Unusual SCD in cancer cells: a phenomenon related to decreased BrdU incorporation?
The simultaneous presence of chromosome segments with and without sister chromatid differentiation (SCD) in the same metaphase was observed in a human melanoma cell line after BrdU incorporation for 48-72 hr. This phenomenon was related to the time of BrdU incorporation by the cells: i.e., it was not observed after exposure to BrdU for only 17-20 hr. It is proposed, therefore, that the unusual staining pattern is caused through reduced BrdU incorporation by the cells after completing the first division.


Brood size and the cost of provisioning nestlings: interpreting Lack's hypothesis.
A recent model of parental provisioning (the "tradeoff model") suggests that the maximum delivery rate of food to nestlings represents a tradeoff between parental residual reproductive value and nestling survival. In contrast, Lack's hypothesis suggests that maximum provisioning rate determines brood size and therefore delivery rates are limited by shortages of food or foraging time, not by tradeoffs of parental investment. Several authors have examined the shape of the per-nestling feeding curves to test the tradeoff model against Lack's hypothesis. We show that Lack's hypothesis can produce per-nestling feeding curves consistent with the tradeoff model. Therefore, the shape of the per-nestling feeding curve cannot be used to distinguish between the models.


Outcomes Following ACL Reconstruction Based on Graft Type: Are all Grafts Equivalent?
Anterior cruciate ligament reconstruction is one of the most common orthopedic procedures performed, accounting for over 200,000 cases annually. Despite the high prevalence, there is still much debate as to the optimal graft choice. The purpose of this review is to evaluate the current literature and discuss the reported outcomes for the most common graft choices. The most common autografts being used include bone-patellar tendon-bone (BPTB), hamstring tendon (HT), and quadriceps tendon (QT). Hamstring tendon might have a slightly higher re-tear rate when compared with BPTB (2.84 versus 2.80). However, BPTB has a higher rate of anterior knee and kneeling pain in the short- and mid-term follow-up. This has not been shown to be the case in long-term follow-up. Allograft is a viable option for revisions and primaries in patients greater than 35 years old; however, re-tear rate increases significantly in younger patients. ACL reconstruction graft choice is a highly studied and yet still exceedingly debated topic. Most large studies report either no significant difference or a small difference in failure rate and outcome scores between the different autograft choices. Allografts have been demonstrated to have an increased risk of failure in younger athletes and should be reserved for revision cases and those aged 35 years and older. Graft choice should ultimately be decided upon based on surgeon comfort and experience and individual patient characteristics.


MIMP: predicting the impact of mutations on kinase-substrate phosphorylation.
Protein phosphorylation is important in cellular pathways and altered in disease. We developed MIMP (http://mimp.baderlab.org/), a machine learning method to predict the impact of missense single-nucleotide variants (SNVs) on kinase-substrate interactions. MIMP analyzes kinase sequence specificities and predicts whether SNVs disrupt existing phosphorylation sites or create new sites. This helps discover mutations that modify protein function by altering kinase networks and provides insight into disease biology and therapy development.


The microbial metabolite Urolithin A reduces C. difficile toxin expression and repairs toxin-induced epithelial damage.
Clostridioides difficile is a gram-positive, anaerobic, spore-forming bacterium that is responsible for antibiotic-associated pseudomembranous colitis. Clostridioides difficile infection (CDI) symptoms can range from diarrhea to life-threatening colon damage. Toxins produced by C. difficile (TcdA and TcdB) cause intestinal epithelial injury and lead to severe gut barrier dysfunction, stem cell damage, and impaired regeneration of the gut epithelium. Current treatment options for intestinal repair are limited. In this study, we demonstrate that treatment with the microbial metabolite urolithin A (UroA) attenuates CDI-induced adverse effects on the colon epithelium in a preclinical model of CDI-induced colitis. Moreover, our analysis suggests that UroA treatment protects against C. difficile-induced inflammation, disruption of gut barrier integrity, and intestinal tight junction proteins in the colon of CDI mice. Importantly, UroA treatment significantly reduced the expression and release of toxins from C. difficile, without inducing bacterial cell death. These results indicate the direct regulatory effects of UroA on bacterial gene regulation. Overall, our findings reveal a novel aspect of UroA activities, as it appears to act at both the bacterial and host levels to protect against CDI-induced colitis pathogenesis. This research sheds light on a promising avenue for the development of novel treatments for C. difficile infection.


Helicobacter pylori picB, a homologue of the Bordetella pertussis toxin secretion protein, is required for induction of IL-8 in gastric epithelial cells.
Approximately 60% of Helicobacter pylori strains are cagA+ and this genotype is more frequently associated with duodenal ulcer disease. Although most wild-type cagA+ strains are both cytotoxigenic and induce enhanced Interleukin-8 (IL-8) secretion in gastric epithelial cells, isogenic cagA- mutants retain full activity in these assays; thus, cagA appears to be a marker of enhanced virulence. Delineation of the nucleotide sequence of a 4 kb region upstream of cagA allowed the identification of 966 bp (picA) and 2655 bp (picB) open reading frames encoding 36 kDa and 101 kDa polypeptides, respectively. picA and picB constitute an operon in opposite orientation to cagA. The deduced picB product showed significant homology (26% identity and 50% similarity) with the Bordetella pertussis toxin secretion protein (PtlC). Of 55 H. pylori clinical isolates, the picA and picB segment was conserved exclusively in cagA+ strains and present in all isolates from patients with duodenal ulceration, versus 59% of isolates from patients with gastritis alone (P = 0.01). Using gene-replacement techniques, we constructed picA and picB mutant H. pylori strains and demonstrated that the picB gene product is involved in the induction of IL-8 expression in gastric epithelial cells. Further, Northern blot hybridization and RT-PCR data showed that picA and picB are co-transcribed and an insertional mutation in picA ablates picB expression. These studies indicate a role of picA and picB in the induction of an inflammatory response in gastric epithelial cells either directly or by enabling secretion of an unidentified product, and suggest a mechanism for the overrepresentation of strains possessing these genes in patients with peptic ulceration.


Relationship between the activity of pancreatic phospholipase A2 and the physical state of the phospholipid substrate.
When pancreatic phospholipase A2 is mixed with small unilamellar vesicles made from dipalmitoyl phosphatidylcholine at or above the phase transition temperature (tm), the time course of hydrolysis exhibits a distinct lag period. On the other hand, if the enzyme is preincubated with substrate for a short period of time below the transition temperature (during which time only limited hydrolysis occurs) and then assayed at high temperature, no lag period is observed. This phenomenon does not appear to be the result of product formation. Instead, the reported results lend substantial support to the hypothesis that a relatively slow substrate-enzyme organizational step is required as part of an activation process and this is most rapid when the substrate is in the gel state. However, the intrinsic activity (after the initial step of activation) is maximal when the substrate is in the liquid-crystalline state.


Early complications of high-dose-rate brachytherapy in soft tissue sarcoma: a comparison with traditional external-beam radiotherapy.
Radiotherapy and surgery are routinely utilized to treat extremity soft tissue sarcoma. Multiple radiation modalities have been described, each with advantages and disadvantages, without one modality demonstrating clear superiority over the others. We determined the overall initial complication rate in patients receiving surgery and radiotherapy, which specific complications were found when comparing different modalities, and whether combination therapy increased the overall rate of complications compared with surgery and single-modality radiotherapy. We retrospectively reviewed the records of 190 patients who received external-beam radiotherapy (141 patients), high-dose-rate brachytherapy (37 patients), or both (12 patients). We evaluated 100 men and 90 women (mean age, 57 years; range, 18-94 years) for tumor size and subtype, comorbidities, stage, grade, margin of resection, type of adjuvant treatment, and complications. Minimum followup was 3 months (mean, 40 months; range, 3-155 months). The most frequent early complications in the high-dose-rate brachytherapy cohort were infection, cellulitis, and seroma and/or hematoma. In the external-beam radiotherapy cohort, chronic edema, fibrosis, and chronic radiation dermatitis were more frequently encountered. The total number of early complications and overall incidence of major complications requiring further surgery were similar among the three cohorts, but a larger number of patients in the high-dose-rate brachytherapy group required subsequent surgery for infection compared with the external-beam radiotherapy group. High-dose-rate brachytherapy decreases radiation exposure and allows shorter duration of treatment compared with traditional external-beam radiotherapy but has a higher perioperative wound complication rate. Level III, therapeutic study. See the Guidelines for Authors for a complete description of levels of evidence.


Meta-analysis Comparing Outcomes in Patients With and Without Cardiac Injury and Coronavirus Disease 2019 (COVID 19).
Current evidence is limited to small studies describing the association between cardiac injury and outcomes in patients with coronavirus disease 2019 (COVID-19). To address this, we performed a comprehensive meta-analysis of studies in COVID-19 patients to evaluate the association between cardiac injury and all-cause mortality, intensive care unit (ICU) admission, mechanical ventilation, acute respiratory distress syndrome, acute kidney injury and coagulopathy. Further, studies comparing cardiac biomarker levels in survivors versus nonsurvivors were included. A total of 14 studies (3,175 patients) were utilized for the final analysis. Cardiac injury in patients with COVID-19 was associated with higher risk of mortality (risk ratio [RR]:7.79; 95% confidence interval [CI]: 4.69 to 13.01; I2=58%), ICU admission (RR: 4.06; 95% CI: 1.50 to 10.97; I2 = 61%), mechanical ventilation (RR: 5.53; 95% CI: 3.09 to 9.91; I2 = 0%), and developing coagulopathy (RR: 3.86; 95% CI:2.81 to 5.32; I2 = 0%). However, cardiac injury was not associated with increased risk of acute respiratory distress syndrome (RR:3.22; 95% CI:0.72 to 14.47; I2 = 73%) or acute kidney injury (RR: 11.52, 95% CI:0.03 to 4,159.80; I2 = 0%). The levels of hs-cTnI (MD:34.54 pg/ml;95% CI: 24.67 to 44.40 pg/ml; I2 = 88%), myoglobin (MD:186.81 ng/ml; 95% CI: 121.52 to 252.10 ng/ml; I2 = 88%), NT-pro BNP (MD:1183.55 pg/ml; 95% CI: 520.19 to 1846.91 pg/ml: I2 = 96%) and CK-MB (MD:2.49 ng/ml;95% CI: 1.86 to 3.12 ng/ml; I2 = 90%) were significantly elevated in nonsurvivors compared with survivors with COVID-19 infection. The results of this meta-analysis suggest that cardiac injury is associated with higher mortality, ICU admission, mechanical ventilation and coagulopathy in patients with COVID-19.


