in phase I and phase II trials. Optimal care may require tailoring treatments according to gender, genetic risk, injury severity and inflammatory status.


Tissue Regeneration from Mechanical Stretching of Cell-Cell Adhesion.
Cell-cell adhesion complexes are macromolecular adhesive organelles that integrate cells into tissues. This mechanochemical coupling in cell-cell adhesion is required for a large number of cell behaviors, and perturbations of the cell-cell adhesion structure or related mechanotransduction pathways can lead to critical pathological conditions such as skin and heart diseases, arthritis, and cancer. Mechanical stretching has been a widely used method to stimulate the mechanotransduction process originating from the cell-cell adhesion and cell-extracellular matrix (ECM) complexes. These studies aimed to reveal the biophysical processes governing cell proliferation, wound healing, gene expression regulation, and cell differentiation in various tissues, including cardiac, muscle, vascular, and bone. This review explores techniques in mechanical stretching in two-dimensional settings with different stretching regimens on different cell types. The mechanotransduction responses from these different cell types will be discussed with an emphasis on their biophysical transformations during mechanical stretching and the cross talk between the cell-cell and cell-ECM adhesion complexes. Therapeutic aspects of mechanical stretching are reviewed considering these cellular responses after the application of mechanical forces, with a focus on wound healing and tissue regeneration. Impact Statement Mechanical stretching has been proposed as a therapeutic option for tissue regeneration and wound healing. It has been accepted that mechanotransduction processes elicited by mechanical stretching govern cellular response and behavior, and these studies have predominantly focused on the cell-extracellular matrix (ECM) sites. This review serves the mechanobiology community by shifting the focus of mechanical stretching effects from cell-ECM adhesions to the less examined cell-cell adhesions, which we believe play an equally important role in orchestrating the response pathways.


Rendez-vous at mitosis: TRRAPed in the chromatin.
Cell cycle progression and cell cycle checkpoints are guided by dynamic changes in gene expression that requires concerted efforts of chromatin modifying/remodeling activities and transcription machinery. Epigenetic modifications including acetylation of specific lysine residues within the amino-terminal tails of core histones play an important role in these processes. In the last few years, a flurry of biochemical studies has identified numerous histone acetyltransferases (HAT) whose activity is dependent on the multiprotein assemblies and responsible for histone acetylation. In addition to their well-known involvement in the control of gene transcription, recent studies implicated HATs and histone acetylation in other important cellular processes, such as DNA replication, cell cycle control, DNA repair and genomic stability. With the exception of catalytic subunits of the HAT assemblies, the role of other components of these large multi-subunit complexes in cellular processes remains largely unknown. Recent genetic and cellular studies have shown that Trrap, a common component of HAT complexes, regulates the mitotic checkpoint function by modulation of mitotic checkpoint genes. This regulation involves a concerted and cell cycle stage-coupled recruitment of HAT activity to promoters of specific checkpoint genes, providing a functional link between specific chromatin modifications and cell cycle control. These findings shed new light on the role of HAT components and histone acetylation in cell cycle control and underscore functional significance of epigenetic modifications in cellular processes.


A translating-bed technique for total-body irradiation.
Total-body irradiation (TBI) is a therapy modality that is being used with increasing frequency, in conjunction with chemotherapy, for patients undergoing bone-marrow transplantation. At the Ottawa Regional Cancer Centre a technique has been developed for the delivery of TBI to patients prior to bone-marrow transplantation. In this technique patients are treated on a mobile couch at approximately 195 cm SSD with a field size of 66.5 cm wide by 57 cm long. A computer-controlled stepping motor drives the patient couch at a user-selectable speed. The total dose delivered to the patient is a function of couch velocity, field size and patient separation. Treatment times are of the order of 10 min for each of the anterior and posterior fields for a 400 cGy fraction. It has been found that the conventional central axis tissue maximum ratio (TMR) and percentage depth dose (PDD) functions are not appropriate for describing dose delivered during dynamic treatment. To this end we have developed dynamic TMR and PDD functions. Extensive measurements have been performed in an anthropomorphic water phantom to determine the dose distributions in three dimensions and the efficacy of polymethyl methacrylate (PMMA) beam spoilers as a replacement for anterior and lateral bolus. It has been found that 2.4 cm PMMA spoilers do provide full skin dose and negate the requirement for lateral bolus. This TBI procedure is simple, rapid and appears to be well tolerated by the patients. 55 patients have been treated since the introduction of this technique in 1991.


