Association of gamma-glutamyl transferase concentrations with all-cause and cause-specific mortality in Chinese adults with type 2 diabetes.
Evidence links gamma-glutamyl transferase (GGT) to mortality in the general population. However, the relationship of GGT with all-cause and cause-specific mortality risk has been little explored in type 2 diabetes mellitus (T2DM) patients. We recruited 20 340 community-dwelling T2DM patients between 2013 and 2014 in Jiangsu, China. Cox regression models were used to assess associations of GGT with all-cause and specific-cause mortality. Restricted cubic splines were used to analyze dose-response relationships between GGT and mortality. Stratified analysis was conducted to examine potential interaction effects by age, sex, smoking status, body mass index (BMI), diabetes duration, and dyslipidemia. During a median follow-up period of 7.04 years (interquartile range: 6.98-7.08), 2728 deaths occurred, including 902 (33.09%) due to cardiovascular disease (CVD), and 754 (27.58%) due to cancer. GGT concentrations were positively associated with all-cause, CVD, and cancer mortality. Multivariable hazard ratios (HRs) for the highest (Q5) vs. the lowest quintile (Q1) were 1.63 (95% confidence intervals [CI]: 1.44-1.84) for all-cause mortality, 1.87 (95% CI: 1.49-2.35) for CVD mortality, and 1.43 (95% CI: 1.13-1.81) for cancer mortality. Effect modification by BMI and dyslipidemia was observed for all-cause mortality (both p for interaction <.05), and HRs were stronger in the BMI <25 kg/m2 group and those without dyslipidemia. Our findings suggest that, in Chinese T2DM patients, elevated serum GGT concentrations were associated with mortality for all-cause, CVD, and cancer, and further research is needed to elucidate the role of obesity, nonalcoholic fatty liver disease, and lipids in this association.


Meet the authors: Adam De Jesus and Hossein Ardehali.
We talk to first author of "Hexokinase 1 cellular localization regulates the metabolic fate of glucose" Adam De Jesus about his pandemic nighttime lab schedule, incidental findings based on the color of cell culture media, and being a first-generation Chicano, as well as group leader Hossein Ardehali about the research in his lab and his experiences as an immigrant.


Survival in Kidney and Bladder Cancers in Four Nordic Countries through a Half Century.
Kidney and bladder cancers share etiology and relatively good recent survival, but long-term studies are rare. We analyzed survival for these cancers in Denmark, Finland, Norway (NO), and Sweden (SE) over a 50-year period (1971-2020). Relative 1- and 5-year survival data were obtained from the NORDCAN database, and we additionally calculated conditional 5/1-year survival. In 2016-2020, 5-year survivals for male kidney (79.0%) and bladder (81.6%) cancers were best in SE. For female kidney cancer, NO survival reached 80.0%, and for bladder cancer, SE survival reached 76.1%. The magnitude of 5-year survival improvements during the 50-year period in kidney cancer was over 40% units; for bladder cancer, the improvement was over 20% units. Survival in bladder cancer was worse for women than for men, particularly in year 1. In both cancers, deaths in the first year were approximately as many as in the subsequent 4 years. We could document an impressive development for kidney cancer with tripled male and doubled female 5-year survival in 50 years. Additionally, for bladder cancer, a steady improvement was recorded. The current challenges are to curb early mortality and target treatment to reduce long-term mortality.


Knee osteoarthritis negatively affects the recovery step following large forward-directed postural perturbations.
The reasons for higher fall risk of people with osteoarthritis (OA) compared to people without OA are not known. It is possible that following a loss of balance OA may negatively affect the recovery stepping response. Stepping responses have not been reported for people with knee OA. Here, we compared recovery step kinematics following laboratory-induced trip and following a large treadmill-delivered perturbation simulating a trip between a group of women with and without self-reported knee OA. We hypothesized that knee OA would significantly impair recovery step kinematics compared to those of a control group. Following the laboratory-induced trip, step length and trunk flexion velocity at recovery step completion of women with OA were significantly impaired and more so for the women who fell. Following the treadmill-delivered perturbation, the recovery step kinematics of women with OA were not significantly impaired. For both perturbations, the women who fell had significantly impaired recovery step kinematics compared to those who did not fall, regardless of OA. The results are consistent with previous work on healthy middle aged and older women and suggest that the same biomechanical risk factors for trip-related falls are shared by middle age and older women regardless of the presence of knee OA. The results support the need to determine whether training protocols which have been shown to improve recovery step kinematics and reduce prospective falls by healthy older women can have similar outcomes for people with knee OA.


Volumetric intravascular ultrasound imaging to illustrate the extent of coronary plaque burden in type 2 diabetic patients.
Although angiography is the gold standard for coronary imaging, its efficacy in outlining diffuse coronary atherosclerosis in diabetic patients remains questionable. We aimed to compare quantitative cineangiographic analysis (QCA) with three-dimensional intravascular ultrasound (IVUS) imaging in type 2 diabetic patients with coronary artery disease. IVUS runs of 104 significant coronary lesions in 88 diabetic patients were performed. Arterial remodeling index was calculated as vessel area at minimal lumen area divided by mean reference vessel area. No difference between the two analysis modes was shown for lesion length and minimal lumen diameter, whereas a significant discrepancy between QCA and IVUS was found for diameter stenosis (10 +/- 9% vs. 41 +/- 8%; P<.001) and vessel diameter (3.01 +/- 0.66 vs. 4.53 +/- 0.70 mm; P<.001). A significant difference on arterial remodeling at lesion site was found between insulin-treated diabetic patients and non-insulin-treated diabetic patients (remodeling index: 0.98 +/- 0.16 vs. 1.07 +/- 0.21; P=.04). Coronary angiographic diagnosis in diabetic patients may be distorted due to a large plaque burden over longer vessel segments and the resulting absence of plaque-free reference segments. This distortion was found to be more pronounced in QCA analysis requiring a reference diameter, whereas volumetric IVUS imaging illustrated coronary artery dimensions more accurately according to anatomic structures. Constrictive arterial remodeling was observed more frequently in type 2 diabetic patients treated with insulin.


Characterization of the effectiveness of a hydrocarbon liquid solidifier.
Solidifiers are dry, granular hydrophobic polymers that form physical bonds with hydrocarbons by molecular interactions (hydrogen bonding, London forces), and are used to immobilize hydrocarbon spill propagation and dispersion. CIAgent© is a non-toxic, proprietary polymer blend listed as an "Oil Solidifier" on the EPA's National Contingency Plan Product Schedule for use on hydrocarbon spills in the navigable waterways of United States. CIAgent solidifies the liquid hydrocarbons through a rapid transformation into a cohesive rubber-like inert mass upon contact and retains the liquid for easier removal and disposal. The objective of this paper is to determine the effectiveness of the solidifier with a variety of hydrocarbon liquids that could be encountered in an oil spill scenario. The effectiveness of the solidifier was characterized in terms of the application rate, temperature change, solubility parameters and solidification time for a variety of hydrocarbon liquids (e. g., gasoline, diesel fuel, crude oil) that could be encountered by measuring the heat of solidification using a solution calorimeter. A thermogram was obtained and the heat of solidification was calculated using the temperature difference upon solidification. The temperature change and the degree of swelling in the solidifier were used to determine the solubility parameter of the solidifier (6.77 Hildebrands). The heat of solidification value was used to determine the ease and speed of the solidification of the hydrocarbon liquids. Solidification times ranged from 40 to 120 s for the liquids tested. The average application ratio in weight of solidifier to weight of hydrocarbon ranged was 3.35.


Improving continuous wound infusion effectiveness for postoperative analgesia after cesarean delivery: a randomized controlled trial.
To evaluate in which anatomical layer (above the fascia or below the fascia) continuous wound infusion of local anesthetic, combined with nonsteroidal antiinflammatory drugs, through a multiorifice catheter has the best effectiveness during the first 48 hours on postoperative pain intensity after elective cesarean delivery. Fifty-six women undergoing elective cesarean delivery under spinal anesthesia were randomly allocated to receive 48-hour continuous wound infusion either above the fascia or below the fascia using ropivacaine and ketoprofene through a multiholed wound catheter. No other systemic analgesics were used, except for rescue patient-controlled intravenous morphine. Evaluation by a blinded investigator included visual analog scale scores at rest and at movement, morphine consumption, patient satisfaction, residual pain at 1 and 6 months, and undesirable side effects. Continuous wound infusion below the fascia resulted in significantly reduced pain at rest and total postoperative morphine consumption (15.7 mg, 95% confidence interval 9.7-20.7 mg) compared with wound administration above the fascia (26.4 mg, 95% confidence interval 18.1-34.7). No undesirable side effects or residual pain requiring treatment were recorded in both groups, whereas analgesia and satisfaction were excellent. After cesarean delivery, continuous wound infusion over 48 hours with ropivacaine and ketoprofene through a multiholed wound catheter inserted below the fascia results in better analgesia when compared with administration above the fascia. ClinicalTrials.gov, www.clinicaltrials.gov, NCT01160913. I.


Real-world pharmacological treatment of patients with postpartum depression in China from 2016 to 2020: A cross-sectional analysis.
To determine the use and tendency of drugs for postpartum depression (PPD) in nine cities across China in 2016-2020 in order to provide a reference for drug use in the clinic. The prescription data of drugs for PPD outpatients in nine cities (Beijing, Chengdu, Guangzhou, Harbin, Hangzhou, Shanghai, Shenyang, Tianjin and Zhengzhou) across China in 2016-2020 were extracted from the Hospital Prescription Analysis Cooperation Project database of the Hospital Pharmacy Professional Committee of Chinese Pharmaceutical Association. A cross-sectional analysis was then conducted of patient age, total prescription quantity, average prescription amount, defined daily doses (DDDs) of drugs, defined daily cost (DDC) of drugs and so on. In 2016-2020, more than half of PPD patients in these nine cities were distributed in first-tier cities (Beijing, Shanghai and Guangzhou), and were mainly aged 30-39 years. During the five-year period, the prescription quantity, total prescription amount and average prescription amount increased by 20.95%, 35.41% and 11.02%, respectively. In terms of prescription frequency and prescription amount, selective serotonin reuptake inhibitors (SSRIs) ranked first, followed by serotonin-norepinephrine reuptake inhibitors (SNRIs). With regard to DDDs, escitalopram, sertraline, paroxetine and venlafaxine ranked high, and sertraline was top year by year. The DDC of vortioxetine and milnacipran was greatest, while that of escitalopram, olanzapine and quetiapine declined sharply after being included in the China centralized drug-procurement program. The number of PPD patients and the total prescription amount in the nine cities across China increased in 2016-2020. SSRIs and SNRIs were the main drugs for PPD treatment. Due to the national centralized drug-procurement policy, there has been a great reduction in the DDC, lightening the economic burden on patients.