Percutaneous closure of moderate to large perimembranous ventricular septal defect in small children using left ventricular mid-cavity approach.
To report mid-term follow-up result of transcatheter closure of perimembranous Ventricular septal defect (VSD) in children weighing less than 10 kg using Amplatzer Duct Occlude-I (ADO-I) by left ventricular (LV) mid-cavity approach. This is retrospective review of 35 children weighing less than 10 kg with moderate to large perimembranous VSD who were selected for transcatheter closure of VSD using ADO-I in between October 2016 to September 2018. Mean age was 2.08 ± 0.67 years (mean ± SD) and mean weight was 7.2 ± 1.2 kg (mean ± SD). Procedure was done by crossing the VSD from right ventricular side instead of using the standard approach by forming arterio-venous loop. Average fluoroscopic time was 9.2 ± 2.9 min (mean ± SD) and mean procedure time was 34.1 ± 13.1 min (mean ± SD). Mean follow-up period was 8.7 months (3-12 months) RESULT: Device closure was successfully done in 32 patients. There was device induced aortic regurgitation (AR) in one case who was sent for surgery. One child was found to have complete heart block on first post-procedure day requiring permanent pace-maker implantation. During follow up none of them had any residual VSD, rhythm disturbance, AR and left or right ventricular outflow obstruction. Device closure can be successfully done in moderate to large perimembranous VSD using left ventricular mid cavity approach in small children. LV mid-cavity approach has advantages in terms of lesser hemodynamic instability, lesser fluoroscopy and lesser chance of device induced AR than standard technique particularly in smaller children.


Gender Difference in Offspring's Alcohol Use Disorder by 21 Years: A Longitudinal Study of Maternal Influences.
There is little known about the extent to which maternal alcohol consumption influences offspring's alcohol use disorder. This study aims to examine whether different maternal alcohol consumption trajectories predict gender difference in adolescent alcohol use disorder at child age 21 years. Data are from a prospective cohort, the Mater-University of Queensland Study of Pregnancy (MUSP) and its outcomes. The study involves 2531 mother-child pairs for whom data are available at the 21-year follow-up survey. Maternal alcohol consumption trajectories were determined by group-based trajectory modelling. Offspring's lifetime ever alcohol use disorder was assessed using DSM-IV diagnostic criteria. Over 14 years of follow-up after the birth of a child, three distinct alcohol consumption trajectories were identified (abstainer, low-stable. and moderate-escalating drinker). A maternal trajectory of moderate-escalating alcohol consumption independently predicted offspring's lifetime ever alcohol use disorder at 21 years after adjustment for a range of potential confounders. "Cross-gender influence" is observed in the study. A maternal life course pattern of alcohol consumption may have an independent effect on offspring alcohol consumption, with male offspring being more vulnerable to the effects of maternal alcohol use than are female offspring. Programs intended to address alcohol consumption by adolescents and young adults need to focus on the behaviors of both parents but acknowledging that maternal patterns of alcohol consumption may be particularly important for male offspring.


Sickle cell disease: acute clinical manifestations in early childhood and molecular characteristics in a group of children in Rio de Janeiro.
To describe clinical events of sickle cell disease and the correlation with β-globin haplotypes and α-thalassemia in under 6-year-old children. A retrospective study was conducted of under 6-year-old children from the neonatal screening program in Rio de Janeiro. Forty-eight male and 48 female children were enrolled in this study, 79 with sickle cell anemia and 17 with hemoglobin SC. The mean age was 29.9 (standard deviation = 20.9) months, 62 (16.2 ± 8.6) were aged between 0-3 years old and 34 (54.9 ± 11.3) were from 3-6 years old. Painful events, acute splenic sequestration, hemolytic crises, hand-foot and acute chest syndromes and infections were evaluated. The events were more frequent in under 3-year-old children, 94% of children had at least one episode. Infection was the most common event affecting 88.5% of children. Acute splenic sequestration took place earlier, while painful crises and acute chest syndromes in under 6-year-old children. Thal-α 3.7 was observed in 20.9% of cases. Bantu was the most frequent haplotype found, followed by Benin. No correlation was observed between clinical events and β-globin haplotypes. Children with sickle cell anemia and α-thalassemia have less infectious events. No correlation was found among these polymorphisms and clinical events, however, the majority of children with Bantu/Bantu and without α-thalassemia had more clinical events.


Analysis of size characterized manganese species from liver extracts using capillary zone electrophoresis coupled to inductively coupled plasma mass spectrometry (CZE-ICP-MS).
Mn is of toxicological concern because overexposure can lead to progressive, permanent neurodegenerative damage. Monomethyl-Mn-pentadienyl-tricarbonyl (MMT) is used as an anti-knock agent in fuel. Exhausted Mn compounds are absorbed in the lung and transported to the liver. Extended exposure causes an overflow of the liver with Mn species moving e.g. to the brain, causing irreversible central nervous system (CNS) disorders like Manganism. This paper focuses on experiments for getting more information on Mn species in liver extracts. The investigations are performed with respect to (1) a size characterization and (2) a subsequent identification of the Mn species in liver extracts using preparative size exclusion chromatography (SEC) followed by capillary zone electrophoresis coupled to inductively coupled plasma mass spectrometry (CZE-ICP-MS). First, extracts were analyzed using a mass calibrated SEC column coupled to ICP-MS detection. The chromatogram showed the 55 Mn-trace and proved main Mn elution between ca. 60-150 kDa. Second, liver extracts were fractionated on the same SEC column, however, now the effluent was directed to a fraction collector. This resulted in fractions containing pre-purified, size characterized Mn species per fraction. It turned out that the Mn concentrations per fraction reflected roughly the previous on-line Mn trace. Third, the fractions were subject to CZE-ICP-MS, where the MS was operated additionally with dynamic reaction cell (DRC) technique. From size characterization (with SEC coupled on-line to ICP-MS or connected to a fraction collector and subsequent Mn determination in fractions) it was shown that most Mn species from liver extract were of high molecular mass (HMM) nature as they eluted mostly between 50 and 80 min, corresponding to ca. 60-150 kDa. With the two-dimensional speciation approach employing first SEC and then CZE-ICP-DRC-MS together with standard addition method, a series of Mn species was identified. Mn species predominantly were Mn-enzymes e.g. arginase, isocitric dehydrogenase, galactosyltransferase, prolidase, pyruvate carboxylase and oxalate oxidase. A typical Mn-transporter--Mn-albumin-- was also seen, whilst Mn-transferrin obviously was degraded during SEC separation. This Mn-compound (independent whether as a standard or from liver extract) was not stable during SEC even at the finally chosen physiological conditions.


Ultraconservative dentistry.
Ultraconservative techniques are providing dentists with new opportunities for the rehabilitation of the debilitated dentition. These techniques involve the use of bonded, reinforced ceramics that restore function and esthetics. The new parameters of ultraconservative treatment include the routine use of supragingival margins, color gradients, and the alteration of the occlusal vertical dimension. The minimally invasive nature of these new techniques makes them more readily acceptable to, and more easily maintainable by, patients.


Radiocesium immobilization to leaf litter by fungi during first-year decomposition in a deciduous forest in Fukushima.
Vast forest areas in eastern Japan have been contaminated with radio-isotopes by the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. Radiocesium (radioCs) is known to remain bioavailable in forest ecosystems for a long time, and it is necessary to terminate the cycling process to decontaminate the forest ecosystem. We observed radiocesium concentrations of leaf litter during decomposition on a forest floor where radiocesium ((137)Cs) contamination was ∼155 kBq/m(2). Litter bag experiments were conducted with newly fallen mixed deciduous leaf litter in a deciduous forest (alt. 610 m) about 50 km from the FDNPP. Litter bags were retrieved in April, June, August, October, and December 2012. Fresh litter (137)Cs concentration was ∼3000 Bq/kg in December 2011. During the decomposition process on the forest floor, litter (137)Cs concentration increased rapidly and exceeded 25,000 Bq/kg after 6 months, whereas potassium (K) concentration in the litter was rather stable, indicating that radiocesium and K showed contrasting dynamics during the early decomposition phase. Nitrogen, phosphorus, and (137)Cs contents were positively correlated to fungal biomass, evaluated by phospholipid fatty acids in the litter during decomposition. The increase of radiocesium concentration mainly occurred during from April to October, when fungal growth peaked. Therefore, this suggests fungal translocation of nutrients from outside the litter substrate (immobilization) is the mechanism to increase radiocesium in the decomposing litter. The amount of (137)Cs contained in the 1-year-old decomposed leaf litter was estimated to be 4% per area of the soil-contaminated (137)Cs.


Particulate matter (PM10) exposure induces endothelial dysfunction and inflammation in rat brain.
Epidemiological studies suggest that particulate matter (PM(10)) inhalation was associated with adverse effects on brain-related health, however, existing experimental data lacked relevant evidences. In this study, we treated Wistar rats with PM(10) at different concentrations (0.3, 1, 3 and 10 mg/kg body weight (bw)), and investigated endothelial dysfunction and inflammatory responses in the brain. The results indicate that mild pathological abnormal occurred after 15-day exposure (five times with 3 days each), followed by the changes of endothelial mediators (ET-1 and eNOS) and inflammatory markers (IL-1β, TNF-α, COX-2, iNOS and ICAM-1). Also, the sample up-regulated bax/bcl-2 ratio and p53 expression, and induced neuronal apoptosis. It implicates that PM(10) exerted injuries to mammals' brain, and the mechanisms might be involved in endothelial dysfunction and inflammatory responses.


Inflammatory Bowel Disease.
Inflammation response plays an important role in host survival, and it also leads to acute and chronic inflammatory diseases such as rheumatoid arthritis, bowel diseases, allergic rhinitis, asthma, atopic dermatitis and various neurodegenerative diseases. During the course of inflammation, the ROS level increases. In addition to ROS, several inflammatory mediators produced at the site lead to numerous cell-mediated damages. Inflammatory bowel disease (IBD), including ulcerative colitis and Crohn's disease, is a chronic intestinal disorder resulting from a dysfunctional epithelial, innate and adaptive immune response to intestinal microorganisms. The methods involving indomethacin-induced enterocolitis in rats with macroscopic changes of IBD, myeloperoxidase assay, microscopic (histologic) characters and biochemical parameters are discussed.


Conformational effects of coenzyme binding to porcine lactic dehydrogenase.
Changes induced on addition of the coenzyme, NADH or NAD+, to porcine lactic dehydrogenase isoenzymes H4 and M4 have been studied by hydrodynamic and spectroscopic methods. As shown by ultracentrifugal analysis, the native subunit structure remains unchanged on holoenzyme formation; an approximately 5% increase of the sedimentation coefficient, parallelled by a slight decrease of the partial specific volume (less than 1%) indicate a significant change in the native tertiary and/or quaternary structure of the enzymes, corroborating earlier calorimetric data (Hinz and Jaenicke, 1975). The binding constant for the enzyme from skeletal muscle (M4) and NADH are found to be in agreement with KD-values obtained from equilibrium dialysis, as well as spectroscopic and thermal titration experiments (8 muM). Far UV circular dichroism measurements do not show significant changes on ligand binding, indicating unchanged helicity or compensatory conformational effects. In the near UV, ligand binding is reflected by an extrinsic Cotton effect around 340 nm; in the range of aromatic absorption no changes are detectable. The experimental results suggest that there are gross structural changes on coenzyme binding to lactic dehydrogenase which do not affect the intrinsic spectral properties normally applied to analyze transconformation reactions in protein molecules.