Resident training in pediatric critical care transport medicine: a survey of pediatric residency programs.
The Accreditation Council for Graduate Medical Education (ACGME) Program Requirement for Pediatrics includes specific objectives that pediatric residents participate in both the pre-hospital care of acutely ill or injured patients and the stabilization and transport of patients to critical care areas. Previously, residents were often included as the physician component for many pediatric critical care transport teams. Subsequent regionalization of transport services and development of nurse-only transport teams prompted us to determine the current level of resident participation in pediatric critical care transport as well as how individual residency programs were meeting the educational objectives. A questionnaire was mailed to each pediatric residency program listed in the 1996-1997 GME Directory. Information was obtained regarding the size of the hospital and the residency program, the presence of a pediatric critical care transport team, the number of annual transports, and transport team leader. In addition, the use of pediatric residents for transports was ascertained, as well as their specific role, training requirements, and method of evaluation. Data were received from 138 programs for a return rate of 65%. Eighty percent of programs offered a pediatric critical care transport service. Nurse-led teams were used for 51% of NICU and 44% of PICU transports. Of the 82 NICU and 84 PICU teams that used residents, the majority used them as team leaders (60% and 70%; respectively) with only the minority requiring that they be at the PL-3 year or greater. The training and/or certification required for resident participation in transports varied among programs, with 85% requiring completion of a NICU or PICU rotation, and 94% requiring NRP or PALS certification. Programs that did not allow resident participation provided exposure to Transport Medicine by various mechanisms, including lectures and emergency department (ED) rotations. Pediatric resident participation in critical care transport varies widely among pediatric critical care transport teams. The degree to which residents participate in the transport team would appear to have diminished in comparison to previous studies. Transport teams often use other resources, such as nurses, fellows, or attendings, to lead their transport teams. Pediatric resident exposure to and participation in Transport Medicine varies among programs, as do the methods used to prepare residents for their experience.


Long-term depression of presynaptic cannabinoid receptor function at parallel fibre synapses.
Inhibition of synaptic responses by activation of presynaptic cannabinoid type-1 (Cb1) receptors is reduced at parallel fibre synapses in the cerebellum following 4 Hz stimulation. Activation of adenylyl cyclase is necessary and sufficient for down-regulation of Cb1 receptors induced by 4 Hz stimulation. 4 Hz stimulation reduces Cb1 receptor function by (i) increasing the rate of endocannabinoid clearance from the synapse and (ii) decreasing expression of Cb1 receptors. Cannabinoid type-1 receptors (Cb1R) are expressed in the presynaptic membrane of many synapses, including parallel fibre-Purkinje cell synapses in the cerebellum, where they are involved in short- and long-term plasticity of synaptic responses. We show that Cb1R expression itself is a plastic property of the synapse regulated by physiological activity patterns. We made patch clamp recordings from Purkinje cells in cerebellar slices and assessed Cb1R activity by measuring depolarization-induced suppression of excitation (DSE). We find that DSE is normally stable at parallel fibre synapses but, following 4 Hz stimulation, DSE is persistently reduced and recovers more rapidly. Using a combination of electrophysiology, pharmacology and biochemistry, we show that changes in DSE are a result of the reduced expression of Cb1Rs and increased degradation of endocannabinoids by monoacylglycerol lipase. Long-term changes in presynaptic Cb1R expression may alter other forms of Cb1R-dependent plasticity at parallel fibre synapses, priming or inhibiting the circuit for associative learning.


Observation of Oscillatory Radial Electric Field Relaxation in a Helical Plasma.
Measurements of the relaxation of a zonal electrostatic potential perturbation in a nonaxisymmetric magnetically confined plasma are presented. A sudden perturbation of the plasma equilibrium is induced by the injection of a cryogenic hydrogen pellet in the TJ-II stellarator, which is observed to be followed by a damped oscillation in the electrostatic potential. The waveform of the relaxation is consistent with theoretical calculations of zonal potential relaxation in a nonaxisymmetric magnetic geometry. The turbulent transport properties of a magnetic confinement configuration are expected to depend on the features of the collisionless damping of zonal flows, of which the present Letter is the first direct observation.


Quality in anesthesia care: lessons from industry and a proposal for valid measurement and improvement.
Quality anesthetic care is a goal fundamental to our tradition and our training, but defining and measuring quality in anesthesia presents special challenges. Industrial models of quality, especially those so fundamental to the re-emergence of post-war Japan, deserve careful study and are discussed at some length, but they clearly have limitations in understanding quality in anesthesiology. We suggest that most current quality efforts are inherently flawed. Whether or not they rigorously attempt to define quality, they are hampered by lack of data concerning outcomes and alternatives, as well as lack of distinction between quality and efficacy. Quality efforts in American medicine and anesthesiology seem mired in a "criterion of potential benefit," which is still central to many of our prescriptions for individual medical care. Current quality improvement efforts do not seem well suited to correct these flaws. Anesthetic care, and that of American medicine in general, is fragmented, enormously costly, and sometimes inappropriate or poor. Anesthesiologists are suspicious of current quality efforts to improve this care. The system often seems more geared to eliminate bad apples than to improve patient care. Because anesthesia is a specialty that facilitates care but seldom "cures," we face greater challenges in studying and defining quality than do other specialties. Because of this, it is imperative that several principles govern future quality improvement efforts in anesthesiology. First, a reasonable balance must be attained between study of outcomes and processes of anesthesia care. Second, anesthesia-specific severity of illness indexing must be developed. Third, and perhaps most important, anesthetic processes and outcomes must be reported on a national level. Fundamental to future quality efforts in our specialty, we believe, is the establishment of a protected National Anesthesia Outcome Registry. This article reviews the industrial and medical history of quality, its measurement and improvement, and attempts to apply principles learned over many decades to anesthesiology.


