Incorporation and binding of estrogens into melanin: comparison of mushroom and mammalian tyrosinases.
The activities of mushroom and melanoma tyrosinases towards the estrogens were compared. While the fungal enzyme is capable of hydroxylating estradiol to the 2-hydroxy compound and to oxidize the latter to the quinone, the mammalian enzyme does not have this ability. With dopa as substrate and an estrogen present in the reaction mixture, both enzyme reactions yield melanin with the steroid firmly incorporated into the pigment, although with the mammalian enzyme the incorporation is small. The steroid appears to be incorporated by covalent linkage. It is suggested that the incorporation of estrogens into melanin produced by mammalian tyrosinase is via their oxidation by oxidized intermediates of the dopa to melanin transformation. Melanin itself may function as oxidant for the estrogens. Whole melanoma cells are capable of binding estrogens and incorporating small amounts into melanosomes. Similarly, fresh melanosomes in isolation can incorporate estrogens into their structure, presumably by covalent bonding to their melanin.


O brother how art thou: Propensity to report self-assessed unmet need.
Research investigating self-assessed unmet need (SUN) has taken the reports from surveys as given and subsequently attempted to discover patterns in inequality of access to healthcare. This requires the yet untested assumption that, given a certain level of care and demand, the likelihood of reporting unmet need does not vary across socioeconomic/demographic status (SEDS), be satisfied. Using an administrative dataset spanning 2001 to 2011 comprised of sufferers of a set of conditions that suggest unmet need (n = 3300) we evaluate the proposition that, given health status and care received, the propensity to report unmet need does not vary along SEDS. The results are further validated using the Canadian Community Health Surveys between 2001 and 2013 (n = 237,483). We find that the assumption of independence between reporting SUN and SEDS is not satisfied. Many of the groups found to have less access in previous studies may simply be more prone to interpret/answer the survey questions about unmet need in a certain way. The results of this research suggest that, in its present incarnation, survey data on self-assessed unmet need does not accurately measure what much of the academic literature has assumed it does.


Performance evaluation of a high-throughput separation system for circulating tumor cells based on microcavity array.
Circulating tumor cells (CTCs) are widely known as useful biomarkers in the liquid biopsies of cancer patients. Although single-cell genetic analysis of CTCs is a promising diagnostic tool that can provide detailed clinical information for precision medicine, the capacity of single-CTC isolation for genetic analysis requires improvement. To overcome this problem, we previously developed a multiple single-cell encapsulation system for CTCs using hydrogel-encapsulation, which allowed for the high-throughput isolation of single CTCs. However, isolation of a single cell from adjacent cells remained difficult and often resulted in contamination by neighboring cells due to the limited resolution of the generated hydrogel. We developed a novel multiple single-cell encapsulation system equipped with a high magnification lens for high throughput and a more accurate single-cell encapsulation. The multiple single-cell encapsulation system has sufficient sensitivity to detect immune-stained CTCs, and could also generate a micro-scaled hydrogel that can isolate a single cell from adjacent cells within 10 µm, with high efficiency. The proposed system enables high throughput and accurate single-cell manipulation and genome amplification without contamination from neighboring cells.


Sleep modulation of neuroendocrine function: developmental changes in gonadotropin-releasing hormone secretion during sexual maturation.
To assess sleep-associated changes in gonadotropin-releasing hormone secretion during sexual maturation, we studied nighttime and daytime patterns of LH and FSH secretion in two groups with qualitatively similar sex steroid levels: girls with central precocious puberty and young adult women in the early follicular phase of an ovulatory menstrual cycle. In the girls with central precocious puberty, all indices of LH secretion were significantly higher at night than during the day (mean LH levels, 12 +/- 2 versus 5 +/- 1 IU/L, p less than or equal to 0.01; LH pulse amplitude 16 +/- 2 versus 7 +/- 1 IU/L, p less than or equal to 0.01; and LH pulse frequency 0.70 +/- 0.05 versus 0.35 +/- 0.08 pulse/patient-h, p less than or equal to 0.01). Girls with a history of menses, who were presumably the most mature, lacked this diurnal variability. Mean nocturnal FSH levels were only slightly higher than daytime levels (7.6 +/- 0.5 versus 7.2 +/- 0.5 IU/L, p less than or equal to 0.05) resulting in alternating periods of LH (nighttime) and FSH (daytime) predominance in this pubertal population.(ABSTRACT TRUNCATED AT 250 WORDS)


MAFLD might be better in identifying subjects with sarcopenia or cardiovascular risk than NAFLD: A nationwide study.
Clinical features of non-alcoholic fatty liver disease (NAFLD), but not fulfilling the diagnostic criteria of metabolic dysfunction-associated fatty liver disease (MAFLD), remain unclear. We investigated the risk of sarcopenia and cardiovascular disease (CVD) in MAFLD and non-metabolic risk (MR) NAFLD. Subjects were selected from the Korean National Health and Nutrition Examination Surveys 2008-2011. Liver steatosis was assessed using fatty liver index. Significant liver fibrosis was defined using fibrosis-4 index, categorized by age cut-offs. Sarcopenia was defined as the lowest quintile sarcopenia index. Atherosclerotic CVD (ASCVD) risk score > 10% was defined as high probability. A total of 7248 subjects had fatty liver (137 with non-MR NAFLD, 1752 with MAFLD/non-NAFLD, and 5359 with overlapping MAFLD and NAFLD). In non-MR NAFLD group 28 (20.4%) had significant fibrosis. The risk of sarcopenia (adjusted odds ratio [aOR] = 2.71, 95% confidence index [CI] = 1.27-5.78) and high probability of ASCVD (aOR = 2.79, 95% CI = 1.23-6.35) was significantly higher in MAFLD/non-NAFLD group than in non-MR NAFLD group (all P < 0.05). The risk of sarcopenia and high probability of ASCVD was similar between subjects with and without significant fibrosis in non-MR NAFLD group (all P > 0.05). However, the risk was significantly higher in MAFLD group than in non-MR NAFLD group (aOR = 3.38 for sarcopenia and 3.73 for ASCVD; all P < 0.05). The risks of sarcopenia and CVD were significantly higher in MAFLD group but did not differ according to fibrotic burden in non-MR NAFLD group. The MAFLD criteria might be better for identifying high-risk fatty liver disease than the NAFLD criteria.


Two different conformations of the antitumor beta-D-glucan produced by Sclerotinia sclerotiorum IFO 9395.
The antitumor beta-D-glucan [SSG, (1----3)-linked with a D-glucosyl group attached to position 6 of alternate units] produced by Sclerotinia sclerotiorum IFO 9395 afforded a gel in neutral aqueous solution which gave no 13C-n.m.r. signals. Signals appeared on the addition to the gel of sodium hydroxide to 0.15M and the line-width was narrowest near 0.25M base (gel-to-sol transition). SSG bound to Congo Red and shifted the lambda max in a manner similar to that of curdlan. Neutralisation of the sol regenerated the gel. SSG and regenerated SSG showed different c.p.-m.a.s. 13C-n.m.r. spectra, especially the signals for C-3 which appeared at 86 and 89 p.p.m., respectively. The c.p.-m.a.s. spectrum of regenerated SSG was similar to that of curdlan, suggesting a helical structure. Thus, SSG and regenerated SSG appear to have different conformations.


It's not that kind of a job.
The duties of the corporation president and the hospital administrator are less publicly visible than those of the senator and the physician, but that doesn't make them less valuable.


Laparoscopic repeated pancreatectomy for isolated local recurrence in remnant pancreas following laparoscopic radical pancreatectomy for pancreatic ductal adenocarcinoma: Two cases report.
Typical treatment for recurrent pancreatic cancer is potent chemotherapy or chemoradiotherapy. However, recent clinical investigations suggest a potential therapeutic role of local resection in recurrent pancreatic cancer. Based on accumulating surgical experience of minimally invasive pancreatectomy, we report cases of two patients who underwent successful laparoscopic re-resection of isolated local recurrence following laparoscopic radical pancreatectomy for pancreatic cancer.


Amphotericin B lipid complex is efficacious in the treatment of Candida albicans biofilms using a model of catheter-associated Candida biofilms.
The purpose of this study was to determine the efficacy of amphotericin B lipid complex (ABLC) against Candida albicans biofilms using a rabbit model of catheter-associated candidal biofilm. A clinical C. albicans isolate was allowed to form biofilms on catheters placed in vivo and was then exposed to lock therapy with ABLC (1.5mg, locked for 4h or 8h for 7 days). Untreated biofilms served as controls. Fungal loads on the proximal and distal sections of catheters were determined by counting colony-forming units (CFUs), whilst surface architecture of formed biofilms was evaluated by scanning electron microscopy (SEM). Studies revealed that all ABLC-treated catheters were sterilised and yielded 0 CFU (P<or=0.0027 for both proximal and distal segments compared with the untreated controls). Furthermore, SEM analyses showed that while catheters retrieved from untreated control animals were overlaid with a thick biofilm, those treated with ABLC lock therapy showed only some debris with no fungal cells. Taken together, these studies demonstrated that using ABLC it was possible to sterilise catheters on which C. albicans biofilms were formed.


Variation in Cnidoscolus texanus in relation to herbivory.
Twelve Oklahoma populations of Cnidoscolus texanus were investigated for morphological variation between and within populations, and between site classes with contrasting histories of use by domesticated livestock. Characters scored were numbers of stinging hairs on petioles and midribs, and depth of leaf lobing. Leaf lobing was not significantly variable within or among populations or grazing categories. Stinging hair numbers showed significant differences within and among populations. Petiole stinging hairs were more numberous in heavily grazed sites, suggesting population differentiation in response to the selective pressures of herbivory.


Preservation and detection of specific antibody--peptide complexes by matrix-assisted laser desorption ionization mass spectrometry.
The direct detection of an antibody-peptide complex is reported by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). Experimental conditions have been found in which specific, noncovalent interactions in solution are maintained throughout the sample preparation and ionization process. Mass measurements based on the ion signals for the intact antibody and 1:1 antibody-peptide complex reveal that specific noncovalent associations between a monoclonal antibody and a peptide, which comprises the determinant of the corresponding antigen, are maintained in the gas phase. These results support the wider application of MALDI-MS to studies of the structure and specificity of macromolecular complexes important to immune and other biological function.