A proposed cause for the hepatic dysfunction associated with parenteral nutrition.
Total parenteral nutrition (TPN) is associated with cholestasis and hepatic steatosis, which can be lethal in infants who cannot be fed orally. It was determined that route of administration was not the critical variable in the development of hepatic steatosis. Two groups of young rats received equivalent amounts of a standard TPN solution either orally or intravenously for an 8- to 10-day period during which they received no other nutrition. Both groups gained equivalent weight and developed marked hepatic steatosis. To test whether the solution was toxic or deficient, three groups of rats were given TPN solution orally and a fraction of their usual daily intake of rat chow. Rats receiving less than 10% of their usual chow intake developed steatosis; rats receiving more than that did not. To determine the solubility of the protective material in chow, two groups of rats were given TPN solution orally and chow that been extracted with either water or the organic solvent chloroform. Rats eating the water-extracted chow developed steatosis, rats eating chloroform-extracted chow did not. Although the protective component in chow was apparently water soluble, addition of a water soluble extract of chow to the TPN solution fed another two groups of rats did not prevent steatosis at 0.1 mg/mL and only partially, if at all, at 10 mg/mL. TPN-related hepatic dysfunction, as measured by the development of hepatic steatosis in this model, may be due to a deficiency in the TPN solution. The missing constituent(s) appears to be present in rat chow and can be extracted with water, but not with an organic solvent.


Multivariate linear regression model based on cross-entropy for estimating disorganisation in drone formations.
Static formations of swarms of rotorcraft drones, used for example in disaster management, are subject to intrusions, and must bear the cost of holding the formation while avoiding collisions which leads to their increased energy consumption. While the behaviour of the intruder is unpredictable, the formation can have its parameters set to try to balance the cost of avoidance with its functionality. The novel model presented in this paper assists in the selection of parameter values. It is based on multivariate linear regression, and provides an estimate of the average disturbance caused by an intruder as a function of the values of the parameters of a formation. Cross-entropy is used as a metric for the disturbance, and the data based are generated through simulations. The model explains up to 54.4% of the variability in the value of the cross-entropy, providing results that are twice as good as the baseline estimator of the mean cross-entropy.


The effect of adult feeding on the fecundity, weight loss and survival of the pine beauty moth, Panolis flammea (D&S).
Adult females of the pine beauty moth, Panolis flammea, fed on a saturated honey solution, laid significantly more eggs than those individuals given water only or not fed at all.At 20°, the longevity of both male and female moths was significantly increased when they were supplied with a food source. At 10° fed male months were not significantly longer lived than unfed moths. Fed female moths at 10° were however, significantly longer lived than unfed individuals. At 20°, fed male moths lived longer than fed females (8.17 days as compared with 5.95 days). At 10°, fed female moths lived longer than fed males (16.00 days compared with 11.41 days).At 20°, weight loss in unfed females over the reproductive period was proportional to the number of eggs laid and the survival time of the individual. In fed females, no significant weight loss occurred over the reproductive period, irrespective of the number of eggs laid or the longevity of the individual.Egg production, even in fed females, over the life span of an individual was only a fifth of the total eggs present in the reproductive tract at eclosion.It is concluded that adult females of P. flammea are likely to require an extra external food source to achieve their full reproductive potential.


The nature of alcoholism in patients with different family histories for alcoholism.
1. The course and consequences of alcohol abuse were examined in male and female patients being treated for alcoholism. 2. Demographic characteristics, family history for alcoholism, psychopathology, drinking history and social and physical consequences of alcohol abuse were assessed. 3. Probands with alcoholism on both the maternal and paternal sides of their pedigree experience greater psychosocial and physical consequences of alcohol abuse than other alcoholic patients. 4. Sex of the proband and type of family pedigree for alcoholism were found not to be interactive but to contribute separate additive effects.


Molecular Cloning and Expression Profiles of Thermosensitive TRP Genes in Agasicles hygrophila.
Global warming has gradually reduced the control efficacy of Agasicles hygrophila against the invasive weed Alternanthera philoxeroides. To better understand the summer collapse of A. hygrophila populations, we cloned the cDNA sequences of the high temperature-sensing TRPA1, Painless, and Pyrexia from A. hygrophila, and analyzed their temporal expressions and the impacts of high temperatures on their expression in eggs, the most vulnerable stage of A. hygrophila to hot temperatures. All the three genes obtained had the signature domains of TRPA channels and were constitutively expressed in eggs, larvae (L1, L2, L3), pupae, and adults, but AhPainless had the highest expression, followed by AhPyrexia, and AhTRPA1. The lowest and highest expression stages were adult and pupae for AhTRPA1, egg and L3 for AhPainless, and pupae/adult and L2 for AhPyrexia. The expressions of AhTRPA1, AhPainless, and AhPyrexia remained low at the preferred temperature range of 25-28 °C, elevated to their peak levels at 37.5, 30, and 30 °C, respectively, then fell to their 25-28 °C levels (AhTRPA1, AhPainless) or a lower level (AhPyrexia) at one or more temperatures >30 or 37.5 °C. These results suggest that their temperature-sensing roles and importance may be different, stage-specific, and their expression may be decoupled from their activation.


Promoting physical activity with self-management support for those with multimorbidity: a randomised controlled trial.
Targeted self-management programmes may improve health and increase physical activity (PA) in people with multimorbidity. To investigate the impact of a structured, theoretically driven, self-management group education programme on habitual PA levels in people with multimorbidity. Individually randomised controlled trial with 12-month follow-up, involving nine primary care practices in Leicestershire, UK. In total, 353 adults with multimorbidity (age 67.8 years [±9 years], 161 male sex) were randomised to intervention (n = 180) or control (n = 173) groups. Intervention participants were invited to attend four group-based self-management sessions, centred primarily on increasing PA, and received motivational text-message support. The primary outcome measure was change in overall volume (time and intensity) of daily PA at 12 months, as measured by the GENEActiv wrist-worn accelerometer device. At baseline, the total sample achieved 22 min of moderate-vigorous intensity PA per day (mean/participant). At 12 months, in the complete-case analysis, a reduction in daily mean PA volume was seen in the intervention group relative to control (-0.80 milligravity [m g]; 95% confidence interval [CI] = -1.57 to -0.03; P = 0.04). Reductions were also seen in the intervention group in time spent in moderate-vigorous PA (-3.86 min per day; 95% CI= -6.70 to -1.03; P = 0.008) and time spent at an intensity equivalent to a slow walk (-4.66 min per day; 95% CI = -8.82 to -0.51; P = 0.028). However, the per-protocol analysis (excluding participants who did not attend at least one education session) found no between-group differences in overall daily PA at 12 months (-0.65 mg; 95% CI = -1.46 to 0.15; P = 0.11). The self-management programme elicited a slight reduction in PA levels in people with multimorbidity. Future research should identify and target subgroups of those with multimorbidity in greatest need of PA promotion in order to maximise potential capacity for benefit, and also focus on refining the intervention in order to increase efficacy in increasing PA.


First trimester depression and/or anxiety disorders increase the risk of low birthweight in IVF offspring: a prospective cohort study.
Are maternal depression and/or anxiety disorders (MDAD) before and during pregnancy associated with IVF outcomes? A total of 5661 women starting their first IVF cycle between 15 August 2014 and 31 December 2015 were pooled from a prospective cohort of IVF-conceived children. The self-rating depression scale (SDS) and self-rating anxiety scale (SAS) were used to determine MDAD. IVF outcomes were compared between MDAD+ and MDAD- groups. A total of 10.3% (572/5556) of women had MDAD before IVF (bMDAD). The fertilization rate was lower in the bMDAD+ group (59.41 ± 22.11% versus 61.72 ± 22.18%, Padjust < 0.05). No difference was found in the other IVF outcomes. Pregnancy and neonatal outcomes in women with singleton live births were similar between the two groups. A total of 17.4% (347) women with singleton live births had MDAD during the first trimester (pMDAD). Birthweight (3383 ± 556 g versus 3447 ± 518 g, Padjust < 0.05) was lower and incidence of low birthweight (LBW) (6.9% versus 3.3%, Padjust < 0.01) was higher in the pMDAD group. After adjustment for potential confounders (gestational age, maternal age, maternal pre-pregnancy body mass index, threatened abortion, hypertensive disorder complicating pregnancy and gestational diabetes mellitus), pMDAD remained significantly associated with LBW (odds ratio [OR] 2.50, 95% confidence interval [CI] 1.16-5.42, Padjust < 0.05). The preconception psychological state in the pMDAD group did not demonstrate any additional impact on neonatal outcomes. MDAD during the first trimester is associated with increased risk of LBW in offspring, whether preconception MDAD exists or not.


Glycosylated Hemoglobin A1c Predicts Intracerebral Hemorrhage with Acute Ischemic Stroke Post-Mechanical Thrombectomy.
Intracerebral hemorrhage, including symptomatic intracerebral hemorrhage, is a serious post-mechanical thrombectomy complication in patients with acute ischemic stroke. We aimed to determine whether glycosylated hemoglobin A1c parameters could predict intracerebral hemorrhage in this patient population. We enrolled patients with acute occlusion of the internal carotid artery or proximal middle cerebral artery and who had undergone mechanical thrombectomy. According to the glycosylated hemoglobin A1c level (%) assessed during the hospital stay, the patients were divided into two groups: > 6.5% and ≤ 6.5%. Intracerebral hemorrhage was evaluated and classified based on cranial computed tomography scans obtained within 24-48 h or when neurological conditions worsened. We assessed the outcome at the end of 90 days using the modified Rankin Scale scores. Among 202 patients, 86 (42.6%) suffered intracerebral hemorrhage, while 25 (12.4%) had symptomatic intracerebral hemorrhage; 35.6% of the patients had a favorable outcome (modified Rankin Scale scores 0-2). Multivariable analysis demonstrated an association of glycosylated hemoglobin A1c > 6.5% with intracerebral hemorrhage. Furthermore, glycosylated hemoglobin A1c > 6.5% was independently associated with symptomatic intracerebral hemorrhage (OR, 2.136; 95% CI, 1.279-3.567; P = 0.004). In addition, glycosylated hemoglobin A1c > 6.5% was significantly associated with increased mortality (OR, 1.511; 95% CI, 1.042-2.191; P = 0.029) and negatively associated with favorable outcome (OR, 0.480; 95% CI, 0.296-0.781; P = 0.003) at 90 days. Glycosylated hemoglobin A1c is an independent predictor of intracerebral hemorrhage (specifically, symptomatic intracerebral hemorrhage) in patients with acute ischemic stroke treated with mechanical thrombectomy. Further studies are needed to validate these findings.