Pripper: prediction of caspase cleavage sites from whole proteomes.
Caspases are a family of proteases that have central functions in programmed cell death (apoptosis) and inflammation. Caspases mediate their effects through aspartate-specific cleavage of their target proteins, and at present almost 400 caspase substrates are known. There are several methods developed to predict caspase cleavage sites from individual proteins, but currently none of them can be used to predict caspase cleavage sites from multiple proteins or entire proteomes, or to use several classifiers in combination. The possibility to create a database from predicted caspase cleavage products for the whole genome could significantly aid in identifying novel caspase targets from tandem mass spectrometry based proteomic experiments. Three different pattern recognition classifiers were developed for predicting caspase cleavage sites from protein sequences. Evaluation of the classifiers with quality measures indicated that all of the three classifiers performed well in predicting caspase cleavage sites, and when combining different classifiers the accuracy increased further. A new tool, Pripper, was developed to utilize the classifiers and predict the caspase cut sites from an arbitrary number of input sequences. A database was constructed with the developed tool, and it was used to identify caspase target proteins from tandem mass spectrometry data from two different proteomic experiments. Both known caspase cleavage products as well as novel cleavage products were identified using the database demonstrating the usefulness of the tool. Pripper is not restricted to predicting only caspase cut sites, but it gives the possibility to scan protein sequences for any given motif(s) and predict cut sites once a suitable cut site prediction model for any other protease has been developed. Pripper is freely available and can be downloaded from http://users.utu.fi/mijopi/Pripper. We have developed Pripper, a tool for reading an arbitrary number of proteins in FASTA format, predicting their caspase cleavage sites and outputting the cleaved sequences to a new FASTA format sequence file. We show that Pripper is a valuable tool in identifying novel caspase target proteins from modern proteomics experiments.


A generalized solid strengthening rule for biocompatible Zn-based alloys, a comparison with Mg-based alloys.
Solid solution strengthening has been widely used in designing various high-performance biocompatible Mg-based alloys, but its transferability to other biocompatible metals such as Zn-based alloys is questionable or nearly absent. In the present study, an ab initio informed Peierls-Nabarro model and Leyson et al.'s strengthening model are used for a systematic investigation on solute strengthening in Zn-based alloys, which is compared with the widely studied Mg-based alloys. Although an inverse relationship was revealed between volume misfit εb and chemical misfit εSFE for both Zn-based and Mg-based alloys, most solutes would however result in positive εb and negative εSFE for Zn-based alloys, differing from Mg-based alloys. With εb and εSFE as two key descriptors, a generalized scaling diagram is finally drawn for a fast evaluation of solid solution strengthening in Zn-based alloys, indicating that the alkaline-earth and rare earth elements are better strengtheners for Zn-based alloys, which provides a general rule in designing novel biocompatible materials.


The timecourse of apoptotic cell death during postnatal remodeling of the mouse cochlea and its premature onset by triiodothyronine (T3).
Apoptosis underlies various forms of tissue remodeling during development. Prior to the onset of hearing, thyroid hormone (T3) promotes cochlear remodeling, which involves regression of the greater epithelial ridge (GER), a transient structure of columnar cells adjacent to the mechanosensory hair cells. We investigated the timecourse of apoptosis in the GER and the influence of ectopic T3 on apoptosis. In saline-treated mice, activated caspase 3-positive cells were detected in the GER between postnatal days 7 and 13 and appeared progressively along the cochlear duct from base to apex over developmental time. T3 given on P0 and P1 advanced the overall program of apoptosis and remodeling by ~4 days. Thyroid hormone receptor β was required for these actions, suggesting a receptor-mediated process of initiation of apoptosis. Finally, T3 given only at P0 or P1 resulted in deafness in adult mice, thus revealing a transient period of susceptibility to long-term damage in the neonatal auditory system.


Brillouin microscopy of collagen crosslinking: noncontact depth-dependent analysis of corneal elastic modulus.
Corneal collagen crosslinking (CXL) is designed to halt the progression of keratoconus and corneal ectasia by inducing corneal stiffening. However, it currently is difficult to monitor and evaluate CXL outcome objectively due to the lack of suitable methods to characterize corneal mechanical properties. We validated noncontact Brillouin microscopy to quantify corneal mechanical properties before and after CXL. CXL was performed on fresh porcine eyes using various presoaking times and light doses, with or without epithelial debridement. From Brillouin maps of corneal elastic modulus, stiffness and average modulus of anterior, middle, and posterior stroma were analyzed. Corneal stiffening index (CSI) was introduced as a metric to compare the mechanical efficacy of a given CXL protocol with respect to the standard protocol (30-minute riboflavin presoak, 3 mW/cm² ultraviolet illumination for 30 minutes). Brillouin corneal stiffness increased significantly (P < 0.001) by epi-off and epi-on CXL. The increase of Brillouin modulus was depth-dependent, indicating that anterior stromal stiffening contributes the most to mechanical outcome. The increase of anterior Brillouin modulus was linearly proportional to the light dose (R² > 0.98). Compared to the standard epi-off procedure, a typical epi-on procedure resulted in a third of stiffness increase in porcine corneas (CSI = 33). Brillouin microscopy allowed imaging and quantifying CXL-induced mechanical changes without contact in a depth-dependent manner at high spatial resolution. This technique may be useful to evaluate the mechanical outcomes of CXL procedures, to compare different crosslinking agents, and for real-time monitoring of CXL in clinical and experimental settings.