A simple and specific competitive protein binding assay for 24,25-dihydroxyvitamin D in human serum.
A sensitive and specific competitive protein binding assay for 24,25-dihydroxyvitamin D was developed using rat kidney cytosol as the binding protein. It also included a chromatographic step on a Sephadex LH 20 column to separate the metabolite specifically. The average value for 24, 25-dihydroxyvitamin D in serum in 19 male and female healthy subjects aged from 20 to 40 was found to be 8.24 +/- 0.34 ng/ml (1.98 +/- 0.08 . 10(-8) mol/l) mean +/- S.D. In hepatic insufficiencies the levels are higher (14.15 +/- 3.59 ng/ml) than in normal subjects. A comparison between 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D levels was made.


Should "handled" prey be considered? Some consequences for functional response, predator-prey dynamics and optimal foraging theory.
Predator-prey models consider those prey that are free. They assume that once a prey is captured by a predator it leaves the system. A question arises whether in predator-prey population models the variable describing prey population shall consider only those prey which are free, or both free and handled prey together. In the latter case prey leave the system after they have been handled. The classical Holling type II functional response was derived with respect to free prey. In this article we derive a functional response with respect to prey density which considers also handled prey. This functional response depends on predator density, i.e., it accounts naturally for interference. We study consequences of this functional response for stability of a simple predator-prey model and for optimal foraging theory. We show that, qualitatively, the population dynamics are similar regardless of whether we consider only free or free and handled prey. However, the latter case may change predictions in some other cases. We document this for optimal foraging theory where the functional response which considers both free and handled prey leads to partial preferences which are not observed when only free prey are considered.


The peptidyl-prolyl isomerase activity of SlyD is not required for maturation of Escherichia coli hydrogenase.
Escherichia coli SlyD, which is involved in the biosynthesis of the metal cluster in the [NiFe]-hydrogenase enzymes, exhibits several activities including that of a peptidyl-prolyl isomerase (PPIase). Mutations that result in deficient PPIase activity do not produce corresponding decreases in the other activities of SlyD in vitro or in hydrogenase production levels in vivo.


Influence of chloroquine on the porphyrin metabolism.
Rats were treated with the well-known porphyrogen hexachlorobenzene (HCB) to induce experimental porphyria. At the same time another group of rats was treated with chloroquine in addition to HCB. The HCB-induced increase of the urinary excretion of porphyrin precursors could thereby be reduced to normal levels and the porphyrin excretion rates were decreased significantly in comparison to those of the other group. The delta-aminolevulinate synthase in the liver of the animals was slightly increased by exclusive treatment with chloroquine, which in the HCB-treated rats chloroquine led to a dramatic decrease in the key enzyme of the porphyrin (heme)-biosynthesis. The influence of chloroquine on the HCB-induced increase of the cytochrome P-450 content and the dependent enzymatic activities were different. The 7-ethoxycumarin deethylase and the arylhydrocarbon hydroxylase activities were not influenced, whereas the increased aminopyrine-N-demethylase activity was reduced to nearly normal levels. Our findings indicate that chloroquine acts by reduction of the delta-aminolevulinate synthase activity, probably by influencing the regulation of the key enzyme of the heme biosynthesis, which is enhanced in human porphyria cutanea tarda, as well as in the HCB-induced porphyria of the rats.


Efficacy and safety of dipeptidyl peptidase-4 inhibitors in combination with metformin.
Use of dipeptidyl peptidase-4 (DPP-4) inhibitors is prevalent for the treatment of type 2 diabetes since they have fewer adverse effects compared with other non-insulin medications currently available; however, as monotherapy, the glycosylated hemoglobin (HbA1c)-lowering power of these agents is moderate. The aim of this article is to evaluate the current literature regarding the safety and efficacy of DPP-4 inhibitors in combination with metformin. A literature search was conducted through MEDLINE (from 1950 to October 2012), PubMed (from 1966 to October 2012), EMBASE (from 1966 to October 2012), and International Pharmaceutical Abstracts (from 1970 to October 2012) using the search terms "sitagliptin," "linagliptin," "alogliptin," "vildagliptin," "saxagliptin," and "metformin." Studies that did not evaluate the DPP-4 inhibitors in combination with metformin and those that were not phase 3, were excluded. Many of the studies evaluated DPP-4 inhibitors in combination with metformin versus glucagon-like peptide-1 (GLP-1) agonists, placebo, DPP-4 inhibitors as monotherapy, thiazolidinediones, and sulfonylureas. The results of these noninferiority trials were that DPP-4 inhibitors as a whole are noninferior to either each other or other agents except for GLP-1 agonists. Also, in superiority studies, GLP-1 agonists proved to have greater HbA1c lowering. In summary, DPP-4 inhibitors play a vital role in the treatment of diabetes. They have relatively limited adverse effects, especially regarding hypoglycemia. DPP-4 inhibitors in combination with metformin are generally well tolerated and are available as combination products to reduce pill burden and enhance compliance. The limitations to DPP-4 inhibitors are the lack of outcomes data and more limited HbA1c lowering than other medications currently approved for the treatment of type 2 diabetes. However, as previously stated, thiazolidinediones, glinides, sulfonylureas, pramlinitide, and GLP-1 agonists are all quite beneficial in HbA1c lowering but are not without major adverse effects. Therefore, DPP-4 inhibitors have a vital role as an oral add-on agent for the treatment of type 2 diabetes.


Border screening for SARS in Australia: what has been learnt?
To assess the effectiveness of the Australian border entry screening program to detect arriving travellers with symptoms of severe acute respiratory syndrome (SARS). Descriptive study of outcomes of screening at Australian airports and seaports between 5 April 2003 and 16 June 2003. To determine the number of international travellers who were symptomatic on arrival in Australia but missed by screening, data were obtained on the number of arrivals screened and the number with symptoms (from the Australian Quarantine and Inspection Service [AQIS]), as well as the number of people investigated for SARS (from the Australian SARS Case Register). There were 1.84 million arrivals into Australia during the study period, and 794 were referred for screening to AQIS staff. Of these, the findings in four travellers were consistent with the World Health Organization case definition for SARS, and they were referred by the Chief Quarantine Officers to designated hospitals for further investigation. None of these people was confirmed to have SARS. One person reported as a probable SARS case acknowledged being symptomatic on arrival, but had been missed by border screening. The low identification rate was attributed to the low prevalence of SARS, the use of exit screening by affected countries, and the subjective measures used in the screening process. With current knowledge about SARS, border screening should focus on educating incoming travellers, especially groups at high risk of transmitting the disease (the elderly and those with underlying chronic illnesses). Objective screening measures should be used during SARS outbreaks to prevent importation of the disease.


Comparative efficacy of three indexes of left ventricular performance derived from pressure-volume loops in heart failure induced by tachypacing.
The purpose of this study was to serially evaluate the response and variability of the end-systolic pressure-volume relation, the left ventricular end-diastolic volume-peak positive first derivative of left ventricular pressure (dP/dt) relation and the left ventricular end-diastolic volume-stroke work relation in the development of progressive left ventricular dysfunction. Evaluation of systolic performance of the failing left ventricle may be enhanced by using relatively load-insensitive measures of left ventricular performance. The end-systolic pressure-volume, left ventricular end-diastolic volume-peak positive dP/dt and left ventricular end-diastolic volume-stroke work relations adequately define left ventricular performance under multiple loading conditions, but efficacy has not been fully assessed in the failing heart, particularly in the intact circulation. Fourteen dogs underwent instrumentation and rapid pacing to heart failure. Variably loaded pressure-volume beats were produced by inferior vena cava occlusion. The dogs were evaluated at baseline and at three progressively more severe levels of left ventricular dysfunction. There was a progressive increase in left ventricular volumes at end-diastole ([mean value +/- SE] 60 +/- 28 to 73 +/- 29 ml, p < 0.001) and end-systole (39 +/- 19 to 61 +/- 27 ml, p < 0.001) during the 3 weeks of rapid pacing and a progressive decline in peak positive dP/dt (1,631 +/- 410 to 993 +/- 222 mm Hg/s, p < 0.001) and ejection fraction (37 +/- 8% to 16 +/- 11%, p < 0.001). There was a corresponding decline in the slope of each of the three relations: for end-systolic pressure-volume, 6.3 +/- 2.2 to 2.8 +/- 0.7 (p < 0.05); for left ventricular end-diastolic volume-stroke work, 61.9 +/- 9.1 to 26.5 +/- 2.4 (p < 0.05); and for left ventricular end-diastolic volume-peak positive dP/dt, 47.1 +/- 13.6 to 20.31 +/- 6.8 (p < 0.05). There was also a corresponding increase in position volumes: for end-systolic pressure-volume, 33.6 +/- 3.9 to 61.2 +/- 6.6 ml (p < 0.05); for left ventricular end-diastolic volume-stroke work, 46.2 +/- 3.6 to 89.3 +/- 7.6 ml (p < 0.05); and for left ventricular end-diastolic volume-peak positive dP/dt, 29.1 +/- 19.1 to 68.6 +/- 25.9 ml (p < 0.05). The relative degree of change in each of the three relations was similar as more severe heart failure developed. The coefficients of variation for position were significantly less than the variation for slopes. The response of volume intercepts was heterogeneous. For left ventricular end-diastolic volume-stroke work, the intercept increased as ventricular performance decreased. The intercept of the end-systolic pressure-volume relation was significantly more variable than the left ventricular end-diastolic volume-stroke work relation and did not change with progressive heart failure. The intercept for left ventricular end-diastolic volume-peak positive dP/dt was highly variable and showed no consistent changes as left ventricular function declined. All three relations consistently describe changes in left ventricular performance brought about by tachypacing. Evolution of left ventricular dysfunction causes a decline in slope and a rightward shift of these relations. The position of the relation is the most sensitive and least variable indicator of left ventricular systolic performance.