Reliability of preparation procedure for sphingosomal vincristine.
The reliability of the preparation procedure for sphingosomal vincristine was studied. The effect of minor variations in the constitution conditions on the integrity of the drug product was also examined. Two studies were conducted. The laboratory study, which had two parts, was conducted to determine the effects of deliberately varying the constitution conditions, including such key parameters as incubation time and incubation temperature. In the field study, 20 pharmacists unfamiliar with sphingosomal vincristine were asked to constitute the product using the written instructions provided in the package insert as their sole guidance. All samples in both studies were evaluated by measuring key product characteristics, including free (un-encapsulated) vincristine sulfate, total vincristine sulfate, vincristine degradation products, and in vitro release rate. Vincristine loading into sphingosomes was considered acceptable if the percentage of free vincristine sulfate did not exceed 10%. In the laboratory study, samples that were incubated at 60-75 degrees C for 5-60 minutes met all the acceptance criteria. However, acceptable loading was not achieved for samples that were incubated at 55 degrees C for 10 minutes or less. In the field study, all the pharmacist-prepared samples met the acceptance criteria, with the results for free vincristine sulfate demonstrating a high degree of statistical confidence in the reliability of the loading procedure. The recommended constitution procedure of sphingosomal vincristine from a three-vial kit can be reliably performed in pharmacies with a high degree of confidence. Small variations in temperature and incubation time had no effects on the quality of the product.


Methylphenidate in hyperactive children: differential effects of dose on academic, learning, and social behavior.
Methylphenidate (Ritalin) has been shown to have differential effects on hyperactive children's behavior as a function of dose level. In the present investigation, a triple-blind, placebo-control, within-subject (crossover) experimental design was employed in which 12 hyperactive boys between 6 and 10 years received three different dosages of methylphenidate (5, 10, and 15 mg) in a randomly assigned sequence. Dosage effects were assessed on clinic-(PAL--Paired Associates Learning test) and school-(percent on task, teacher ratings, work completion rates, and accuracy) related behaviors. For 10 of the children, classified as responders to medication by the PAL using the criteria of Swanson, Kinsbourne, and colleagues, a series of ANCOVAs with repeated measures showed significant dosage effects on teacher ratings (p less than .01), percent on task (p less than .01), academic accuracy (p less than .05), and assignment completion rates (p less than .05). PAL performance was also significantly enhanced (p less than .01) after optimal dose levels were considered. Subsequent trend analysis showed a significant positive linear relationship between dose and each of the dependent variables. A comparison of fixed-dose and miligram-per-kilogram plots showed that children's performance across the different dosages were clearly individualistic and task-specific, even when similar body weights were compared. The implications of using clinic-based testing to determine optimal medication responsivity were discussed.


Pulmonary blood volume measured by cardiovascular magnetic resonance: influence of pulmonary transit time methods and left atrial volume.
Increased pulmonary blood volume (PBV) is a measure of congestion and is associated with an increased risk of cardiovascular events. PBV can be quantified using cardiovascular magnetic resonance (CMR) imaging as the product of cardiac output and pulmonary transit time (PTT), the latter measured from the contrast time-intensity curves in the right and left side of the heart from first-pass perfusion (FPP). Several methods of estimating PTT exist, including pulmonary transit beats (PTB), peak-to-peak, and center of gravity (CoG). The aim of this study was to determine the accuracy and precision for these methods of quantifying the PBV, taking the left atrium volume (LAV) into consideration. Fifty-eight participants (64 ± 11 years, 24 women) underwent 1.5 T CMR. PTT was quantified from (1) a basal left ventricular short-axis image (FPP), and (2) the reference method with a separate contrast administration using an image intersecting the pulmonary artery (PA) and the LA (CoG(PA-LA)). Compared to the reference, PBV for (a) PTB(FPP) was 14 ± 17% larger, (b) peak-peak(FPP) was 17 ± 16% larger, and (c) CoG(FPP) was 18 ± 10% larger. Subtraction of the LAV (available for n = 50) decreased overall differences to - 1 ± 19%, 2 ± 18%, and 3 ± 12% for PTB(FPP), peak-peak(FPP), and CoG(FPP), respectively. Lowest interobserver variability was seen for CoG(FPP) (- 2 ± 7%). CoG(PA-LA) and FPP methods measured the same PBV only when adjusting for the LAV, since FPP inherently quantifies a volume consisting of PBV + LAV. CoG(FPP) had the best precision and lowest interobserver variability among the FPP methods of measuring PBV.


FNR-Dependent RmpA and RmpA2 Regulation of Capsule Polysaccharide Biosynthesis in Klebsiella pneumoniae.
Fumarate nitrate reduction regulator (FNR) is a direct oxygen-responsive transcriptional regulator containing an iron-sulfur (Fe-S) cluster. During anaerobic growth, the [4Fe-4S] cluster in FNR (holo-FNR) binds specifically to DNA, whereas exposure to oxygen results in the loss of its DNA-binding activity via oxidation of the [4Fe-4S] cluster. In this study, we aimed to investigate the role of FNR in regulation of capsular polysaccharide (CPS) biosynthesis, serum resistance, and anti-phagocytosis of K. pneumoniae. We found that the CPS amount in K. pneumoniae increased in anaerobic conditions, compared to that in aerobic conditions. An fnr deletion mutant and a site-directed mutant (fnr 3 CA), with the three cysteines (C20, C23, and C29) replaced with alanines to mimic an FNR lacking the [4Fe-4S] cluster, showed marked increase in CPS amount under anaerobic conditions. A promoter-reporter assay and qRT-PCR confirmed that the transcription of the cps genes was repressed by holo-FNR. In addition, we found that holo-FNR could repress the transcription of rmpA and rmpA2, encoding cps transcriptional activators. Deletion of rmpA or rmpA2 in the Δfnr strain reduced CPS biosynthesis, suggesting that RmpA and RmpA2 participated in the holo-FNR-mediated repression of cps transcription, thereby regulating the CPS amount, serum resistance, and anti-phagocytosis. Taken together, our results provided evidence that RmpA and RmpA2 participated in the holo-FNR-mediated repression of CPS biosynthesis, and resistance to the host defense in response to oxygen availability.


Alternative modes of ventilation. Part I. Disadvantages of controlled mechanical ventilation: intermittent mandatory ventilation.
Controlled mechanical ventilation is an accepted therapy for acute respiratory failure but by virtue of the increase in intrathoracic pressure has a large number of disadvantages. It is to overcome these disadvantages that alternative modes of ventilation have been introduced. These aim to reduce the effects of abnormally high airway pressure on the lung whilst recruiting solid alveoli and at the same time maintaining effective blood volume. Intermittent mandatory ventilation is a mode of ventilation first introduced to aid weaning which may reduce the need for sedation, permit better tolerance of high levels of PEEP and maintain urine osmolar output. High frequency ventilation utilising low airway pressures can maintain pulmonary gas exchange whilst reducing the effects of stretch on the lung. Its major role would seem to be in cases of bronchopleural fistula and necrotising pneumonia where a low mean airway pressure is essential. Low frequency positive pressure ventilation with extra corporeal CO2 removal, whilst a very labour intensive technique, has produced a favourable outcome in patients with terminal respiratory failure. Use of PEEP is associated with further deleterious haemodynamic effects which are largely overcome with use of continuous positive airway pressure during spontaneous respiration. PEEP is widely used. Its effect on pulmonary compliance, dead space and oxygen delivery are unpredictable making haemodynamic monitoring mandatory. Inversed ratio ventilation requires further evaluation whereas differential lung ventilation is logical, complicated but very valuable where the time constants for each lung are significantly different.


Phytofabricated silver nanoparticles: Discovery of antibacterial targets against diabetic foot ulcer derived resistant bacterial isolates.
The present study selected the predominant multi antibiotic-resistant diabetic foot ulcer (DFU) derived bacterial isolates such as Pseudomonas aeruginosa (PA), Escherichia coli (EC), Staphylococcus aureus (SA) and Bacillus subtilis (BS) and evaluated their response against the well-characterized Aerva lanata (AL) reduced multiple phytochemicals fabricated silver nanoparticles (AL-AgNPs). The overnight culture of DFU isolates was processed and subjected to various studies such as antimicrobial activity, growth kinetics, biofilm disruption, reactive oxygen species (ROS), membrane leakage, membrane permeability, and damage and genotoxicity. The molecular docking of AL phytochemicals was also performed with bacterial enzyme DNA gyrase. Interestingly, AL-AgNPs were produced the remarkable antibacterial effect against the resistant DFU isolates, which was closely similar to the effect of AL-AgNPs observed against the reference strains. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of AL-AgNPs against the DFU isolates were found to be 5-15 μg/mL and 10-20 μg/mL, respectively. The AL-AgNPs were depicted a concentration-dependent growth inhibition of DFU bacterial isolates. The MIC and MBC of AL-AgNPs were effectively destroyed the preformed biofilms of DFU isolates. Furthermore, the MBC of AL-AgNPs was displayed the increased intracellular ROS accumulation, membrane leakage, permeability and damage, and genotoxicity in the DFU isolates. Additionally, the in silico study revealed that the AL phytochemicals were fitted over the binding pocket of the DNA gyrase B subunit. The observed results were confirmed that the negative impacts of the AL-AgNPs at the level of the membrane and intracellular components of DFU isolates.


Dendritic Effects of Injectable Biodegradable Thermogels on Pharmacotherapy of Inflammatory Glaucoma-Associated Degradation of Extracellular Matrix.
The development of advanced drug delivery systems with extensively sustained release and multiple functions is highly imperative for effective attenuation of the degradation of ocular extracellular matrix that is associated with inflammatory glaucoma. Here, the generation of amine-terminated polyamidoamine dendrimers in an injectable biodegradable thermogel is demonstrated to be important for achieving prolonged drug release profiles and potent anti-inflammatory effects. Among various generations (Gx, x = 0, 1, 3, 5), third-generation G3 is proved as the most effective material for optimizing the synergistic effects of gelatin and poly(N-isopropylacrylamide) and generating a thermogel with the highest biodegradation resistance, the best drug encapsulation/extended-release performance, and the best ability to reduce the elevated expression of inflammatory molecules. A pharmacotherapy based on intracameral injection of thermogels coloaded with pilocarpine and ascorbic acid results in effective alleviation of progressive glaucoma owing to the anti-inflammatory activity and long-acting drug release (above a therapeutic level of 10 µg mL-1 over 80 days) of thermogels, which simultaneously suppress inflammation and stimulate regeneration of stromal collagen and retinal laminin. These findings on the dendritic effects of rationally designed injectable biomaterials with potent anti-inflammatory effects and controlled drug release demonstrate great promise of their use for pharmacological treatment of progressive glaucoma.