The association between nut consumption and the risk of total and ischemic stroke in a German cohort study.
Nuts have beneficial effects on coronary heart disease and many cardiovascular risk factors. However, their effect on stroke is less established, and no studies on the topic are available in Northern and Central European populations. Therefore, we aimed at investigating the association between nut consumption and the risk of stroke in a German population. We used data from a prospective cohort of 26,285 participants of the European Prospective Investigation into the Cancer and Nutrition Potsdam Study. During a median follow-up time of 8.3 years (interquartile range: 7.5-9.2), 288 incident cases of stroke occurred. Nut consumption (standard portion size of 50 g) was assessed at baseline with a semiquantitative food-frequency questionnaire. The median nut intake was 0.82 g per day, interquartile range: 0.41-4.11. In the multivariable model, an increased risk of stroke was observed among participants who never consumed nuts (hazard ratio (HR): 1.56, 95% confidence interval: 1.17-2.08), compared with those consuming <½ portion/week. However, there was no evidence of a dose-response relationship between nut consumption and stroke. Compared with those who consumed <½ portion/week, the multivariable HR for total stroke was 1.06 (0.75-1.52) among those who consumed ½ to 1 portion/week and 1.37 (0.92-2.05) for those who consumed >1 portion/week. Similar nonsignificant associations were observed in stratified analysis for gender, or for fatal and nonfatal stroke. We could not observe an association between nut consumption and the risk of developing stroke (fatal/nonfatal) in a population with low habitual nut consumption.


HPMA copolymer-anticancer drug-OV-TL16 antibody conjugates. 1. influence of the method of synthesis on the binding affinity to OVCAR-3 ovarian carcinoma cells in vitro.
The influence of different methods of binding the OV-TL16 antibody and its Fab' fragment to N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer--drug (adriamycin [ADR] or meso chlorin e6 mono(N-2-aminoethylamide) (Mce6)) conjugates on the affinity of conjugates to an ovarian carcinoma (OVCAR-3) cell associated antigen was investigated. The binding of the antibody to HPMA copolymer--drug (ADR or Mce6) conjugates via amino groups resulted in conjugates which were heterogeneous in their antigen binding. Coupling, the HPMA copolymer--Mce6 conjugate to the carbohydrate region of the antibody resulted in conjugates with a more homogeneous distribution of affinity constants than conjugates prepared by linking the antibody to the polymer via amino groups. However, both methods resulted in a decrease in the affinity constant compared to the native antibody. Conjugates prepared with the Fab' frgment of the OV-TL16 antibody demonstrated a more homogenous affinity than either conjugate prepared with the whole antibody. To verify the hypothesis that the changes in the binding affinity and homogeneity are a consequence of conformational changes in the antibody structure, a series of physiocochemical methods were employed to characterize the conjugates. The excitation energy transfer between OV-TL16 antibody and drugs (ADR and Mce6) and the spectral properties of Mce6 were used to monitor the interactions between the antibody and drugs. The quenching of the intrinsic fluorescence of the antibody was also employed to study its conformational changes. An attempt has been made to correlate the biorecognition at the cellular surface with the interactions of drug with the antibody molecule and with the changes in antibody conformation.


Acute simvastatin inhibits K ATP channels of porcine coronary artery myocytes.
Statins (3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitors) consumption provides beneficial effects on cardiovascular systems. However, effects of statins on vascular KATP channel gatings are unknown. Pig left anterior descending coronary artery and human left internal mammary artery were isolated and endothelium-denuded for tension measurements and Western immunoblots. Enzymatically-dissociated/cultured arterial myocytes were used for patch-clamp electrophysiological studies and for [Ca(2+)]i, [ATP]i and [glucose]o uptake measurements. The cromakalim (10 nM to 10 µM)- and pinacidil (10 nM to 10 µM)-induced concentration-dependent relaxation of porcine coronary artery was inhibited by simvastatin (3 and 10 µM). Simvastatin (1, 3 and 10 µM) suppressed (in okadaic acid (10 nM)-sensitive manner) cromakalim (10 µM)- and pinacidil (10 µM)-mediated opening of whole-cell KATP channels of arterial myocytes. Simvastatin (10 µM) and AICAR (1 mM) elicited a time-dependent, compound C (1 µM)-sensitive [(3)H]-2-deoxy-glucose uptake and an increase in [ATP]i levels. A time (2-30 min)- and concentration (0.1-10 µM)-dependent increase by simvastatin of p-AMPKα-Thr(172) and p-PP2A-Tyr(307) expression was observed. The enhanced p-AMPKα-Thr(172) expression was inhibited by compound C, ryanodine (100 µM) and KN93 (10 µM). Simvastatin-induced p-PP2A-Tyr(307) expression was suppressed by okadaic acid, compound C, ryanodine, KN93, phloridzin (1 mM), ouabain (10 µM), and in [glucose]o-free or [Na(+)]o-free conditions. Simvastatin causes ryanodine-sensitive Ca(2+) release which is important for AMPKα-Thr(172) phosphorylation via Ca(2+)/CaMK II. AMPKα-Thr(172) phosphorylation causes [glucose]o uptake (and an [ATP]i increase), closure of KATP channels, and phosphorylation of AMPKα-Thr(172) and PP2A-Tyr(307) resulted. Phosphorylation of PP2A-Tyr(307) occurs at a site downstream of AMPKα-Thr(172) phosphorylation.