Trauma patient discharge and care transition experiences: Identifying opportunities for quality improvement in trauma centres.
Challenges delivering quality care are especially salient during hospital discharge and care transitions. Severely injured patients discharged from a trauma centre will go either home, to rehabilitation or another acute care hospital with complex management needs. This purpose of this study was to explore the experiences of trauma patients and families treated in a regional academic trauma centre to better understand and improve their discharge and care transition experiences. A qualitative study using inductive thematic analysis was conducted between March and October 2016. Telephone interviews were conducted with trauma patients and/or a family member after discharge from the trauma centre. Data collection and analysis were completed inductively and iteratively consistent with a qualitative approach. Twenty-four interviews included 19 patients and 7 family members. Participants' experiences drew attention to discharge and transfer processes that either (1) Fostered quality discharge or (2) Impeded quality discharge. Fostering quality discharge was ward staff preparation efforts; establishing effective care continuity; and, adequate emotional support. Impeding discharge quality was perceived pressure to leave the hospital; imposed transfer decisions; and, sub-optimal communication and coordination around discharge. Patient-provider communication was viewed to be driven by system, rather than patient need. Inter-facility information gaps raised concern about receiving facilities' ability to care for injured patients. The quality of trauma patient discharge and transition experiences is undermined by system- and ward-level processes that compete, rather than align, in producing high quality patient-centred discharge. Local improvement solutions focused on modifiable factors within the trauma centre include patient-oriented discharge education and patient navigation; however, these approaches alone may be insufficient to enhance patient experiences. Trauma patients encounter complex barriers to quality discharge that likely require a comprehensive, multimodal intervention.


Manometric evaluation of the pharynx.
Over the past few years, studies of manometric techniques have improved our ability to accurately assess pharyngeal pressure events during swallowing. Solid-state transducers, circumferentially recording transducers, and on-line computer interpretation allow quantitative measurements. Studies in normal subjects will permit better recognition of pathologic states.


Lentivirus delivery of shRNA constructs into osteoblasts.
Osteoblasts are the sole cell responsible for bone formation in vivo (1). Although genetic techniques have been extremely valuable to study the functions of certain genes in these cells in vivo, this approach is time consuming and expensive. An alternative loss-of-function approach that has been validated in many mammalian systems is shRNA-mediated gene silencing. This chapter describes methodology designed to introduce shRNA constructs into primary murine osteoblasts ex vivo in order to quickly assess the function of genes in osteoblast differentiation and extra cellular matrix mineralization. Both the production of shRNA-expressing lentiviruses and the infection of calvarial osteoblasts with these lentiviruses are detailed.


Cytogenetic characterization of diffuse large cell lymphoma using multi-color fluorescence in situ hybridization.
We have employed multi-color fluorescence in situ hybridization (M-FISH) to characterize the cytogenetic changes in 20 diffuse large B-cell lymphomas (DLBCL), that contained complex and partially characterized karyotypes. The M-FISH analysis helped to delineate 94% of the unidentified abnormalities and assisted in redefining some unidentified/misidentified karyotypic changes. Recurrent breakpoints observed in approximately 20% cases included 14q32, 3p21, 3q27, 22q12, 1q25, and 18q21 (in decreasing order), and 1p22, 1q21, 4q31, 6q21, and 8q24 (in four cases each). Numerical gain of chromosomes 7, 9, 12, and X and loss of chromosomes 1, 4, 6, 17, and Y, were noted in approximately 20% of cases. The minimum deleted regions encompassed 6q21-q25, 1p22-p36, 1q32-q44, 2p23-p25, 4q31-q35, 13p13-q14, and 17p11-p13. Two cases presented with a sole structural abnormality, and one contained a der(17)t(9;17)(p21;p13), which has not been reported earlier as a sole abnormality in DLBCL. Upon completely characterizing the karyotypes, we observed with interesting that in 55% of the cases, more than one BCL gene bearing regions was involved in translocations. In the remaining 45%, where only one or none of the BCL gene regions was involved in a rearrangement, we observed the loss of chromosomes 6 and/or 17 or partial deletions of 6q and/or 17p or gain of 7 and/or 12. Our findings suggest that, although BCL2 and BCL6 are most often implicated in DLBCL, the possibility of the disruptions of BCL3, BCL8, BCL9, and BCL10 as a "primary event" in DLBCL cannot be ruled out. Most often, a combination of events may be necessary for the genesis of DLBCL or progression of follicular lymphoma to DLBCL. Overall, M-FISH has enhanced our ability to provide a comprehensive karyotypic analysis, and has helped in defining the importance of BCL3, BCL8, BCL9, and BCL10 carrying breakpoints in DLBCL.


Mesalazine (Mesasal/Claversal) 1.5 g b.d. vs. placebo in the maintenance of remission of patients with Crohn's disease.
INVESTIGATORS: This multicentre study was conducted by 29 principal investigators in 11 countries. To compare the safety and efficacy of oral mesalazine (Mesasal/Claversal, 5-ASA) 1.5 g b.d. in comparison with placebo in the maintenance of remission in 286 patients with Crohn's disease. Patients had to score less than 150 in their Crohn's Disease Activity Index (CDAI), and had to have had one period of clinical activity (CDAI > 150) within 18 months of the study start. Patients were randomized to receive 5-ASA 1.5 g b.d. daily or matching placebo for 12 months. Study visits were scheduled for months 1, 3, 6, 9 and 12, or when symptoms suggested a relapse of the disease. Relapse was defined as a CDAI score greater than 150, with at least a 60-point increase from the baseline index score. None of the patients used glucocorticoids or immunosuppressants during the trial. In the first group, 207 patients with Crohn's colitis or ileocolitis were randomized: there were 101 females and 106 males, in age range 18-71 years. A total of 106 patients (51 in the 5-ASA group and 55 in the placebo group) were withdrawn from the study due to adverse events, insufficient therapeutic effect, or for other reasons. This left 101 patients (51 in the 5-ASA group and 50 in the placebo group) who completed the 12-month trial. In the second group, 79 patients with Crohn's ileitis were randomized to treatment. There were 53 females and 26 males, age range 18-66 years. A total of 41 patients (19 in the 5-ASA group and 22 in the placebo group) were withdrawn from the study. This left 38 patients (17 in the 5-ASA group and 21 in the placebo group) who completed the 12-month trial. The primary efficacy variable was the CDAI. A protocol-eligible analysis and an intent-to-treat analysis were performed. No statistical differences were noted between the two analyses. In patients with Crohn's colitis or ileocolitis, or in those with ileitis, no statistically significant differences were noted with respect to the relapse rates between the 5-ASA and the placebo treatment groups. Adverse events in the gastrointestinal system were the most frequently reported in both treatment groups. Many of the events such as diarrhoea or abdominal pain are symptoms of Crohn's disease. The majority of the events reported were mild or moderate in severity. In neither study was the prevalence of adverse events or the proportion of drop-outs different between patients in the treatment or in the placebo groups. The site of the Crohn's disease had no effect on the frequency of adverse events. The relapse rates of Crohn's disease were similar for up to 12 months in both the 5-ASA 1.5 g b.d. and the placebo treatment groups.


Further studies on the mechanism of inhibition of intestinal chylomicron transport by Pluronic L-81.
This study explored further the hypothesis that intestinal cells have two pathways for producing large triacylglycerol-rich lipoprotein particles. The hydrophobic surfactant Pluronic L-81 (L-81) inhibits formation of chylomicrons (containing triacylglycerol synthesized from dietary fatty acids and monoacylglycerol, through the monoacylglycerol pathway), but not formation of very-low-density lipoproteins. L-81 does not inhibit lymphatic lipid transport during infusion of egg phosphatidylcholine, whose fatty acid is processed through the alpha-glycerol phosphate pathway and is transported in lymph in very-low-density lipoproteins. Thus, the first part of this study tested whether L-81 cannot inhibit the alpha-glycerol phosphate pathway, and thus L-81 can only affect chylomicron lipid secretion. Intestinal lymph fistula rats were infused with a lipid emulsion containing [1-14C]oleic acid, but no monoacylglycerol, to ensure that the oleic acid will be channeled to the alpha-glycerol phosphate pathway. Experimental rats received 1 mg/h of L-81 in their emulsion whereas control rats lacked L-81. Lymphatic triacylglycerol output, measured both chemically and radioactively, was markedly suppressed in the experimental rats as compared to the controls. Thus, these data indicate that the reason why lipid transport was unaffected by L-81 when egg phosphatidylcholine was infused was not because of the pathway used for the resynthesis of triacylglycerol from phosphatidylcholine. In the second part of this study, we measured the appearance time for chylomicron (in control rats) and for very-low-density lipoprotein (in L-81-treated rats). The appearance time is defined as the time between placement of radioactive fatty acid into the intestinal lumen and the appearance of radioactive lipid in the central lacteal. The average appearance time for the control rats was 10.8 min, which was significantly shorter than the 16.2 min in the L-81-treated experimental rats. This difference in appearance time further supports the hypothesis that chylomicron and very-low-density lipoprotein are packaged separately in the enterocytes and only the formation of chylomicron is inhibited by L-81.


Interobserver variability in the detection of spontaneous echo contrast, left atrial thrombi, and left atrial appendage thrombi by transoesophageal echocardiography.
To assess the interobserver variability between two observers from different echocardiographic laboratories. Two observers reviewed video recordings blinded to the other's diagnosis. In part I (n = 88), they determined interobserver variability for spontaneous echo contrast, left atrial thrombi, and appendage thrombi. No diagnostic criteria for thrombi were defined. In part II (n = 85), diagnostic criteria for thrombi were defined. Part I: Both observers agreed in diagnosing spontaneous echo contrast in 97%, left atrial thrombi in 90%, left atrial appendage thrombi in 94%. Part II: With predefined criteria no disagreement occurred in diagnosing left atrial thrombi. In the diagnosis of left atrial appendage thrombi both observers agreed in 89%. The mean diameters of the 10 thrombi on which the observers agreed were greater than of the nine appendage thrombi on which they disagreed. Interobserver variability in the diagnosis of spontaneous echo contrast is low. Defined criteria decrease interobserver variability for left atrial and appendage thrombi, although one third of the thrombi diagnosed by one observer were not confirmed by the other. Interobserver variability is high in the assessment of small structures (< 15 mm) within the left atrial appendage.