High-Dose Once-Weekly Semaglutide: A New Option for Obesity Management.
To review the pharmacology, efficacy, and safety of high-dose once-weekly semaglutide for chronic weight management. PubMed/MEDLINE and ClinicalTrials.gov were searched (inception to September 8, 2021) using keywords "semaglutide" and "obesity," "weight," "high dose," "high-dose," or "2.4." Clinical trials with published results were included. Publications studying the oral or <2.4 mg formulation of semaglutide were excluded. Four phase 3, multicenter, randomized, double-blind trials demonstrated efficacy of high-dose once-weekly semaglutide compared with placebo for weight loss. Study populations included patients with overweight or obesity (STEP 1, STEP 3, and STEP 4) or patients with diabetes and with overweight or obesity (STEP 2). Lifestyle interventions for diet and exercise were included for all participants. Weight loss from baseline was significant for all studies, and secondary outcomes demonstrated cardiometabolic improvements including waist circumference, systolic blood pressure, and lipid profiles. Gastrointestinal adverse effects were common, but the medication was otherwise well tolerated. High-dose semaglutide offers significant weight-lowering potential and favorable effects on cardiometabolic risk factors and glycemic indices. Clinicians and patients should consider the route and frequency of administration, adverse effect profile, and cost when choosing an antiobesity medication. The importance of concomitant lifestyle interventions should be emphasized. High-dose once-weekly semaglutide can significantly reduce weight, and although gastrointestinal adverse effects were common, it is generally well tolerated.


[Analysis of MECP2 gene variants in three pedigrees affected with Rett syndrome].
To detect potential variants of MECP2 gene in three pedigrees affected with Rett syndrome (RTT). All exons and their flanking regions of the MECP2 gene were subjected to Sanger sequencing and multiplex ligation-dependent probe amplification assay. The probands of pedigrees 1 and 2 have respectively carried a c.965C>G and a c.1157_1197del41 variant of the MECP2 gene, while the proband of pedigree 3 carried a heterozygous deletional variant in exon 4 of the MECP2 gene. Variants of the MECP2 gene probably underlay the RTT in the three pedigrees. Above finding has enriched the spectrum of MECP2 gene variants, and provided a guidance for the patients upon preimplantation genetic testing and prenatal diagnosis.


[Efficacy of Levetiracetam as an Add-on Therapy in Children with Localization-related Epilepsy].
To clarify the efficacy and safety of levetiracetam (LEV) in children with localization-related epilepsy, we performed a retrospective study in our hospital. We reviewed the cases of 70 pediatric patients (mean age, 9.4 years, range 4-15 years) who had undergone LEV add-on therapy between October 2010 and September 2014. The subjects were followed for a period of 24 weeks. The mean starting dose was 9.9 mg/kg/day, and the mean final dose was 39.9 mg/kg/day. The LEV continuation rate was estimated to be 87%. The responder rate (based on ≥ 50% reduction in seizure frequency) was 57%. Twenty-four of the 70 patients experienced side effects, such as drowsiness, fatigue, and aggressiveness. Although most of the adverse events were mild and tolerable, it was necessary to discontinue the medication in four patients. We evaluated the effects of gender, age, duration of illness, secondarily generalized seizure, baseline seizure frequency, MRI abnormality, number of previously administered drugs, number of concomitant drugs, and final dose. MRI abnormality and final dose of LEV had a statistically significant influence on the efficacy. LEV can be used as an effective and safe add-on treatment in children with localization-related epilepsy.


Streamlined approval of biosimilars: moving on from the confirmatory efficacy trial.
Licensing of biosimilars is essential to promote patient access to 21st-century biological medicines. Regulatory approval of biosimilars is based on the totality of evidence from a head-to-head comparison with reference products (RPs). A clinical efficacy trial is usually required, but this is increasingly questioned. Based on a thorough review of biosimilar applications in the European Union (EU), we conclude that in-depth knowledge of the reference product, allied with high-performing analytical tools, largely predicts clinical comparability, subject to confirmation by a comparative pharmacokinetic (PK) trial. We provide a blueprint for a biosimilar pathway that reduces the need for clinical efficacy trials in exceptional cases, together with qualifying criteria and requirements for streamlined assessment to expedite wider access to affordable biological medicines.


A functional XPNPEP2 promoter haplotype leads to reduced plasma aminopeptidase P and increased risk of ACE inhibitor-induced angioedema.
Angiotensin I-converting enzyme inhibitors (ACEi) are widely used antihypertensive agents that are associated with a potentially life-threatening reaction, ACEi-angioedema. Impaired metabolism of bradykinin and des-Arg(9) -bradykinin by aminopeptidase P (APP) is a key contributor to ACEi-angioedema. This study aimed to characterize the genetic regulation of the XPNPEP2 gene and identify the genetic factors contributing to variance in plasma APP activity and ACEi-angioedema. Additive genetic factors accounted for 47.3% of variance in plasma APP activity in healthy individuals. Nested deletion analysis identified the minimal promoter (-338 bp to -147 bp) and an enhancer region (-2,502 bp to -2,238 bp). Three polymorphisms (c.-2399C>A, c.-1612G>T, and c.-393G>A) were significantly associated with plasma APP activity. Haplotype ATG was significantly associated with reduced reporter gene activity and with reduced plasma APP activity. The c.-2399C>A polymorphism was located in an enhancer region and was predicted to differentially bind hepatic nuclear factor 4 (HNF4). Over expression of HNF4 increased the activation of haplotype ATG compared with haplotype CGG. In a case control study of subjects with a history of ACEi-angioedema, haplotype ATG was significantly associated with ACEi-angioedema (OR 4.87 [1.78-13.35] P = 0.002). The ATG haplotype is functional and contributes to ACEi-angioedema through a reduction in APP.


Unexpected Pneumonia Diagnosis From Pediatric Abdominal Pain: A Case Report.
Abdominal pain in the context of fever, tachypnea, or cough is a known presentation of pneumonia in preschool-aged children. We describe a 4-year-old male who presented to a pediatric emergency department with abdominal pain and decreased appetite. During his abdominal pain workup, he was found to have pneumonia complicated by pleural effusion and pneumothorax. It is critical for pediatric providers to be aware of age-based differentials.


Beta-galactosidase-deficient human fibroblasts: uptake and processing of the exogenous precursor enzyme expressed by stable transformant COS cells.
COS-1 cells were transfected by electroporation with a cDNA for human acid beta-galactosidase cloned in our laboratory and stable transformants expressing the enzyme activity were selected. The precursor form of the enzyme was secreted in large quantities into the culture medium. The fibroblasts from patients with GM1-gangliosidosis or Morquio B disease showed a remarkable increase of enzyme activity, up to the normal level, after culture in this medium for 2 days; the amount of uptake was essentially the same as that for the precursor form in human fibroblasts. After endocytosis, the precursor molecules were processed normally to the mature form and remained as stable as those produced by human fibroblasts. On the other hand, cells from galactosialidosis patients did not show any increase of enzyme activity in a similar experiment. It was concluded that the transformants are useful as the source of precursor proteins for the study of intracellular turnover of enzyme molecules in mutant cells.


Decreased bone turnover in children and adolescents with well controlled type 1 diabetes.
Osteoporosis is a common long-term complication of type 1 diabetes (T1DM). We aimed to determine whether bone mineral density (BMD) and turnover are already altered during childhood. We recruited 27 T1DM children and 32 controls (age 10.5 +/- 2.5 yr.) and measured BMD (dual-energy x-ray absorptiometry); bone biomarkers levels (osteocalcin: OC; procollagen type 1 propeptides amino-terminal: PINP; crosslinking telopeptides of type 1 collagen C-terminal: CTX), glycated hemoglobin (HbA1c), dietary intake and physical activity. Patients with T1DM had lower levels of OC (70.3 +/- 3.3 vs 105.3 +/- 6.8), PINP (556.4 +/- 47.6 vs 716.3 +/- 53.8), CTX(0.97 +/- 0.07 vs 1.20 +/- 0.08), physical activity, and calcium intake. Biomarkers were negatively correlated with HbA1c. Though, BMD was similar among groups and not related to HbA1c, disease duration, physical activity or dietary intakes. Bone turnover is altered in T1DM children, whereas BMD remains normal during growth. Physical activity and optimal calcium intakes may improve bone metabolism and delay osteoporosis.


In Vitro Studies on Hemodynamics of Type B Aortic Dissection: Accuracy and Reliability.
1. Hemodynamic of TBAD is important to improve the long-term outcome of TEVAR.2. This review provides an overview of the in-vitro for the hemodynamic study of TBAD.3. The accuracy and validity of in-vitro TBAD experiments should be further studied.


Unusual locations for cavernous hemangiomas: report of two cases and review of the literature.
Cavernous hemangiomas are most commonly found within the subcortical neural parenchyma near the fissura Rolandi, in the basal ganglia, or in the brain stem. Because of advancing neuro-imaging technology and thus resulting in a higher incidence of cavernous hemangiomas they have rising impact in neurosurgery. We present two unusual cases of extra-axial cavernous hemangiomas: one located at the frontal falx, the other within the bone of the right frontal bone. We discuss these and other cases in the literature with respect to the more common differential diagnoses and the appropriate therapy regimen for cavernous hemangiomas in these locations.


Blind versus open weighing from an eating disorder patient perspective.
Weighing is a key component in the treatment of eating disorders. Most treatment protocols advocate for open weighing, however, many clinicians choose to use blind weighing, especially during the early phase of treatment. Despite considerable debate about this issue in the literature, there is no empirical evidence supporting the superiority of one weighing approach over the other. In addition, little is known about patients' perspectives of open and blind weighing and which weighing practice they view as more acceptable and/or beneficial for their treatment. Semi-structured qualitative interviews were conducted with 41 women with a current or past diagnosis of Anorexia or Bulimia Nervosa: 26 were undergoing specialist inpatient treatment (n = 13 being blind weighed; n = 13 being open weighed) and 15 were community members who have recovered from an eating disorder. Interviews were audiotaped, transcribed verbatim and analysed thematically using framework methods. Participant demographics, clinical characteristics, weighing anxiety and weight concerns were also assessed. Qualitative analyses yielded five themes: (1) therapy engagement and progress; (2) Control and tolerance of weight uncertainty; (3) treatment team relationships and autonomy; (4) life outside of treatment; and (5) weighing practice preferences and rationale. Participants stated that blind weighing decreased anxiety and eating disorder psychopathology (e.g., weight preoccupation) and increased treatment responsivity. For many, relinquishing control over their weight facilitated body trust and was a necessary step towards recovery. Participants found that not knowing their exact weight helped challenge their overconcern with weight. Lack of support post-discharge was identified as a major difficulty of blind weighing. Overall, the majority of participants preferred blind weighing, particularly at the early, acute stage of treatment, whereas open weighing was viewed as more suitable at later stages of recovery. Quantitative analyses found current blind-weighed patients felt significantly less anxiety around being weighed and had greater tolerance of weight uncertainty than current open-weighed patients. This study provided in-depth patient insights into open versus blind weighing practices. The next step for future research will be to supplement these insights with treatment outcomes gained from randomised controlled trials comparing the two weighing practices.