Triple antiplatelet therapy after PCI--should or could it be done?
In case of coronary stenting in patients taking long-term oral anticoagulants (OAC), addition of both aspirin and clopidogrel in the drug regimen (so-called triple antiplatelet therapy) is recommended. However such triple therapy increases the risk of serious bleeding events. Comparative data on the efficacy and safety of the triple therapy versus therapy with a single antiplatelet agent and oral anticoagulants in case of coronary stenting are very rare. Most studies show a decreased stroke and myocardial infarction risk, but an increased risk of bleeding events in case of triple therapy. There is general agreement that, when possible, the duration of triple therapy should be shortened followed by clopidogrel and an oral anticoagulant to minimize bleeding risk without increasing other adverse events. In patients with a high risk of bleeding, BMS should be used and triple therapy should be applied for only 1 month, followed by one antiplatelet agent and oral anticoagulant. The WOEST study was the first study showing that therapy with clopidogrel and OAC is safe and reduces bleeding and mortality more effectively than triple therapy including aspirin in patients undergoing coronary stenting. Although the risk of bleeding increases with triple therapy as compared to OAC plus a single antiplatelet agent, the second treatment regimen cannot be recommended to all patients. However for those at the highest risk of bleeding it is not unreasonable to consider. Additional randomized studies are needed for the implementation of future treatment guidelines in patients with high risk for bleeding and thrombotic complications.


Comparative Flavivirus-Host Protein Interaction Mapping Reveals Mechanisms of Dengue and Zika Virus Pathogenesis.
Mosquito-borne flaviviruses, including dengue virus (DENV) and Zika virus (ZIKV), are a growing public health concern. Systems-level analysis of how flaviviruses hijack cellular processes through virus-host protein-protein interactions (PPIs) provides information about their replication and pathogenic mechanisms. We used affinity purification-mass spectrometry (AP-MS) to compare flavivirus-host interactions for two viruses (DENV and ZIKV) in two hosts (human and mosquito). Conserved virus-host PPIs revealed that the flavivirus NS5 protein suppresses interferon stimulated genes by inhibiting recruitment of the transcription complex PAF1C and that chemical modulation of SEC61 inhibits DENV and ZIKV replication in human and mosquito cells. Finally, we identified a ZIKV-specific interaction between NS4A and ANKLE2, a gene linked to hereditary microcephaly, and showed that ZIKV NS4A causes microcephaly in Drosophila in an ANKLE2-dependent manner. Thus, comparative flavivirus-host PPI mapping provides biological insights and, when coupled with in vivo models, can be used to unravel pathogenic mechanisms.


Zingerone (4-(4-hydroxy-3-methylphenyl) butan-2-one) protects against alloxan-induced diabetes via alleviation of oxidative stress and inflammation: Probable role of NF-kB activation.
Diabetes is considered as the most common metabolic disease affecting millions of people all around the world. Use of natural herbal medicines can be effective in treating diabetes. Zingerone (4-(4-hydroxy-3-methylphenyl) butan-2-one) a polyphenolic alkanone extracted from ginger has a broad spectrum of pharmacological properties and thus can be used as a promising candidate against various ailments. In the current study we aimed at demonstrating the protective effect of zingerone against diabetes mellitus and elucidating its possible mechanism. Five groups of animals (I-V) were made with ten animals each. Group I (control) was given normal saline orally. Group II (diabetic positive control) was given alloxan at the dose rate of 100 mg/kg bwt once. Group III and IV was given alloxan once at the dose rate of 100 mg/kg bwt. and received oral treatment of zingerone at a dose rate of 50 and 100 mg/kg bwt respectively daily for 21 days. Group V was given alloxan at the dose of 100 mg/kg bwt. and was treated with standard drug glibenclamide at the dose rate of 4.5 mg/kg bwt. daily for 21 days. According to our findings we confirmed that zingerone restrained the alloxan induced oxidative stress by increasing the activity of reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and reducing the peroxidative damage. We also confirmed that zingerone suppressed the level of redox sensitive transcription factor NFκB and downregulated other downstream inflammatory cytokines like interleukins (IL1-β IL-2, IL-6) and tumor necrosis factor alpha (TNF-α). Moreover, the experimental findings suggested that zingerone improved the insulin levels. Taken together our results indicated that zingerone effectively ameliorated the diabetes induced complications which provide a strong theoretical basis for zingerone to be used clinically for treatment of diabetes.