Intraflagellar transporter protein (IFT27), an IFT25 binding partner, is essential for male fertility and spermiogenesis in mice.
Intraflagellar transport (IFT) is an evolutionarily conserved mechanism essential for the assembly and maintenance of most eukaryotic cilia and flagella. In mice, mutations in IFT proteins have been shown to cause several ciliopathies including retinal degeneration, polycystic kidney disease, and hearing loss. However, little is known about its role in the formation of the sperm tail, which has the longest flagella of mammalian cells. IFT27 is a component of IFT-B complex and binds to IFT25 directly. In mice, IFT27 is highly expressed in the testis. To investigate the role of IFT27 in male germ cells, the floxed Ift27 mice were bred with Stra8-iCre mice so that the Ift27 gene was disrupted in spermatocytes/spermatids. The Ift27: Stra8-iCre mutant mice did not show any gross abnormalities, and all of the mutant mice survived to adulthood. There was no difference between testis weight/body weight between controls and mutant mice. All adult homozygous mutant males examined were completely infertile. Histological examination of the testes revealed abnormally developed germ cells during the spermiogenesis phase. The epididymides contained round bodies of cytoplasm. Sperm number was significantly reduced compared to the controls and only about 2% of them remained significantly reduced motility. Examination of epididymal sperm by light microscopy and SEM revealed multiple morphological abnormalities including round heads, short and bent tails, abnormal thickness of sperm tails in some areas, and swollen tail tips in some sperm. TEM examination of epididymal sperm showed that most sperm lost the "9+2″ axoneme structure, and the mitochondria sheath, fibrous sheath, and outer dense fibers were also disorganized. Some sperm flagella also lost cell membrane. Levels of IFT25 and IFT81 were significantly reduced in the testis of the conditional Ift27 knockout mice, and levels of IFT20, IFT74, and IFT140 were not changed. Sperm lipid rafts, which were disrupted in the conditional Ift25 knockout mice, appeared to be normal in the conditional Ift27 knockout mice. Our findings suggest that like IFT25, IFT27, even though not required for ciliogenesis in somatic cells, is essential for sperm flagella formation, sperm function, and male fertility in mice. IFT25 and IFT27 control sperm formation/function through many common mechanisms, but IFT25 has additional roles beyond IFT27.


Vector analysis of myopic astigmatism corrected by femtosecond refractive lenticule extraction.
To evaluate the visual outcome of refractive lenticule extraction in eyes with myopic astigmatism using the Visumax femtosecond laser. Department of Ophthalmology, HELIOS Klinikum Erfurt, Erfurt, Germany. Nonrandomized clinical trials. The data set encompassed treatments of sphere and astigmatism. Vector analysis was performed to study the astigmatic results at each follow-up visit at 1 week and 1, 3, and 6 months. Visual outcomes were analyzed for the 6-month follow-up period. The data evaluated was a complete set of refractive data of 182 treatments (87 left eyes, 95 right eyes). Efficacy, predictability, and safety with regard to astigmatic correction were comparable to the results with U.S. Food and Drug Administration-approved excimer lasers. At 6 months, the mean error ratio was 0.68 ± 0.75 (SD) and the mean correction ratio was 1.11 ± 0.69. There was slight astigmatic undercorrection (approximately 10%) over time. The residuum of the linear regression analysis of the predictability plot was approximately 0.4 diopter. Ninety-six percent of the eyes had a corrected distance visual acuity (CDVA) of 20/20 or better; no eye had a CDVA worse than 20/32. Femtosecond refractive lenticule extraction is an effective procedure for correcting myopic astigmatism.


Impact of Opioid Use on Duration of Time Loss After Work-Related Lower Limb Injury.
Purpose This study sought to determine patterns of opioid use among workers with a compensated lower limb injury, factors associated with opioid use, and how opioid use is associated with time loss duration. Methods Claims and medication data were provided by the workers' compensation regulator of Victoria, Australia, for claims lodged 2008-2018 from workers aged 15+ years with a lower limb injury. Descriptive statistics showed the number and prevalence of each opioid type (weak/strong) by demographic, claim and injury predictors. Binary and multinomial logistic regression determined the likelihood of any opioid use, and use of strong, weak or a combination of strong and weak opioids by predictors. Cox regression determined the effect of each opioid type on duration of time loss, controlling for predictors. Results There were 51,334 claims and of these 23.6% were dispensed opioids (9.2% for strong opioids only, 6.6% for weak opioids only and 7.8% for a combination). Weak opioids, on average, were dispensed 15 days earlier than strong opioids. Time loss claims and workers with fractures or hip injuries were most likely to be dispensed opioids. All opioids were associated with increased duration of time loss, with those dispensed both weak and strong opioids having the longest duration of time loss. Conclusions Any opioid use was associated with longer time loss duration, with increasing opioid strength having a greater effect. Review of pain management methods should be undertaken to reduce opioid use, which may have a positive impact on duration of time loss and long-term function.


Targeting HOTAIRM1 ameliorates glioblastoma by disrupting mitochondrial oxidative phosphorylation and serine metabolism.
Serine hydroxymethyltransferase 2 (SHMT2), which catalyzes the conversion of serine to glycine and one-carbon transfer reactions in mitochondria, is significantly upregulated in glioblastoma (GBM). However, the mechanism by which the stability of SHMT2 gene expression is maintained to drive GBM tumorigenesis has not been clarified. Herein, through microarray screening, we identified that HOXA Transcript Antisense RNA, Myeloid-Specific 1 (HOTAIRM1) modulates the SHMT2 level in various GBM cell lines. Serine catabolism and mitochondrial oxidative phosphorylation activities were decreased by HOTAIRM1 inhibition. Mechanistically, according to our mass spectrometry and eCLIP-seq results, HOTAIRM1 can bind to PTBP1 and IGF2BP2. Furthermore, HOTAIRM1 maintains the stability of SHMT2 by promoting the recognition of an m6A site and the interaction of PTBP1/IGF2BP2 with SHMT2 mRNA. The stability of HOTAIRM1 can also be enhanced and results in positive feedback regulation to support the progression of GBM. Thus, targeting HOTAIRM1 could be a promising metabolic therapy for GBM.


Bioavailability of phenytoin in dogs: effect of crystal form and particle size.
Significant differences in bioavailability were observed in dogs when phenytoin crystals of different particle size were administered. No significant differences in bioavailability were observed when different crystal forms were compared.


Effect of MRI on preterm infants and their families: a randomised trial with nested diagnostic and economic evaluation.
We tested the hypothesis that routine MRI would improve the care and well-being of preterm infants and their families. Parallel-group randomised trial (1.1 allocation; intention-to-treat) with nested diagnostic and cost evaluations (EudraCT 2009-011602-42). Participants from 14 London hospitals, imaged at a single centre. 511 infants born before 33 weeks gestation underwent both MRI and ultrasound around term. 255 were randomly allocated (siblings together) to receive only MRI results and 255 only ultrasound from a paediatrician unaware of unallocated results; one withdrew before allocation. Maternal anxiety, measured by the State-Trait Anxiety inventory (STAI) assessed in 206/214 mothers receiving MRI and 217/220 receiving ultrasound. Secondary outcomes included: prediction of neurodevelopment, health-related costs and quality of life. After MRI, STAI fell from 36.81 (95% CI 35.18 to 38.44) to 32.77 (95% CI 31.54 to 34.01), 31.87 (95% CI 30.63 to 33.12) and 31.82 (95% CI 30.65 to 33.00) at 14 days, 12 and 20 months, respectively. STAI fell less after ultrasound: from 37.59 (95% CI 36.00 to 39.18) to 33.97 (95% CI 32.78 to 35.17), 33.43 (95% CI 32.22 to 34.63) and 33.63 (95% CI 32.49 to 34.77), p=0.02. There were no differences in health-related quality of life. MRI predicted moderate or severe functional motor impairment at 20 months slightly better than ultrasound (area under the receiver operator characteristic curve (CI) 0.74; 0.66 to 0.83 vs 0.64; 0.56 to 0.72, p=0.01) but cost £315 (CI £295-£336) more per infant. MRI increased costs and provided only modest benefits. ClinicalTrials.gov NCT01049594 https://clinicaltrials.gov/ct2/show/NCT01049594. EudraCT: EudraCT: 2009-011602-42 (https://www.clinicaltrialsregister.eu/).


Hippocampal stimulation promotes intracellular Tip60 dynamics with concomitant genome reorganization and synaptic gene activation.
Genomic reorganizations mediating the engagement of target genes to transcription factories (TFs), characterized as specialized nuclear subcompartments enriched in hyperphosphorylated RNA polymerase II (RNAPII) and transcriptional regulators, act as an important layer of control in coordinating efficient gene transcription. However, their presence in hippocampal neurons and potential role in activity-dependent coregulation of genes within the brain remains unclear. Here, we investigate whether the well-characterized role for the histone acetyltransferase (HAT) Tip60 in mediating epigenetic control of inducible neuroplasticity genes involves TF associated chromatin reorganization in the hippocampus. We show that Tip60 shuttles into the nucleus following extracellular stimulation of rat hippocampal neurons with concomitant enhancement of Tip60 binding and activation of specific synaptic plasticity genes. Multicolor three-dimensional (3D) DNA fluorescent in situ hybridization (DNA-FISH) reveals that hippocampal stimulation mobilizes these same synaptic plasticity genes and Tip60 to RNAPII-rich TFs. Our data support a model by which external hippocampal stimulation promotes intracellular Tip60 HAT dynamics with concomitant TF associated genome reorganization to initiate Tip60mediated synaptic gene activation.


Arterial stiffness acutely decreases after whole-body vibration in humans.
Increased arterial stiffness is a well-established cardiovascular risk factor. Mechanical stimuli to artery, such as compression, elicit vasodilation and acutely decrease arterial stiffness. As whole-body vibration (WBV)-induced oscillation is propagated at least to lumbar spine, WBV mechanically stimulates abdominal and leg arteries and may decrease arterial stiffness. WBV is feasible in vulnerable and immobilized humans. Therefore, it is worthwhile to explore the possibility of WBV as a valuable adjunct to exercise training. The aim of this study was to investigate the acute effects of WBV on arterial stiffness. Ten healthy men performed WBV and control (CON) trials on separate days. The WBV session consisted of 10 sets of vibration (frequency, 26 Hz) for 60 s with an inter-set rest period of 60 s. Subjects maintained a static squat position with knees bent on a platform. In the CON trial, WBV stimulation was not imposed. Blood pressure, heart rate and brachial-ankle pulse wave velocity (baPWV), an index of arterial stiffness, were measured before and 20, 40 and 60 min after both trials. Heart rate and blood pressure did not change from baseline after both trials. Although baPWV did not change in the CON trial (baseline vs. after 20, 40 and 60 min; 1144 +/- 35 vs. 1164 +/- 41, 1142 +/- 39, and 1148 +/- 34 cm s(-1)), baPWV decreased 20 and 40 min after the WBV trial and recovered to baseline 60 min after the trial (1137 +/- 28 vs. 1107 +/- 30, 1108 +/- 28, and 1128 +/- 25 cm s(-1)). These results suggest that WBV acutely decreases arterial stiffness.