Profiling genes related to mitochondrial function in mice treated with N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine.
Since mitochondrial dysfunction plays an important role in the pathogenesis of dopaminergic neurodegeneration in Parkinson's disease, we determined the expression of genes related to mitochondrial function in the substantia nigra of mice treated with N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) using a cDNA array. MPTP treatment significantly depleted striatal dopamine, but did not result in apparent neuronal loss in the substantia nigra at 3 and 18 days post-treatment. We also examined changes in genes in the hypothalamus, a region containing dopaminergic neurons that are relatively resistant to MPTP. Finally, we confirmed those genes identified by microarrays as differentially expressed in the substantia nigra but not in the hypothalamus using in situ hybridization. Our results demonstrated that MPTP significantly changed the expressions of six genes in nigral neurons, four of which were related to the mitochondrial electron transport chain: the NADH-ubiquinone oxidoreductase 13 kDa B subunit, the NADH-ubiquinone oxidoreductase MNLL subunit, cytochrome c, and the cytochrome c oxidase Va subunit. Two other differentially expressed genes were the dihydropyridine-sensitive L-type calcium channel alpha-2 subunit precursor and type III alpha-1 procollagen. None of these six genes are encoded by mitochondrial DNA. The potential significance of these gene alterations in the context of Parkinson's disease is discussed.


Number and location of radiolabeled, intraoperatively identified sentinel nodes in 48 head and neck cancer patients with clinically staged N0 and N1 neck.
The value of sentinel node (SN) biopsy for squamous cell carcinoma of the head and neck (HNSCC) has not been determined yet. A critical evaluation of this concept seems to be mandatory with regard to the increasing acceptance of SN biopsy in other tumor entities. Against the background of the results of 48 previously untreated patients, a reproducible technique for SN biopsy in the head and neck level, which has been adjusted to the special topographic conditions of this anatomic region, is presented. Methods included intraoperative SN biopsy, which was performed in 48 previously untreated patients suffering from squamous cell carcinoma (2x lower lip, 8x oral cavity, 20x oropharynx, 15x larynx, 3x hypopharynx). Using ultrasound imaging, 43 patients were staged as N0 necks, and 5 patients were staged as N1 necks. Fine-needle aspiration cytology (FNAC) was performed in cases of doubt. Surgery on the neck was carried out according to the suspected stage of lymphogenic spread once the SN1 as well as one or two further hot nodes (SN2, SN3) had been identified. Numbers and distribution of the intraoperatively excised nodes SN1-3 were documented according to their relation to the tumor location. Post-operatively, the histologic results of the intraoperatively excised nodes SN1-3 were compared to the histologies of the neck dissection specimen. Results showed that in all 48 patients, a SN1 could be identified intraoperatively. In 20 cases an additional SN2 and in 6 cases a SN3 was diagnosed. In carcinomas of the lower lip and oral cavity, the SN1 was found in 4 cases in level I (2x lower lip, 2x floor of the mouth) and in 6 cases in level II (6x lateral tongue). In carcinomas of the oropharynx, the respective nodes were found in 17 of 20 cases in level II (carcinomas of the tonsil) and in 3 cases in level III (carcinomas of the base of the tongue). In supraglottic carcinomas the SN1 was identified in 8 of 10 cases in level II and in 2/5 patients with glottic carcinomas, while in 3/5 glottic carcinomas as well as in all hypopharyngeal carcinomas, the SN1 was found in level III. In relation to the predictiveness of the detected SN, it has to be remarked that in 38 patients a SN1 free of tumor was representative for the regional lymph node status (pN0). An isolated metastasis (pN1) was diagnosed in the SN (9x SN1, 1x SN2) in 10 patients. In conclusion, the results of a SN biopsy modified to a strictly intraoperative method of detection are encouraging. Critical indications showed that a thorough and standardized technical performance of the injection as well as a mandatory, so far unchanged, neck dissection form the basis for the development of a SN concept for SCCs of the upper aerodigestive tract. The value of the SN concept, however, currently remains unclear for patients suffering from HNSCC.


Spectrum of ectopic ureters in children.
We reviewed the clinicopathological features, diagnosis, and surgical treatment of ectopic ureters (EU) in children in order to evaluate the anatomy, renal function, outcome, and the existence of any racial or regional difference in this abnormality. The records of 27 consecutive patients with 33 EU managed over a period of 7 years were analyzed. There were 25 female and two male patients. Their age ranged from 1 to 13 years, with an average of 4.5 years. The presenting features included history of continuous dribbling of urine with otherwise normal voiding in toilet-trained female patients, incontinence of urine, urinary tract infection, abdominal pain, and renal failure. The diagnostic work-up included ultrasonography (US) in all 27 patients, intravenous pyelography (IVP) in 23, micturating cystourethrogram (MCU) in 19, nuclear imaging (NI) in 16, and computed tomogram (CT) in five patients. All patients underwent examination under anesthesia (EUA) and cystourethrovaginoscopy immediately before the operation. Single-system ectopic ureter (SSEU) was present in 19 patients with 25 ureters (six bilateral), and duplex with ectopic ureter was present in eight cases. The left side was involved in 14 cases, the right side in seven, and the anomaly was bilateral in six cases. Abnormalities noted in the imaging studies were USG 23/27, IVP 23/23, MCU 11/19, NI 13/16, and CT 5/5 cases. EUA and cystourethrovaginocsopy helped in localization of the ectopic site and evaluation of the bladder. The surgical treatment included nephroureterectomy (NUT) for non-functioning kidney in 7 patients, heminephroureterectomy (HNUT) for non functioning upper pole of duplex kidney with EU in 6 patients, ureteric reimplantation (UTR) in 13 patients (19 ureters, 6 bilateral), and ureteropyelostomy (UTP) in one patient. In the follow-up period ranging from 1 to 5 years, 20 patients achieved continence; however, six cases with bilateral SSEU and one case of unilateral SSEU, who also had a patulous bladder neck, continued to dribble urine. SSEU was more common than duplex with ectopic ureter. A large number of functioning renal units associated with SSEU deserved preservation. The success of surgical treatment in terms of achievement of continence was high and depended on the integrity of the bladder neck.


Gonococcal pericarditis with tamponade - use of molecular technology to improve diagnosis and management.
We report a case of gonococcal pericarditis, which was unexpected due to its extremely unusual occurrence. A 42-year-old man presented with fever, chest pain, dyspnea, and tachycardia. He was initially stable but rapidly deteriorated, developing pericardial effusion with tamponade requiring a pericardial window. Incompletely decolorized gram stain of the pericardial fluid initially suggested the presence of gram-positive diplococci, which wrongly directed treatment toward possible pneumococcal infection. Because cultures were negative, identification of the causative organism was attempted by molecular and genotyping analysis. These techniques identified Neisseria gonorrhoeae-multi-antigen sequence type 14994 (por 5136/tbpB 33) as the etiology, which has been associated with disseminated gonococcal disease. Real-time polymerase chain reaction showed no evidence of mutations within the N. gonorrhoeae penA gene responsible for causing ceftriaxone resistance. This was crucial in guiding antibiotic treatment, in light of the high prevalence of multi-drug-resistant N. gonorrhoeae. This case highlights the utility of diagnostic molecular techniques in identifying N. gonorrhoeae as the etiology of an exceedingly rare case of pericarditis.


Workplace incivility and its influence on professional quality of life among nurses from multicultural background: A cross-sectional study.
To investigate the workplace incivility of nurses working in two Saudi hospitals and analyse its influence on the nurses' professional quality of life (ProQOL). The prevalence and economic impact of workplace incivility cannot be overstated and disregarded. To the current authors' knowledge, no extensive study on this topic has been conducted in Saudi Arabia. The influence of workplace incivility to ProQOL of nurses from different cultural backgrounds has never been thoroughly investigated. Descriptive, cross-sectional design. A sample of 378 nurses working in two government hospitals in Saudi Arabia were surveyed using the Nursing Incivility Scale and the ProQOL Scale version 5 from February to May 2018. A multivariate multiple regression analysis was conducted to analyse the multivariate effect of workplace incivility on the nurses' ProQOL. The study adhered to the STROBE guideline (See Supporting Information File 1). The nurses perceived a moderate level of workplace incivility from the different sources of uncivil acts measured in this study. Among the five sources of incivility explored in this study, the nurses reported the majority of workplace incivility experienced from patients/visitors (M = 2.44, SD = 0.80), while the lowest was from supervisors (M = 1.90, SD = 0.66). The mean scores of the respondents in the compassion satisfaction, burnout and secondary traumatic stress subscales were 36.50 (SD = 6.30), 26.43 (SD = 4.81) and 26.47 (SD = 6.06), respectively. General incivility, supervisor incivility, physician incivility and patient/visitor incivility showed a significant multivariate effect on the three ProQOL subscales. Nurses' experience of workplace civility and its sources were associated with ProQOL. The findings of this study can be used as guide in establishing human resource policies towards achieving nurses' needs, reducing workplace incivility and improving ProQOL.


Amyloid-like fibril formation of co-chaperonin GroES: nucleation and extension prefer different degrees of molecular compactness.
The molecular chaperone GroES, together with GroEL from Escherichia coli, is the best characterized protein of the molecular chaperone family. Here, we report on the in vitro formation of GroES amyloid-like fibrils and the mechanism of formation. When incubated for several weeks at neutral pH in the presence of the denaturant guanidine hydrochloride, GroES formed a typical amyloid fibril; unbranched, twisted, and extended filaments stainable by thioflavin T and Congo red. GroES fibril formation was accelerated by the addition of preformed fibril seeds, in accordance with a nucleation-extension mechanism. Interestingly, whereas the spontaneous formation of GroES fibrils was favored in the structural transition region of GroES dissociation/unfolding, the extension of fibrils from preformed fibril seeds was favored in the region corresponding to an expanded molecular state. We concluded that the two stages of GroES fibril formation prefer different molecular states of the same protein. The significance of this preference is discussed.