Validity and reliability of the Italian version of the ECOS-16 questionnaire in postmenopausal women with prevalent vertebral fractures due to osteoporosis.
To investigate the reliability and validity of the Italian version of ECOS-16 (Assessment of health related quality of life in osteoporosis) in comparison to other questionnaires in postmenopausal women with osteoporosis. A cross-sectional multicentre study was carried out among postmenopausal women with osteoporosis who were attending primary care centres and hospital outpatient clinics. The patient group included 234 females (mean age 69 years, range 48-89) who presented vertebral fractures due to osteoporosis. The control group consisted of 244 asymptomatic osteoporotic subjects matched for age with the patient group. The psychometric properties of the questionnaires were evaluated in terms of feasibility, validity (construct validity and discriminant validity) and internal consistency. Test-retest reliability was analysed for 196 outpatients who reported that their general health status due to osteoporosis had not changed after one week. In all patients the ECOS-16, the SF-36 (Medical Outcomes Study Short Form-36), EUROQoL (EQ-5D), mini-OQLQ (mini-Osteoporosis Quality of Life Questionnaire), and RMDQ (Roland-Morris Disability Questionnaire) were administered, and all clinical variables and sociodemographic variables were taken into account. Construct and discriminant validity were assessed by Spearman's correlations, the Wilcoxon rank sum test, the Kruskal Wallis test and by receiver operating characteristic (ROC) curves. Internal consistency was evaluated using Cronbach's alpha and the test-retest reliability was evaluated by intra-class correlation coefficients (ICCs). 96.9% of the patients answered all items of the ECOS-16 questionnaires. The mean administration time was 10 minutes. Factor analysis yielded two factors that accounted for 88.4% of the explained variance in the ECOS-16 questionnaire. The first factor was the ECOS-16 Physical Component Score (PCS) (45.9% of the explained variance) and the second factor was the ECOS-16 Mental Component Score (MCS) (42.4% of the explained variance). The inter-item correlation between the two factors was 0.48. Significant correlations were found between the scores of similar domains or subscales of the ECOS-16 and SF-36, EQ-5D and mini-OQLQ, supporting the concept of convergent construct validity. The total ECOS-16 score progressively increased with the number of prevelant vertebral fractures (p<0.001) and the effect of the first fracture was already statistically significant (p<0.01). On ROC curve analysis the total ECOS-16 score showed the highest performance among the different questionnaires in discriminating between patients with vertebral fractures and controls with no fractures. In the reliability study, internal consistency within the domain of ECOS-16 was generally good, with Chronbach's alpha values ranging from 0.81 to 0.89. Test-retest reliability was 0.87 for the total ECOS-16 score. The Italian version of the ECOS-16 questionnaire was demonstrated to have good psychometric properties and could offer a useful tool in research and routine clinical practice to evaluate HRQoL in post-menopausal women with osteoporosis. A full validation of the psychometric properties will require data on its sensitivity to change.


Preoperative cystoscopic findings in resectable rectal adenocarcinoma.
Forty-nine patients underwent preoperative cystoscopy prior to surgical resection for adenocarcinoma of the rectum. There were 30 men and 19 women ranging in age from 26 to 78 years, with a median age of 59 years. Surgical resections consisted of abdominoperineal resection, posterior exenteration, or total pelvic exenteration. Twenty patients had positive findings on preoperative cystoscopy while 29 had negative findings. Of the 20 patients with positive cystoscopic findings, eight (40%) had either urologic symptoms or abnormal microscopic findings on urinalysis; however, only one of these eight patients had tumor invasion of the bladder wall at cystoscopy, while the remaining seven had associated benign conditions. In the 29 cystoscopy procedures in which findings were negative, only three patients (10%) had urologic symptoms or abnormal results of urinalysis, none of which were secondary to the rectal carcinoma. We conclude that urinary symptoms can be misleading and unreliable when predicting neoplastic involvement of the bladder by carcinoma of the rectum because of the high incidence of associated benign disorders of the bladder. Therefore, cystoscopy does not have a useful role in staging patients undergoing surgery for rectal carcinoma in terms of local tumor extension.


The glycan code of the endoplasmic reticulum: asparagine-linked carbohydrates as protein maturation and quality-control tags.
The majority of proteins that traverse the secretory pathway receive asparagine (Asn)-linked glycosylations. Glycans are bulky hydrophilic modifications that serve a variety of structural and functional roles within the cell. Here, we review the recent growing knowledge on the role of Asn-linked glycans as maturation and quality-control protein tags in the early secretory pathway. The carbohydrate composition encodes crucial information about the structure, localization and age of glycoproteins. The "glycan code" is encoded by a series of glycosidases and carbohydrate transferases that line the secretory pathway. This code is deciphered by carbohydrate-binding proteins that possess distinct carbohydrate binding properties and act as molecular chaperones or sorting receptors. These glycosidases and transferases work in concert with resident secretory pathway carbohydrate-binding proteins to form a network that assists in the maturation and trafficking of both native and aberrant glycoproteins within the cell.