Assessment of calf muscle contraction by diffusion tensor imaging.
The goal of this study was to assess the changes of water diffusion during contraction and elongation of calf muscles using diffusion tensor (DT) MRI in normal volunteers. Twenty volunteers (mean age, 29+/-4 years) underwent DT MRI examination of the right calf. Echo planar imaging sequence was performed at rest, during dorsal flexion and during plantar flexion. The three eigenvalues (lambda1, lambda2, and lambda3), apparent diffusion coefficient (ADC) and fractional anisotropy (FA) of the diffusion tensor were calculated for medial gastrocnemius (mGM) and tibialis anterior (TA). A fiber tractography was performed on both muscles. Non-parametric Wilcoxon and Mann Whitney tests were used for statistical evaluation. At rest, lambda1, lambda2 and ADC of mGM were higher than their counterparts of TA (P<0.01). During dorsal flexion, the three eigenvalues and ADC of TA significantly increased (P<0.05) as their counterparts of mGM slightly decreased (P=NS). Opposite variations were detected during plantar flexion of the foot. Visual analysis evidenced a relationship between 3D representations of MRI fibers and physiological state of muscles. Contraction of calf muscles produces changes in DT parameters, which are related to the physiological state of the muscle.


Standard and Customized Alloplastic Facial Implants Refining Orthognathic Surgery: Outcome Evaluation.
Conventional orthognathic osteotomies provide appropriate functional outcomes but might be unable to correct midface deficiency, achieve a satisfactory outcome in asymmetrical cases, or allow sufficient chin advancement. We evaluated the outcome of both standard and customized facial high-density porous polyethylene implants used to refine the cosmetic outcome of orthognathic surgery. We implemented a retrospective study. The sample was composed of all patients who underwent facial alloplastic augmentation between June 2011 and October 2018 in our department. The complication rate was recorded after a mean follow-up period of 41 months postoperatively, and patient satisfaction was assessed through a qualitative evaluation based on an 11-item questionnaire. The sample was composed of 24 implants placed in 14 patients: 13 mandibular angle implants, among which 4 were customized; 8 malar implants; and 3 chin implants. No physical complications such as hematoma, infection, migration, or hypoesthesia were observed. Two implants had to be removed because of early unsatisfactory esthetic outcomes. Of 14 patients, 11 answered our questionnaire. Eighty-two percent strongly agreed that the overall outcome was satisfactory. The results of this study confirm the low physical complication rate described in the literature, and the esthetic complication rate remains lower than the rates observed in previous reports. A high satisfaction rate was found among patients. The lowest mean satisfaction score was noted for appropriate implant symmetry (3.5), whereas the highest mean satisfaction score (3.8) was achieved when using customized implants. If standard high-density porous polyethylene implants appear to be relevant adjuncts to orthognathic surgery, customized implants seem to achieve higher satisfaction, although their prohibitive cost should be considered.


Effect of Antioxidants on the Production of MCP-1 Chemokine by EA.hy926 Cells in Response to IL-6.
An elevated level of circulatory interleukin 6 (IL-6) is a biomarker for cytokine storm of various etiologies, including COVID-19, and contributes to poor prognosis. Vascular endothelial cells are one of the main targets of pathological action of IL-6. IL-6 activates the trans-signaling pathway via the formation of the IL-6/sIL-6Ra/gp130 receptor complex and subsequent activation of the JAK/STAT3 signaling pathway and PI3K/AKT and MEK/ERK kinases in some cases. Previously, it was shown by the authors' group and other researchers that reactive oxygen species (ROS), including mitochondrial ROS (mito-ROS), contribute to the induction of IL-6 expression in the endothelium, mainly due to increased activation of the transcription factor NF-kB. We have also shown that the mitochondria-targeted antioxidant SkQ1 (Plastoquinolyl-10(6'-decyltriphenyl)phosphonium) prevented tumor necrosis factor (TNF)-induced cytokine storm and death in mice. In the aortas of these animals, SkQ1 also prevented the increase in the expression of NF-kB-dependent genes, including the cytokine IL-6 and the chemokine MCP-1. In the current work, the hypothesis of mito-ROS involvement in the IL-6-signaling-mediated proinflammatory gene expression in endothelial cells is tested. SkQ1 suppressed the expression and secretion of the MCP-1 chemokine, induced by IL-6 in combination with sIL-6-Ra, but not the expression of ICAM-1 adhesion molecules in EA.hy926 human endothelial cells. Using specific inhibitors, the authors have shown that, in EA.hy926 cells, IL-6-induced expression of MCP-1 and ICAM-1 depends on the signaling protein and transcription activator STAT3 and, in some cases, on JNK, PI3K, and MEK1/2 kinases and is independent of p38 kinase. In this model, IL-6 induced rapid STAT3 activation, while ERK1/2 activation was less pronounced, and there was no IL-6 effect on Akt and JNK activation. SkQ1 partially suppressed STAT3 and ERK1/2 activation. Thus, we have shown that SkQ1 suppresses not only NF-kB-dependent expression of IL-6 and other proinflammatory genes but also IL-6-induced activation of JAK/STAT3 and STAT3-dependent expression of MCP-1, which probably contributes to the overall therapeutic effect of SkQ1.


Clinical M2 macrophages-related genes to aid therapy in pancreatic ductal adenocarcinoma.
Increasing evidence supports that infiltration M2 Macrophages act as pivotal player in tumor progression of pancreatic ductal adenocarcinoma (PDAC). Nonetheless, comprehensive analysis of M2 Macrophage infiltration and biological roles of hub genes (FAM53B) in clinical outcome and immunotherapy was lack. The multiomic data of PDAC samples were downloaded from distinct datasets. CIBERSORT algorithm was performed to uncover the landscape of TIME. Weighted gene co-expression network analysis (WGCNA) was performed to identify candidate module and significant genes associated with M2 Macrophages. Kaplan-Meier curve and receiver operating characteristic (ROC) curves were applied for prognosis value validation. Mutation data was analyzed by using "maftools" R package. Gene set variation analysis (GSVA) was employed to assign pathway activity estimates to individual sample. Immunophenoscore (IPS) was implemented to estimate immunotherapeutic significance of risk score. The half-maximal inhibitory concentration (IC50) of chemotherapeutic drugs was predicted by using the pRRophetic algorithm. Finally, quantitative real-time polymerase chain reaction was used to determine FAM53B mRNA expression and TIMER database was utilized to uncover its possible role in immune infiltration of PDAC. Herein, 17,932 genes in 234 samples (214 tumor and 20 normal) were extracted from three platforms. Taking advantage of WGCNA, significant module (royalblue) and 135 candidate genes were considered as M2 Macrophages-related genes. Subsequently, risk signature including 5 hub genes was developed by multiple analysis, which exhibited excellent prognostic performance. Besides, comprehensive prognostic nomogram was constructed to quantitatively estimate risk. Then, intrinsic link between risk score with tumor mutation burden (TMB) was explored. Additionally, risk score significantly correlated with diversity of tumor immune microenvironment (TIME). PDAC samples within different risk presented diverse signaling pathways activity and experienced significantly distinct sensitivity to administering chemotherapeutic or immunotherapeutic agents. Finally, the biological roles of FAM53B were revealed in PDAC. Taken together, comprehensive analyses of M2 Macrophages profiling will facilitate prognostic prediction, delineating complexity of TIME, and contribute insight into precision therapy for PDAC.


The Influence of Storage in Saline or Irradiation by Ultraviolet on Surface Hydrophilicity of Implant and Osseointegration: An Experimental Study in Rabbits.
Storage in aqueous solution or ultraviolet (UV) irradiation can retain or regain the hydrophilicity of titanium implant surface. In this study, 3 types of commercial titanium implants were used: ZBL (ZDI Bone Level), CEL (C-tech Esthetic Line), and modSLA (Straumann SLActive). ZBL and CEL implants were treated with UV irradiation for 4 hours. Surface characterization of the 4 groups (ZBL, ZBL-UV, CEL-UV, and modSLA) was evaluated by scanning electron microscopy and contact angle measurements. The in vivo bone response was evaluated by removal torque (RTQ) tests and histomorphometric analysis at 3, 6, and 12 weeks postimplantation. A total of 144 implants and 36 rabbits were used for experiments according to a previously established randomization sequence. The ZBL-UV, CEL-UV, and modSLA groups were hydrophilic, and nanostructures were observed on the modSLA implant surface. ModSLA achieved better RTQ value than ZBL at 12 weeks (P < .05). For histomorphometric analysis, ZBL-UV and CEL-UV implants showed higher bone area values in the cancellous bone zone at 6 weeks than did modSLA and ZBL implants (P < .05). In the cortical bone zone, all groups showed comparable bone-to-implant contact at all healing time points (P > .05). Both storage in saline and UV irradiation could retain or provoke hydrophilic surfaces and improve osseointegration. Compared with storage in saline, UV irradiation displayed slight advantages in promoting new bone formation in cancellous bone zone at an early stage.


MRI of the tectorial and posterior atlanto-occipital membranes in the late stage of whiplash injury.
Our aim was to characterise and classify permanent structural changes in the tectorial and posterior atlanto-occipital membranes several years after a whiplash injury, and to evaluate the reliability of our classification. We obtained sagittal proton density-weighted images of the craniovertebral junction of 92 whiplash-injured and 30 uninjured individuals. Structural abnormalities in the two membranes were classified as grades 1-3 independently by three radiologists blinded for clinical information. Grading criteria were based on reduced tectorial membrane thickness, and elongation or rupture of the posterior atlanto-occipital membrane/dura mater complex. The same images were reassessed 4 months later. Image quality was graded good in 104 cases, slightly reduced in 13 and unsatisfactory in five. Of 117 tectorial membranes 31 (26.5%) showed grade 2 or 3 lesions, in the uninjured group none were grade 3 and only three were grade 2. Pair-wise interobserver agreement (weighted kappa) was moderate (0.47-0.50), while the intraobserver agreement was moderate to good (0.51-0.70). Of 117 posterior atlanto-occipital membranes 20 (17.1%) had grade 2 or 3 lesions; there was no grade 3 and only one grade 2 lesion in the uninjured group. Inter- and intraobserver agreement was good (0.61-0.74 and 0.65-0.86, respectively). Reduced image quality was the main reason for disagreement, but partial thinning and lateral tapering, as normal tectorial membrane variations, created difficulties in some cases. This study strongly indicates that whiplash trauma can damage the tectorial and posterior atlanto-occipital membranes; this can be shown on high-resolution MRI. Better knowledge of normal anatomical variations and improved image quality should increase the reliability of lesion classification.


Network-Driven Drug Discovery.
We describe an approach to early stage drug discovery that explicitly engages with the complexities of human biology. The combined computational and experimental approach is formulated on a conceptual framework in which network biology is used to bridge between individual molecular entities and the cellular phenotype that emerges when those entities interact in a network. Multiple aspects of early stage discovery are addressed including the data-driven elucidation of biological processes implicated in disease, target identification and validation, phenotypic discovery of active molecules and their mechanism of action, and extraction of genetic target support from human population genetics data. Validation is described via summary of a number of discovery projects and details from a project aimed at COVID-19 disease.