Effect of the uterus on induced cystic ovarian follicles in the sow.
Large multiple cystic ovarian follicles were induced in 16 sows and small multiple cystic ovarian follicles were induced in five sows by daily injections of adrenocorticotrophic hormone during the follicular phase of the oestrous cycle. Large ovarian cysts persisted until euthanasia at 35 to 51 days after complete bilateral hysterectomy between days 5 and 10 in four sows. Atresia of large cysts was seen about days 15-18 in a sham-hysterectomy experiment. Unilateral hysterectomy of three sows resulted in persistence of large cysts on the ovary adjacent to uterine horn removal and atresia in the contralateral ovary. Lutein patches developed on the follicular wall of persistent cysts between the time of hysterectomy and autopsy. Atresia did not occur after hysterectomy in three sows with small cystic follicles, nor did sham hysterectomy of a sow with small ovarian cysts result in cyst atresia. A pronounced reduction in the size of large cysts occurred in three of four sows after the intrauterine injection of prostaglandin F2a. When a prostaglandin inhibitor, indomethacin, was injected into one uterine horn, the adjacent ovarian cysts persisted while those on the ovary next to the uterine horn receiving saline solution underwent atresia. Daily urinary oestrogen values from a hysterectomised sow with large cysts were initially high but subsequently fell to values between 30 and 60 microng/ml, except for several high values when signs of oestrus were observed. Oestrogen values in urine from a hysterectomised sow with small cysts were comparatively lower, and rose to high values only once in the 50-day period studied. Plasma oestrogen values in a hysterectomised sow with large cysts were high, although considerable day-to-day variation occurred. Plasma progesterone values were also high in this sow. In a sow with small cystic follicles for a 50-day period, oestrogen values in plasma were relatively low, but higher values were obtained occasionally. Plasma progesterone levels remained consistently low in this sow.


UDP-glucuronosyltransferase 1A4 (UGT1A4) polymorphisms in a Jordanian population.
Glucuronidation is one of the most important phase II metabolic pathways. It is catalyzed by a family of UDP-glucuronosyltransferase enzymes (UGTs). One of the subfamilies is UGT1A. Allele frequencies in UGT1A4 differ among ethnic groups. The aim of this study was to determine the allelic frequency of two most common defective alleles: UGT1A4*2 and UGT1A4*3 in a Jordanian population. A total of 216 healthy Jordanian Volunteers (165 males and 51 females) were included in this study. Genotyping for UGT1A4*1, UGT1A4*2 and UGT1A4*3 was done using a well established polymerase chain reaction-restriction fragment length polymorphism test. Among 216 random individuals studied for UGT1A4*2 mutation there were 26 individuals who were heterozygous, giving a prevalence of 12% and an allele frequency of 6.5%. Only one individual was homozygous for UGT1A4*2. The UGT1A4*3 mutation was detected as heterozygous in 9 of 216 individuals indicating a prevalence of 4.2% and allele frequency of 3.5%. Three individuals were homozygous for the UGT1A4*3 indicating a prevalence of 1.4%. The prevalence of UGT1A4*2 is similar to the Caucasians but different from other populations whilst the UGT1A4*3 prevalence in the Jordanian population is distinct from other populations. Our results provide useful information for the Jordanian population and for future genotyping of Arab populations in general.


The effect of stimulus contrast on the latency and amplitude of the pattern electroretinogram.
Our studies have verified that a linear relationship exists between the amplitude of the pattern onset ERG and retinal contrast, but indicate that there is no systematic variation in peak latency with contrast. The optical degradation of high spatial frequency patterns cannot therefore provide an explanation for the variation in peak latency with spatial frequency.


Effect of gastroenteritis during pregnancy on neonatal outcome.
Previous studies have shown that gastrointestinal disease in women during pregnancy may be a risk factor for low-birthweight infants. In the present study, the prevalence of gastroenteritis during pregnancy and its effect on neonatal outcome was examined for each gestational month in 10,597 single-birth mother-infant pairs in the ABIS Project (All Babies in Southeast Sweden). After exclusion of mothers with inflammatory bowel disease, celiac disease, lactose intolerance, or cow's milk allergy, data on the remaining 10,229 mother-infant pairs were compiled. Overall, 32.5% of the mothers suffered from gastroenteritis during pregnancy (95% confidence interval [CI], 32.5-32.5%). Risk factors included young maternal age (P for trend, <0.001), previous infants (P<0.001), work in a pediatric day-care facility (P=0.004), and experience of a major life event (P=0.027). Binary logistic and multiple linear regression analyses were adopted for the following variables of neonatal outcome: birth week, preterm birth (<37 weeks), birthweight, low birthweight (< or = 2,499 g), birth length, cesarean section, and hospitalization in a neonatal care unit. Maternal gastroenteritis during month 4 (-0.18 week; 95%CI=-0.36, -0.01 week), month 5 (-0.30 week; 95%CI=-0.49, -0.11 week), or month 7 (-0. 18 week; 95%CI=-0.35, -0.01 week) of pregnancy was associated with a shorter pregnancy (adjusted for confounders). Gastroenteritis during part of the pregnancy was associated with a shortened pregnancy, but it had no other adverse effects on neonatal outcome. The reduction in the duration of pregnancy is probably of little clinical relevance. These findings should be confirmed in a prospective study.


Investigations of the discriminative properties of the frog's olfactory mucosa using a photoactivable odorant.
The photoactivable compound phenylazide interacts reversibly with the frog's olfactory epithelium when delivered as an olfactory stimulus. Its application together with UV irradiation resulted in a differential reduction of the responses (electro-olfactogram) to several odorants. It was found that this chemical modification of the sensory membrane affected the responses in relative ratios which could be predicted from a classification of the odorants based upon independent studies of receptor cell odor sensitivity. The responses to chemicals representative of the 'aromatic group' were clearly more reduced than those to compounds of the 'camphoraceous group'. It is suggested that the olfactory discrimination mechanisms can be approached by the use of this method.


A pilot trial of S-1 plus irinotecan chemotherapy for esophageal adenocarcinoma.
This study investigated the clinical efficacy and toxicity of the combination chemotherapy using S-1 plus irinotecan for esophageal adenocarcinoma. This study included 10 patients with histologically confirmed adenocarcinoma of the esophagus or esophagogastric junction between April 2005 and August 2011. S-1 was administered orally at a dose of 80 mg/m²/day from day 1 to 14 and irinotecan was given intravenously on day 1 and 8 at a dose of 80 mg/m². A total of 65 cycles were administered and the response rate was 62.5%. The 50% progression-free survival period and the 50% overall survival period for all of the patients was 8.4 months and 19.1 months, respectively and 5.9 months and 16.3 months for the 8 patients with unresectable or recurrent tumors, respectively. The 2 patients that received adjuvant chemotherapy demonstrated a prophylactic effect for the post-operative recurrence. On the other hand, this therapy showed no severe non-hematological toxicity and only 20% experienced grade 3 neutropenia. As a result, the treatment regimen could generally be performed in an outpatient basis. The combination chemotherapy using S-1 and irinotecan showed tolerable clinical efficacy in terms of the response rate, survival and toxicity for esophageal adenocarcinoma.


Expression analysis of recombinant herpes simplex virus type 1 DNase.
Expression of recombinant herpes simplex virus type 1 (HSV-1) deoxyribonuclease (DNase) was analyzed in BHK-21 cells, a standard cell line for virus propagation, by using mammalian cell expression systems based on vaccinia virus and on Semliki Forest virus (SFV)1. Although the establishing of recombinant vaccinia virus failed due to the apparent toxicity of the herpesviral enzyme, soluble and functional HSV-1 DNase was efficiently expressed in BHK-21 cells by the vaccinia virus/T7 RNA polymerase hybrid system as well as by recombinant Semliki Forest virus. Using rabbit antiserum ExoC, directed against the C-terminal residues 503-626, or mouse monoclonal antibody (MAb) Q1, raised against the type 2 enzyme, a major 85-kDa protein with the identical size of the enzyme from HSV-1-infected cells was identified to be induced in both expression systems. With recombinant SFV functional HSV-1 DNase coincided with the overproduction of a single major 85-kDa protein reaching an optimum between 16 h and 36 h after infection. At later times of infection the enzymatic activity vanished. Thus, recombinant SFV may be an appropriate expression vector for biochemical studies of the enzyme when (i) packaged recombinant virus particles are used for infection and (ii) infection does not exceed 24 h. Due to the limitations of transient expression systems, the vaccinia/T7 RNA polymerase hybrid system is suited for expression analysis on a small scale, and for studying intracellular interactions of the enzyme as demonstrated by immunofluorescence microscopy studies. Using vector pTM1, recombinant HSV-1 DNase was efficiently overproduced in BHK-21 cells at 6 h after transfection and was shown to colocalize with the cellular chromatin at sites apparently distinct from the bulk of the herpesviral replication sites the way it is observed for the enzyme of lytically infected cells. The deleting of the 123 C-terminal amino acid residues did not alter this nuclear localization of HSV-1 DNase, suggesting that the latter sequences and other herpesviral factors are not required for the chromatin association.


Direct electron-transfer of native hemoglobin in blood: kinetics and catalysis.
A novel approach that uses nature biological tissues, fish blood, for the study of the direct electron-transfer of hemoglobin and its catalytic activity for H(2)O(2) and NO(2)(-) is observed. The direct electron-transfer of hemoglobin in red blood cells in fish blood on glassy carbon electrode was observed for the first time. By simply casting fish blood on GC electrode surface and being air-dried, a pair of well-defined redox peaks for HbFe (III)/HbFe (II) appeared at about -0.36 V (vs SCE) at the fish blood film modified GCE in a pH 7.0 phosphate buffer solution. Ultraviolet visible (UV/VIS) characterization and the enhancement of the redox response of Hb by adding pure Hb in fish blood suggested that Hb preserved the native second structures in the fish blood film. Optical micrographs showed that the RBCs retained its integrity in blood. Hb in blood/GCE maintained its activity and could be used to electrocatalyze the reduction H(2)O(2) and NO(2)(-).


Rituximab dosing and monitoring strategies in neuromyelitis optica patients: creating strategies for therapeutic success.
Neuromyelitis optica (NMO) is an autoimmune condition that predominantly causes severe optic neuritis and transverse myelitis. Rituximab therapy has dramatically improved patient care, but standardized dosing regimens and guidelines are lacking. The objective of this study was to define a rituximab dosing strategy for NMO patients that achieves the lowest rate of relapses. This was a retrospective chart review of patients treated with various doses of rituximab. Combining data from the NMO and multiple sclerosis (MS) patients, identified that the mean number of days after a 100 mg dose of rituximab until the CD19 population was greater than 2% was 99 days (standard deviation 36, range 43-172). When allowed to rise, the mean number of days after a 1000 mg dose of rituximab until the CD19 population was greater than 2% was 184 (standard deviation 72, range 52-288). The median number of days until a CD19 percentage of 2% was achieved was 133 days in the 100 mg dosing arm and 259 days in the 1000 mg dosing arm. Analysis of the survival curves via both the Mantel-Cox log-rank test and the Wilcoxon test determined that the difference between medial survival for 100 and 1000 mg doses was statistically significant with p-values <0.0001. Low doses of rituximab have a high rate of early B-cell repopulation. Any NMO patient treated with rituximab should be followed with monthly CD19 counts in order to identify the rare, but clinically significant, early repopulators.