Conjugation to gold nanoparticles enhances polyethylenimine's transfer of plasmid DNA into mammalian cells.
Branched polyethylenimine (PEI) chains with an average molecular mass of 2 kDa (PEI2) have been covalently attached to gold nanoparticles (GNPs), and the potency of the resulting PEI2-GNPs conjugates as vectors for the delivery of plasmid DNA into monkey kidney (COS-7) cells in the presence of serum in vitro has been systematically investigated. The transfection efficiencies vary as a function of the PEI/gold molar ratio in the conjugates, with the best one (PEI2-GNPII) being 12 times more potent than the unmodified polycation. This potency can be further doubled by adding amphiphilic N-dodecyl-PEI2 during complex formation with DNA. The resulting ternary complexes are at least 1 order of magnitude more efficient than the 25-kDa PEI, one of the premier polycationic gene-delivery vectors. Importantly, although unmodified PEI2 transfects just 4% of the cells, PEI2-GNPII transfects 25%, and the PEI2-GNPII/dodecyl-PEI2 ternary complex transfects 50% of the cells. The intracellular trafficking of the DNA complexes of these vectors, monitored by transmission electron microscopy, has detected the complexes in the nucleus <1 h after transfection.


Correlation of secondary structures of bradykinin B1 receptor antagonists with their activity.
The secondary structure of a bradykinin B(1)receptor antagonist B-10324 (F5C-Lys-(1)- Lys(0)-Arg(1)-Pro(2)- Hyp(3)-Gly(4)-CpG(5)- Ser(6)-DTic(7)-CpG(8)) was determined by NMR at 800MHz. The conformational data are compared with those obtained previously for two bradykinin B(1) receptor antagonists, namely B-9858 (Lys-(1)- Lys(0)-Arg(1)-Pro(2)- Hyp(3)-Gly(4)-Igl(5)- Ser(6)-DIgl(7)-Oic(8)) and B-10148 (Lys-(1)-Lys(0)-Arg(1)- Pro(2)-Hyp(3)-Gly(4)- Igl(5)-Ser(6)-DF5F(7)- Oic(8)). The abnormal amino acids are: Hyp, trans-4- hydroxyproline; Tic, 1, 2, 3, 4-tetrahydroisoquinoline-3-carboxylic acid; Oic, (2S, 3aS, 7aS)-octahydroindole-2-carboxylic acid; Igl, alpha(2- indanyl)glycine; F5F, 2,3,4,5,6-pentafluorophenylalanine; CpG, alpha- cyclopentylglycine. F5C, pentafluorocinnamoyl, is the N-terminal protecting group and is not involved in the peptide secondary structure. B-10324 contains an N-terminal Pro(2)- CpG(5) distorted type II beta-turn whereas the rest of the peptide is random. A salt bridge is not observed between the carboxylate group at the C-terminal end and the Arg(1) side chain, in contrast to that previously observed for B-9858 and B- 10148. The conformations are correlated with the measured B(1) receptor antagonist activities (J.-F. Larrivée, L. Gera, S. Houle, J. Bouthillier, D. R. Bachvarov, J. M. Stewart and F. Marc au, Br. J. Pharmacol. 131, 885-892 (2000)). The importance of the N-terminal beta-turn is highlighted.


A histochemical study of the enzyme profile of Krabbe's cells.
A histochemical study of the enzyme profile of Krabbe's cells revealed a high activity of lysosomal enzymes, including the tartrate-resistant isoenzyme of acid phosphatase, and considerable activity of extralysosomal nonspecific esterase and mitochondrial and extramitochondrial dehydrogenases. The presence of lysozyme was demonstrated. This generally high metabolic activity was found closely reminiscent of that in Gaucher's cells and in epitheloid transformed macrophages though quite unlike the exclusive predominance of lysosomal enzyme activity typical of classical foamy storage cells in Niemann-Pick and Wolman's disease.


Recurrent podocytopathy in a patient with systemic lupus erythematosus.
Podocytopathy in systemic lupus erythematosus is characterised by diffuse foot process effacement without significant peripheral capillary wall immune deposits as seen on electron microscopy. Lupus podocytopathy falls outside the scope of the current International Society of Nephrology and the Renal Pathology Society classification of lupus nephritis. We present a case of relapsing podocytopathy with nephrotic syndrome occurring simultaneously with two extra-renal and serological disease flares, which makes it likely that podocytopathy was related to systemic lupus erythematosus activity. This case adds to the growing body of evidence that lupus podocytopathy must be considered in the differential diagnosis of systemic lupus erythematosus patients presenting with nephrotic syndrome.


Understanding the sorption and biotransformation of organic micropollutants in innovative biological wastewater treatment technologies.
New technologies for wastewater treatment have been developed in the last years based on the combination of biological reactors operating under different redox conditions. Their efficiency in the removal of organic micropollutants (OMPs) has not been clearly assessed yet. This review paper is focussed on understanding the sorption and biotransformation of a selected group of 17 OMPs, including pharmaceuticals, hormones and personal care products, during biological wastewater treatment processes. Apart from considering the role of "classical" operational parameters, new factors such as biomass conformation and particle size, upward velocity applied or the addition of adsorbents have been considered. It has been found that the OMP removal by sorption not only depends on their physico-chemical characteristics and other parameters, such as the biomass conformation and particle size, or some operational conditions also relevant. Membrane biological reactors (MBR), have shown to enhance sorption and biotransformation of some OMPs. The same applies to technologies bases on direct addition of activated carbon in bioreactors. The OMP biotransformation degree and pathway is mainly driven by the redox potential and the primary substrate activity. The combination of different redox potentials in hybrid reactor systems can significantly enhance the overall OMP removal efficiency. Sorption and biotransformation can be synergistically promoted in biological reactors by the addition of activated carbon. The deeper knowledge of the main parameters influencing OMP removal provided by this review will allow optimizing the biological processes in the future.


The molecular and cellular signatures of the mouse eminentia thalami support its role as a signalling centre in the developing forebrain.
The mammalian eminentia thalami (EmT) (or thalamic eminence) is an embryonic forebrain structure of unknown function. Here, we examined the molecular and cellular properties of the mouse EmT. We first studied mRNA expression of signalling molecules and found that the EmT is a structure, rich in expression of secreted factors, with Wnts being the most abundantly detected. We then examined whether EmT tissue could induce cell fate changes when grafted ectopically. For this, we transplanted EmT tissue from a tau-GFP mouse to the ventral telencephalon of a wild type host, a telencephalic region where Wnt signalling is not normally active but which we showed in culture experiments is competent to respond to Wnts. We observed that the EmT was able to induce in adjacent ventral telencephalic cells ectopic expression of Lef1, a transcriptional activator and a target gene of the Wnt/β-catenin pathway. These Lef1-positive;GFP-negative cells expressed the telencephalic marker Foxg1 but not Ascl1, which is normally expressed by ventral telencephalic cells. These results suggest that the EmT has the capacity to activate Wnt/β-catenin signalling in the ventral telencephalon and to suppress ventral telencephalic gene expression. Altogether, our data support a role of the EmT as a signalling centre in the developing mouse forebrain.


Psychosomatic aspects of patients on hemodialysis. 4. The relationship between quality of life and alexithymia.
The relationship between quality of life (QOL) and alexithymia was examined in dialysis patients. Hemodialysis (HD) patients with diabetic nephropathy had poor QOL in medical dimension. Although they had good QOL in psychological and social dimensions, a high prevalence of alexithymia associated with self-control ability was observed. These results suggest that their inability to care for themselves was reflected in poor medical dimension, and that good psychological and social dimensions were merely an outward appearance. Therefore, identification of alexithymia in dialysis patients is important along with a three-dimensional evaluation of QOL. Multidisciplinary comprehensive assessment of QOL including alexithymia will lead to a real improvement of dialysis patients' QOL.


Impact of the COVID-19 Pandemic on Urologists in Germany.
In order to contain the coronavirus disease 2019 (COVID-19) pandemic, Germany has implemented drastic restrictions on public or social life, while health institutions are invoked to postpone elective procedures. Although urologists are less involved in the direct treatment of COVID-19 patients, the current situation strongly affects the urological work routine. To analyze the impact of the COVID-19 pandemic on various aspects of work and personal life among urologists in Germany. A total of 589 urologists in Germany participated in an online survey between March 27 and April 11, 2020. Participants were stratified into subgroups according to professional characteristics. Most urologists rated Germany as "well prepared" and the increasing restrictions of social life as "very positive." Routine operation was more restricted in hospitals than in the outpatient sector (p = 0.046). Moreover, urologists from the outpatient sector felt significantly less prepared for the COVID-19 pandemic (p = 0.001), reported a higher shortage of protective medical equipment (p < 0.001), and described a tendency toward a higher level of threat (p = 0.054). Although restrictions regarding telemedicine approaches were reported by 60% of participants, the outpatient sector used telehealth more frequently than hospitals (25.5% vs 17.0%, p < 0.001). Limitations include the national design and the restricted survey period. This survey systematically evaluates the impact of the COVID-19 pandemic on personal and professional aspects of German urologists. We identified several issues, such as a higher shortage of medical protective equipment in the outpatient sector that could trigger specific measures to further improve the quality of urological care in Germany. We evaluated a potential impact of the coronavirus disease 2019 (COVID-19) pandemic on professional and personal aspects of the urologists in Germany. Our results suggest that the outpatient sector should receive specific attention as, for example, shortage of protective equipment was more common.


Endosulfan application to a stream mesocosm: studies on fate, uptake into passive samplers and caged toxicity test with the fish M. ambigua.
A model mesocosm system was used to simulate the effect of endosulfan entering a waterway from episodic events such as accidental overspray or in runoff water containing contaminated sediment following storm events. The fate of technical endosulfan applied to a 24-stream mesocosm was compared in experiments where the pesticide was applied either directly as water contamination or after being pre-bound to sediment. The flow of water through the streams was discontinued for a 10-12 h period following the pesticide application. Following the water application, only approximately 3% of endosulfan remained in the streams after 4 days and then was not detectable after 7 days. In contrast, after application pre-bound to sediment, approximately 33% of the endosulfan remained in the streams after 4 days and 14% after 7 days. Additionally, with the sediment-bound application, the proportion of endosulfan was higher in the substrate (11%) after 7 days than in the overlying water (3%), and approximately 1% was oxidised to the sulphate form. The dissipation of endosulfan in the water column of both experiments followed a two-parameter exponential decay model characterised by a relatively fast first-order single-phase process. In sediment of both experiments and the gravel of the sediment-dosing experiment, the dissipation of endosulfan followed more closely a four-parameter bi-exponential decay model characterised by first-order kinetics of two fractions: the first fraction dissipates quickly, and in the longer term the second fraction dominates the overall dissipation with a slower rate. In the gravel section of the water-dosing experiment, endosulfan dissipation was characterised by relatively very slow two-parameter exponential decay. The overall dissipation rates of the alpha- and beta-endosulfan isomers were significantly higher in the water-dosing than in the sediment-dosing experiment, except in the gravel section of the mesocosm. The uptake of the endosulfan into passive samplers constructed from polyethylene membrane bags containing trimethylpentane solvent (TRIMPS) placed in the overlying water was linear. In contrast, TRIMPS buried in sediment failed to uptake endosulfan from the sediment substrate, indicating that short-term deployment of passive samplers can only be used to determine time-weighted average concentrations of bioavailable chemicals in the water column. A 34-h LC50 of 2.8 microg/l [95% confidence interval (CI) 1.5-4.2 microg/l] for juveniles of the native fish Macquaria ambigua was obtained when exposed during the water-dosing experiment. This study demonstrated that the pulse entry of sediment contaminated with endosulfan into a receiving waterway was more persistent compared with direct aqueous contamination and the endosulfan would be bioavailable to pelagic organisms following a gradual partitioning to the water column.