Cell Therapy Drug Product Development: Technical Considerations and Challenges.
Cell therapy uses living cells as a drug to treat diseases. To develop a cell therapy drug product (DP), cryopreservation plays a central role in extending the shelf life of these living medicines by pausing their biological activities, especially preventing degradation, at a temperature as low as liquid nitrogen. This helps overcome the temporal and geographical gaps between centralized manufacturing and clinical administration, as well as allowing sufficient time for full release testing and flexibility in scheduling patients for administration. Cryopreservation determines or influences several key manufacturing, logistical, or clinical in-use processes, including formulation, filling, controlled rate freezing, cryogenic storage and transportation, thawing, and dose preparation. This article overviews the key technical aspects of cell therapy DP development and elucidates fundamental principles of cryobiology that should be considered when we design and optimize the relevant processes. This article also discusses the challenges that motivate continued innovation for cell therapy drug product development.


Valorization of thermochemical conversion of lipid-extracted microalgae to levulinic acid.
Scenedesmus obliquus, a green microalga of the class Chlorophyceae, has been used to produce biofuels. However, limited research has been reported on platform chemicals that use microalgae as biomass to replace fossil sources. This paper reports on the investigation of levulinic acid (LA) production from lipid-extracted S. obliquus with an acid-catalyzed thermochemical conversion using a statistical experimental approach. For the reaction factors, the highest effect on LA yield resulted from catalyst concentration. The optimized LA yield of 45.63 wt% (70.7 mol%) was achieved with 5 wt% lipid-extracted microalgae and reaction factors of 0.85 M HCl as a catalyst at 180 °C for 10 min. Also, the LA yield as a function of the combined severity factor followed a sigmoid curve. High LA yield resulted from combined severity factors greater than 3.4. These results indicate that the production of platform chemicals may be possible using microalgae feedstocks and thermochemical conversion.


Antiviral therapy after curative treatment of hepatitis B/C virus-related hepatocellular carcinoma: A systematic review of randomized trials.
Available published work on the benefit of adjuvant antiviral therapy after curative treatment of hepatocellular carcinoma (HCC) reports controversial results. The objective of this systematic review was to evaluate the effect of adjuvant antiviral therapy on recurrence and survival after curative treatment of HCC. We conducted an extensive search strategy. All randomized controlled trials comparing adjuvant antiviral therapy versus placebo or no treatment were considered for this review. Results were expressed as hazard ratio for time-to-event outcomes with 95% confidence intervals using RevMan 5. We included nine trials (three of low risk of bias and six of unclear risk of bias) with 954 patients. All the included studies used conventional interferon (IFN) as adjuvant antiviral therapy; none of them used pegylated IFN or nucleoside analogs. There were significant improvements for recurrence-free survival and overall survival in the adjuvant IFN group compared with the control group. Subgroup analysis also showed a significant difference favoring IFN therapy in hepatitis C virus (HCV)-related HCC patients, but for hepatitis B virus (HBV)-related patients, the difference failed to reach statistical significance. A dose reduction was needed in 28.3% of patients and discontinuation of IFN therapy happened in 8.2% of patients due to moderate to severe side-effects. Our study suggested potential benefits of adjuvant IFN therapy following curative treatment of HCC, especially for HCV-related HCC. Further high-quality randomized controlled trials of more effective adjuvant antiviral regimens, either used alone or in combination, for virus-related HCC, especially HBV-related HCC, are needed.


Factors influencing mortality in atrial fibrillation. Post hoc analysis of an observational study in outpatients.
Whether patients with atrial fibrillation (AF) have an increased mortality compared with the general population is controversially assessed. The mortality of 409 outpatients with AF (36% female, mean age 62 years, 39% paroxysmal AF, 27% lone AF), who were included in a prospective observational study, was compared with the mortality of the general population. The mortality was 4%/year and constant during the 10 years of follow-up. Age (p<0.0001), heart failure (p=0.0013) and echocardiographically detected reduced left ventricular systolic function (p=0.0353) were independent predictors. The mortality of AF patients was 1.4-fold higher than in the general population, of female 1.5 and of male 1.3. Patients with permanent AF had a 1.5-fold higher and patients with heart failure a 1.7-fold higher mortality than the general population. Patients with lone AF did not differ in the mortality from the general population. The mortality of AF patients was not influenced by the duration of atrial fibrillation. The prognosis of AF patients is dependent on the cardiovascular co-morbidity, especially the presence or absence of heart failure.


Synthetic metabolic transducers in Saccharomyces cerevisiae as sensors for aromatic permeant acids and bioreporters of hydrocarbon metabolism.
Yeast-based biosensors have great potential for various applications, although the present range of detectable chemicals is still very minimal. This work provides an enlargement of the knowledge on detectable chemicals and creates an additional basis for engineering modular yeast biosensors. Bacterial allosteric transcription factors, such as MarR and PdhR, were recruited to build transducer circuits in Saccharomyces cerevisiae. MarR-based biosensors were designed for the detection of aromatic permeant acids (benzoate and salicylate), whereas the PdhR-expressing yeast cells were engineered for responding to pyruvate. In general, all our engineered strains showed a fast response time and a strong fluorescent output signal to chemical concentrations ranging from 5 mM down to 2 fM. They exhibited versatile dynamic range and were capable of operating in a variety of complex media that might contain any of these compounds. A new milestone in biosensor design is the engineering of inter/intracellular metabolic biosensors that would allow real-time monitoring of either the metabolism of particular compounds, or the detection of their intermediate/end products. Our synthetic cells are applicable to different areas, from adequate real-time detection of aromatic permeant acids to regulation/monitoring of different hydrocarbon metabolisms. The new strains engineered in this study could be of great importance because of the ecological significance of aromatic permeant acids from their formations during either hydrocarbon degradation or metabolism of different chemicals to their involvement in different biological and non-biological systems.


Contractile proteins: possible targets for the cardiotonic action of MCI-154, a novel cardiotonic agent?
The effect on the contractile protein system of MCI-154, a novel and potent cardiotonic agent, was examined by using thin bundles of chemically skinned fibers from the guinea-pig papillary muscles. MCI-154 increased the Ca2+ sensitivity of the contractile protein system of the cardiac skinned fibers. Moreover, MCI-154 enhanced the maximum tension developed at pCa 4.4. These results suggest that an increase in responses of the contractile protein system to Ca2+ ion is, at least in part, responsible for the positive inotropic action of MCI-154.


Lost in transmission? Self- and other-annotation of emotional words.
It is generally assumed that someone's affective state can be correctly detected and interpreted by other people, nowadays even by computer algorithms, in their writing. However, it is unclear whether these perceptions match the actual experience and communicative intention of the author. Therefore, we investigated the relation between affect expression and perception in text in a two-part study. In Part 1, participants (authors) wrote about emotional experiences according to four combinations of two appraisals (High/Low Pleasantness, High/Low Control), rated the valence of each text, and annotated words using 22 emotions. In Part 2, another group of participants (readers) rated and annotated the same texts. We also compare the human evaluations to those provided by computerized text analysis. Results show that valence differed across conditions and that authors rated and annotated their texts differently than readers. Although the automatic analysis detected levels of positivity and negativity across conditions similar to human valence ratings, it relied on fewer and different words to do so. We discuss implications for affective science and automatic sentiment analysis.


Biologic effects of equilin sulfate in postmenopausal women.
In order to determine the relative potency of equilin sulfate (EqS), a major constituent of conjugated equine estrogens, 15 women received oral doses of EqS (0.15, 0.31, and 0.625 mg) for 25 days. Doses of 0.31 and 0.625 mg significantly stimulated hepatic globulins. This stimulatory effect ranged from being 1.5 to 8 times greater than the effects of comparable doses of estrone sulfate and conjugated equine estrogens. A significant stimulation in high-density lipoprotein-cholesterol occurred with as little as 0.15 mg of EqS. Elevations in the high-density lipoprotein/low-density lipoprotein-cholesterol ratio occurred with EqS, which resulted in an approximately 4-fold greater response than that achieved with comparable doses of conjugated equine estrogens. The fasting urinary calcium/creatinine ratio was only significantly lowered with 0.625 mg of EqS and was less potent than conjugated equine estrogens in this regard. It is concluded that EqS is a potent estrogen that contributes significantly to the hepatic stimulatory effects of conjugated equine estrogens. These data also provide support for the suggestion that there may be a dissociation in potency between estrogenic effects on liver and bone.


Malignant extraovarian endometriosis: two case reports and review of the literature.
Two cases of adenocarcinoma arising in extraovarian endometriosis 19 and 8 years following abdominal hysterectomy and bilateral salpingo-oophorectomy are described. Both patients presented with hydronephrosis. One had been on chronic estrogen therapy. The literature is reviewed in reference to frequency, tumor type, and sites of occurrence.


The biological relationship of mouse hepatitis virus (MHV) strains and interferon: in vitro induction and sensitivities.
Five prototype strains of mouse hepatitis virus (MHV) -1, -3, -S, -A59 and -JHM were analyzed for their ability to induce interferon (IFN) in seven cell lines of rodent origin. Induction of IFN by all of the prototype MHV strains was infrequent and unpredictable, while IFN was produced consistently by five cell lines treated with known inducers. Priming and/or aging of cells did not enhance IFN induction by the MHV strains except in the case of MHV-A59 which consistently induced moderate levels of IFN on L-cells which were both primed and aged. Kinetic studies of MHV-A59-induced IFN on primed and aged L-cells demonstrated that detectable levels of IFN were not produced until 24 hours post-inoculation (p.i.). Peak levels were attained at 30 hours p.i. with no additional IFN produced through 48 hours p.i. MHV-induced IFN was similar in composition and properties to Newcastle disease virus-induced IFN. The sensitivities of the five MHV strains to eight concentrations of preformed L-cell IFN were also assessed. All strains except MHV-S fit a linear model with MHV-3, MHV-A59 and MHV-JHM having similar slopes. At most concentrations MHV-3 was less sensitive than MHV-1, -A59 or -JHM to IFN. The response curve for MHV-S was non-linear. This strain was more sensitive to the antiviral effects of the pre-formed IFN except at the highest concentrations of IFN used.


Isolation of natural products by low-pressure column chromatography.
This chapter describes the isolation of natural products using low-pressure column chromatography (LPLC) and provides a brief summary of the adsorption and size exclusion processes involved in LPLC. Different types of stationary phases used in both adsorption and size exclusion LPLC are also presented with specific examples. Various operational parameters in relation to column packing (both wet and dry), column equilibration, sample application, and column development, and a summary of generic procedures for adsorption and size exclusion LPLC are also incorporated in this chapter. Throughout the chapter, several specific examples of the application of LPLC for the isolation of various natural products including alkaloids, coumarins, flavones, and saponins from various plants are given. Many of these examples include a combination of adsorption and size exclusion chromatography to obtain specific compounds.