Respiration characteristics of mitochondria in parental and giant transformed cells of the murine Nemeth-Kellner lymphoma.
Respiration characteristics of mitochondria of the parental and giant cells of murine NK/Ly (Nemeth-Kellner lymphoma) were studied. The giant cell-enriched ascites were obtained by serial intraperitoneal injections of vinblastine in tumour-bearing mice. Ascites containing >70% giant cells were used. Their diameter of was over 17 μm (~2800 μm(3)), while the diameter of the parental cells was 12.7 μm (1100 μm(3)). The respiration rate of mitochondria in situ was measured by oxygen consumption in intact and digitonin-permeabilized NK/Ly cells. Endogenous respiration of intact giant NK/Ly cells was three times higher compared to the parental ones, roughly in agreement with the volume change. The giant NK/Ly cells were far more resistant to permeabilization with digitonin than the parental cells, as shown by Trypan Blue and LDH (lactate dehydrogenase) release tests. After digitonin permeabilization, oxygen consumption was reduced to a minimal level (0.06 ng atom O/(s × 106 cells) in both types of cells. Addition of α-ketoglutarate or succinate to the incubation medium increased oxygen consumption in the parental cells by 46 and 164% respectively. In the giant NK/Ly cells, the corresponding increases were 164 and 276%. Addition of ADP to α-ketoglutarate- or succinate-supplemented medium further stimulated oxygen consumption of the permeabilized NK/Ly cells; however, the effect of ADP was more pronounced in the giant cells. In addition, indices of respiratory control were significantly higher in the giant cells. Oligomycin suppressed considerably the respiration of the intact giant cells but had a much weaker effect on parental cells. Thus, giant NK/Ly cells possess much higher respiration rates and show tighter coupling between the respiration and oxidative phosphorylation compared with parental cells.


Unplugging of prolene IOL loops.
From end 1981 until early 1984, 415 Severin-type lenses were implanted in our two centres. In 10 instances i.e. 2.4%, we experienced unplugging of the anterior 12 o'clock positioned loop, apparently a result of the loop being held with the insertions forceps during insertion. The unplugging of the loop possibly affected the visual outcome in one instance.


Unlocking Charge Transfer Limitations for Extreme Fast Charging of Li-Ion Batteries.
Extreme fast charging (XFC) of high-energy Li-ion batteries is a key enabler of electrified transportation. While previous studies mainly focused on improving Li ion mass transport in electrodes and electrolytes, the limitations of charge transfer across electrode-electrolyte interfaces remain underexplored. Herein we unravel how charge transfer kinetics dictates the fast rechargeability of Li-ion cells. Li ion transfer across the cathode-electrolyte interface is found to be rate-limiting during XFC, but the charge transfer energy barrier at both the cathode and anode have to be reduced simultaneously to prevent Li plating, which is achieved through electrolyte engineering. By unlocking charge transfer limitations, 184 Wh kg-1 pouch cells demonstrate stable XFC (10-min charge to 80 %) which is otherwise unachievable, and the lifetime of 245 Wh kg-1 21700 cells is quintupled during fast charging (25-min charge to 80 %).


Nascent Integrin Adhesions Form on All Matrix Rigidities after Integrin Activation.
Integrin adhesions assemble and mature in response to ligand binding and mechanical factors, but the molecular-level organization is not known. We report that ∼100-nm clusters of ∼50 β3-activated integrins form very early adhesions under a wide variety of conditions on RGD surfaces. These adhesions form similarly on fluid and rigid substrates, but most adhesions are transient on rigid substrates. Without talin or actin polymerization, few early adhesions form, but expression of either the talin head or rod domain in talin-depleted cells restores early adhesion formation. Mutation of the integrin binding site in the talin rod decreases cluster size. We suggest that the integrin clusters constitute universal early adhesions and that they are the modular units of cell matrix adhesions. They require the association of activated integrins with cytoplasmic proteins, in particular talin and actin, and cytoskeletal contraction on them causes adhesion maturation for cell motility and growth.


Identification of land reclamation stages based on succession characteristics of rehabilitated vegetation in the Pingshuo opencast coal mine.
Land reclamation is a dynamic ecological restoration process, and rehabilitated vegetation requires a certain amount of time to develop, stabilize and mature. The development characteristics of rehabilitated vegetation at different land reclamation stages are significantly different, and these differences can be used to identify the key stages of land reclamation. In this study, normalized differential vegetation index (NDVI) time series data from the western dump of the Pingshuo opencast coal area on the Loess Plateau from 1989 to 2018 were collected and analyzed by fluctuation analysis and filter processing with the Best Index Slope Extraction and Wavelet Transform (BISE-WT) filter to reveal the succession characteristics of rehabilitated vegetation. Then, the key periods of land reclamation under different vegetation types (arbor, shrub, arbor-shrub, and grass) in the dump were determined by S-logistic function fitting and derivative analyses. The NDVI time series changes in land parcels reclaimed in different years exhibited different interannual change characteristics. Based on the number of years required for the rehabilitated vegetation to reach a stable state, the average development period of land reclamation in eligible units in the mining dumps was thirteen years, including ten years in the rapid development period and three years in the steady development period. The differences in land reclamation periods among the different vegetation types were significant (α = 0.05), and the number of years required for each period mainly followed the order of arbor > arbor-shrub > shrub > grass. Analyzing the vegetation succession characteristics and identifying the key periods of land reclamation for different vegetation types is conducive to dynamically evaluating land reclamation effects and is expected to provide a basis for strengthening the implementation of manual intervention measures in reclaimed mining areas.


Renal transplantation of the infant and young child and the use of pediatric cadaver kidneys for transplantation in pediatric and adult recipients.
Transplantation of infants less than 1 year of age with kidneys from live-related parental donors has recently led to good results, whereas cadaver donor renal transplantation in this recipient age group has led to a high mortality rate (11/13). Similarly, the results of cadaver donor renal transplantation in infants and young children less than 5 years of age has been suboptimal in the past, although recent data are more encouraging. With recent availability of long-term peritoneal dialysis for the infant and young child with end-stage renal disease (ESRD), it is possible to defer transplantation until an optimal donor becomes available. Because of the possible immunologic hyperactivity of such recipients, the immunosuppressive regimen may need to be modified if improved cadaver donor survival rates are to be obtained. The use of anencephalic kidneys for transplantation has been associated with a high incidence of primary nonfunction and few recipients with long-term functioning grafts. Harvesting of kidneys from anencephalic donors declared "brain-dead" at birth may reduce the incidence of primary nonfunction and increase the availability of anencephalic kidneys for transplantation. Reports of the use of pediatric cadaver kidneys for transplantation into pediatric and adult recipients yields discrepant results. Analysis of the data indicates that if pediatric cadaver kidneys from donors less than 6 years of age are used, the potential for decreased graft survival rates and an increased incidence of technical complications exists. However, the use of pediatric cadaver kidneys can provide adequate graft function in both pediatric and adult recipients and the use of such kidneys should increase the number of kidneys available for transplantation.


Identification and interrogation of the gene regulatory network of CEBPA-double mutant acute myeloid leukemia.
Acute myeloid leukemia (AML) is a heterogeneous hematological malignancy caused by mutations in genes encoding transcriptional and epigenetic regulators together with signaling genes. It is characterized by a disturbance of differentiation and abnormal proliferation of hematopoietic progenitors. We have previously shown that each AML subtype establishes its own core gene regulatory network (GRN), consisting of transcription factors binding to their target genes and imposing a specific gene expression pattern that is required for AML maintenance. In this study, we integrate gene expression, open chromatin and ChIP data with promoter-capture Hi-C data to define a refined core GRN common to all patients with CEBPA-double mutant (CEBPAN/C) AML. These mutations disrupt the structure of a major regulator of myelopoiesis. We identify the binding sites of mutated C/EBPα proteins in primary cells, we show that C/EBPα, AP-1 factors and RUNX1 colocalize and are required for AML maintenance, and we employ single cell experiments to link important network nodes to the specific differentiation trajectory from leukemic stem to blast cells. Taken together, our study provides an important resource which predicts the specific therapeutic vulnerabilities of this AML subtype in human cells.


Validity and practical utility of accelerometry for the measurement of in-hand physical activity in horses.
Accelerometers are valid, practical and reliable tools for the measurement of habitual physical activity (PA). Quantification of PA in horses is desirable for use in research and clinical settings. The objective of this study was to evaluate a triaxial accelerometer for objective measurement of PA in the horse by assessment of their practical utility and validity. Horses were recruited to establish both the optimal site of accelerometer attachment and questionnaire designed to explore owner acceptance. Validity and cut-off values were obtained by assessing PA at various gaits. Validation study- 20 horses wore the accelerometer while being filmed for 10 min each of rest, walking and trotting and 5 mins of canter work. Practical utility study- five horses wore accelerometers on polls and withers for 18 h; compliance and relative data losses were quantified. Accelerometry output differed significantly between the four PA levels (P < 0•001) for both wither and poll placement. For withers placement, ROC analyses found optimal sensitivity and specificity at a cut-off of <47 counts per minute (cpm) for rest (sensitivity 99.5 %, specificity 100 %), 967-2424 cpm for trotting (sensitivity 96.7 %, specificity 100 %) and ≥2425 cpm for cantering (sensitivity 96.0 %, specificity 97.0 %). Attachment at the poll resulted in optimal sensitivity and specificity at a cut-off of <707 counts per minute (cpm) for rest (sensitivity 97.5 %, specificity 99.6 %), 1546-2609 cpm for trotting (sensitivity 90.33 %, specificity 79.25 %) and ≥2610 cpm for cantering (sensitivity 100 %, specificity 100 %) In terms of practical utility, accelerometry was well tolerated and owner acceptance high. Accelerometry data correlated well with varying levels of in-hand equine activity. The use of accelerometers is a valid method for objective measurement of controlled PA in the horse.


[Hereditary pheochromocytoma and paraganglioma: screening and follow-up strategies in asymptomatic mutation carriers].
The management of pheochromocytoma and paraganglioma has deeply evolved over the last years due to the discovery of novel genes of susceptibility, especially SDHx, MAX and TMEM127. While the modalities of diagnosis and management of patients presenting with hereditary pheochromocytoma and paraganglioma are now well defined, screening and follow-up strategies for asymptomatic mutation carriers remain a matter of debate. This raises major questions as these asymptomatic patients will require a lifelong follow-up. The aim of this review is an attempt to give insights on the optimal screening and follow-up strategies of asymptomatic carriers of SDHx, MAX and TMEM127 mutations, with additional thoughts on the forensic and psychological aspects of the management of such patients with rare diseases.