Adipose stem cell sheets improved cardiac function in the rat myocardial infarction, but did not alter cardiac contractile responses to β-adrenergic stimulation.
Adipose stem cells (ASCs) are a source of regenerative cells available for autologous transplantation to hearts. We compared protective actions of ASC sheets on rat myocardial infarction (MI) in comparison with those of skeletal myoblast cell sheets. Their effects on infarcted hearts were evaluated by biological, histochemical as well as physiological analyses. ASC sheets secreted higher concentrations of angiogenic factors (HGF, VEGF, and bFGF; P < 0.05) under normoxic and hypoxic conditions than those of myoblast cell sheets, associated with reduction of cell apoptosis (P < 0.05). Like myoblast cell sheets, ASC sheets improved cardiac function (P < 0.05) and decreased the plasma level of ANP (P < 0.05) in MI hearts. ASC sheets restored cardiac remodeling characterized by fibrosis, cardiac hypertrophy and impaired angiogenesis (P < 0.05), which was associated with increases in angiogenic factors (P < 0.05). In isolated perfused rat hearts, ASC sheets improved both systolic and diastolic functions, which was comparable to cardiac functions of myoblast cell sheets, while both cell sheets failed to restore cardiac contractile response to either isoproterenol, pimobendan or dibutyryl cAMP. These results indicated that ASC sheets improved cardiac function and remodeling of MI hearts mediated by their paracrine action and this improvement was comparable to those by myoblast cell sheets.


Atomic resolution imaging using the real-space distribution of electrons scattered by a crystalline material.
We present an alternative atomic resolution incoherent imaging technique derived from scanning transmission electron microscopy (STEM) using detectors in real space, in contrast to conventional STEM that uses detectors in diffraction space. The images obtained from various specimens have a resolution comparable to conventional high-angle annular dark-field (HAADF) STEM with good contrast, which seems to be very robust with respect to thickness, focus and imaging conditions. The results of the simulations are consistent with the experimental results and support the interpretation of the real-space STEM image contrast as being a result of aberration-induced displacements of the high-angle scattered electrons.


Genetic polymorphism of thiopurine methyltransferase and its clinical relevance for childhood acute lymphoblastic leukemia.
Thiopurine methyltransferase (TPMT) catalyses the S-methylation of thiopurines, including 6-mercaptopurine and 6-thioguanine. TPMT activity exhibits genetic polymorphism, with about 1/300 inheriting TPMT deficiency as an autosomal recessive trait. If treated with standard doses of thiopurines, TPMTdeficient patients accumulate excessive thioguanine nucleotides in hematopoietic tissues, leading to severe hematological toxicity that can be fatal. However, TPMT-deficient patients can be successfully treated with a 10- to 15-fold lower dosage of these medications. The molecular basis for altered TPMT activity has been defined, with rapid and inexpensive assays available for the three signature mutations which account for the majority of mutant alleles. TPMT genotype correlates well with in vivo enzyme activity within erythrocytes and leukemic blast cells and is clearly associated with risk of toxicity. The impact of 6-mercaptopurine dose intensity is also being clarified as an important determinate of event-free survival in childhood leukemia. In addition, there are emerging data that TPMT genotype may influence the risk of secondary malignancies, including brain tumors and acute myelogenous leukemia. Ongoing studies aim to clarify the influence of TPMT on thiopurine efficacy, acute toxicity, and risk for delayed toxicity. Together, these advances hold the promise of improving the safety and efficacy of thiopurine therapy.


[Features of topical diagnosis of postembolic lesions of the pulmonary trunk and main pulmonary arteries].
Radiopaque and radio-isotope findings in the pulmonary arterial channel in 43 patients with postembolic lesions of the pulmonary trunk and its major branches are reported. Angiographic and scanographic symptoms are described. The significance of detected systolic-pressure gradient in the probing of pulmonary vessels is demonstrated. Possible sources of diagnostic errors are analysed, and ways of eliminating them are suggested. Total and selective multipositional angiopulmonography is believed to be the principal method of diagnosing this condition, while pulmonary scanning only allows topical diagnosis in cases of markedly stenosed major pulmonary arteries.


Short-term dynamics of a mixed chemical and electrical synapse in a rhythmic network.
In the rhythmically active pyloric circuit of the spiny lobster, the synapse between the lateral pyloric (LP) neuron and pyloric constrictor (PY) neuron has an inhibitory depressing chemical and an electrical component. To understand how the dynamics of the LP-->PY synapse affect the relative firing times between these two neurons in an ongoing rhythm, we characterized the dynamics of the LP-->PY synapse after a pharmacological block of ongoing activity. When a train of voltage pulses was applied to the voltage-clamped LP neuron, the inhibitory chemical component of the postsynaptic potential (PSP) in the PY neuron rapidly depressed. Thus, after the first few pulses, the PSP was either hyperpolarizing or depolarizing, depending on the interpulse duration, with shorter interpulse durations producing depolarizing PSPs. To characterize the synaptic response during rhythmic activity, we played back prerecorded realistic waveforms in the voltage-clamped LP neuron. After an initial transient, the resulting PSP in PY was always depolarizing, suggesting that in an ongoing rhythm, the electrical component of the synapse is dominant. However, our results indicate that the chemical component of the synapse acts to delay the peak time of the PSP and to reduce its amplitude, and that these effects become more important at slower cycle periods.