Competitive relationships existing between two closely related species of Argemone living in the same area.
The present paper deals with the reaction of two closely related species of Argemone L., A. mexicana L. and A. ochroleuca Sweet, to its own density in pure stands and a comparison of this with its reaction to the density of the associated species. The results of the pure culture studies indicate that whilst each of the two species reacted to an increase in its own density by higher mortality and a striking reduction in vegetative and reproductive growth, A. mexicana was more susceptible to intraspecific effect than A. ochroleuca. On the other hand, mixed culture studies indicated that A. mexicana was more successful when in competition with A. ochroleuca. Further, that species which was in a majority in the mixture suffered most compared to the minority species. Such a density dependent differential behaviour in pure stands and frequency dependent differential behaviour in mixed cultures along with the reversal in superiority of the two species in pure and mixed cultures was considered to be responsible for the cohabitation of the two species and their relative population sizes in nature.


Precordial electrocardiographic mapping in the identification of patients with left main stem narrowing.
This study was designed to determine if exercise testing using 16-lead precordial mapping can be used to identify patients with left main stem narrowing. In a group of 235 consecutive patients undergoing coronary angiography there were 35 patients with left main stem narrowing. The patients with left main stem disease differed from the others in that not only did they develop extensive S-T segment depression, but more specifically these changes occupied a characteristic position high on the precordium above the usual site of the precordial leads of the 12-lead electrocardiogram. This finding was then tested prospectively in a second group of 100 patients. Fourteen of the 100 patients had left main stem narrowing; the sensitivity and specificity of S- T segment changes high on the precordium in identifying patients with left main stem disease were 82 and 84% respectively. Thus precordial mapping and exercise testing is valuable in the diagnosis of patients with left main stem narrowing. The technique is simple and inexpensive and provides data not available using the conventional precordial leads of the 12-lead electrocardiogram.


An update on coronary risk factors.
Epidemiologic research continues to produce additional independent risk factors, including small dense LDL, Lp(a), isolated systolic hypertension, the insulin-resistance syndrome, leucocyte count, plasma fibrinogen, homocystinemia, and sleep disturbance. Alcohol in moderation appears protective. Because the hazard posed by any particular risk factor is markedly influenced by other metabolically linked risk factors with which it tends to cluster, multivariate risk assessment provides a substantially better risk estimate than that based on single risk factors.


Odontogenic tumors. A demographic study of 759 cases in a Chinese population.
Seven hundred fifty-nine cases of odontogenic tumors retrieved from the files of College of Stomatology, West China University of Medical Sciences were classified according to the World Health Organization's Histological Classification of Odontogenic Tumors and compared with similar reports from other countries. Among these cases, 93.9% of the tumors were benign and 6.1% were malignant. Ameloblastomas predominated (58.6%) with a predilection for the mandible, while odontomas, generally regarded as the most frequent odontogenic tumor in North America, only accounted for 6.7%, the fourth most common tumor in this series. The mandible and the maxilla were divided into eight anatomic regions, and the distribution of each odontogenic tumor type amongst these regions was recorded. The relative incidence of each tumor type, patient age and gender were also compared with data from other selected large series. Geographic differences were noted in the relative incidence of ameloblastoma, odontoma and malignant odontogenic tumors among the Chinese/African, North American and Turkish/German groups. Ameloblastoma and malignant odontogenic tumors are not considered rare in a Chinese population.


The in vitro development of the pupal integument and the effects of ecdysteroids in Tenebrio molitor (Insecta, Coleoptera).
In order to study the pupal-adult metamorphosis of Tenebrio in vitro, pupal sternites of different ages were cultured in Landureau's medium and their development systematically observed by electron microscopy. In hormone-free medium, explants taken from young pupae do not secrete pupal postecdysial cuticle in vitro, and the epidermis spontaneously detaches from the pupal cuticle. On the contrary, explants taken from pharate adults continue to secrete adult preecdysial cuticle in vitro, and the epidermis never detaches from the cuticle. Ecdysterone in physiological concentrations (0.2 to 4 micrograms/ml) induces the secretion of a new cuticle in explants from young pupae but the epidermis remains undifferentiated. Ecdysone is necessary for the induction of some adult differentiation. Moreover, the quality of the cuticle secreted in vitro is increased by the addition of 2% foetal calf serum; the best results have thus far been obtained in a medium containing 0.2 microgram/ml ecdysone, 1 microgram/ml ecdysterone, and 2% foetal calf serum.


Expression of Streptomyces melC and choA genes by a cloned Streptococcus thermophilus promoter STP2201.
Streptococcus thermophilus (ST) chromosomal DNA (chr DNA) fragments having promoter activity were cloned and selected in Escherichia coli using a chloramphenicol acetyltransferase- (cat-) based promoter-probe vector pKK520-3. Insertion of a promoterless streptomycete melanin biosynthesis operon (melC) downstream from the promoters of the library further identified clone STP2201 as a strong promoter in E. coli. Subcloning of a STP2201-melC DNA fragment into the pMEU-series S. thermophilus-E. coli shuttle vectors yielded pEU5xML2201x plasmids that conferred Mel+ phenotype to E. coli. The pEU5aML2201a was further shown to afford a high level of tyrosinase pro-anti-tyrosinase antiserum in S. thermophilus. Substituting melC with a streptomycete cholesterol oxidase gene (choA) in the same orientation yielded pEU5aCH2201a that conferred ChoA activity to an E. coli transformant at a level of (1.06 +/- 0.15) x 10(-7) units mg-1 protein. Introduction of this plasmid into S. thermophilus by electrotransformation yielded ChoA+ transformant that produced the enzyme at about 25% of the level found in E. coli.


Giant Juvenile Xanthogranuloma of face in an adult: A rare complexity.
Juvenile xanthogranuloma, a form of non-Langerhans cell histiocytosis can be defined as the proliferation of cells with macrophage like characteristics. It has been described as a benign, asymptomatic and common self-healing disorder of non-Langerhans cell histiocytosis (LCH), affecting mostly infants, children and rarely adults. We have documented a case of a 40 year old male who presented to us with extensive insidious papulonodulous growth over the face. The facial disfigurement caused was catastrophic. An extensive surgical excision and reconstruction was performed followed by histopathological evaluation. Microscopic study and immune histochemistry revaled Juvenile Xanthogranuloma of adult. The article highlights the presentation, diagnosis and management of this mammoth, rare disease.


Emission factors and emission inventory of diesel vehicles in Nepal.
A comprehensive emission inventory of the transport sector through fuel-based emission factors (EFs) was developed for the first time in Nepal. This study estimates air pollutants emission from diesel vehicles between the years 1989 and 2018 based on national statistical data, average vehicle kilometers travelled, fuel mileage, and measurement-based EFs for each vehicle category during idle and moving conditions. The consumption of diesel by vehicle category was also estimated and total consumption was compared with national sales data. The Monte Carlo was used to estimate uncertainties. Nationally, total diesel consumption was estimated as 892,770 kL (85-115%) in 2017/18, 13.4 times higher than 1989/90. Ratnoze1 and Microaeth were used to conduct the tail pipe emission measurements. The fuel-based EFs of CO2, CO, BC, and PM2.5 were calculated through the carbon mass balance method. Of all diesel vehicles measured (n = 29) during idling, the average EFs were estimated as CO2 2600 (99-101%), CO 33.3 (44-156%), BC 0.6 (25-101%), and PM2.5 5.2 (0-235%) in unit of g L-1. For moving conditions (n = 5), the average EFs were estimated to be CO2 2476 (90-110%), CO 97.3 (0-232%), BC 1.7 (46-110%), and PM2.5 20.7 (0-255%), all in g L-1. Multiplying fuel consumption by EFs, national air pollutant emissions were estimated as 2214 (90-110%) to 2781(85-115%) for CO2, 27.7 (42-158%) to 88.8 (0-232%) for CO, 0.51 (23-177%) to 3.55 (46-110%) for BC and 3.42 (0-236%) to 23.47 (0-255%) for PM2.5 in 2017/18 in unit of Gg. This paper recommends revising national vehicle mass emission standards based on the findings of this study and including and enhancing sustainable low-carbon transport through amendment of transport policy.


The BeachLitter dataset for image segmentation of beach litter.
This dataset consists of 3500 images of beach litter and 3500 corresponding pixel-wise labelled images. Although performing such pixel-by-pixel semantic masking is expensive, it allows us to build machine-learning models that can perform more sophisticated automated visual processing. We believe this dataset may be of significance to the scientific communities concerned with marine pollution and computer vision, as this dataset can be used for benchmarking in the tasks involving the evaluation of marine pollution with various machine learning models. The beach litter images were obtained from coastal environment surveys conducted between 2011 and 2019 by the Yamagata Prefectural Government, Japan. These images were originally obtained owing to the reporting guidelines concerning regular coastal-environmental-cleanup and beach-litter-monitoring surveys. Based on these images, the Japan Agency for Marine-Earth Science and Technology created 3500 images comprising eight classes of semantic masks for beach litter detection [1].


High order approximation on non-uniform meshes for generalized time-fractional telegraph equation.
This paper presents a high order approximation scheme to solve the generalized fractional telegraph equation (GFTE) involving the generalized fractional derivative (GFD). The GFD is characterized by a scale function σ ( t ) and a weight function ω ( t ) . Thus, we study the solution behavior of the GFTE for different σ ( t ) and ω ( t ) . The scale function either stretches or contracts the solution while the weight function dramatically shifts the numerical solution of the GFTE. The time fractional GFTE is approximated using quadratic scheme in the temporal direction and the compact finite difference scheme in the spatial direction. To improve the numerical scheme's accuracy, we use the non-uniform mesh. The convergence order of the whole discretized scheme is, O ( τ 2 α - 3 , h 4 ) , where τ and h are the temporal and spatial step sizes respectively. The outcomes of the work are as follows: •The error estimate for approximation of the GFD on non-uniform meshes is established.•The numerical scheme's stability and convergence are examined.•Numerical results for four examples are compared with those obtained using other method. The study shows that the developed scheme achieves higher accuracy than the scheme discussed in literature.


Impact of Combined Lifestyle Factors on All-Cause and Cause-Specific Mortality and Life Expectancy in Chinese: The Singapore Chinese Health Study.
To examine the impact of combined lifestyle factors on premature mortality and life expectancy in Chinese adults. A total of 44,052 Chinese adults aged 45-74 years free of cardiovascular disease (CVD), cancer, and diabetes were followed from recruitment (1993-1998) to the end of 2016 in the Singapore Chinese Health Study. A composite score (0-5 scale) was calculated based on five baseline healthy lifestyle factors including healthy diet, nonsmoking status, light to moderate alcohol drinking, being physically active and optimal body mass index. Mortality cases were identified through linkage with the nationwide death registry. Adopting five healthy versus none was associated with a lower risk of all-cause and cause-specific mortality, and the hazard ratio (95% confidence interval [CI]) was 0.38 (0.29, 0.51) for all-cause mortality, 0.26 (0.13, 0.52) for CVD mortality, and 0.59 (0.37, 0.92) for cancer mortality. Nonadherence to 4-5 healthy lifestyle factors accounted for 34.9% (95% CI = 29.2, 40.2) in population attributable fraction for all-cause mortality, 35.1% (23.7, 44.9) for CVD mortality, and 18.0% (6.5, 28.0) for cancer mortality. Conversely, adherence to 4-5 healthy lifestyle factors versus none could achieve a gain of 8.1 years in women and 6.6 years in men for the life expectancy at 50 years. A healthier lifestyle is associated with a substantially reduced risk of mortality and a longer life expectancy in the Chinese population. Our findings highlight the necessity of coordinated actions targeting combined lifestyle factors in reducing the overall burden of diseases and premature deaths.