Cell differentiation to "mating bodies" induced by an integrating and conjugative element in free-living bacteria.
Lateral gene transfer (LGT) is one of the most important processes leading to prokaryotic genome innovation. LGT is typically associated with conjugative plasmids and bacteriophages, but recently, a new class of mobile DNA known as integrating and conjugative elements (ICE) was discovered, which is abundant and widespread among bacterial genomes. By studying at the single-cell level the behavior of a prevalent ICE type in the genus Pseudomonas, we uncover the remarkable way in which the ICE orchestrates host cell differentiation to ensure horizontal transmission. We find that the ICE induces a state of transfer competence (tc) in 3%-5% of cells in a population under nongrowing conditions. ICE factors control the development of tc cells into specific assemblies that we name "mating bodies." Interestingly, cells in mating bodies undergo fewer and slower division than non-tc cells and eventually lyse. Mutations in ICE genes disrupting mating-body formation lead to 5-fold decreased ICE transfer rates. Hence, by confining the tc state to a small proportion of the population, ICE horizontal transmission is achieved with little cost in terms of vertical transmission. Given the low transfer frequencies of most ICE, we anticipate regulation by subpopulation differentiation to be widespread.


An improved balloon snake for HIFU image-guided system.
Target segmentation in ultrasound images is a key step in the definition of the intro-operative planning of high-intensity focused ultrasound therapy. This paper presents an improvement for the balloon snake in segmentation. A sign function, designed by the edge map and the moving snake, is added to give the direction of the balloon force on the moving snake separately. Segmentation results are demonstrated on ultrasound images and the effectiveness and convenience shown in applications.


Ethanol locks to prevent catheter-related bloodstream infections in parenteral nutrition: a meta-analysis.
Patients with pediatric intestinal failure (IF) depend on parenteral nutrition for growth and survival, but are at risk for complications, such as catheter-related bloodstream infections (CRBSIs). CRBSI prevention is crucial, as sepsis is an important cause of IF-associated liver disease and mortality. We aim to estimate the pooled effectiveness and safety of ethanol locks (ELs) in comparison with heparin locks (HLs) with regard to CRBSI rate and catheter replacements for pediatric IF patients with chronic parenteral nutrition dependence. A systematic review without language restriction was performed on Medline (1948-2010), Embase (1980-2010), and conference programs and trial registries up to December 2010. Search terms included "Catheter-Related Infections," "Catheter," "Catheters, Indwelling," "alcohol," "ethanol," and "lock." Two authors identified 4 retrospective studies for the pediatric IF population. Double, independent data extraction using predefined data fields and risk of bias assessment (Newcastle-Ottawa scale) was performed. In comparison with HLs, ELs reduced the CRBSI-rate per 1000 catheter days by 7.67 events and catheter replacements by 5.07. EL therapy decreased the CRBSI rate by 81% and replacements by 72%. One hundred eight to 150 catheter days of EL exposure were necessary to prevent 1 CRBSI and 122 to 689 days of exposure avoided 1 catheter replacement. Adverse events were rare and included thrombotic events. In pediatric patients with IF, EL is a more effective alternative to HL. Adverse events include thrombotic events.


Life events in Xhosas in Cape Town.
A life events scale for research into Xhosa speaking people of Cape Town was developed. It was standardised on 131 residents of Cape Town's three major Black suburbs. Measures of perceived impact and rate of occurrence were elicited. The Spearman Rank correlation between prevalence and impact scores was not significant (r = 0.04). Items were ranked according to composite impact scores. Cultural and social factors were considered to play a part in explaining differences between the present findings and those of other studies. Limitations on the applicability of the scale and methodological issues were discussed. The problems in direct comparison between different cultural groups were emphasized, with particular stress on constraints on life events research in an African setting.


Injection of human mesenchymal stem cells improves healing of vocal folds after scar excision--a xenograft analysis.
Using a xenograft model the aim was to analyze if injection of human mesenchymal stem cells (hMSC) into the rabbit vocal fold (VF), after excision of an established scar, can improve the functional healing of the VF. Prospective design with an experimental xenograft model. The VFs of 12 New Zealand rabbits were injured by a bilateral localized resection. After 9 weeks the scar after the resection was excised and hMSC were injected into the VFs. After another 10 weeks 10 VFs were dissected and stained for histology. Lamina propria thickness and relative content of collagen type I were measured. Viscoelasticity of 14 VFs at phonatory frequencies was quantified by a simple-shear rheometer. The hMSC survival was determined using a human DNA specific reference probe, that is, FISH analysis. The viscoelastic measurements, that is, dynamic viscosity and elastic shear modulus for the hMSC-treated VFs, were found to be similar to those of normal controls and were significantly lower than those of untreated controls (P < .05). A significant reduction in lamina propria thickness was also shown for the hMSC treated VFs compared with the untreated VFs (P < .05). This histologic finding corresponded with the viscoelastic results. No hMSC survived 10 weeks after the injection. Human mesenchymal stem cells injected into the rabbit VF following the excision of a chronic scar, were found to enhance the functional healing of the VF with reduced lamina propria thickness and restored viscoelastic shear properties.


Optimization of voriconazole therapy for treatment of invasive aspergillosis: Pharmacogenomics and inflammatory status need to be evaluated.
Cytochrome 2C19 genotype-directed dosing of voriconazole (VRC) reduces the incidence of insufficient VRC trough concentrations (Cmin ) but does not account for CYP3A polymorphisms, also involved in VRC metabolism. This prospective observational study aimed to evaluate the utility of a genetic score combining CYP2C19 and CYP3A genotypes to predict insufficient initial VRC Cmin (<1 mg/L). The genetic score was determined in hematological patients treated with VRC. The higher the genetic score, the faster the metabolism of the patient. The impact of the genetic score was evaluated considering initial VRC Cmin and all VRC Cmin (n = 159) determined during longitudinal therapeutic drug monitoring. Forty-three patients were included, of whom 41 received VRC for curative indication. Thirty-six patients had a genetic score ≥2, of whom 11 had an initial insufficient VRC Cmin . A genetic score ≥2 had a positive predictive value of 0.31 for having an initial insufficient VRC Cmin and initial VRC Cmin was not associated with the genetic score. The lack of association between the genetic score and VRC Cmin may be related to the inflammatory status of the patients (C-reactive protein [CRP] levels: median [Q1-Q3]: 43.0 [11.0-110.0] mg/L), as multivariate analysis performed on all VRC Cmin identified CRP as an independent determinant of the VRC Cmin adjusted for dose (P < .0001). The combined genetic score did not predict low VRC exposure in patients with inflammation, which is frequent in patients with invasive fungal infections. Strategies for the individualization of VRC dose should integrate the inflammatory status of patients in addition to pharmacogenetic variants.


Drug-Drug Interactions between Direct Oral Anticoagulants and Hepatitis C Direct-Acting Antiviral Agents: Looking for Evidence Through a Systematic Review.
Direct oral anticoagulants (DOACs), as substrates of cytochrome P450 (CYP) 3A4 and/or P-glycoprotein, are susceptible to drug-drug interactions (DDIs). Hepatitis C direct-acting antiviral agents (DAAs), via P-glycoprotein or CYP3A4 inhibition, may increase DOAC exposure with relevant bleeding risk. We performed a systematic review on DDIs between DOACs and DAAs. Two reviewers independently identified studies through electronic databases, until 7 July 2020, supplementing the search by reviewing conference abstracts and the ClinicalTrials.gov website. Of 1386 identified references, four articles were finally included after applying the exclusion criteria. Three phase I clinical studies in healthy volunteers assessed interactions between dabigatran and glecaprevir/pibrentasvir, odalasvir/simeprevir, or sofosbuvir/velpatasvir/voxilaprevir, showing an increase in the dabigatran area under the concentration-time curve (AUC) by 138%, 103%, and 161%, respectively. DOACs and DAAs are under-investigated for DDI risk. Real-world studies are needed to assess the clinical relevance of the pharmacokinetic interactions with dabigatran and describe the actual spectrum of possible DDIs between DAAs and other DOACs.


The opisthobranch gastropods (Mollusca: Heterobranchia) from Venezuela: an annotated and illustrated inventory of species.
The Caribbean waters of Venezuela are composed by a large variety of habitats, with over 2800 km of coastline, islands, and islets. This area is a transitional zone between two main biogeographic provinces, the Caribbean and the Brazilian, separated by the fresh water outflows of the Orinoco and Amazon rivers, and is therefore expected to be an area of high species diversity. However, concerning the study of molluscs, Venezuela is probably the poorest known region in the Caribbean. The best compilation of opisthobranch species known in Venezuela was produced almost a decade ago, mentioning the occurrence of 57 species, plus seven determined only to genus level. In this work, 134 species are reported for Venezuela (71 are illustrated), representing about 40 % of the entire diversity of opisthobranchs known in the Caribbean. Among the species occurring in Venezuela, 49 have here the southern limit of their distribution range and only one the northern limit. Forty-six species are recorded for the first time to the country and one is a new record for the Caribbean Sea, namely Placida cremoniana. In addition, the distribution and ecology of the species are given based in literature and new data.


Delivery Room Respiratory Stabilization of Preterm Neonates: A Randomized, Controlled Trial.
To determine whether heated humidified high-flow nasal cannula (HHHFNC) is noninferior to NCPAP to provide DR respiratory support to preterm neonates of gestational age (GA) 28-36 wk. This randomized, controlled, noninferiority trial included 124 spontaneously breathing preterm neonates who developed respiratory distress soon after birth and/or had a FiO2 requirement > 0.3. Primary outcome measure was treatment failure within 24 h. The absolute risk difference with 95% confidence interval (CI) were calculated with a noninferiority margin of 10%. Secondary outcome variables were temperature at admission, time to treatment failure, treatment failure at 72 h, need for surfactant, intubation, duration of respiratory support, and incidences of adverse events including mortality. Intention-to-treat analysis was done in Stata software. Both the groups were similar in baseline characteristics. There was no statistically significant difference between the treatment failure rates with HHHFNC (13.1%, n = 61) and NCPAP (11.1%, n = 63) (risk difference 2.0%, 95% CI - 9.9% to 14.07%, p = 0.73). However, noninferiority of HHHFNC to NCPAP could not be conclusively proved as the 95% CI crossed both 0 and the noninferiority margin of 10%. There were no significant differences in secondary outcomes. HHHFNC showed similar efficacy and safety as NCPAP irrespective of gestational age, though its noninferiority to NCPAP remained inconclusive. Clinical Trial Registry of India, Registration no: CTRI/2019/10/021633, https://ctri.icmr.org.in/.