Nanodiscs for structural and functional studies of membrane proteins.
Membrane proteins have long presented a challenge to biochemical and functional studies. In the absence of a bilayer environment, individual proteins and critical macromolecular complexes may be insoluble and may display altered or absent activities. Nanodisc technology provides important advantages for the isolation, purification, structural resolution and functional characterization of membrane proteins. In addition, the ability to precisely control the nanodisc composition provides a nanoscale membrane surface for investigating molecular recognition events.


Data-driven method based on deep learning algorithm for detecting fat, oil, and grease (FOG) of sewer networks in urban commercial areas.
The content of fat, oil and grease (FOG) in the sewer network sediments is the key indicator for diagnosing sewer blockage and overflow. However, the traditional FOG detection is time-consuming and costly, and the establishment of mathematical models based on statistical methods to predict the content of FOG fail to provide satisfactory accuracy. Herein, a deep learning algorithm used a data-driven FOG content prediction model is proposed to achieve a more accurate prediction of FOG content. Meanwhile, global sensitivity analysis (GSA) is exploited to evaluate the contribution of input indicators to the output indicator (FOG) in the model, so that some input indicators that have less impact on the prediction performance can be screened out, the best combination of input indicators can be determined, and the operation cost of the model can be reduced. To evaluate the effectiveness of the proposed model, a case study was conducted in a city in southern China. The experimental results indicate that the prediction model obtains good FOG estimations and performs well from a single site to multiple sites with a mean R2 of 0.922, showing a good generalization performance. Through GSA, the key input indicators in the model were identified as pH, water temperature (T), relative humidity (RH), sewage flow (Flow), drinking water supply (DWS), velocity (V) and conductivity (σ), and the input indicators such as air pressure (AP), population (Pop.), and liquid level (LV) can be reduced without affecting the prediction accuracy of the model.


Vaccination response following aerobic exercise: can a brisk walk enhance antibody response to pneumococcal and influenza vaccinations?
High intensity acute exercise at the time of vaccination has been shown to enhance the subsequent antibody response. This study examines whether an acute moderate intensity aerobic intervention prior to vaccination can enhance antibody response to pneumonia and half dose influenza vaccination. Sixty young (age (SD)=22.0 (6.1) years) and 60 older (age (SD)=57.5 (6.5) years) adults attended the laboratory on two separate occasions. At the first session, baseline antibody titres were determined, before participants completed either a brisk walk around campus at >55% of their age-predicted heart rate maximum, or a resting control condition, for 45 min. After the intervention, all participants received a full-dose pneumococcal vaccination and a half-dose influenza vaccination. Four weeks later, participants returned for a follow up blood sample. Multivariate ANOVA revealed an increase in total antibody titres against the influenza vaccine (F((12,106))=25.76, p<.001, η(2)=.75) and both the IgM (F((12,106))=17.10, p<.001, η(2)=.66) and IgG (F((12,106))=25.76, p<.001, η(2)=.75) antibody titres against the pneumococcal vaccine. However, there were no significant Time×Group interactions (p's all >.15), indicating that a 45 min brisk walk prior to vaccination did not affect antibody response to either the influenza or pneumonia vaccine. The results suggest that higher intensity exercise is necessary to augment antibody response to vaccination.


[Study on quantitative assessment of Siraitia grosvenorii germplasms by general index].
To establish a weight-marking method and evaluate Siraitia grosvenorii germplasms. The characters of 21 kinds of S. grosvenorii germplasms in the field and the lab were analyzed, seven important characters were selected to weight the marks. A general evaluation index was made and used to evaluate S. grosvenorii germplasms. The evaluation result of 21 kinds of S. grosvenorii germplasms by the weight-marking method was consistent with production practice. Meanwhile, the new variety Yongqing No. 1 and major cultivars were the superior germplasms. A rational technique system of evaluating S. grosvenorii germplasms was established, and the superior germplasms were selected.


Using microwave-accelerated digestion instead of dry ashing during sodium analysis of low-moisture, part-skim mozzarella.
Mineral analysis of cheese products is a labor- and time-intensive process. Pretreatment using dry or atmospheric wet ashing to decompose organic matter before measurement on a spectrometer can take more than 24 h to complete. Samples of low-moisture, part-skim mozzarella (LMPS) cheese were digested using microwave-accelerated digestion (MAD) and conventional dry ashing. A comparison of 23 samples, in duplicate, was performed to determine equivalency. A two one-sided test (TOST) analysis showed that the confidence intervals were within acceptable differences for standard method variation (less than 70 mg/100 g per International Dairy Federation method 119:2007). Bias was observed for the quantification of sodium in microwave-digested samples, averaging 14 mg/100 g higher. Based on the TOST criteria, we concluded that the methods are equivalent.


Formation of cadmium sulfide nanoparticles mediates cadmium resistance and light utilization of the deep-sea bacterium Idiomarina sp. OT37-5b.
Heavy metal is one of the major factors threatening the survival of microorganisms. Here, a deep-sea bacterium designated Idiomarina sp. OT37-5b possessing strong cadmium (Cd) tolerance was isolated from a typical hydrothermal vent. Both the Cd-resistance and removal efficiency of Idiomarina sp. OT37-5b were significantly promoted by the supplement of cysteine and meanwhile large amount of CdS nanoparticles were observed. Production of H2 S from cysteine catalysed by methionine gamma-lyase was further demonstrated to contribute to the formation of CdS nanoparticles. Proteomic results showed the addition of cysteine effectively enhanced the efflux of Cd, improved the activities of reactive oxygen species scavenging enzymes, and thereby boosted the nitrogen reduction and energy production of Idiomarina sp. OT37-5b. Notably, the existence of CdS nanoparticles obviously promoted the growth of Idiomarina sp. OT37-5b when exposed to light, indicating this bacterium might grab light energy through CdS nanoparticles. Proteomic analysis revealed the expression levels of essential components for light utilization including electron transport, cytochrome complex and F-type ATPase were significantly up-regulated, which strongly suggested the formation of CdS nanoparticles promoted light utilization and energy production. Our results provide a good model to investigate the uncovered mechanisms of self-photosensitization of nonphotosynthetic bacteria for light-to-chemical production in the deep biosphere.


Cholesterol oxidase immobilized inulin based nanocomposite as the sensing material for cholesterol in biological and food samples.
In the present study, inulin based nanocomposite viz., TiO2-MWCNT@Inulin was prepared by embedding Inulin (a biopolymer extracted from Allium sativum L.) with TiO2 and MWCNTs. The morphology of the prepared nanocomposite was characterized by High Resolution transmission electron microscopy (HRTEM). Cholesterol oxidase (ChOx) enzyme was then immobilized into the nanocomposite and the immobilization was examined by UV-vis and FT-IR spectral studies. The ChOx immobilized nanocomposite was integrated into carbon paste (CP) matrix to prepare the working electrode for the sensing of cholesterol. Electrochemical characterization of the modified CP/TiO2-MWCNT@Inulin/ChOx electrode was done by cyclic voltammetric (CV) and electrochemical impedance spectroscopic (EIS) studies. Differential pulse voltammetric (DPV) studies were carried out to determine the concentration of cholesterol at the interface of the newly fabricated electrode. The fabricated electrode demonstrated a linear range from 83 μM to 14.28 mM, low limit of detection (35 μM), good sensitivity (21.26 μA mM-1 cm-2), low Km (0.49 mM), high stability (120 days) and good selectivity. The presence of Inulin biopolymer played a vital role in attaching ChOx enzyme firmly to the nanocomposite thereby enhancing the stability and electron transfer efficiency of the electrode. The analysis of product that was formed within the electrochemical cell during the electrochemical oxidation of cholesterol was performed by using sodium nitroprusside. This resulted in a deep purple coloured solution which suggested the electrochemical conversion of cholesterol to cholestenone. The practical applicability of the fabricated electrode was also assessed by the determination of cholesterol in spiked blood serum and milk samples.


UTR dinucleotide simple sequence repeat evolution exhibits recurring patterns including regulatory sequence motif replacements.
New genome sequence information was used to study evolution of 22 dinucleotide simple sequence repeat (diSSR) sites whose upstream flanking sequences were shown to be conserved comparing Homo sapiens with the marsupial, Monodelphis domestica. Among mammals, most of these diSSR sites were conserved both upstream and downstream of the diSSR. However, individual diSSRs were frequently replaced by alternative repeats. Conserved among mammals examined, the Vsnl1 gene's 3' UTR-localized (AC)n repeat replaced an A-rich tract in non-mammalian vertebrates examined. The Sema6D gene's (GT)n was also well conserved among mammals examined. Such conservation provides evidence of a functional role. The UTR-localized diSSRs of other genes evolved by replacing alternative diSSRs, by replacing mononucleotide-rich tracts and, in fewer cases, by expansion from short repeating sequences. Extension of the study to less conserved diSSR sites revealed that some diSSRs replaced post-transcriptional regulatory motifs, such as AU-rich elements (AREs) and C-rich tracts. The Mtap2 gene's UTR-localized (AC)n was located within a known dendritic targeting element. These evolutionary replacements suggest that some diSSRs belong to a broader group of weak-folding repetitive sequences with potential regulatory roles.