Acculturation, nutrition, and health disparities in Latinos.
Latinos have become the largest minority group in the United States and will represent 25% of the US population by 2050. Latinos experience a disproportionate burden of poverty and poor health outcomes. We critically examined the evidence for a link between acculturation and health disparities in Latinos with a focus on type 2 diabetes (T2D) and nutrition-related risk factors and illustrated how acculturation principles can help design a culturally appropriate T2D self-management intervention in Latinos. Evidence presented in this article was drawn from 1) systematic reviews identified through PubMed searches, 2) backward searches that were based on articles cited, 3) experts in the field, and 4) the author's personal files. The preponderance of the evidence supported an association of acculturation with poor dietary quality and obesity. These associations appeared to be modified by several socioeconomic and demographic factors and were not always linear. The association between acculturation and T2D is unclear. Longitudinal studies and more sophisticated analytic approaches are needed to better understand if and how acculturation affects health-disparity outcomes in Latinos. Tailoring interventions to the acculturation level of individuals is likely to help reduce health disparities in Latinos.


DNA content, rDNA loci, and DAPI bands reflect the phylogenetic distance between Lathyrus species.
The nuclear DNA content, the proportion of A+T base pairs, the chromosomal positions of 5S and 25S rRNA genes, as well as of DAPI (4,6-diamidino-2-phenylindole) bands are described for seven species belonging to three different sections of the genus Lathyrus. These data and chromosome measurements allowed to establish precise idiograms and to discriminate most of the chromosome pairs of the seven species. The karyotypic features correlate well with the phylogenetic distances between these species.


Cystic fibrosis, atopy, asthma and ABPA.
The role of atopy on cystic fibrosis (CF) progression remains unclear but evidence suggests that it may influence the appearance of co-morbid conditions such as CF asthma or allergic bronchopulmonary aspergillosis (ABPA). Recognising asthma in patients with CF is not always easy but the identification of atopic markers favours the diagnosis. Physicians should be aware of this fact in order to achieve a better control of respiratory symptoms in patients with CF. Bronchial mucosa inflammation and abnormal mucus predispose to mould colonisation. These patients are at higher risk of allergic sensitisation, especially when atopic susceptibility is present. In the particular case of A. fumigatus, allergic sensitisation precedes ABPA development, which occurs in up to 10% of CF patients. Progression of lung function deterioration is most strikingly pronounced in patients with ABPA. Therefore, sensitisation with A. fumigatus should be regularly tested in patients with CF, especially those at higher risk. Recombinant allergens constitute an important advance in differentiating Aspergillus sensitisation from ABPA itself.


Enteric bacterial catalysts for fuel ethanol production.
The technology is available to produce fuel ethanol from renewable lignocellulosic biomass. The current challenge is to assemble the various process options into a commercial venture and begin the task of incremental improvement. Current process designs for lignocellulose are far more complex than grain to ethanol processes. This complexity results in part from the complexity of the substrate and the biological limitations of the catalyst. Our work at the University of Florida has focused primarily on the genetic engineering of Enteric bacteria using genes encoding Zymomonas mobilis pyruvate decarboxylase and alcohol dehydrogenase. These two genes have been assembled into a portable ethanol production cassette, the PET operon, and integrated into the chromosome of Escherichia coli B for use with hemicellulose-derived syrups. The resulting strain, KO11, produces ethanol efficiently from all hexose and pentose sugars present in the polymers of hemicellulose. By using the same approach, we integrated the PET operon into the chromosome of Klebsiella oxytoca to produce strain P2 for use in the simultaneous saccharification and fermentation (SSF) process for cellulose. Strain P2 has the native ability to ferment cellobiose and cellotriose, eliminating the need for one class of cellulase enzymes. Recently, the ability to produce and secrete high levels of endoglucanase has also been added to strain P2, further reducing the requirement for fungal cellulase. The general approach for the genetic engineering of new biocatalysts using the PET operon has been most successful with Enteric bacteria but was also extended to Gram positive bacteria, which have other useful traits for lignocellulose conversion. Many opportunities remain for further improvements in these biocatalysts as we proceed toward the development of single organisms that can be used for the efficient fermentation of both hemicellulosic and cellulosic substrates.


Identification of human immunodeficiency virus sequences by using in vitro enzymatic amplification and oligomer cleavage detection.
Human immunodeficiency virus (HIV) has been associated with acquired immunodeficiency syndrome and related disorders. Assays to detect antibodies to HIV proteins have been developed and used to screen sera for the identification of individuals who have been exposed to the virus. Although these serological tests have significant sensitivity and specificity for detecting exposure to the virus, they do not provide direct identification of HIV. We report here the application of recently developed nucleic acid amplification and oligonucleotide-based detection procedures for the identification of HIV sequences in established infected cell lines and in cells cultured from infected individuals.