Evaluation of blood-warming devices with the apparent thermal clearance.
Hypothermia (less than 35 C) associated with massive transfusions is a serious clinical problem. Maintenance of normal temperatures in this setting is essential and requires the use of rapid and efficient blood-warming devices. The optimal design of such units should include a low priming volume, large heat transfer area, low pressure drop, the ability to operate at high flow rates, and the capability of warming blood from 4 to 32 C, or higher, without untoward effects. We have previously formulated a theoretic model for blood warmers and described a measure of the intrinsic efficiency of these devices, the apparent thermal clearance (VT,cl). In this study, we tested four different types of commercially available blood warmers (the single-coil immersion heater; the single-channel dry wall electric heater; the multichannel countercurrent heat exchanger; and the single-channel, countercurrent heat exchanger) and determined the apparent thermal clearance values using saline and packed red blood cells. The results indicate that only the single-channel countercurrent unit is efficient enough to meet the established requirements. The apparent thermal clearance provides a useful and rigorous means for comparing and evaluating blood-warming devices.


Enhanced BCAT1 activity and BCAA metabolism promotes RhoC activity in cancer progression.
Increased expression of branched-chain amino acid transaminase 1 or 2 (BCAT1 and BCAT2) has been associated with aggressive phenotypes of different cancers. Here we identify a gain of function of BCAT1 glutamic acid to alanine mutation at codon 61 (BCAT1E61A) enriched around 2.8% in clinical gastric cancer samples. We found that BCAT1E61A confers higher enzymatic activity to boost branched-chain amino acid (BCAA) catabolism, accelerate cell growth and motility and contribute to tumor development. BCAT1 directly interacts with RhoC, leading to elevation of RhoC activity. Notably, the BCAA-derived metabolite, branched-chain α-keto acid directly binds to the small GTPase protein RhoC and promotes its activity. BCAT1 knockout-suppressed cell motility could be rescued by expressing BCAT1E61A or adding branched-chain α-keto acid. We also identified that candesartan acts as an inhibitor of BCAT1E61A, thus repressing RhoC activity and cancer cell motility in vitro and preventing peritoneal metastasis in vivo. Our study reveals a link between BCAA metabolism and cell motility and proliferation through regulating RhoC activation, with potential therapeutic implications for cancers.


[Possible effects of pharmacologic prophylaxis for Mendelson's syndrome on newborns].
Forty-eight newborn infants whose mothers underwent both elective and urgent caesarean section were studied. Mothers were randomly assigned to one of the treatment groups for the prophylaxis of Aspiration Pneumonitis: group A was treated with ranitidine, 50 mg i.v.; group B with ranitidine, 50 mg, and metoclopramide, 10 mg i.v.; group C no medication. Neonatal assessment included Apgar score and various haematological laboratory tests: haemogasanalysis, glycemia, electrolites, hematocrit, hemoglobin, bilirubinemia, SGOT, SGPT. Nor clinical or statistic differences were noted among the three infant groups.


Transanal endoscopic operation for rectal lesions using two-dimensional visualization and standard endoscopic instruments: a prospective cohort study and comparison with the literature.
The transanal endoscopic operation (TEO) has proved to be an effective alternative to conventional surgery for the treatment of rectal lesions. The TEO procedure offers reduced morbidity, faster recovery and equivalent oncologic outcome. Currently, two instrument sets are available: one with three-dimensional (Wolf) and one with two-dimensional (Storz) optic capacities. The three-dimensional (3D) instrument set is considered the golden standard. Although the advantages of TEO are imposing, the procedure with the 3D armamentarium has certain technical and financial drawbacks. This study therefore aimed to compare results for the TEO 2D alternative with recently published results for 3D TEO. All consecutive patients with benign or malignant pT1 or pT2 rectal lesions undergoing TEO were prospectively followed. All procedures were performed with the 2D armamentarium using standard endoscopic instruments, a rectoscope (diameter, 4 cm; working length, 7.5-15 cm), and 5-mm Ligasure and Ultracision. Operating times, complications, hospital stay, and oncologic outcome were gathered and compared with published data. Between 2004 and 2006, 31 patients with a median age of 75 years (range, 33-87 years) underwent 31 TEOs for a total of 36 rectal lesions (29 tubulovillous adenomas and 7 adenocarcinomas). The median distance of the lesion from the anal verge was 7.5 cm (range, 5-15 cm). The median lesion diameter was 2.3 cm (range, 0.5-5.0 cm). The locations of the lesions were as follows: 18 on the dorsal, 5 on the ventral, and 5 on the lateral rectal wall. The median operating time was 55 min (range, 25-165 min), compared with 105 min reported in the literature. All the lesions except one could be radically excised. No intraoperative complications occurred. Postoperative complications occurred for three patients, all due to hemorrhage. The median hospital stay was 3 days (range, 1-21 days). During a median follow-up period of 15 months (range, 1-35 months), two recurrences took place. The study findings showed that for rectal tumors located up to 15 cm from the anal verge with a maximal diameter of 5 cm, TEO using standard laparoscopic instruments with a 2D view is feasible and provides results comparable with those associated with a 3D view and dedicated instruments. Furthermore, the 2D procedure can be performed with improved ergonomics due to movable monitors and is considerably less expensive.


Fas-mediated apoptosis of melanoma cells and infiltrating lymphocytes in human malignant melanomas.
In a rodent system, melanoma cells expressing Fas ligand (FasL) could kill Fas-positive lymphocytes, suggesting that FasL expression was an essential factor for melanoma cell survival in vivo. These findings led us to investigate apoptosis, and to histochemically analyze involvement of Fas and FasL in the induction of apoptosis, in human malignant melanoma tissues. The percentages of terminal deoxynucleotidyl transferase-mediated biotin-dUTP nick end-labeling (TUNEL)-positive melanoma cells and of proliferating cell nuclear antigen (PCNA)-positive melanoma cells in melanoma tissues (n = 22) were greater than those in melanocytes in uninvolved skin (n = 6) and nevus cells in nevi tissues (n = 9). The infiltrating lymphocytes around melanomas were also TUNEL positive. Immunohistochemistry revealed expression of Fas and FasL in melanoma cells and lymphocytes, whereas no Fas or FasL expression was detected in normal skin melanocytes and nevus cells. There was significant correlation between Fas-positive indices and TUNEL indices in melanoma tissues. Moreover, TUNEL-, Fas-, and FasL-positive indices of melanoma cells from patients with Stage 3 melanomas were significantly lower than those with Stage 2 melanomas. The PCNA index of Stage 1 melanoma was significantly lower than that of the other stages, although the difference of PCNA index was insignificant among Stages 2 to 4. Among Stages 1 to 4, there was no difference in the PCNA, TUNEL-, and Fas-positive indices of lymphocytes, although the FasL-positive index of lymphocytes from Stage 3 melanomas was significantly lower than in that from Stage 2. These data reveal that melanoma cells and infiltrating lymphocytes have the potential to induce their own apoptosis regulated by Fas and FasL in an autocrine and/or paracrine fashion and that the decline of Fas-mediated apoptosis of melanoma cells, rather than the apoptosis of infiltrating lymphocytes, may affect the prognosis of melanoma patients, possibly through the accumulation of more aberrant cells acquiring metastatic activity.


Comorbid conditions associated with Parkinson's disease: a population-based study.
The burden of comorbidity in Parkinson's disease (PD) remains unclear. All Olmsted County, Minnesota, residents with incident PD in 1976-1995 (n = 197) plus one age- and sex-matched non-PD referent subject per case were followed for all clinical diagnoses from 5 years before through 15 years after index (i.e., year of PD onset for each case and same year for the referent subject). Both members of a case-referent pair were censored at death or emigration of either member to ensure equivalent follow-up. Cases and referent subjects were compared for summary comorbidity (Charlson index) and for the likelihood of having one or more diagnoses within each International Classification of Diseases chapter/subchapter. Before index, the groups were similar for all comparisons. After index, cases had a higher likelihood of diagnoses within the chapters "Mental Disorders" and "Diseases of the Genitourinary System," and within the subchapters "Organic Psychotic Conditions," "Other Psychoses," "Neurotic/Personality/Other Nonpsychotic Disorders," "Hereditary/Degenerative Diseases of Central Nervous System," "Symptoms," "Other Diseases of Digestive System," "Other Diseases of Urinary System," "Diseases of Veins/Lymphatics/Other Circulatory System Diseases," "Fractures of Lower Limb," "Other Diseases of Skin/Subcutaneous Tissue," "Osteopathies/Chrondropathies/Acquired Musculoskeletal Deformities," and "Pneumonia and Influenza." The excess morbidity and mortality observed for persons with PD are consistent with recognized PD sequelae.


Signal regulatory protein α negatively regulates mast-cell activation following FcεRI aggregation.
Mast cells elicit allergic reaction through degranulation and release of proinflammatory mediators after aggregation of the IgE receptor FcεRI. Here we provide evidence to show that signal regulatory protein α (SIRPα), an ITIM-containing receptor, is an endogenous regulator of IgE-Ag induced mast-cell activation. SIRPα expression is promptly reduced in mast cells in response to FcεRI aggregation. Impaired expression of SIRPα in mast cells facilitates FcεRI-evoked degranulation and de novo synthesis of cytokines (IL-4, IL-13, IL-6, and TNF-α). We further demonstrate that SIRPα knockdown in mast cells accelerates calcium mobilization and affects cytoskeletal rearrangement (F-actin disassembly and polymeric tubulin formation) after FcεRI aggregation. Mechanistic studies highlight the prolonged activation of NF-κB and MAPKs as well as PLC-γ after FcεRI stimulation as a consequence of the inhibition of SIRPα expression in mast cells. Immunoprecipitation analysis shows that SIRPα knockdown markedly increases IgE-induced SHP2 interaction with PI3K regulatory subunit PI3Kp85 or IKK-β in mast cells, indicating that SIRPα may accomplish this through its association and sequestration of SHP2. Collectively, our results strongly indicate that SIRPα is a biological important regulator of FcεRI signaling.