Evolution of photosystem I and the control of global enthalpy in an oxidizing world.
Life on earth is governed by light, chemical reactions, and the second law of thermodynamics, which defines the tendency for increasing entropy as an expression of disorder or randomness. Life is an expression of increasing order, and a constant influx of energy and loss of entropic wastes are required to maintain or increase order in living organisms. Most of the energy for life comes from sunlight and, thus, photosynthesis underlies the survival of all life forms. Oxygenic photosynthesis determines not only the global amount of enthalpy in living systems, but also the composition of the Earth's atmosphere and surface. Photosynthesis was established on the Earth more than 3.5 billion years ago. The primordial reaction center has been suggested to comprise a homodimeric unit resembling the core complex of the current reaction centers in Chlorobi, Heliobacteria, and Acidobacteria. Here, an evolutionary scenario based on the known structures of the current reaction centers is proposed.


Temperature responses of developmental processes have not been affected by breeding in different ecological areas for 17 crop species.
• Rates of tissue expansion, cell division and progression in the plant cycle are driven by temperature, following common Arrhenius-type response curves. • We analysed the genetic variability of this response in the range 6-37°C in seven to nine lines of maize (Zea mays), rice (Oryza spp.) and wheat (Triticum aestivum) and in 18 species (17 crop species, different genotypes) via the meta-analysis of 72 literature references. • Lines with tropical or north-temperate origins had common response curves over the whole range of temperature. Conversely, appreciable differences in response curves, including optimum temperatures, were observed between species growing in temperate and tropical areas. • Therefore, centuries of crop breeding have not impacted on the response of development to short-term changes in temperature, whereas evolution over millions of years has. This slow evolution may be a result of the need for a synchronous shift in the temperature response of all developmental processes, otherwise plants will not be viable. Other possibilities are discussed. This result has important consequences for the breeding and modelling of temperature effects associated with global changes.


Effect of oral melatonin administration on melatonin, 5-hydroxyindoleacetic acid, indoleacetic acid, and cyclic nucleotides in human cerebrospinal fluid.
Melatonin was given orally to patients undergoing diagnostic pneumoencephalography and various compounds were measured in the lumbar and cisternal CSF. Melatonin markedly increased plasma and CSF melatonin concentrations. The plasma: CSF melatonin ratios were similar in patients who received, and in those who did not receive, melatonin. This supports the idea that melatonin is released from pineal to blood and gets into the CSF via the blood. Melatonin did not affect CSF 5-hydroxyindoleacetic acid, which indicates that it has no effect on 5-hydroxytryptamine metabolism. Melatonin increased CSF indoleacetic acid significantly, indicating increased metabolism of tryptamine. Melatonin did not affect CSF cAMP levels, but increased cGMP levels. The effect on indoleacetic acid and cGMP was seen in both lumbar and cisternal CSF, suggesting that melatonin can have generalized actions throughout the CNS.


Biocatalytic conversion of aloeresin A to aloesin.
Leaf exudates from Aloe species, such as the Southern African Aloe ferox, are used in traditional medicines for both humans and livestock. This includes aloesin, a skin bleaching product that inhibits the synthesis of melanin. Aloesin, (a C-glycoside-5-methylchromone) can be released from aloeresin A, an ester of aloesin, through hydrolysis. The objective of the current study was to identify an enzymatic hydrolysis method for converting aloeresin A to aloesin, resulting in increased concentrations of aloesin in the aloe bitters extract. More than 70 commercially available hydrolytic enzymes were screened for the conversion of aloeresin A. An esterase (ESL001-02) from Diversa, a lipase (Novozym 388) and a protease (Aspergillus oryzae) preparation were identified during screening as being capable of providing conversion of pure aloeresin A, with the protease giving the best conversion (~100%). It was found that a contaminating enzyme in Novo 388 was responsible for the conversion of aloeresin A to aloesin. This contaminating enzyme, possibly a protease, was able to give almost complete conversion using crude aloe bitters extract, doubling the concentration of aloesin in aloe bitters extract via the hydrolysis of aloeresin A.


Dietary exposure to diesel exhaust particles and oxidatively damaged DNA in young oxoguanine DNA glycosylase 1 deficient mice.
Pulmonary exposure to diesel exhaust particles (DEP) has been associated with high levels of oxidized DNA in lung cells, whereas long-term oral DEP exposure appears to induce the DNA repair system with concomitant unaltered levels of oxidized DNA in the colon and liver of rats. Here we studied the generation of oxidatively damaged DNA in young wild type (WT) and oxoguanine DNA glycosylase 1 (OGG1) deficient mice after dietary exposure to 0mg/kg, 0.8 mg/kg, or 8 mg/kg Standard Reference Material 1650 in the feed for 21 days. The ingestion of DEP did not increase the levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine and comet assay endpoints in terms of strand break, endonuclease III, and formamidopyrimidine glycosylase (FPG) in the colon, liver, and lung tissue of WT or Ogg1(-/-) mice. The level of OGG1 mRNA could only be measured in WT mice and it was not increased by DEP feeding. On the contrary, the level of FPG sites was twofold higher in the liver and lung of Ogg1(-/-) mice compared to the levels in the WT mice tissues. In conclusion, although Ogg1(-/-) mice have high levels of oxidized guanine lesions, they do not appear to be markedly vulnerable to the genotoxicity by oral administration of DEP.


Targeting TEAD-ious resistance.
Recent advances in targeting mutant KRAS are limited by resistance. A recent study in Nature Cancer by Hagenbeek et al. utilizes a novel inhibitor that targets the TEAD transcription factor, GNE-7883, to overcome resistance to KRAS inhibitors. Thus, TEAD inhibitors may maximize the durability of KRAS inhibitors in patients with cancer.


Role of immunoreactions and mast cells in pathogenesis of human endometriosis--morphologic study and gene expression analysis.
To investigate the pathophysiology of human endometriosis, we examined by morphological and molecular biological methods. Samples of ovarian endometriosis and normal ovarian tissues were obtained laparoscopically after informed consent. A morphological study by toluidine blue staining, immunohistochemistry of c-kit and electron microscopy demonstrated the localization of mast cells in the stromal lesions of endometriosis. Oligonucleotide microarrays were used for gene expression analysis. Infiltration of numerous mast cells and development of fibrosis was observed throughout the stromal lesions. Gene expression analysis by oligonucleotide microarrays indicated inflammatory immunoreactions in the lesions. Expressions of the FCER1G and PGDS, which are considered to be mast cell-specific genes, were upregulated in the ovarian endometriotic lesions as compared to the normal ovarian tissues. Furthermore, expressions of genes associated with immunological inflammation, such as IL-8, GRO1, GRO2, CXCR4, MCP1, and those related to tissue remodeling (MMP, COL4A2, and COL5A2) were also higher in endometriotic lesions than in the normal ovarian tissue. Thus it is likely that mast cells and their related inflammatory immunoreactions via chemokines play important roles in producing fibrosis and adhesions in endometriotic lesions.


[Surgical management of anus incontinence using free-transplanted autologous, endogenous intestinal musculature].
A new treatment for anal incontinence is presented. Smooth muscle from the large intestine is transplanted to form a new sphincter. The advantage of using smooth muscle as a sphincter plastic id discussed.


Construction of the survival nomograms for colon cancer patients of different ages based on the SEER database.
Three nomograms for predicting the outcomes of early- and late-onset colon cancer (COCA) among patients not stratified by age were constructed using data in the Epidemiology and End Results (SEER) database (1975-2019). The accuracy of the nomogram was then assessed. Clinical data of 6107 patients with COCA were obtained from the SEER database. The patients were randomly divided into training and validation cohorts in a ratio of 7:3. Univariate and multivariate COX analyses of factors that could independently impact the prognosis of COCA were performed, and the corresponding nomograms for early-onset and late-onset COCA were constructed. Calibration curves, ROC curves, and C-index were used to determine the predictive accuracy. The discriminatory ability of the nomograms to assess their clinical utility, which was compared with the TNM staging system of the 8th edition of AJCC, was verified using survival analysis. Tumor primary site, ethnicity, and serum carcinoembryonic antigen (CEA) level significantly impacted the prognosis of colon cancer. Race, brain metastasis, and CEA were independent factors for predicting COCA prognosis. C-index, ROC, and calibration curves demonstrated that the three nomograms were accurate and superior to the traditional TNM staging system. Among the three nomograms, the early-onset COCA nomogram had the highest predictive accuracy, followed by that of colon cancer not stratified by age. Three nomograms for patients not stratified by age, early-onset colon cancer, and late-onset colon cancer were constructed. The accuracies of the nomograms were good and were all superior to the conventional TNM staging system. The early- and late-onset COCA nomograms are useful for clinical management and individualized treatment of COCA patients at different ages.


Rotational States of Bose gases with attractive interactions in anharmonic traps.
A rotated and harmonically trapped Bose gas with attractive interactions is expected to either remain stationary or escape from the trap. Here we report that, on the contrary, in an anharmonic trapping potential the Bose gas with attractive interactions responds to external rotation very differently, namely, through center-of-mass motion or by formation of vortices.


Algal wastewater treatment systems for seasonal climates: application of a simple modelling approach to generate local and regional design guidelines.
Algal waste stabilisation ponds (WSP) provide a means of treating wastewater, and also a potential source of water for re-use in irrigation, aquaculture or algal biomass cultivation. The quantities of treated water available and the periods in which it is suitable for use or discharge are closely linked to climatic factors. This paper describes the application, at a continent-wide scale, of a modelling approach based on the use of readily available climate datasets to provide WSP design and performance guidelines linked to geographical location. Output is presented in regionally-based contour maps covering a wide area of Russia and central Asia and indicating pond area, earliest discharge date, discharge duration, wastewater inflow:outflow ratio and salinity under user-specified conditions. The results confirm that broad-brush discharge guidelines of the type commonly used in North America can safely be applied; but suggest that a more detailed approach is worthwhile to optimise operating regimes for local conditions. The use of long-series climate data can also permit tailoring of designs to specific sites. The work considers a simple 2-pond system, but other configurations and operating regimes should be investigated, especially for the wide range of locations across the world that are intermediate between the 'one short discharge per year' mode and year-round steady-state operation.


Survival and recurrence of resectable tongue cancer: Resection margin cutoff value by T classification.
The purpose of this study was to determine an appropriate cutoff value for the resection margin according to the initial T classification. The medical records of 151 patients treated by surgery for tongue cancer were retrospectively reviewed to identify the significant perioperative parameters and appropriate cutoff value associated with disease-specific survival (DSS) and recurrence-free survival (RFS). The posterior resection margin was significantly correlated with survival and local recurrence (P = .020 and .016, respectively), whereas the deep resection margin was correlated with overall recurrence (P = .047). The cutoff values of the posterior and deep resection margins for survival and recurrence were different and larger in the advanced-stage T classification group (0.45 cm vs 0.95 cm and 0.25 cm vs 0.80 cm, respectively). Posterior and deep resection margins are significant prognosticators in tongue cancer. A larger resection margin may be needed in advanced-stage T classification.