Identification, synthesis and characterization of a novel antimicrobial peptide HKPLP derived from Hippocampus kuda Bleeker.
A novel gene encoding 55 amino-acid residues has been identified from the brooding pouch cDNA library of Hippocampus kuda Bleeker. The deduced amino-acid sequence is highly homologous to several pleurocidin-like peptides from the winter flounder and comprises a signal peptide, a pro-peptide and a mature peptide. The glycine-rich mature peptide, designated HKPLP, contains 24 amino-acid residues and has been synthesized by solid-phase peptide synthesis. The purified HKPLP exhibits antimicrobial activity against several Gram-positive and Gram-negative bacterial strains at low concentrations (MIC 1.5-7.5 μM). Thermal stability assay data show good heat stability. CD spectroscopy experiments indicate that the dominant contents are anti-parallel and parallel sheets, which may have β-sheet or β-strand motif. It is inferred that HKPLP participates in the host defense during egg fertilization and embryo development as an antimicrobial peptide in brooding pouch.


Circularly polarised luminescence from planar-chiral Phanephos/Tb(iii)(hfa)3 hybrid luminophores.
Planar-chiral Phanephos, containing the coordinatable P(iii), formed P(iii)/Tb(iii)(hfa)3 hybrid luminophores that successfully emitted characteristic circularly polarised luminescence (CPL) due to 5D4→7F5 transitions in solution. On the other hand, BINAP, containing P(iii)[double bond, length as m-dash]O as axially chiral ligand, exhibited no detectable CPL with Tb(iii)(hfa)3.


Evaluation of Single-Nucleotide Polymorphisms of Transcription Factor 7-Like 2 and ATP2B1 Genes as Cardiovascular Risk Predictors in Chronic Kidney Disease.
Cardiovascular disease (CVD) is the primary cause of morbidity and premature mortality in chronic kidney disease (CKD). The transcription factor 7-like 2 (TCF7L2) gene product TCF4 is a transcription factor that acts as a downstream effector in the canonical Wnt signaling pathway and may be important in the development of both type 2 diabetes and renal development and disease. It is, therefore, plausible that mutations in this gene could manifest themselves in reduced kidney function or kidney disease through their effects on hyperglycemia as well as independent of this mechanism. The ATP2B1 gene encodes the plasma membrane calcium ATPase isoform 1, which removes bivalent calcium ions from eukaryotic cells against very large concentration gradients and is responsible for controlling the contraction and dilation of vascular smooth muscles. The aims of this study are (1) to evaluate single-nucleotide polymorphisms (SNPs) of TCF7L2 gene as cardiovascular risk predictors in CKD and (2) to evaluate SNPs of ATP2B1 gene as cardiovascular risk predictors in CKD. Fifty clinically diagnosed CKD patients in the age group between 20 and 60 years of both genders were selected as cases and fifty healthy participants from the master health checkup department were selected as controls. Genomic DNA was extracted based on the spin column kit method. The DNA samples were stored at -20°C until analysis. Genotyping for TCF7L2 gene rs7903146 (C/T) and ATP2B1 gene rs11105354 (A/G) was carried out through polymerase chain reaction. T allele frequency was observed in 12 controls and 23 cases (odds ratio [OR] = 2.2, 95% confidence interval [CI]: 1.0-4.7). CC genotype was observed in 38 controls and 27 cases and CT genotype in 22 cases and 12 controls. A allele was found in 38 cases and 23 controls (OR = 2, 95% CI: 1.1-3.8). The mean values of cholesterol, low-density lipoprotein, triglycerides, glucose, insulin, urea, and creatinine were high in cases when compared to controls. T allele of TCF7L2 gene rs7903146 (C/T) and A allele of ATP2B1 (A/G) gene rs11105354 (A/G) are associated with CVD in CKD patients.


Instruments measuring behavioral disturbance in relatives with schizophrenia.
This article presents a review of 16 instruments measuring behavioral disturbance of persons with schizophrenia as perceived by their family members. Information about the domain, the types of rating scales, and the psychometric properties of these instruments are provided. Future directions in the study of behavioral disturbance are proposed.


Compliance with parents' rules: between-person and within-person predictions.
The current study focused on the childhood to adolescence transition and sought to determine why some children are more compliant than others as well as why children comply more often with some of their parents' rules than with others. Indices of parents' agency and children's agency were tested as predictors of compliance. Parent-based decision-making and parents' responses to expressed disagreement served as indices of parents' agency while children's beliefs regarding the legitimacy of parents' rules and felt obligation to obey rules served as indices of children's agency. Parent-child dyads (n = 218; 51% female, 49% European American, 47% African American) were interviewed during the summers following the children's 5th (M adolescent age = 11.9 years) and 6th grade school years. Children who felt that their parents' rules were more legitimate were more compliant overall than were children who felt that the rules were less legitimate. Children compiled more with rules governing topics perceived to be legitimately regulated by parents, when parents made more decisions regarding the topic and when parents responded to disagreement by standing strong. Results were generally consistent across parents' and children's reports of compliance and across cross-sectional and longitudinal analyses. At the transition from childhood to adolescence, only children's agency explained why some children are more compliant than others, but parents' and children's agency helped to explain why children complied with some rules more than others.


Utero-ovarian low grade endometrial stromal sarcoma, a case report.
Endometrial stromal sarcoma (ESS) is a rare malignancy of uterine in middle aged women. There are numerous subtypes for ESS which share the same clinical picture of uterine bleeding and pelvic pain. Consequently, diagnosis and treatment modalities of LG-ESS with metastasis are challenging. However, both molecular and immunological study of samples can be useful. In this case study, we report a 52-year-old woman presenting with the chief complaint of unusual uterine bleeding. There was no specific finding in her past medical history. The CT study revealed enlarged bilateral ovary with a significantly large left ovarian mass and suspicious mass in uterus. By the diagnosis of ovarian mass, patient went under total abdominal hysterectomy with bilateral salpingo-oophorectomy (BSO), greater omentectomy, and appendectomy followed by post-op hormone therapy. Her follow-up was uneventful. The IHC and pathological study of samples revealed incidental LG-ESS uterus mass with metastasis to ovaries despite her primary diagnosis. LG-ESS has low metastasis rate. Surgical modalities and neoadjuvant therapies are recommended base on the stage of ESS. In the following study, we represent a case of incidental LG-ESS with bilateral ovarian invasion who was initially diagnosed as an ovarian mass. Our patient was successfully managed by surgical intervention. Despite scarcity of LG-ESS, it is advised to consider LG-ESS as a differential diagnosis in management of patients with a uterus mass with bilateral ovarian involvement.


Nerve endings isolated from chick embryonic optic tectum. 2. Developmental aspects of synaptosomal membrane.
Fractions enriched in nerve endings (synaptosomes) have been isolated from chick embryonic optic tectum during development. After osmotic shock, these fractions appeared to be enriched in membranes which during development acquire typical features of mature synaptosomal membranes.


Relationship between columnar cell changes and low-grade carcinoma in situ of the breast--a cytogenetic study.
Columnar cell lesions of the breast include columnar cell changes without atypia and columnar cell changes with atypia. The latter frequently coexist and share molecular changes with low-grade carcinoma in situ and invasive carcinoma, suggesting that columnar cell changes may be precursors to progression of low-grade advanced lesions. In this study, we assessed chromosomal aberrations at 16q, hallmark for low-grade lesions, in columnar cell changes with or without atypia and their adjacent carcinoma in situ by fluorescent in situ hybridization using 3 region-specific probes spanning the entire chromosomal arm. The results were correlated with the histomorphological features of the corresponding lesions. Forty-four percent of low-grade carcinoma in situ and 31% of high-grade carcinoma in situ were associated with columnar cell changes with atypia, suggesting a link between columnar cell changes with atypia and low-grade carcinoma in situ. For the genetic aberrations, heterozygous deletion of 16q was present in 56% of low-grade carcinoma in situ but only in 19% of high-grade carcinoma in situ. Conversely, aneuploidy was found mostly in high-grade carcinoma in situ (88%). Twenty percent of columnar cell changes with atypia but none of the columnar cell changes without atypia showed heterozygous deletion of 16q. Interestingly, the same changes in 16q were observed in the columnar cell changes and their associated low-grade carcinoma in situ lesions. These findings demonstrated a genetic commonality between columnar cell changes with atypia and low-grade carcinoma in situ and substantiated the precursor role of columnar cell changes with atypia for low-grade carcinoma in situ but not high-grade carcinoma in situ of the breast.


Qualitative investigation of the meanings of eating fruits and vegetables for adult couples.
The purpose of this study was to develop a substantive theory expressing the meanings couples associated with eating fruits and vegetables. This inductive qualitative study was based on a grounded theory approach and employed the constant comparison method of data analysis. Data were collected using semistructured individual interviews and a life history approach. Ten adult couples, aged 20 to 60 years, with and without children, all of whom were born in North America, were recruited using modified snowball sampling. Two overarching themes emerged. The "should syndrome" describes a morality concerning fruit and vegetable consumption arising from a tension between the low status of these foods in participants' childhood homes and their contemporary idealized status. The creation of couple gastronomies expresses couples'efforts to construct their own food norms and practices within a context of changes in social norms and fruit and vegetable availability. The substantive theory, making choices that balance their lives, conveys the dynamic processes involved in participants' fruit and vegetable choices. Future research will determine the transferability of the "should syndrome" and new couples' receptiveness to trying new fruits and vegetables. Understanding the changing contexts of food choice may help nutrition professionals better support healthful eating.


Associations of handgrip strength with morbidity and all-cause mortality of cardiometabolic multimorbidity.
Cardiometabolic multimorbidity (CM) is an increasing public health and clinical concern. However, predictors for the development and prognosis of CM are poorly understood. The aims of this study were to investigate the relation between handgrip strength (HGS) and the risk of CM and to examine the association of HGS with all-cause mortality risk among patients with CM. This prospective cohort study involved 493,774 participants from the UK Biobank. CM was defined as the simultaneous occurrence of two or more of the following conditions: type 2 diabetes, stroke, and coronary heart disease (CHD). Cox proportional hazards models were performed to estimate hazard ratios (HRs) and 95% confidence intervals (95% CIs). During a median follow-up of 12.1 years, 4701 incident CM cases were documented among participants with none cardiometabolic disease at baseline. Compared with the fourth quartile (Q4), the multivariable adjusted HR (95% CI) value of Q1 of HGS for developing CM was 1.46 (1.34-1.60). In participants with one cardiometabolic disease at baseline, participants in Q1 of HGS also possessed higher risk of CM than those in Q4, with HRs (95% CIs) being 1.35 (1.23-1.49) in patients with type 2 diabetes, 1.23 (1.04-1.46) in patients with stroke, and 1.23 (1.11-1.36) in patients with CHD. For participants with CM at recruitment, HGS was also associated with the risk of all-cause mortality (Q1 vs. Q4 HR: 1.57, 95% CI: 1.36-1.80). Our study provided novel evidence that HGS could be an independent predictor of morbidity and all-cause mortality of CM.