Effect of polyunsaturated fatty acids of the alpha-linolenic series on the development of rat testicles.
The effect of 22:6 omega 3 acid provided by dietary fish oil on the development of germinal tissue of rat testes, fatty acid composition of lipids, and linoleic or alpha-linolenic acid delta 6 desaturation capacity was investigated. Results were compared to those obtained in animals fed methyl palmitate and sunflower seed oil (linoleate). At 7 and 9 weeks of age, development of germinal tissue of animals fed fish oil was normal. The fatty acid composition showed a decrease in 22:5 omega 6 acid content and an increase in 22:6 omega 3 acid in triacylglycerol, phosphatidylcholine, and phosphatidylethanolamine. The fatty acid delta 6 desaturation capacity of testicules microsomes was increased. It is suggested that 22:6 omega 3 acid may functionally replace 22:5 omega 6 acid in germinal tissue.


Relationship between the initial dose of micafungin and its efficacy in patients with candidemia.
Micafungin, the first licensed echinocandin in Japan, has shown excellent in vitro and in vivo activity against all Candida species. However, the appropriate dose for the initial treatment of candidemia remains to be determined. In this study, we retrospectively examined the relationship between the clinical outcome of candidemia and the initial dose of micafungin. Patients were divided into two groups according to the initial dose of micafungin administered: group I (<2.25 mg/kg/day) and group II (>or=2.25 mg/kg/day). Micafungin produced an excellent 30-day clinical response in patients with candidemia, including Candida parapsilosis; the overall 30-day clinical response was 86%. The administration of higher doses of micafungin accelerated the clinical response and duration until the clinical response in group II was significantly shorter than that in group I (P = 0.021). However, no significant differences were observed in the 30-day mortality attributable to the fungal infection between the two groups. Considering these results, we recommend the administration of 2.25 mg/kg/day or more of micafungin in the initial treatment of patients with candidemia.


The structure of the PDZ3-SH3-GuK tandem of ZO-1 protein suggests a supramodular organization of the membrane-associated guanylate kinase (MAGUK) family scaffold protein core.
Membrane-associated guanylate kinases (MAGUKs) are a large family of scaffold proteins that play essential roles in tethering membrane receptors, adhesion molecules, and macromolecular signaling complexes for tissue developments, cell-cell communications, and intracellular signal transductions. The defining feature of the MAGUK family scaffolds is that each member contains a conserved core consisting of a PSD-95/Dlg/ZO-1 (PDZ) domain, an Src homology 3 (SH3) domain, and a catalytically inactive guanylate kinase (GuK) domain arranged in tandem, although the structural features and functional implications of the PDZ-SH3-GuK tandem arrangement are unclear. The structure of the ZO-1 PDZ3-SH3-GuK tandem solved in this study reveals that the PDZ domain directly interacts with the SH3-GuK module, forming a structural supramodule with distinct target binding properties with respect to the isolated domains. Structure-based sequence analysis suggests that the PDZ-SH3-GuK tandems of other members of the MAGUK family also form supramodules.


HLA genotype as a marker of multiple sclerosis prognosis: A pilot study.
The identification of a biomarker with prognostic value is an unmet need in multiple sclerosis (MS). The objective of this study was to investigate a possible association of HLA genotype with disease status and progression in MS, based on comprehensive and sensitive clinical and magnetic resonance imaging (MRI) parameters to measure disease effects. A total of 118 MS patients (79 females, 39 males) underwent HLA typing. Patient MS status was assessed at two time points in a 2-year interval, based on clinical scores (including EDSS, MSSS, T25FW, 9-HPT, SDMT, BVMT, CVLT-II) and MRI evaluations. Quantitative brain MRI values were obtained for whole brain atrophy, FLAIR lesion volume change and number of new lesions using MSmetrix. Predefined HLA patient groups were compared as of disease status and progression. Global assessment was achieved by an overall t-statistic and assessment per measurement by a Welch test and/or Mann Whitney U test. The effects of multiple covariates, including age, gender and disease duration as well as scan parameters, were also evaluated using a regression analysis. The HLA-A*02 allele was associated with better outcomes in terms of MSSS, EDSS and new lesion count (Welch test p-value<0.05). The HLA-B*07 and HLA-B*44 alleles showed a global negative effect on disease status, although none of the measurements reached significance (p-value<0.05). Results for the HLA-DRB1*15, HLA-DQB1*06 and HLA-B*08 alleles were inconclusive. The influence of the confounding variables on the statistical analysis was limited.


Naturally-occurring neoplasms in the Mongolian gerbil, Meriones unguiculatus.
10 spontaneous tumours were observed. Several had not been previously reported for this species and 1 occurred in the youngest gerbil yet reported to have a spontaneous tumour. 7 of the 10 tumours originated from the female reproductive tract.


Hypothesis: is viral transformation mediated by alterations in chromosomal proteins?
Efforts to clarify the mechanisms of viral carcinogenesis have been hampered, no doubt, by the lack of understanding of the complex gene-regulatory processes in the eukaryotic genome. However, numerous investigators have produced enlightening information concerning potential gene-regulatory, nuclear phosphorylation reactions. In addition, equally interesting studies involving transforming viruses have revealed that some viral products responsible for transformation have an associated protein kinase activity. These similar biochemical processes may suggest how some transforming viruses could subvert normal gene-regulatory processes.


Methacarn fixation for genomic DNA analysis in microdissected cells.
We have found methacarn, a non-crosslinking protein-precipitating fixative, to be useful for the analysis of DNA from microdissected specimens of wax-embedded tissue. In this chapter, we present the procedure regarding genomic DNA analysis in methacarn-fixed wax-embedded microdissected rat tissue. Using nested polymerase chain reaction (PCR), and a rapid extraction procedure, fragments of DNA up to 2.8 kb in size can be amplified from a 1 x 1 mm area of a 10-microm-thick tissue section. Target fragments of about 500 bp can be amplified from a single cell, but 10-20 cells are necessary for practical detection by nested PCR. Although tissue staining with hematoxylin and eosin inhibits the PCR, amplification of about 500-bp fragments is successful with 150-270 cells by single-step PCR. Immunostaining results in a substantial decrease of yield and degradation of extracted DNA. However, even after immunostaining, fragments of about 180 bp can be amplified with 150-270 cells by single-step PCR. These features demonstrate the suitability of methacarn-fixed wax-embedded tissue for practical genomic DNA analysis in terms of tissue handling, extraction efficiency, and satisfactory PCR results.


Biodegradation of the neonicotinoid insecticide Acetamiprid by bacterium Pigmentiphaga sp. strain AAP-1 isolated from soil.
The Acetamiprid-degrading bacterium AAP-1 was isolated from contaminated soil, and identified as Pigmentiphaga sp. combined traditionary categorization method with modern molecule method. The strain could utilize Acetamiprid as the sole carbon, nitrogen and energy source for growth and metabolized 100 mgL(-1) Acetamiprid within 2.5h. During the degradation of Acetamiprid, one N-deacetylation metabolite, was characterized by FT-IR, GC-MS and NMR analysis. A novel microbial biodegradation pathway for Acetamiprid was proposed on the basis of the metabolite. Compared with uninoculated soils, the addition of the AAP-1 strain into soils treated with Acetamiprid gained a higher degradation rate, and the bacteria community analysis by T-RFLP in contaminated soil recovered after inoculation of the AAP-1 strain. On the basis of these results, strain AAP-1 has the potential to be used in the bioremediation of Acetamiprid-contaminated environments. This is the first report of Acetamiprid-degrading isolate from the genus of Pigmentiphaga.


An RNA polymerase II holoenzyme responsive to activators.
RNA polymerase II requires multiple general transcription factors to initiate site-specific transcription. These proteins can assemble in an ordered fashion onto promoter DNA in vitro, and such ordered assembly may occur in vivo (Fig. 1a). Some general transcription factors can interact with RNA polymerase II in the absence of DNA, however, suggesting that RNA polymerase II may also assemble into a multi-component complex containing a subset of initiation factors before binding to promoter DNA (Fig. 1b). Here we present evidence from the yeast Saccharomyces cerevisiae for such an RNA polymerase II holoenzyme, a multi-subunit complex containing roughly equimolar amounts of RNA polymerase II, a subset of general transcription factors, and SRB regulatory proteins. Transcription by this holoenzyme is stimulated by the activator protein GAL4-VP16, a feature not observed with purified RNA polymerase II and general transcription factors alone. We propose that the holoenzyme is a form of RNA polymerase II readily recruited to promoters in vivo.


Determinants of ochratoxin A exposure--a one year follow-up study of urine levels.
Dietary exposure to the ochratoxin A (OTA) occurring in Portugal is characterized by a high frequency of contamination of the consumed foodstuffs, although at low levels. The exposure bears significance for the total food consumed, and not for a particular one. Biomonitoring studies are thus fundamental in simplifying the evaluation of exposure, with no need to examine the entire range of consumed foodstuffs. Biomonitoring studies further allow the identification of host factors as predictors of OTA exposure in epidemiologic studies, the results of which are merited for targeting intervention strategies by public health authorities and advising official regulatory decisions. Using a longitudinal approach, this study examined factors related to OTA exposure in the adult population over a one-year period. Anthropometric measures, season of the year and region were the selected factors correlated with OTA exposure biomarker. Urine samples from 95 inhabitants from six Portuguese main geographical areas were assayed through spectrofluorimetric detection. Exposure to OTA proved to markedly increase in winter, and gender differences were observed only in summer, which might be related to different dietary patterns not only between seasons, but also between genders. The same rationale may also serve the observed statistically significant differences between some regions. No other strong association upon the remaining determinants under testing was observed. These observations reinforce the need for OTA exposure evaluation, possibly specifically targeting the staple foods or dietary habits that sustain potential predictors or determinants of exposure.


Crystal structure of the erythromycin polyketide synthase dehydratase.
The dehydratases (DHs) of modular polyketide synthases (PKSs) catalyze dehydrations that occur frequently in the biosynthesis of complex polyketides, yet little is known about them structurally or mechanistically. Here, the structure of a DH domain, isolated from the fourth module of the erythromycin PKS, is presented at 1.85 A resolution. As with the DH of the highly related animalian fatty acid synthase, the DH monomer possesses a double-hotdog fold. Two symmetry mates within the crystal lattice make a contact that likely represents the DH dimerization interface within an intact PKS. Conserved hydrophobic residues on the DH surface indicate potential interfaces with two other PKS domains, the ketoreductase and the acyl carrier protein. Mutation of an invariant arginine at the hypothesized acyl carrier protein docking site in the context of the erythromycin PKS resulted in decreased production of the erythromycin precursor 6-deoxyerythronolide B. The structure elucidates how the alpha-hydrogen and beta-hydroxyl group of a polyketide substrate interact with the catalytic histidine and aspartic acid in the DH active site prior to dehydration.