Pyrroloquinoline quinone, a method for its isolation and identification by mass spectrometry.
Procedures for the unambiguous detection and for the isolation and mass spectrometric identification of pyrroloquinoline quinone (PQQ) are presented. The procedure involved acid hydrolysis of protein in the presence of phenylhydrazine and successive isolation and identification of the formed adduct using mass spectrometry. In HPLC the phenylhydrazone of PQQ gave many methylated products, of which the predominant compound was the pentamethylated derivative. After reaction of the phenylhydrazone derivative of PQQ (PHPQQ) with ammonia, a product was obtained which did not contain phenylhydrazine and which formed a pentamethylated derivative as the main methylation product. The HPLC profiles of the methylated products of PHPQQ and of its ammonia derivative were very characteristic and could be used for identification in addition to mass spectrometry. However, prolonged treatment of proteins with phenylhydrazine during hydrolysis can result in the formation of a material that resembles PQQ in some aspects of its behaviour. Thus, analysis by MS is essential for unambiguous identification. This analytical procedure was applied to pig plasma benzylamine oxidase, pig aorta lysyl oxidase, pig kidney diamine oxidase and bovine serum albumin with negative results. However, samples of pronase contained variable quantities of non-covalently bound PQQ: this can lead to erroneous identification of PQQ in enzyme after pronase digestion.


Methods of conserving carbon dioxide in artificially ventilated patients: a clinical investigation.
A technique is described for maintaining the PaCO2 within, or close to, the normal range in patients hyperventilated using the Mapleson D Circuit or the Circle system with the soda-lime excluded. Control is achieved by limiting the fresh gas inflow to 4-5 litres/minute.


Genetic loci and metabolic states associated with murine epigenetic aging.
Changes in DNA methylation (DNAm) are linked to aging. Here, we profile highly conserved CpGs in 339 predominantly female mice belonging to the BXD family for which we have deep longevity and genomic data. We use a 'pan-mammalian' microarray that provides a common platform for assaying the methylome across mammalian clades. We computed epigenetic clocks and tested associations with DNAm entropy, diet, weight, metabolic traits, and genetic variation. We describe the multifactorial variance of methylation at these CpGs and show that high-fat diet augments the age-related changes. Entropy increases with age. The progression to disorder, particularly at CpGs that gain methylation over time, was predictive of genotype-dependent life expectancy. The longer-lived BXD strains had comparatively lower entropy at a given age. We identified two genetic loci that modulate epigenetic age acceleration (EAA): one on chromosome (Chr) 11 that encompasses the Erbb2/Her2 oncogenic region, and the other on Chr19 that contains a cytochrome P450 cluster. Both loci harbor genes associated with EAA in humans, including STXBP4, NKX2-3, and CUTC. Transcriptome and proteome analyses revealed correlations with oxidation-reduction, metabolic, and immune response pathways. Our results highlight concordant loci for EAA in humans and mice, and demonstrate a tight coupling between the metabolic state and epigenetic aging.


Conventional DNA-Damaging Cancer Therapies and Emerging cGAS-STING Activation: A Review and Perspectives Regarding Immunotherapeutic Potential.
Many traditional cancer treatments such as radiation and chemotherapy are known to induce cellular DNA damage as part of their cytotoxic activity. The cGAS-STING signaling axis, a key member of the DNA damage response that acts as a sensor of foreign or aberrant cytosolic DNA, is helping to rationalize the DNA-damaging activity of these treatments and their emerging immunostimulatory capacity. Moreover, cGAS-STING, which is attracting considerable attention for its ability to promote antitumor immune responses, may fundamentally be able to address many of the barriers limiting the success of cancer immunotherapy strategies, including the immunosuppressive tumor microenvironment. Herein, we review the traditional cancer therapies that have been linked with cGAS-STING activation, highlighting their targets with respect to their role and function in the DNA damage response. As part of the review, an emerging "chemoimmunotherapy" concept whereby DNA-damaging agents are used for the indirect activation of STING is discussed as an alternative to the direct molecular agonism strategies that are in development, but have yet to achieve clinical approval. The potential of this approach to address some of the inherent and emerging limitations of cGAS-STING signaling in cancer immunotherapy is also discussed. Ultimately, it is becoming clear that in order to successfully employ the immunotherapeutic potential of the cGAS-STING axis, a balance between its contrasting antitumor and protumor/inflammatory activities will need to be achieved.


Methods for genetic transformation of filamentous fungi.
Filamentous fungi have been of great interest because of their excellent ability as cell factories to manufacture useful products for human beings. The development of genetic transformation techniques is a precondition that enables scientists to target and modify genes efficiently and may reveal the function of target genes. The method to deliver foreign nucleic acid into cells is the sticking point for fungal genome modification. Up to date, there are some general methods of genetic transformation for fungi, including protoplast-mediated transformation, Agrobacterium-mediated transformation, electroporation, biolistic method and shock-wave-mediated transformation. This article reviews basic protocols and principles of these transformation methods, as well as their advantages and disadvantages.


Long-term results of laparoscopic totally extraperitoneal inguinal herniorrhaphy.
The short-term advantages of laparoscopic totally extraperitoneal mesh herniorrhaphy (TEP) are well documented, but the long-term results remain unclear. The purpose of the present paper is to clarify this issue with particular reference to hernia recurrence and chronic pain. A retrospective case series of 100 consecutive patients undergoing TEP 5 years ago by a single surgeon were followed up prospectively with a focused physical examination and interview. Data collected included recurrence, chronic groin pain, tenderness, sensory disturbance, activity or occupational limitation, and personal satisfaction. During the period between 1997 and 1999, 100 consecutive patients underwent 110 TEP hernia repairs. The mean age was 56 years. Two patients were excluded from the study because they had had open repairs for technical reasons. Median follow up was 64 months. Follow up was complete (interview and physical examination) in 88% and partial (telephone interview only) in a further 5%. There was no major morbidity or mortality. Hernia recurrence rate was 1%. Chronic pain occurred in 14 patients (14%), which was mild in 13 patients and moderate in one. Ninety-eight per cent of patients were satisfied with their repair and would or had recommended TEP to others. Long-term results of TEP demonstrate it to be an effective and safe procedure with a low recurrence and low prevalence of chronic pain that is generally of a mild, infrequent nature.


Benchmarks for Electronically Excited States: A Comparison of Noniterative and Iterative Triples Corrections in Linear Response Coupled Cluster Methods: CCSDR(3) versus CC3.
CCSDR(3) calculations of vertical excitation energies are reported for a set of 24 molecules and 121 excited valence singlet states from a recently published benchmark of organic molecules. The same geometries (MP2/6-31G*) and basis set (TZVP) were employed as in our previous linear response CC2, CCSD, and CC3 calculations. The CCSDR(3) results are compared against the CCSD and CC3 results. Statistical evaluation of all CCSDR(3) excitation energies gives mean absolute deviations of 0.09 eV from CC3 and 0.30 eV from CCSD. For excited states, which are dominated by single excitations, the absolute mean deviation from CC3 is reduced to 0.02 eV and the maximum deviation is 0.09 eV. CCSDR(3) is thus a very cost-effective accurate alternative to CC3.


Genetics of sex-ratio variation inferred from parent-offspring regressions and sib correlations in the apple snail Pomacea canaliculata.
The brood sex ratio in the apple snail Pomacea canaliculata varies almost continuously from all male to all female, but the population sex ratio is nearly 1:1. In this study, regressions of the offspring sex ratio on the sex ratios of the parents' siblings as well as correlations in the brood sex ratios between sisters or brothers were investigated, in order to infer the genetic system that produces the sex-ratio pattern. There were significant positive relationships between the offspring sex ratio and the sex ratio of the mother's siblings (slope=0.28), and between the offspring sex ratios of two sisters (r=0.41). On the other hand, the father-offspring regression (slope=0.10), and the correlations between two brothers (r=-0.13) or between the brother and the sister (r=0.17) were not significant. These patterns differed from predictions using typical cytoplasmic sex factors, sex-ratio genes or sex-determining polygenes. Thus, the results suggest the involvement of either a small number of sex-determining genes or a more complicated system such as sex-ratio or sex-determining polygenes that act nonadditively.


Elevated blood pressure and risk of aortic valve disease: a cohort analysis of 5.4 million UK adults.
To test two related hypotheses that elevated blood pressure (BP) is a risk factor for aortic valve stenosis (AS) or regurgitation (AR). In this cohort study of 5.4 million UK patients with no known cardiovascular disease or aortic valve disease at baseline, we investigated the relationship between BP and risk of incident AS and AR using multivariable-adjusted Cox regression models. Over a median follow-up of 9.2 years, 20 680 patients (0.38%) were diagnosed with AS and 6440 (0.12%) patients with AR. Systolic BP (SBP) was continuously related to the risk of AS and AR with no evidence of a nadir down to 115 mmHg. Each 20 mmHg increment in SBP was associated with a 41% higher risk of AS (hazard ratio 1.41, 95% confidence interval 1.38-1.45) and a 38% higher risk of AR (1.38, 1.31-1.45). Associations were stronger in younger patients but with no strong evidence for interaction by gender or body mass index. Each 10 mmHg increment in diastolic BP was associated with a 24% higher risk of AS (1.24, 1.19-1.29) but not AR (1.04, 0.97-1.11). Each 15 mmHg increment in pulse pressure was associated with a 46% greater risk of AS (1.46, 1.42-1.50) and a 53% higher risk of AR (1.53, 1.45-1.62). Long-term exposure to elevated BP across its whole spectrum was associated with increased risk of AS and AR. The possible causal nature of the observed associations warrants further investigation.


Bicarbonate in diabetic ketoacidosis - a systematic review.
This study was designed to examine the efficacy and risk of bicarbonate administration in the emergent treatment of severe acidemia in diabetic ketoacidosis (DKA). PUBMED database was used to identify potentially relevant articles in the pediatric and adult DKA populations. DKA intervention studies on bicarbonate administration versus no bicarbonate in the emergent therapy, acid-base studies, studies on risk association with cerebral edema, and related case reports, were selected for review. Two reviewers independently conducted data extraction and assessed the citation relevance for inclusion. From 508 potentially relevant articles, 44 were included in the systematic review, including three adult randomized controlled trials (RCT) on bicarbonate administration versus no bicarbonate in DKA. We observed a marked heterogeneity in pH threshold, concentration, amount, and timing for bicarbonate administration in various studies. Two RCTs demonstrated transient improvement in metabolic acidosis with bicarbonate treatment within the initial 2 hours. There was no evidence of improved glycemic control or clinical efficacy. There was retrospective evidence of increased risk for cerebral edema and prolonged hospitalization in children who received bicarbonate, and weak evidence of transient paradoxical worsening of ketosis, and increased need for potassium supplementation. No studies involved patients with an initial pH < 6.85. The evidence to date does not justify the administration of bicarbonate for the emergent treatment of DKA, especially in the pediatric population, in view of possible clinical harm and lack of sustained benefits.